volume III | INTERNATIONAL COURT OF JUSTICE

Document Number

17552

Parent Document Number

17548

Document File

17552.pdf

INTERNATIONAL COURT OF JUSTICE
____________________________________________

CASE CONCERNING
AERIAL HERBICIDE SPRAYING

(ECUADOR v. COLOMBIA)

COUNTER-MEMORIAL OF THE

REPUBLIC OF COLOMBIA

VOLUME III

ANNEXES 112 - 156

29 MARCH 2010 LIST OF ANNEXES

VOLUME III

O.A.S. DOCUMENTS

Annex 112 Annual Report of the Inter-American Drug Abuse

Control Commission (CICAD) to the General
Assembly nd the Organization of American States
at its 32 Regular Session, 12-15 November
2001, Caracas, Venezuela, 3 April 2002.
OEA/SER.L/XIV.2.30 CICAD/doc.1123/01 rev.1

Annex 113 2004 Memorandum of Understanding between
the Government of Colombia and the
Organization of American States (OAS) for the
execution of the study on the effects of the

Program for the Eradication of Illicit Crops by
aerial spraying with Glyphosate Herbicide
(PECIG) and of illicit crops, on human health and
the environment, 4 February 2004.

Annex 114 Annual Report of the Inter-American Drug Abuse
Control Commission (CICAD) to the General
Assembly of the Organization of American States
at its 34thRegular Session, 17-20 November

2003, Montreal, Canada, 3 May 2004.
OEA/Ser.L/XIV.2.34 CICAD/doc.1264/03 rev. 2
Annex 115 35 Regular Session of CICAD, Washington,

D.C., 27-30 April 2004, Session Documents.
Annex 116 CICAD, K.R. Solomon et al. “Environmental and

Human Health Assessment of the Aerial Spray
Program for Coca and Poppy Control in
Colombia”, OAS, Wash ington, D.C., 31 March
2005, (CICAD I).

iAnnex 117 37 Regular Session of CICAD, Santo Domingo,
Dominican Republic, 26-29 April 2005,
Document 1421.

Annex 118 CICAD, R.A. Brain et al., “The Toxicology of
Substances Used in th e Production and Refining
of Cocaine and Heroin: A Tier-Two Hazard

Assessment”, OAS, Washington, D.C., 31 July
2005, (CICAD I).
Annex 119 2006 Memorandum of Understanding between

the General Secretariat of the Organization of
American States (SG/OAS) and the Government
of Colombia for the execution of the study on the
effects of the Program for the Eradication of
Illicit Crops by aerial spraying with Glyphosate
Herbicide (PECIG) on human health and the

environment, 23 May 2006.
Annex 120 Annual Report of the Inter-American Drug Abuse
Control Commission (CICAD) to the General
Assembly of the Organization of American States

at its 36th Regular Session, Santo Domingo,
Dominican Republic, 4-6 June 2006.
Annex 121 Annual Report of the Pe rmanent Council to the

General Assembly of the Organization of
American States 2006-2007. OAS,
AG/doc.4698/07.

Annex 122 Annual Report of the Inter-American Drug Abuse
Control Commission (CICAD) to the General
Assembly of the Organization of American States
at its 39 thRegular Session, San Pedro Sula,
Honduras, 2-3 June 2009. OEA/Ser.G,
CP/doc.4395/09 corr. 1, 26 May 2009.

SCIENTIFIC AND TECHNICAL REPORTS

Annex 123 SGS (Societé Génerale de Surveillance, S.A)
Colombia S.A., “Report of Contamination

ii Control for glyphosate application at the Sierra of
Santa Marta”, 1987.

Annex 124 J.P. Giesy, S. Dobson S & K.R. Solomon,
“Ecotoxicological risk assessment for Roundup
herbicide”, Reviews of Environmental
Contamination and Toxicology 167: 35-120,
2000.

Annex 125 G. M. Williams et al., “Safety evaluation and risk
assessment of the herbicide Roundup® and its
active ingredient, glyphosate, for humans” in
Regulatory Toxicology aPndarmacology

31:117-165, 2000.
Annex 126 Embassy of the United States of America -
Clínica de Toxicología ‘Uribe Cualla’, Alleged
effects of Glyphosate on Human Health , Bogotá,

December 2001.
Annex 127 R.E. Ramos C., J.P. Ramos B., Environmental
Assessment of the Impact of coca crops and the
processing of coca leaf, Universidad de los

Andes, Bogotá, Colombia, 2002.
Annex 128 O. Saavedra, Laboratorio Inmunopharmos Ltda.,
Toxicity Study on Laboratory Animals for two

concentrations of Glyphosate 44% + Cosmoflux
1% + Water 55%, Bogotá, 15 February 2002.
Annex 129 M. San Sebastián & A-K. Hurtig, “Oil
exploitation in the Amazon Basin of Ecuador: a

public health emergency”, Pan Am J Public
Health, 15(3):205-211, 2004.
Annex 130 K.R. Solomon et al., A Response to Tomás León

Sicard et al., CICAD, 2005.
Annex 131 Journal of Toxicology and Environmental Health,
Part A: Current Issues, Volume 72, Numbers 15-
16, August 1 and August 15 2009, Special Issue:

Production of Illicit Drugs , the Environment and
Human Health, Taylor & Francis, London, 2009.
As follows:

iii• Annex 131-A: K.R. Solomon et al., “Human
Health and Environmental Risks from the Use
of Glyphosate Formulations to Control the
Production of Coca in Colombia: Overview

and Conclusions”, in Journal of Toxicology
and Environmental Health , Part A, 72:914-
920, 2009.

• Annex 131-B: A.J. Hewitt et al., “Spray
Droplet Size, Drift Potential, and Risks to
Nontarget Organisms from Aerially Applied
Glyphosate for Coca Control in Colombia”, in
Journal of Toxicology and Environmental
Health, Part A, 72:921-929, 2009.

• Annex 131-C: E.J.P. Marshall et al, “Coca
(Erythroxylum coca ) Control is Affected by
Glyphosate Formulations and Adjuvants”, in
Journal of Toxicology and Environmental

Health, Part A, 72:930-936, 2009.
• Annex 131-D: R.A. Brain et al., “Comparison
of the Hazards Posed to Amphibians by the

Glyphosate Spray Control Program Versus the
Chemical and Physical Activities of Coca
Production in Colombia”, in Journal of
Toxicology and Environmental Health, Part A,
72:937-948, 2009.

• Annex 131-E: L.H. Sanin et al., “Regional
Differences in Time to Pregnancy Among
fertile Woman from Five Colombian Regions
with Different use of Glyphosate”, in Journal
of Toxicology and Environmental Health , Part

A 72:949-960, 2009.
• Annex 131-F: M.H. Bernal et al., “Toxicity of
Formulated Glyphosate (Glyphos) and Cosmo-

Flux to Larval Colombian Frogs 1. Laboratory
Acute Toxicity”, in Journal of Toxicology and
Environmental Health , Part A 72:961-965,
2009.

iv • Annex 131-G: M.H. Bernal et al., “Toxicity
of Formulated Glyphosate (Glyphos) and
Cosmo-Flux to Larval and Juvenile
Colombian Frogs 2. Fi eld and Laboratory

Microcosm Acute Toxicity”, in Journal of
Toxicology and Environmental Health , Part A
72:966-973, 2009.

• Annex 131-H: J. D. Lynch et al., “Risks to
Colombian amphibian fauna from cultivation
of coca (Erythroxylum coca ): A geographical
analysis” in Journal of Toxicology and
Environmental Health , Part A, 72:974-985,
2009.

• Annex 131-I: C. Bolognesi et al.,
“Biomonitoring of Genotoxic Risk in
Agricultural Workers from Five Colombian
Regions: Association to Occupational

Exposure to Glyphosate”, in Journal of
Toxicology and Environmental Health , Part A
72:986-997, 2009.
Annex 132 Centre for Toxicology and Environmental Health,

L.L.C., University of Arkansas for Medical
Sciences, GLYPHOSATE Frequently Asked
Questions, 2009.

OTHER DOCUMENTS

Annex 133 Safety Data Sheet for Roundup SL.

Annex 134 Label and Safety Data Sheet for GLY-41.
Annex 135 Andean Regulation for the Registration and
Control of Chemical Pesticides for Agricultural

Use, Andean Community, Decision 436 of 2000,
Arts. 57-59; Annex 1 - Glossary, “Mean Lethal
Dose, LD 50”.

vAnnex 136 Report of the Fifth Meeting of Vice-Ministers of
Foreign Affairs of the Andean Community,
Caracas, 16-17 October 2000.

Annex 137 Andean Cooperation Plan for the Control of
Illegal Drugs and Related Offenses, Andean
Community, Decision 505 of 2001.

Annex 138 COSMO-FLUX® 411-F Tec hnical Data Sheet,
Cosmoagro, June 2002.

Annex 139 Press item: “Ecuador accepts sprayings to
continue”, El Comercio (Ecuadorian newspaper),
Quito, 4 August 2004.

Annex 140 US Department of State, Bureau for International
Narcotics and Law Enforcement Affairs,
Memorandum of Justific ation Concerning the
Secretary of State’s 2002 Certification of
Conditions Related to the Aerial Eradication of
Illicit Coca in Colombia, Washington D.C., 2002.

Annex 141 United States Environmental Protection Agency
(EPA), Addendum to memorandum. Subject:
Description of Glyphosat e Use in the U.S. as a
Basis for Comparison to Glyphosate Use in
Colombia for Coca Eradication , From: Virginia

Werling and Timothy Kiely (EPA, BEAD
Biological and Economic Analysis Division)
to Jay Ellenberger (EPA, BEAD Biological and
Economic Analysis Division), 21 August 2002.

Annex 142 United States Environmental Protection Agency
(EPA), Details of the Consultation for
Department of State: Use of Pesticides for Coca
and Poppy Eradication Program in Colombia ,
August 2002.

Annex 143 US Department of State, Bureau for International
Narcotics and Law Enforcement Affairs,
Memorandum of Justific ation Concerning the
Secretary of State’s 2003 Certification of
Conditions Related to the Aerial Eradication of

Illicit Coca in Colombia, Washington D.C., 2003.
viAnnex 144 US Department of State, Bureau for International
Narcotics and Law Enforcement Affairs,
Memorandum of Justific ation Concerning the
Secretary of State’s 2004 Certification of

Conditions Related to the Aerial Eradication of
Illicit Coca in Colombia, Washington D.C., 2004.
Annex 145 Andean Regulation for the Control of chemical
substances used in the illegal manufacture of

narcotic drugs and ps ychotropic substances,
Andean Community, Decision 602 of 2004.
Annex 146 US Department of State, Bureau for International

Narcotics and Law Enforcement Affairs,
Memorandum of Justific ation Concerning the
Secretary of State’s 2005 Certification of
Conditions Related to the Aerial Eradication of
Illicit Coca in Colombia, Washington D.C., 2005.

Annex 147 US Department of State, Bureau for International
Narcotics and Law Enforcement Affairs,
Memorandum of Justific ation Concerning the
Secretary of State’s 2006 Certification of
Conditions Related to the Aerial Eradication of
Illicit Coca in Colombia, Washington D.C., 2006.

Annex 148 Press item: “Defence Minister welcomes the new
Super Tucano airplanes of the Air Force”,
14 December 2006.

Annex 149 US Department of State, Bureau for International
Narcotics and Law Enforcement Affairs,
Memorandum of Justific ation Concerning the
Secretary of State’s 2007 Certification of

Conditions Related to the Aerial Eradication of
Illicit Coca in Colombia, Washington D.C., 2007.
Annex 150 Press item: “Ecuador will sue Colombia over
sprayings in the border”, Revista Caminos, 2 July

2007.
Annex 151 Colombian Association of Autonomous Regional
Corporations (ASOCARS), “Shared

Responsibility: The World Drug Problem from a
vii Green Perspective”, Periódico Virtual, Issue
Nº 14, 2008.

Annex 152 Embassy of the United States in Bogotá, Fact
Sheet 2008, “Cocaine production and cultivation:
Colombia”, 6 November 2009, enclosure to Press
Item: “Official U.S. Colombia Survey Shows
Sharp Drop in Coca Cultivation and Cocaine
Production”.

Annex 153 Arias v. Dyncorp , Plaintiffs’ Motion to dismiss
Three Individual Plaintiffs, 23 December 2009.

Annex 154 Arias v. Dyncorp , Defendants’ Response,
6 January 2010.

Annex 155 Arias v. Dyncorp , --- F. Supp. 2d ---, 2010 WL
94563 (DDC, 2010, Roberts J).
Annex 156 Arias v. Dyncorp , Defendants’ Motion for

Sanctions Against the Aria s/Quinteros Plaintiffs
for Violations of Discovery Orders, 26 January
2010.

viii Annex 112

ANNUAL REPORT OF THINTER-AMERICAND RUGA BUSECONTROL COMMISSION
(CICAD)TO THEGENERAL ASSEMBLY OF THO RGANIZATION OAMERICAN
STATES AT I32NDREGULAR SESSIO,12-15 OVEMBER 2001,CARACAS,
VENEZUELA,3APRIL2002

(OEA/SER.L/XIV.2.30 CICAD/doc.1123/01 rev.1, pp. 7-8, 13)

1Annex 112

THIRTIETH REGULAR SESSION OEA/Ser.L/XIV.2.30

November 12-15, 2001 CICAD/doc.1123/01 rev.1
Caracas, Venezuela 3 April 2002
Original: Spanish

ANNUAL REPORT OF THE
INTER-AMERICAN DRUG ABUSE CONTROL COMMISSION (CICAD) TO THE
GENERAL ASSEMBLY OF THE ORGANIZATION OF AMERICAN STATES (OAS) AT

ITS THIRTY-SECOND REGULAR SESSION

2 Annex 112

Standards of Care in Drug Abuse Treatment

CICAD continues to help member states develop and put into practice standards of care
in drug treatment. This initiative, underway in CICAD since 1997, is designed to help
governments develop a consensus in the national health care community around the

desirability of establishing standards of care, and then to implement practices and
guidelines. A long-term objective, recommended by CICAD’s Expert Group on Demand
Reduction (March 1998, and again in August 2001), is to develop systems of
accreditation for treatment providers.

During 2001, CICAD organized and financed a workshop on the establishment of
standards of care in Uruguay. A Spanish-language manual documenting the different
experiences of each country in the implementation and adaptation of legislation was

published by the Executive Secretariat in October 2000. The English version, published
in July 2001, was used as a reference during the meeting of the CICAD Expert Group in
Demand Reduction held in August 2001 in Montego Bay, Jamaica.

Caribbean Sub-Regional Group of Demand Reduction Experts

CARICOM and CICAD cosponsored a Sub-Regional Demand Reduction Forum in
Georgetown, Guyana on September 17-18. The purpose of the meeting was to bring

together individuals from the Caribbean member states to discuss the findings of an
assessment relating to a regional demand reduction enhancement program, as well as to
develop a strategic approach for carrying out future activities and assessments. The
recommendations put forth at the forum were submitted to the Fifth Meeting of the Council

for Human and Social Development (COHSOD V) in October 2001, and [to a preparatory
meeting for a Caribbean Summit of Heads of Government in December 2001].

Support for the Consultative Group process in Ecuador

As part of its support for Ecuador’s Consultative Group process on drugs, the Executive
Secretariat provided technical assistance on the development of the demand reduction

project portfolio.

C. SUPPLY REDUCTION AND APPLICATION OF CONTROL MEASURES

Expert Group on Chemical (Pharmaceutical Products)

At CICAD XXVIII, the Delegation of Colombia raised concerns regarding the control of

pharmaceutical products. The Commission directed the Expert Group on Chemicals to
examine this issue. The Group met August 13 - 15 in Washington with representatives
from Argentina, Belize, Brazil, Chile, Colombia, Costa Rica, Ecuador, Guatemala,
Jamaica, Mexico, Peru, Dominican Republic, St. Kitts and Nevis, Trinidad and Tobago,

United States, Uruguay, and Venezuela.

The experts identified 7 general problem areas in controlling pharmaceuticals: the
existence and application of adequate legislation, national control systems, the

availability and timely exchange of information and the availability of sufficient financial,
technical and human resources. The Group developed 10 recommendations for
consideration by the Commission and 7 more that were directed to the member states.

3Annex 112

The report and recommendations were presented to the Commission by Dr. Maria

Cristina Chirolla of Colombia, the Chair of the Expert Group.

The Group presented its final report at the thirtieth regular session. As one of the
recommendations the group suggested the creation of a separate Experts Group on

Pharmaceutical Products. The Commission decided that the group will meet for the first
time In Washington DC, in March 2002, and will be chaired by Colombia.

Chemical Control Software

In response to requests by member states the Executive Secretariat continued its work
in the development of a uniform chemical control database designed to assist countries
in registering and reporting on precursor imports and exports, maintaining company

records and generating pre-export notifications. The software was designed by the
chemical control unit of the Ministry of Industry, Tourism, Integration and International
Commercial Negotiations (MITINCI) of Peru.

During the first quarter of 2001, MITINCI performed a number of modifications on the
software based on consultations with CICAD. These modifications created a general
platform that countries could adopt and integrate into their national control systems.
Each country may also perform modifications to the program in order to meet their

individual needs. In March 2001, CICAD and the DEA hosted a detailed training
seminar at MITINCI’s headquarters in Lima for representatives from Venezuela’s
national drug commission (CONACUID) and from the Ministry of Production and
Commerce (MPC). Argentina, Bolivia and Panama have also expressed formal interest

in acquiring the software and are currently undergoing the preparations
for installation.

Study of Maritime Drug Trafficking

In August 2001, the Secretariat conducted a maritime drug trafficking study in Colombia
within the framework of its Maritime Cooperation Strategy in the Southeastern Pacific. A
previous study was undertaken in Peru in 2000. The study in Colombia, which was
conducted in coordination with the Office of Naval Intelligence and the Joint Interagency

Task Force – East, focused on drug trafficking activities around coastal areas and
waters, including port facilities. It also examined systems, resources, procedures, and
capacity to monitor and interdict drug trafficking within the ports and adjacent to the
coastline. A final report published in December 2001 made a series of

recommendations to the Government of Colombia for consideration in its ongoing
national maritime counter drug strategy. A third study will be conducted in cooperation
with the Government of Chile in January 2002; the fourth and final study will focus on
Ecuador. A regional assessment will also be completed in 2002. The goal of the project

is to promote multinational coordination and cooperation.

Maritime Cooperation and Port Security Project

There is an increasing recognition of the role of the private sector in facilitating drug
control. This is particularly important in the case of commercial companies active in
airports and maritime ports. Governmental port authorities have traditionally been
responsible for the administration of maritime ports, but the trend is toward private sector

companies assuming these responsibilities.

4 Annex 112

(GIS) and a computerized image analysis system (IA), using commercial satellite
images, aerial photographs, and on-site verification to that end. The GLEAM tool makes

it possible to map and evaluate the environmental repercussions of land use, identifies
the potential for improvement or development of new infrastructure, evaluates and/or
proposes alternative land-use options by identifying the crops with the most potential to
bring economic development to a given area, and contributes to the construction of a

land-titling database

As a result of the project launched in 1998, in 2001 CICAD delivered the GLEAM project
to the Government of Peru, including a work station, field equipment, and training for

CONTRADROGAS personnel in the use of the system. With this technology the
government of Peru can effectively determine the viability of proposed projects as well
as develop new projects in production areas, and those with potential to be used for illicit
cultivation.

In Bolivia, the Vice Ministry for Alternative Development, through its Monitoring Office,
has been working, in cooperation with CICAD, to compile the necessary information to
implement GLEAM over 500,000 hectares of traditional coca production areas identified

by the government in the North and South Yungas region. This was done based on a
request from the Bolivian Government.

Alternative Development Projects in Indigenous Communities

CICAD, in conjunction with the Foundation ZIO-A'I “Unión de Sabidu
ría” and the National
Alternative Development Plan of Colombia (PNDA), seeks to develop the economy and
production component of the Life Plan (Plan de Vida) of the Cofán People and the

Indigenous Councils of Valle Guamuez and San Miguel, as a socioeconomic
development plan that offers an alternative to coca cultivation for these indigenous
communities in the Department of Putumayo, southern Colombia. The project was
designed by the communities through the Life Plan and it has become the instrument to

ensure the physical and cultural survival of these indigenous communities and to bring
social and economic development to their peoples and to the region they inhabit, by
seeking, at the same time, to ensure licit alternative development and eradication of
coca cultivation from their lands. The purpose of this project is to strengthen the culture,

organizations, businesses, and economic development of indigenous communities;
identify lands suitable for farming and lands that require environmental restoration;
implement a crop, livestock, and animal breeding program, as well as sustainable
production systems to enable the recovery of traditional crops in order to provide food

security and produce marketable surpluses. The overall objective is to create a strong
and sustainable licit economy.

Support for Regional Initiatives

CICAD has been providing financing and participating as technical adviser, along with
other Inter-American and International agencies such as the Inter-American Institute for
Cooperation in Agriculture (IICA), to the Andean Committee for Alternative Development

(CADA). Through this consultation and coordination forum for multilateral dialogue and
decision making, the Andean countries exchange information and experiences to
develop and apply a regional strategy to enable them to implement joint measures,
strengthen bargaining capacity, and promote marketing of alternative products, taking

into consideration all sectors of society and agreements among governments. This

56 Annex 113

2004M EMORANDUM OF U NDERSTANDING BETWEEN THEGOVERNMENT OF

COLOMBIA AND THEO RGANIZATION OA MERICAN STATES(OAS)FOR THE
E XECUTION OF THSTUDY ON THEEFFECTS OF THP ROGRAM FOR THE
ERADICATION OFILLICIC ROPS BYAERIALSPRAYING WITHG LYPHOSATE
H ERBICID(PECIG) AND OFILLICITCROPS,ONH UMAN HEALTH AND THE

ENVIRONMENT ,4 EBRUARY 2004

(Archives of the Ministry of Foreign Affairs of Colombia)

MEMORANDUM OF UNDERSTANDING BETWEEN THE GOVERNMENT OF
COLOMBIA AND THE ORGANIZATION OF AMERICAN STATES (OAS) FOR
THE EXECUTION OF THE STUDY ON THE EFFECTS OF THE PROGRAM FOR
THE ERADICATION OF ILLICIT CROPS BY AERIAL SPRAYING WITH

GLYPHOSATE HERBICIDE (PECIG) AND OF ILLICIT CROPS, ON HUMAN
HEALTH AND THE ENVIRONMENT

[PAGE 2]

The Parties to this Memorandum of Understanding, the Government of
Colombia through the Ministry of Foreign Affairs, represented by the Minister of
Foreign Affairs, Carolina Barco, and the General Secretariat of the Organization of

American States (“SG/OEA”), through the Inter-American Drug Abuse Control
Commission (hereinafter, CICAD), represented by its Assistant Executive Secretary,
Abraham Stein, sign the following Memorandum of Understanding:

CONSIDERING

That the General Secretariat of the Organization of American States (hereafter,

“SG/OAS”), is the main and permanent organ of the Organization of American States
(hereafter, “OAS”), and is authorized to establish and promote relations of cooperation
with member States pursuant to Article 112(h) of the OAS Charter and with its General
Assembly resolution AG/RES. 57 (l-O/71).

That CICAD is an agency of the Organization of American States, established
by Article 52 of the OAS Charter. This agency is technically autonomous and carries
out its duties within the context and scope of the Rio de Janeiro Action Plan against

Consumption, Production, and Illicit Trafficking on Drugs and Psychotropic

7Annex 113

Substances, the mandates of the General Assembly, and the decisions internally adopted
by CICAD.

That the purpose of CICAD is to contribute to eliminate illicit trafficking and

drug abuse. Pursuant to its Statutes, it has attributions with regard to the field of
prevention, assistance and social rehabilitation of drug-addicts, as well as to that of the
prevention, control and punishment of the production and illicit trafficking of drugs and
psychotropic substances.

That within the framework of its Hemispheric Strategy, CICAD promotes
actions against the illicit crops of raw materials destined for the production of illicit

drugs, while always taking into account the preservation of the environment, through
the promotion of programs and/or projects to encourage the development of lawful
economies in the areas of illicit drug production in Member States.

That the Program for the Eradication of Illicit Crops by Aerial Spraying with
Glyphosate Herbicide (PECIG) was implemented in accordance with paragraph g) of
Article 91 of Law 20 of 1986, whereby Colombia adopted the National Anti-Narcotics

Statute that assigns to the National Narcotics Council the duty to “provide for the
destruction of marihuana, coca and other crops from which substances causing
dependency may be extracted, using the most adequate means, following a favourable
opinion of the agencies entrusted with protecting the health of the population and the
preservation and balance of the ecosystem in the country”. [The Program] is regulated
through resolution 0013 of 2003 and operates in all the regions in the country the
presence of illicit crops is evidenced.

That for the Colombian State, the adoption and implementation of the Program for
the Eradication of Illicit Crops by Aerial Spraying with Glyphosate Herbicide (PECIG)
has become an inexorable necessity in view of the fact of the extended presence of
illicit crops in the national territory and the security problems that, in many cases,
preclude resort to other eradication methods.

That the Government of Colombia understands the PECIG as the plan of the

State for the mitigation of the adverse environmental impact caused by illicit crops and
the subsequent processing of illicit drugs.

[PAGE 3]

That in view of the growing domestic and international concern as to the alleged
effects of the Program for the Eradication of Illicit Crops by Aerial Spraying with

Glyphosate Herbicide (PECIG), the Governments of Colombia and the United States of
America, based on the existing cooperation on the matter between both nations,

8Substances, the mandates of the General Assembly, and the decisions internally adopted
by CICAD.

That the purpose of CICAD is to contribute to eliminate illicit trafficking and

That within the framework of its Hemispheric Strategy, CICAD promotes
actions against the illicit crops of raw materials destined for the production of illicit

That the Government of Colombia understands the PECIG as the plan of the

State for the mitigation of the adverse environmental impact caused by illicit crops and
the subsequent processing of illicit drugs.

[PAGE 3]

That in view of the growing domestic and international concern as to the alleged
effects of the Program for the Eradication of Illicit Crops by Aerial Spraying with

Glyphosate Herbicide (PECIG), the Governments of Colombia and the United States of
America, based on the existing cooperation on the matter between both nations,Annex 113

Eradication of Illicit Crops by Aerial Spraying with Glyphosate Herbicide (PECIG) and
of illicit crops, on human health and the environment”.

SECOND CLAUSE: Framework for cooperation

Cooperation and assistance provided in pursuance of the present project will be
carried out in observance of the respect for national sovereignty, confidentiality,
transparency and veracity of conclusions.

[PAGE 4]

THIRD CLAUSE: Study areas

The study will focus on the areas where the Program for the Eradication of Illicit
Crops by Aerial Spraying with Glyphosate Herbicide (PECIG) is implemented and, by
mutual agreement between the Parties, other areas the assessment of which is

considered relevant may be included.

FOURTH CLAUSE: Responsibilities of the Parties

A. CICAD undertakes to:
1. Supervise and follow-up on the works carried out by the Scientific

Assessment Team (SAT) and the Permanent Technical Group for Mobile
Monitoring (PTGMM [shortened form PTG]).
2. Contract, by mutual agreement with the Colombian Government, and
supervise the required personnel for conducting the study that is the object of
the present Memorandum of Understanding.
3. Conduct and follow-up on the study that is the object of the present

Memorandum of Understanding.
4. Coordinate and supervise the adequate progress of the activities foreseen in
the Proposal for Monitoring of the Aerial Spraying Program in Colombia.
5. Provide the funds for conducting the corresponding activities in accordance
with the established Schedule of Activities. Therefore, the project will not
entail any disbursements from the Colombian treasury.

6. Review and approve periodical reports on the progress of the established
work plan.
7. Periodically inform the Government of Colombia on the progress of the
completion of the study that is the object of the present Memorandum of
Understanding.
8. Publicly present the results of the study and widely publicize the

corresponding final report that will have been previously presented to the
Government of Colombia for its information. The results of the study and

10Eradication of Illicit Crops by Aerial Spraying with Glyphosate Herbicide (PECIG) and
of illicit crops, on human health and the environment”.

SECOND CLAUSE: Framework for cooperation

[PAGE 4]

THIRD CLAUSE: Study areas

FOURTH CLAUSE: Responsibilities of the Parties

A. CICAD undertakes to:
1. Supervise and follow-up on the works carried out by the Scientific
Assessment Team (SAT) and the Permanent Technical Group for Mobile
Monitoring (PTGMM [shortened form PTG]).
2. Contract, by mutual agreement with the Colombian Government, and

supervise the required personnel for conducting the study that is the object of
the present Memorandum of Understanding.
3. Conduct and follow-up on the study that is the object of the present
Memorandum of Understanding.
4. Coordinate and supervise the adequate progress of the activities foreseen in
the Proposal for Monitoring of the Aerial Spraying Program in Colombia.
5. Provide the funds for conducting the corresponding activities in accordance
with the established Schedule of Activities. Therefore, the project will not

entail any disbursements from the Colombian treasury.
6. Review and approve periodical reports on the progress of the established
work plan.
7. Periodically inform the Government of Colombia on the progress of the
completion of the study that is the object of the present Memorandum of
Understanding.
8. Publicly present the results of the study and widely publicize the

corresponding final report that will have been previously presented to the
Government of Colombia for its information. The results of the study andAnnex 113

The Parties to this Memorandum of Understanding undertake to preserve the
strictest confidentiality while the study is being developed. Neither Party may, without

the express prior consent of the other, publish partial results of the study under way.

Once the Parties have learned, under reserve, the results of the study, the final
report will be made public and will be widely publicized.

SEVENTH CLAUSE: Termination

The present Memorandum of Understanding may be terminated by mutual
agreement or by either Party, through written advance notice of at least three months to
the other.

EIGHTH CLAUSE: Settlement of disputes

The Parties undertake to settle controversies that may arise of the interpretation
or application of the present Memorandum of Understanding, preferably by mutual
agreement. In case a satisfactory solution is not reached, recourse will be had to the
exceptional arbitration procedure mutually agreed by the CICAD and the Government
of Colombia. If there is no agreement on the procedure, arbitration will be conducted
pursuant to UNCITRAL Rules. The arbitral tribunal constituted in accordance with

those Rules will rule as amiable mediator or ex aequo et bono and its decision will be
final and binding.

None of the provisions in this Memorandum of Understanding signifies or shall
be construed as a relinquishment of the privileges and immunities enjoyed by the
Parties in accordance with international law and practice.

[PAGE 6]

NINTH CLAUSE: Entry into Force, Duration and Amendments

The present Memorandum of Understanding shall enter into force on the date of

its signature and shall be in force for a year that may be extended by mutual agreement
between the Parties.

Addition or amendment to this Memorandum of Understanding will be made by
mutual agreement between the Parties, following compliance with legal requirements.
The instruments registering those modifications will be appended as annexes to the

present Memorandum of Understanding and shall become part thereof.

12 The Parties to this Memorandum of Understanding undertake to preserve the
strictest confidentiality while the study is being developed. Neither Party may, without

the express prior consent of the other, publish partial results of the study under way.

Once the Parties have learned, under reserve, the results of the study, the final
report will be made public and will be widely publicized.

SEVENTH CLAUSE: Termination

The present Memorandum of Understanding may be terminated by mutual
agreement or by either Party, through written advance notice of at least three months to
the other.

EIGHTH CLAUSE: Settlement of disputes

those Rules will rule as amiable mediator or ex aequo et bono and its decision will be
final and binding.

[PAGE 6]

NINTH CLAUSE: Entry into Force, Duration and Amendments

The present Memorandum of Understanding shall enter into force on the date of

its signature and shall be in force for a year that may be extended by mutual agreement
between the Parties.

present Memorandum of Understanding and shall become part thereof.Annex 113

Aerial spraying began with marijuana crops in the late 70’s, then in 1991 poppy crops
started to be sprayed and, lastly, in 1994, aerial spraying of coca began. Over time, the
issue of aerial spraying as an element of the strategy for reducing crops has polarized

the international community. The use of chemical herbicides such as glyphosate has
provoked several verbal reproaches and severe criticism on behalf of peasants, peasant
organizations and environmental watch groups.

In accordance with Resolution 012 of 2003, the Program for the Eradication of Illicit
Crops by Aerial Spraying with Glyphosate Herbicide is to have an Environmental

Management Plan, a provision that was regulated by the Ministry for the Environment,
Housing and Territorial Development, through Resolution 1054 of 30 September 2003.

This Plan has not assuaged the doubts of the opponents to aerial spraying, wherefore an
independent study on such effects is necessary. The results of the studies conducted to
date have not provided solid evidence and have been perceived as subjective and
unilateral studies. In sum, the controversy continues and it is necessary to have a

publicly known study that provides certain results on the matter.

[…]

In light of the growing domestic and international concern as to the environmental and
health effects of the Program for the Eradication of Illicit Crops by Aerial Spraying with

Glyphosate Herbicide that is conducted in Colombia, the Governments of Colombia and
the United States requested the collaboration of CICAD in order to conduct a study
documenting such effects.

[PAGE 8]

To that effect, it is deemed necessary to set out on an independent, objective and
impartial assessment of the current aerial eradication program with the purpose of
providing verifiable and empirical scientific research. This proposal is an attempt to
establish the framework within which to carry out this work, as well as a response to the
need to produce an assessment that is scientifically unquestionable in general terms and
convincing in the way in which it is publicly perceived.

OBJECTIVES

The objectives of this program are the following:

1) To conduct a scientific study of the aerial spraying program in Colombia, that is

notoriously independent and of high quality, on the essential impacts of
glyphosate spraying on individuals, fauna, flora and the environment.

14Aerial spraying began with marijuana crops in the late 70’s, then in 1991 poppy crops
started to be sprayed and, lastly, in 1994, aerial spraying of coca began. Over time, the
issue of aerial spraying as an element of the strategy for reducing crops has polarized
the international community. The use of chemical herbicides such as glyphosate has

provoked several verbal reproaches and severe criticism on behalf of peasants, peasant
organizations and environmental watch groups.

In accordance with Resolution 012 of 2003, the Program for the Eradication of Illicit
Crops by Aerial Spraying with Glyphosate Herbicide is to have an Environmental
Management Plan, a provision that was regulated by the Ministry for the Environment,
Housing and Territorial Development, through Resolution 1054 of 30 September 2003.

[…]

In light of the growing domestic and international concern as to the environmental and
health effects of the Program for the Eradication of Illicit Crops by Aerial Spraying with
Glyphosate Herbicide that is conducted in Colombia, the Governments of Colombia and
the United States requested the collaboration of CICAD in order to conduct a study
documenting such effects.

[PAGE 8]

convincing in the way in which it is publicly perceived.

OBJECTIVES

The objectives of this program are the following:

1) To conduct a scientific study of the aerial spraying program in Colombia, that is

notoriously independent and of high quality, on the essential impacts of
glyphosate spraying on individuals, fauna, flora and the environment.Annex 113

to the experience of international research institutions, such as the Tropical Agronomy
Centre for Research and Teaching (CATIE), the International Atomic Energy Agency
(IAEA), the International CAB and the International Centre for Tropical Agriculture

(CIAT), as well as to leading international universities in the field. Most of [the team’s]
work will be conducted from their current locations, through computer or telephone
links, or by joint work meetings at a convenient location. Once the assessment has been
completed and submitted, the team will be constantly at hand in order to reply to the
different queries that may arise.

The Permanent Technical Group for Mobile Monitoring (PTGMM): This group will be
in charge of daily operations once the Scientific Assessment Team (SAT) has
completed the initial review and operational design. The main work of the PTGMM is
to compile and analyse data, according to SAT’s instructions on specific project
requirements, as well as the occasional complaints or controversies. The PTGMM will
be made up of third-country and/or Colombian nationals, under the coordination of a
Colombian technician. Personnel selected will submit to the corresponding security

studies. The PTGMM will have its seat in Bogotá and be permanently available to
travel around the country.

ACTIVITIES

The initial period for the execution of the program will be a year. During this period,

CICAD proposes that the team should complete its work on the impacts of aerial
spraying and its protocol, and with the information provided by the PTGMM, activities
aimed at random monitoring and constant monitoring of aerial spraying activities be
carried out.

Likewise, the investigation on the issues relating to the environmental impact of

herbicides and fungicides used in the production of illicit crops will be conducted.

The activities foreseen in the project are listed hereafter…

16 Annex 114

ANNUAL REPORT OF THINTER-AMERICAND RUGA BUSECONTROL COMMISSION
(CICAD)TO THEGENERAL ASSEMBLY OF THO RGANIZATION OAMERICAN
STATES AT I34THR EGULARSESSIO,17-20 OVEMBER 2003,MONTREAL,
CANADA,3M AY2004

(OEA/Ser.L/XIV.2.34 CICAD/doc.1264/03 rev. 2, p. 26)

17Annex 114

ORGANIZATION OF AMERICAN STATES

INTER-AMERICAN DRUG ABUSE CONTROL COMMISSION

THIRTY-FOURTH REGULAR SESSION OEA/Ser.L/XIV.2.34
November 17-20, 2003 CICAD/doc.1264/03 rev. 2
Montreal, Canada 3 May 2004
Original: English

ANNUAL REPORT OF THE

INTER-AMERICAN DRUG ABUSE CONTROL COMMISSION (CICAD) TO THE
GENERAL ASSEMBLY OF THE ORGANIZATION OF AMERICAN STATES
AT ITS THIRTY-FOURTH REGULAR SESSION

GENERAL SECRETARIAT OF THE ORGANIZATION OF AMERICAN STATES, WASHINGTON, D.C. 20006

18 Annex 114

consensus on the nature of and responsibilities for the development of an initial system that can
be tested among all participating university sites. Researchers have visited specific ports of
entry in both the Dominican Republic and Belize to familiarize themselves with the on-the-
ground reality of each country so that they can produce a prototype appropriate for field

deployment by national immigration and other governmental agencies. A second technical
meeting, held in September 2003 at Carnegie Mellon University in Pittsburgh, served to
integrate the separate research components of the project being conducted by the participating
universities. This meeting prepared the ground for a technical meeting and initial prototype field

test in Belize in December 2003.

PROJECT ON INSTITUTION-BUILDING IN ANDEAN NATIONAL DRUG COMMISSIONS

The main objective of this project, financed by the Spanish Government, is to strengthen the

national drug commissions of the Andean countries, which will make possible the development
of national drug information and research systems (national observatories), which are the basic
tools for strategic planning of national drug policies. The project has continued to move forward
despite the region's social and political instability and constant changes of officials and technical
staffs of the national institutions involved.

• Bolivia
The project seeks to incorporate national demand reduction data into the existing National
Drug Information System (SINALTID). An early 2003 coordination mission introduced new

data sources into the system (Vice Ministry of Prevention and Rehabilitation, Ministry of
Education, Ministry of Health, and National Statistics Institute, as well as NGOs, universities,
and research institutes), to identify the equipment needed by participating institutions, and to
define the process for purchasing it. Delivery will take place once the Bolivian Government
makes staffing decisions and selects a locale for the Observatory's technical team. A

development workshop f or the Bol ivian Observatory is scheduled for the near future to
define a national map of stable indicators along the lines of the SIDUC drug use and
CICDAT supply control systems of the Inter-American Observatory on Drugs, pending a
positive reply from the Bolivian Government.

• Colombia
Under this project, support is being provided to strengthen the Colombian Drug Observatory,
which was organized by the National Drug Council around SIDCO (Drug Information System
of Colombia). The project will enable SIDCO to be updated and improved and to have an

Internet site. Proposals are also being considered for the development of a national
epidemiological surveillance system, put together by the Social Protection Ministry’s Office
of Public Health.

• Ecuador
Throughout 2002, the project consolidated the development of the Ecuadorian Drug
Observatory. In 2003, CICAD has carefully followed the Observatory's progress, primarily
because of senior management position changes in the National Drug Commission

(CONSEP).

• Peru
Following the definition of the Peruvian Drug Observatory’s work plan at a workshop held in
November 2002, the process was finalized for procurement and delivery of computer

equipment to national institutions participating in the Observatory. Moreover, the technical

1920 Annex 115

35TH REGULAR SESSION OFCICAD,W ASHINGTON,D.C.,27-30 APRIL2004,SESSION
DOCUMENTS

(Available at: http://www.cicad.oas.org/apps/sessions.aspx?lang=ENG&ie=us007
(last visited 7 March 2010))

21Annex 115

OAS/CICAD: CICAD Sessions Page 1 of 2

THIRTY-FIFTH REGULAR SESSION OF CICAD
Washington, D.C. United States
April 27 - 30, 2004

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http://www.cicad.oas.org/apps/sessions.aspx?lang=ENG&ie=us007 27/02/2010

22 Annex 115

OAS/CICAD: CICAD Sessions Page 2 of 2

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http://www.cicad.oas.org/apps/sessions.aspx?lang=ENG&ie=us007 27/02/2010

2324 Annex 116

CICAD, K.R.SOLOMON ET A. “NVIRONMENTAL AND HUMAN HEALTH
ASSESSMENT OF THA ERIALSPRAYPROGRAM FOR COCA ANDPOPPY CONTROL IN
C OLOMBIA”, OAS,WASHINGTON, D.C.,31ARCH 2005,(CICADI)

(Available at: http://www.cicad.oas.org/en/glifosateFinalReport.pdf
(last visited 10 March 2010))

25Annex 116

ENVIRONMENTAL AND HUMAN HEALTH

ASSESSMENT OF THE AERIAL SPRAY

PROGRAM FOR COCA AND POPPY

CONTROL IN COLOMBIA

Dr. Keith R Solomon
Centre for Toxicology and Department of Environmental Biology,
University of Guelph, Guelph, ON, N1G 2W1, Canada

Dr. Arturo Anadón
Departamento de Toxicología y Farmacología
Facultad de Veterinaria,

Universidad Complutense de Madrid
Avda. Puerta de Hierro, s/n
Madrid 28040
Spain

Dr. Antonio Luiz Cerdeira
EMBRAPA, Ministry of Agriculture
Jaguariuna, SP 13820-000, Brazil

Dr. Jon Marshall
Marshall Agroecology Limited
2 Nut Tree Cottages, Barton, Winscombe,

Somerset, BS25 1DU, UK
Dr. Luz-Helena Sanin
Department of Public Health Sciences,
Faculty of Medicine, University of Toronto, Toronto, ON, M5S 1A8, Canada,
Autonomous University of Chihuahua,

and National Institute of Public Health, Mexico

A report prepared for the Inter-American Drug Abuse Control Commission
(CICAD) section of the Organization of American States (OAS)

Washington, DC, USA

March 31, 2005

Page 1 of 121

26 Annex 116

PREFACE
This report was prepared for the Inter-American Drug Abuse Control Commission
(CICAD) section of the Organization of American States (OAS) in response to requests

from the Governments of Colombia, the United Kingdom, and the United States of
America. The request was to conduct a science-based risk assessment of the human
health and environmental effects of the herbicide, glyphosate, used for the control of the
illicit crops, coca and poppy in Colombia.

The initial step in the process was to establish an international panel of experts in
human and environmental toxicology, in epidemiology, in agronomic practices, and in
ecology (SAT). Because both Colombia and the United States were actively involved in
the program for eradication of illicit crops, members of the panel were specifically
selected from other countries.

Initially, the panel met to formulate a framework to conduct this risk assessment.
The framework was based on those commonly used for risk assessment in a number of
jurisdictions and consisted of a problem formulation, characterization of the human
health and environmental effects of the substances used in the eradication program,

characterization of human and environmental exposures, and the drawing together of
these in a risk characterization. During this process, extensive use was made of the
scientific and other literature but, where data gaps and uncertainties related to the
specific uses in Colombia were identified, studies were initiated to assemble additional
data for use in the risk assessment. Some of these studies were carried out in

Colombia. The Colombian team (PTG) were contracted specifically to CICAD and
worked under the direction of the SAT to collect data in the Colombian Environment.
During the conduct of our study, members of the SAT made a number of visits to
Colombia to view, at first hand, all aspects of the program, to gather local information
and data, and to oversee the local studies of the PTG.

We recognize that the illicit crop eradication program in Colombia has generated
considerable local and international interest and is the subject of intense debate for
political, social, and other reasons. We have specifically excluded all social, political,

and economic issues from our study and the final report is strictly based in science and
scientifically based arguments. We believe that the report of the study and its scientific
recommendations will be useful in decision making to protect human health and the
environment.

After the initiation of this project, additional information on other substances used
in the production of coca and poppy and the refining of cocaine and heroin was
requested. This request culminated in two separate detailed reports, a Tier-1 and Tier-2
hazard assessment of 67 and 20 substances used for these purposes, respectively.
These substances are briefly discussed in the Problem Formulation of this report. We

believe that these reports will be useful in comparative hazard assessment and in risk
management decision making.

Page 2 of 121

27Annex 116

ACKNOWLEDGEMENTS
With an international panel of experts and activities in several countries, a study
of this nature requires good co-ordination and organization. We are deeply indebted to

Mr. Jorge Rios and Ms. Adriana Henao of the CICAD office for their excellent work in
organizing meetings, teleconferences, and field trips. They served the Panel well and
frequently worked well beyond the call of duty. We are also very grateful for the
contributions of the Colombian Field Team, the PTG. Unfortunately, we cannot name
these individuals; however, we extend our most grateful thanks to all of you for all the

hard work and the personal risks that you took on behalf of data collection for this
project.
Field visits to Colombia by members of the SAT were facilitated and coordinated
by the staff of the Ministry of Foreign Affairs and the team was afforded protection by

the National Police (Antinarcoticos). We offer our grateful thanks to Brigadier General
Luis Gómez, his staff, the pilots, technicians, and the “Junglas” commandos for aiding
us in our observations and sampling and for tolerating our scientific curiosity in the face
of other priorities. At all times, we were given free and unfettered access to information,
we were allowed to take photographs freely, and we were always treated with respect

and in a most professional manner.
The SAT members are indebted to Drs. Lesbia Smith, Angus Crossan, Richard
Brain, and also to the many students in the Toxicology Program at the University of
Guelph for their work on the separate reports on Tier-1 and Tier-2 hazard assessment

of other substances used in the production and refining of cocaine and heroin. These
data are presented in separate reports.

Page 3 of 121

28 Annex 116

TABLE OF CONTENTS
1 INTRODUCTION................................................................................................13

1.1 BACKGROUND.............................................................................................13
1.2 IMPACTS OF ILLICIT DRUG PRODUCTION IN COLOMBIA.......................14

1.3 THE PROGRAM TO CONTROL ILLICIT DRUG PRODUCTION AND
DISTRIBUTION IN COLOMBIA.....................................................................17
2 PROBLEM FORMULATION...............................................................................19

2.1 STRESSOR CHARACTERIZATION .............................................................19
2.1.1 Glyphosate...............................................................................................20

2.1.1.1 Structure and chemical properties...................................................20
2.1.1.2 Mechanism of action of glyphosate .................................................21

2.1.1.3 Global and local registration and use ..............................................21
2.1.1.4 Environmental fate...........................................................................22
2.1.2 Formulants and adjuvants........................................................................23

2.1.2.1 Surfactants in the glyphosate formulation........................................24
2.1.2.2 Cosmoflux 411F ..............................................................................24

2.1.3 Coca and poppy control programs ...........................................................24
2.1.3.1 Receiving environment....................................................................25

2.1.3.2 Method of application ......................................................................25
2.1.3.3 Frequency of application .................................................................30

2.1.3.4 Exposure pathways in soil, air, water, and other media...................32
2.1.3.5 Off-target deposition........................................................................32

2.2 Framework for risk assessment.....................................................................33
2.2.1 Context of the risks...................................................................................34
2.2.1.1 Human health risks..........................................................................34

2.2.1.2 Ecological risks................................................................................34
2.2.2 Conceptual model ....................................................................................35

2.2.3 Risk hypotheses.......................................................................................36
3 EXPOSURE CHARACTERIZATION ..................................................................37

3.1.1 Human exposure groups.......................................................................... 37
3.1.2 Applicator exposure..................................................................................37

3.1.3 Bystander exposure .................................................................................38
3.1.3.1 Bystanders directly over-sprayed ....................................................38

3.1.3.2 Re-entry...........................................................................................40
3.1.3.3 Inhalation.........................................................................................40
3.1.3.4 Dietary and drinking water...............................................................41

3.1.4 Environmental exposures.........................................................................42
3.1.4.1 Air....................................................................................................42

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29Annex 116

3.1.4.2 Water...............................................................................................42

3.1.4.3 Soil ..................................................................................................47
4 EFFECTS CHARACTERIZATION......................................................................49

4.1 GLYPHOSATE..............................................................................................49
4.1.1 Effects of glyphosate on mammals...........................................................49

4.1.1.1 Laboratory toxicity studies...............................................................49
4.1.1.2 Cases of human poisoning..............................................................52

4.1.1.3 Human epidemiology studies...........................................................53
4.1.2 Human health epidemiology study in Colombia........................................56
4.1.3 Effects of glyphosate in non-target organisms in the environment. ..........61

4.1.3.1 Effects in non-target terrestrial animals ...........................................61
4.1.3.2 Effects in aquatic animals................................................................65

4.1.3.3 Effects of glyphosate on plants........................................................70
4.2 SURFACTANTS............................................................................................71

4.2.1 Effects on glyphosate and Cosmo-Flux® on non-target aquatic organisms
72
4.2.2 Effects of glyphosate and Cosmo-Flux® on mammals.............................72

4.2.2.1 Analysis of the formulation...............................................................72
4.2.2.2 Acute oral toxicity ............................................................................73

4.2.2.3 Acute Inhalation toxicity...................................................................74
4.2.2.4 Acute dermal toxicity .......................................................................75

4.2.2.5 Skin irritation....................................................................................75
4.2.2.6 Eye irritation.....................................................................................76

4.2.2.7 Skin sensitization.............................................................................77
4.2.2.8 General conclusions on the mammalian acute toxicity of glyphosate
and Cosmo-Flux® ...............................................................................78

4.3 EFFECTS IN THE FIELD ..............................................................................78
4.3.1 Duration of effects in the field...................................................................78

4.3.1.1 Forest clearance and soils...............................................................79
4.3.1.2 Effects on associated fauna.............................................................79

4.3.1.3 Interactions with surfactants............................................................80
4.3.2 Recovery from effects...............................................................................81
4.3.2.1 Principles.........................................................................................81

4.3.2.2 Tropical situations............................................................................81
4.3.2.3 Temperate situations.......................................................................82

4.3.2.4 Conclusions.....................................................................................83
5 RISK ASSESSMENT..........................................................................................85

5.1 HUMAN HEALTH..........................................................................................85
5.2 ENVIRONMENT............................................................................................86

Page 5 of 121

30 Annex 116

6 CONCLUSIONS.................................................................................................90

6.1 HUMAN HEALTH RELEVANCE....................................................................90
6.2 ECOLOGICAL RELEVANCE ........................................................................90

6.3 STRENGTHS AND UNCERTAINTIES IN THE ASSESSMENT....................91
6.3.1 Exposures ................................................................................................91

6.3.1.1 Environmental exposures................................................................91
6.3.1.2 Human exposures ...........................................................................92

6.3.2 Effects ......................................................................................................92
6.3.2.1 Environmental effects......................................................................92

6.3.2.2 Effects in humans............................................................................93
6.3.3 Confounding risks.....................................................................................93

6.4 RECOMMENDATIONS .................................................................................94
7 REFERENCES...................................................................................................97

8 GLOSSARY......................................................................................................118

Page 6 of 121

31Annex 116

LIST OF TABLES
Table 1. Pesticides used in the production of coca ..................................................... 15
Table 2. Identity and amounts of substances seized in Colombia as a result of

counter-drug operations .................................................................................... 16
Table 3. Use glyphosate in eradication spraying in Colombia 2000 to 2004............... 21
Table 4. Application rates of glyphosate and Cosmo-Flux® for control of coca and
poppy................................................................................................................. 30
Table 5. Estimates of areas affected by off-target deposition of glyphosate in the

spraying of coca in Colombia............................................................................. 33
Table 6. Protective measures used to reduce exposure of applicators to glyphosate
and formulants as used in poppy and coca eradication programs in Colombia. 37
Table 7. Estimates of human exposure to glyphosate during a spray application....... 39
Table 8. Estimates of human exposure to glyphosate during re-entry to treated fields40

Table 9. Worst-case daily human exposure estimates for glyphosate (mg/kg/day)..... 41
Table 10. Estimates of concentrations of glyphosate in surface water after a spray
application ......................................................................................................... 42
Table 11. Summary data on glyphosate concentration in Midwestern US streams..... 44
Table 12. Characteristics of sampling sites for glyphosate, AMPA and other

pesticides in surface waters and sediments in regions of Colombia.................. 45
Table 13. Concentrations of glyphosate (AE) and AMPA in samples of surface water
collected in Colombia between October 2004 and March 2005 ........................ 46
Table 14. Concentrations of other pesticides in samples of surface water and
sediments taken in Colombia between October 2004 and March 2005............. 47

Table 15. Estimates of glyphosate concentration in the top 25 mm of soil following a
spray application................................................................................................ 48
Table 16. Acute toxicity of glyphosate in selected mammals ...................................... 49
Table 17. Characteristics of the areas used in the epidemiology study....................... 56
Table 18. Causes of fecundability adjusted for the relationship between time to

preganacy (TTP) and region based on an alternative model............................. 60
Table 19. Toxicity values obtained from toxicity tests conducted on a mixture of
glyphosate and Cosmo-Flux®. .......................................................................... 69
Table 20. Summary of reasonable worst-case estimated exposures of humans to
glyphosate resulting from use in the eradication of coca and poppy in

Colombia and margins of exposure................................................................... 86
Table 21. Recommendations for the continuance of current practices in the coca and
poppy eradication program in Colombia............................................................ 94
Table 22. Recommendations for the collection of new data and information in the
coca and poppy eradication program in Colombia............................................. 95

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32 Annex 116

LIST OF FIGURES
Figure 1 Coca plant.....................................................13Error! Bookmark not defined.

Figure 2 Diagrammatic representation of potential impacts of coca production,
refining, and spraying.............................................................................................19
Figure 3 The structure of glyphosate and its major metabolic and breakdown

products.
Figure 4 Binding of glyphosate to soil particles ............................................................20
Figure 5 Penetration of an herbicide such as glyphosate through plant cuticular

waxes in the absence (left) and presence of surfactants (right).............................24
Figure 6 Map showing production of coca in Colombia in 2005. ..................................26

Figure 7 Map showing areas of poppy production in 2005...........................................27
Figure 8 Areas planted with coca and poppy in Colombia from 1994 to 2002 as ha
(above) and as a percent of the total land area of Colombia (below). ...................28

Figure 9 Map showing the region of Colombia identified as part of the Andean
Biodiversity Region. ...............................................................................................29
Figure 10 Photographs of aspects of the spray operation............................................31

Figure 1 Diagram showing exposure routes for various environmental compartments
when glyphosate is used for the control of illicit crops............................................32
Figure 22 Potential human health impacts of the cycle of coca or poppy production

Scores for eradication spraying are specifically omitted.........................................34
Figure 13 Potential environmental impacts of the cycle of coca or poppy production.
Scores for eradication spraying are specifically omitted.........................................35

Figure 14 Illustration of human exposure scenarios.....................................................39
Figure 35 Photograph of coca plants near Caucasia, Colombia, replanted from
cuttings in a field sprayed with glyphosate 56 days previously ..............................48

Figure 46 Summary of the results of the time to pregnancy study. ..............................60
Figure 57 Distribution of toxicity values for glyphosate technical, formulated
glyphosate (Roundup®) in all aquatic organisms and in fish and the glyphosate

and Cosmo-Flux® 411 mixture as used in Colombia.............................................70
Figure 68 Illustration of acute toxicity values in laboratory mammals for glyphosate
plus Cosmo-Flux®, the NOEL from the most sensitive chronic study in

laboratory animals, and the RfD (glyphosate) and the estimated worst-case
acute exposures that may be experiences under conditions of use in Colombia...87
Figure 79 Distribution of toxicity values for glyphosate technical, formulated
glyphosate (Roundup®) in all aquatic organisms and in fish and the toxicity

values in four aquatic species for glyphosate and Cosmo-Flux® 411 mixture as
used in Colombia. ..................................................................................................88
Figure 20 Potential human health impacts of the cycle of coca or poppy production
and the spray eradication program. .......................................................................90

Figure 21 Potential environmental impacts of the cycle of coca or poppy production
and the spray eradication program ........................................................................91

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EXECUTIVE SUMMARY
This report was prepared for the Inter-American Drug Abuse Control Commission
(CICAD) section of the Organization of American States (OAS) in response to requests

from the Governments of Colombia, the United States, and the United Kingdom. The
request was to conduct a science-based risk assessment of the human health and
environmental effects of the use of glyphosate for the control of the illicit crops, coca
and poppy, in Colombia. This became the purpose of the study, which was conducted
in a number of steps.

The initial step in the process was to establish an international Panel of experts
in human, animal, and environmental toxicology, in epidemiology, in agronomic
practices, and in ecology (the Scientific Advisory Team - SAT). In the second step, the
SAT formulated a framework to conduct this risk assessment. The framework is similar

to those commonly used for assessing risks in a number of jurisdictions and consisted
of a problem formulation, characterization of the human health and environmental
effects of the substances used in the eradication program, characterization of human
and environmental exposures, and the drawing together of these in a risk
characterization. During the process of conducting the risk assessment, the SAT used

scientific literature and government reports but, where data gaps and uncertainties
related to the specific uses in Colombia were identified, studies were initiated to
assemble additional data for use in the risk assessment. Several of these studies were
carried out in Colombia. The Colombian Team (PTG) were contracted specifically to
CICAD and worked under the direction of the SAT to collect data in the Colombian

environment. During the conduct of our study, members of the SAT made a number of
visits to Colombia to view, at first hand, all aspects of the program, to gather local
information and data, and to oversee the local studies of the PTG.
The SAT recognized that the growing and production of illicit drugs in Colombia

has significant political, social, and economic, implications. However, this study was
focused specifically on the human health and environmental significance of the
production and eradication of coca and poppy through the use of aerially applied
herbicide. The production of coca and poppy as well as the processing and production
of cocaine and heroin also involves significant environmental impacts. Both coca and

poppy are grown intensively in a process that involves the clearing of land, the planting
of the crop and protection against pests such as weeds, insects, and pathogens. All of
these activities can impact human health and the environment and some, such as clear-
cutting, do so to a significant extent. The total land area used for these activities is
small relative to the entire country. However, much of the production takes place in

remote areas that are close to or part of the Andean Biodiversity Hotspot.
In Colombia, the herbicide glyphosate is widely used in agriculture and for
purposes other than eradication of coca and poppy. Only 10-14% of the total use in
Colombia is in the eradication program. Similarly many of the pesticides and other

substances used in the production of coca and poppy are also widely used in
agriculture. The aerial eradication spray program in Colombia is conducted with
modern state-of-the-art aircraft and spray equipment. The spray equipment is similar to
that used for forest spraying in other parts of the world and produces large droplets
which minimize drift of spray. Identification of target fields and electronic documentation

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of locations and areas sprayed is conducted with high precision. As a result of the use

of best available spray and navigation technology, the likelihood of accidental off-target
spraying is small and is estimated to be less than 1% of the total area sprayed.
The physical, chemical, and biological properties of glyphosate and an adjuvant

(Cosmo-Flux®) added to the spray mix were characterized through the scientific
literature and through new studies specifically conducted for this risk assessment.
Glyphosate is a widely-used herbicide that is well characterized in terms of physical,
chemical, and biological properties. Glyphosate is not highly mobile in the environment
and is rapidly and tightly bound on contact with soil and aquatic sediments. Glyphosate

has a very short biological activity in soils and water, does not biomagnify or move
through the food chain, and does not leach into groundwater from soil.
Exposures of humans to glyphosate under the conditions of use could not be

measured directly in the growers of illicit crops and thus were estimated from literature
values with adjustments for the rates of application used in the eradication program in
Colombia. Estimated exposures resulting from direct overspray, contact with treated
foliage after re-entry to fields, inhalation, diet, and drinking water were small and
infrequent. In a special study in five watersheds, weekly analyses of surface waters and

sediments over a period of 24 weeks showed that, on most occasions, glyphosate was
not present at measurable concentrations; only two samples had residues above the
method detection limit of 25 µg/L. As most of the glyphosate used in Colombia is in
agriculture, this confirms that, regardless of use pattern, glyphosate is not mobile in
environment and it will not move from the treated fields in significant amounts. In

analyses of water samples taken from the same five watersheds, several other
pesticides were found, including the herbicide 2,4-D and the insecticide endosulfan, the
latter a product that is banned in Colombia.

Concentrations of glyphosate in several environmental matrices resulting from
the eradication spray program were estimated. Concentrations in air were predicted to
be very small because of negligible volatility. Glyphosate in soils that are directly
sprayed will be tightly bound and biologically unavailable. Based on observations in
other temperate and tropical areas, no residual activity is expected in soil and even the

most sensitive organisms, plants, will not be prevented from re-establishing themselves.
In Colombia, this is evidenced by the rapid recovery of sprayed fields through
successful replanting of coca and/or colonization by invasive species of plants.
Concentrations of glyphosate plus Cosmo-Flux® will be relatively large in shallow

surface waters that are over-sprayed (maximum instantaneous concentration of 1,052
μg AE/L in water 300 mm deep); however, no information was available on the number
of fields in close proximity to surface waters and it was not possible to estimate the
likelihood of such contamination.

The toxicity of glyphosate has been rigorously assessed in a number of
jurisdictions and in the published literature. Glyphosate itself has low toxicity to non-
target organisms other than green plants. It is judged to have low acute and chronic
toxicity, carcinogenic, mutagenic, or a reproductive toxicant. With respect to humans, is

not considered hazardous, except for the possibility of eye and possibly skin irritation
(from which recovery occurs). The toxicity of the formulation as used in the eradication
program in Colombia, a mixture of glyphosate and Cosmo-Flux®, has been
characterized in specific tests conducted in laboratory animals. The mixture has low

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toxicity to mammals by all routes of exposure, although some temporary eye irritation

may occur. By extrapolation, the spray mixture is also not expected to be toxic to
terrestrial mammals and vertebrates. Epidemiology studies conducted in a number of
jurisdictions around the world have not suggested a strong or consistent linkage
between glyphosate use and specific human health outcomes. A preliminary

epidemiology study was conducted in Colombia to assess any linkage between
glyphosate and the reproductive outcome, time to pregnancy, in humans. This study
did not show any association between time to pregnancy and the use of glyphosate in
eradication spraying.

New data from the environmental literature on the toxicity of some formulations of
glyphosate suggest that amphibians may be the most sensitive group of aquatic
organisms. Special tests of the spray mixture as used in Colombia were conducted
using standardized environmental test organisms. These tests revealed that the

mixture of glyphosate and Cosmo-Flux® was not toxic to honey bees. The mixture was,
however, more toxic to aquatic organisms than formulated glyphosate alone. Extensive
studies on the use of glyphosate in agriculture and forestry in temperate and tropical
areas have been published in the literature. These have shown that direct effects on
non-target organisms other than plants are unlikely to occur. Indirect effects on

terrestrial arthropods and other wildlife have, however, been observed. These are the
result of habitat alteration and environmental change brought about by the removal of
target plants through the effects of glyphosate. Similar effects would be expected
regardless of the type of method used to control plants and also occur as a result of
clear-cutting, burning, and conversion of natural areas into agricultural lands. Because

of the lack of residual activity, recovery of glyphosate-treated areas will be dependent
only on the nature of the recolonizing species and the local conditions. Given
experience in other tropical regions and in Colombia, this process will be rapid because
of good conditions for plant growth. However, return to the conditions of tropical old-

growth forest that existed prior to clear-cutting and burning may take hundreds of years.
It is important to recognize that the impact here is not the use of glyphosate but the
original act of clear-cutting and burning that is the primary cause of the effects on the
environment.

The risk assessment concluded that glyphosate and Cosmo-Flux® as used in the
eradication program in Colombia did not present a significant risk to human health.
Estimated acute worst-case exposures in humans via all routes were less than doses of
concern, even for chronic responses. In the entire cycle of coca and poppy production
and eradication, human health risks associated with physical injury during clear-cutting

and burning and the use of pesticides for protection of the illicit crops were judged to be
more important than those from exposure to glyphosate.
For the environment, risks from the use of glyphosate and Cosmo-Flux® to

terrestrial animals were judged to be small to negligible. Moderate risks could occur in
aquatic organisms in shallow surface waters that are over-sprayed during the
eradication program. However, the frequency of occurrence and extent to which this
happens are unknown as data on the proximity of surface waters to coca fields were not
available. Considering the effects of the entire cycle of coca and poppy production and

eradication, clear-cutting and burning and displacement of the natural flora and fauna

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were identified as the greatest environmental risks and are considerably more important

than those from the use of glyphosate.
Strengths and uncertainties in the assessment were identified and used to
develop recommendations which were then prioritized. It is recommended that the

current application practices for eradication spraying be retained but that additional data
be gathered over a longer time period to better characterize the impacts of coca and
poppy production in the Andean Biodiversity Hotspot and the possibility of non-target
effects in surface waters located close to fields. If shallow waters are routinely found
close to fields, it is recommended that other formulants be tested for the purposes of

selecting products that present a lower risk to aquatic organisms. Although no
association was observed between eradication spraying and reproductive outcomes in
humans, additional studies to identify possible risk factors associated with other human
activities or environmental factors should be considered.

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1 INTRODUCTION

1.1 BACKGROUND
It is estimated that some 200 million people worldwide use illicit drugs. Most of these
drugs have natural origins, such as cannabis, cocaine, and the opiates, however, the

synthetic drugs such as the amphetamines also comprise a significant proportion of
these uses (United Nations 2002). In response to the socio-economic impacts of the
production and distribution of illicit drugs, a number of individual nations, as well as
multinational organizations, have initiated programs to reduce and eventually eliminate
production and distribution (United Nations 2002). While it is recognized that the
political, social, and economic impacts of the production, distribution, and use of all of

these drugs is significant, the focus of this report is on issues related to the program for
reduction and eradication of production of coca and opium poppy and their derivatives,
cocaine and the opiates in Colombia, South America.
Coca (Erythroxylum coca and

related species, Figure 1) are commonly
associated with the tropical mountainous
regions of South America. However, it
has been reported to be grown in Africa,
Sri Lanka, Taiwan, and Indonesia (Bray

and Dallery 1983). A number of species
of coca are found in South America and
various varieties grow in the wild or are
cultivated in different climatic conditions.
It is primarily found in tropical regions with
temperatures above 25°C and moderate

to high rainfall >1000 mm per year.
Currently, it is widely cultivated in Figure 8 Coca plant

Colombia, Bolivia, and Peru, with some
cultivation in Ecuador, Venezuela, Brazil, and Argentina as well.
Historically, coca played an important role in culture of the Incas, Quechuas, and
many other Andean peoples. Coca also played an important role in the conquest of
Latin America by the Spanish when it was used as an incentive and payment for work

on railroads, in agriculture, and in mines. More recently, cocaine, derived from the coca
plant, has become widely used in many countries. Initially used as a medicinal drug, it
was introduced to Europe as cocaine in 1860 as an ingredient of a wine-coca drink
which was apparently used by the likes of Sarah Bernhardt, Queen Victoria of England,
Thomas Edison, and Pope Leo the XIII. It was also used as a local anesthetic. In 1886,

John Pemberton introduced the tonic drink CocaCola® which contained cocaine until
1904 (Gottlieb 1976). Cocaine is now widely used as an illicit addictive drug; global
production between 1995 and 2002 was estimated to range from 640 to 950 tonnes
used by an estimated 14 million people (United Nations 2002). The illicit growing of
coca and its processing into cocaine has become a large and profitable industry that

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has had significant impacts on social and economic order in a number of producer as

well as in consumer nations.
Opium, morphine, and its derivative, heroin, are produced from the poppy,
Papaver somniferum, which is primarily grown in Asia. Global production of opium in
2002 was estimated to be 1,586 tonnes, of which about 160 tonnes were produced in

South America (United Nations 2002). It is estimated that, globally, about 15 million
people use opiates and that about 10 million of these use heroin (United Nations 2002).
Like coca, the use of opium and morphine has historical roots in the traditional society
of the producer regions but became more widely used as a medicinal drug when
introduced to other parts of the world. While morphine is still used for medicinal

purposes, heroin use is largely illegal and its production and distribution has significant
socio-economic impacts in producer and consumer nations.

1.2 IMPACTS OF ILLICIT DRUG PRODUCTION IN COLOMBIA

The growing and production of illicit drugs in Colombia has significant political,
social, economic, and environmental impacts. While recognizing the importance of the
political, social, and economic aspects of the issue, this report is focused on the human
health and environmental significance of the eradication of coca and poppy through the
use of aerially applied herbicide.

Although the focus of this study is on the coca and poppy eradication program, it
is important to recognize that the actual production of coca and poppy as well as the
processing and production of cocaine and heroin involves significant environmental

impacts. Both coca and poppy are grown intensively in a process that involves the
clearing of land, the planting of the crop and its protection against pests such as weeds,
insects, and pathogens.
Depending on the region, the clearing of the land for production purposes may

have large and only slowly reversible effects on the environment. As for other forms of
agricultural production, the clear-cutting of forests for the purposes of coca and poppy
production reduces biodiversity, contributes to the release of greenhouse gases,
increases the loss of soil nutrients, and promotes erosion of soils. As production is
illegal, it normally takes place in remote locations. As a result, the clearing of land is

done with little apparent consideration for the biological and aesthetic value of the
ecosystem.
A number of pesticides are used in the production of illicit drugs (Table 1).
Herbicides may be used in the initial clearing of the land and later in the suppression of

weeds. Similarly, insecticides and fungicides may be used to protect the illicit crops
from pests and diseases. To increase yields, fertilizers and other nutrients may also be
used. Large quantities of agrochemicals have been seized and confiscated as part of
the program to control the production of illicit drugs (Direccion Nacional de
Estupefacientes 2002). Although some of these agrochemicals are highly toxic to

mammals and may have significant environmental impacts, accurate information on the
amounts used, their frequency of use, and the conditions of their use is not available.
Because of this, it is not possible to conduct a detailed human health and ecological risk
assessment. However, the relevant toxicological and environmental properties of these

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substances are summarized in two separate reports and several of these are significant

potential hazards to human health and the environment (CICAD/OAS 2004a, 2005).

Table 1. Pesticides used in the production of coca

Active ingredient Toxicologicala Estimated % Chemical class
classification of use

Paraquat II 61.3 Bipyrilidinium herbicide
Glyphosate IV 19.1 Phosphate herbicide

2,4-D I 9.7 Phenoxy herbicide

Atrazine III 4.8 Triazine herbicide

Diuron III 2.6 Urea herbicide
Carbaryl II NA Carbamate insecticide

Carbendazim III NA Benzimidazole carbamate fungicide

Carbofuran I NA Carbamate insecticide
NA
Chlorpyrifos II Organophosphorus insecticide
Copper oxychloride III NA Metal fungicide

Cypermethrin II NA Pyrethroid insecticide
NA
Diazinon III Organophosphorus insecticide
Endosulfan I NA Organochlorine insecticide

Lambda cyhalothrin III NA Pyrethroid insecticide

Malathion III NA Organophosphorus insecticide
NA
Mancozeb III Carbamate fungicide
Methamidophos I NA Organophosphorus insecticide

Methomyl I NA Carbamate insecticide

Methyl parathion I NA Organophosphorus insecticide
NA
Monocrotophos I Organophosphorus insecticide
Prophenophos II NA Organophosphorus insecticide
a
As classified by the Instituto Colombiano Agropecuaria (ICA) as follows: I (very toxic), II
(toxic), III (slightly toxic). Data from (Direccion Nacional de Estupefacientes 2002)

In addition to the use of agrochemicals in the production of coca and poppy,
large amounts of chemicals are used in the processing of the raw product into refined
cocaine and heroin (Table 2). Processing of the illicit drugs is conducted in remote

locations and in the absence of occupational health and environmental regulations and
controls. During and after use, these substances may be released into the environment
and have significant impacts on human health and the ecosystem. The toxicological

and environmental properties of these substances are summarized in a separate Tier-1
Hazard Assessment Report (CICAD/OAS 2004a). Some of these substances have

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40 Annex 116

potentially large environmental and human health hazards and a subset of these are

dealt with in more detail in Tier-2 Hazard Assessment Report (CICAD/OAS 2005).

Table 2. Identity and amounts of substances seized in Colombia as a result of
counter-drug operations a

Year 1999 2000 2001 2002
Solid substances (units in Kg)

Activated charcoal 36,681 49,323 84,141 93,057
Ammonium chloride 480 7 450 350

Ammonium nitrate - - 2,390 9,350
Ammonium sulfate - - - 900

Calcium carbonate 500 150 255 1,570
Calcium chloride 7,371 33,073 56,985 146,040
Cement, grey 142,818 197,646 502,857 1,053,372

Cement, white - - - 18,700
Lime 24,807 49,783 155,507 220,259

Potassium chloride 2,290 4,766 1,456 34,750
Potassium hydroxide 375 1,425 - 4,700

Potassium nitrate 2 - 2,150 2,390
Potassium permanganate (sum) 71,284 171,798 51,641 80,639
Sodium bicarbonate 52 4,827 8,538 9,939

Sodium carbonate 531,095 248,136 59,521 128,571
Sodium chloride 28,154 17,046 31,594 35,161

Sodium hydroxide 73,776 69,100 111,540 122,619
Sodium hypochlorite - 16 4,208 1,720

Sodium sulfate 5,755 970 1,852 8,667
Urea 62,685 37,995 226,394 360,237

Liquid substances (units in L)
Butyl Acetate 23,732 469 13,089 11,908
Ethyl Acetate 97,723 76,156 23,289 15,336

Acetone 1,666,474 894,070 1,546,651 1,841,860
Hydrochloric Acid 144,804 62,303 126,884 140,650

Sulfuric Acid 303,732 200,404 241,903 277,538
Isopropyl Alcohol 59,379 6,938 16,408 19,330

Ammonia 131,104 154,180 102,512 431,485
Acetic Anhydride 9,938 284 10,855 1,045

Chloroform 465 1,457 1 273
Ethyl Ether 205,984 67,704 53,989 110,098
Gasoline 621,686 1,034,880 2,013,650 2,612,820

Hexane 35,963 4,497 16,991
Kerosene 127,316 90,855 159,818 210,408

Methyl ethyl ketone MEK 88,402 69,209 10,674 41,332
Methanol 269,027 14,107 2,961 3,512

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Table 2. Identity and amounts of substances seized in Colombia as a result of
counter-drug operationsa

Year 1999 2000 2001 2002
Methyl isobutyl ketone MIBK 55,943 2,086

Thinner 226,657 78,156 100,829 203,459
Toluene 3,630 208 19 6,469
Acetic acid 11 14 208 212

Nitric acid 59 6 1 5,300
Isobutyl alcohol 170 3 1,136

Petroleum ether 35,579
Methylene chloride 416 4 45 4,182
Fuel oil 32,082 325,250 346,460 948,083

Solvent No 1 203,603 116,498 435,816 280,921
Solvent No 2 6,505 3,819 5,621 11,942
aThese substances are mainly used in the refining of cocaine, opium, and heroin. It is
estimated that only 20% of the total amounts used are seized. Therefore, total use may be

as much as 5-times greater than indicated in the table. Data from (Direccion Nacional de
Estupefacientes 2002)

1.3 THE PROGRAM TO CONTROL ILLICIT DRUG PRODUCTION AND
DISTRIBUTION IN COLOMBIA

The growing of coca and poppy and the distribution of cocaine and opium/heroin
in Columbia has been the focus of a national control and eradication program starting in
the 1970s. The program involves a number of Departments and Agencies of the

Colombian Government and is coordinated by the Direccion Nacional de
Estupefacientes (DNE), an agency of the Ministry of the Interior and Justice. The
program has three main foci; the control of production of coca and poppy; the control of
the processing, purification, and transport of the cocaine and heroin; and the seizure

and forfeiture of the profits of illicit drug production (Direccion Nacional de
Estupefacientes 2002).

The aerial eradication program in Colombia is the responsibility of the
Antinarcotics Directorate of the Colombian National Police (DIRAN-CNP), supported by
data gathering from other nations such those in North America and Europe. The DIRAN
conducts regular flights with aircraft that spray coca and opium poppy crops with

herbicide. The DIRAN reviews satellite imagery and flies over growing regions on a
regular basis to search for new coca and opium poppy growth and to generate
estimates of the illicit crops through high resolution low-altitude imagery and visual
observation. The DIRAN selects the locations of the illicit crops that are to be sprayed

with input from the DNE or the Government of Colombia's Plan Colombia Office. For
example, at this time, certain existing or future alternative development projects or
national parks may not be sprayed as a matter of policy.

Several concerns have been raised about the use of glyphosate and adjuvants in
the eradication of coca and poppy plants. These concerns range from damage to other
crops to adverse effects on the environment and human health. In response to this, the

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Government of Colombia appointed an independent environmental auditor who reviews

the spray and no-spray areas with the DIRAN, and regularly monitors the results of
spraying through field checks and analysis of data from the computer system.
The objectives of this assessment and report are to provide a science- and data-

based study of the eradication program with a key focus on the environment and human
health, to collect data for use in the assessment, to address specific concerns that have
been raised, and to make the results known to the public and the scientific community.
As with all risk assessments, we have followed a framework based on those used in
other jurisdictions (NRC 1986, USEPA 1992, 1998). This framework consists of a

Problem Formulation, Effects and Exposure Assessment, and Risk Characterization for
both humans and the environment.

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2 PROBLEM FORMULATION

The problem formulation is a key step in the process of the risk assessment and
places the use of the
IMPACTS OF IMPACTS OF
substances being SPRAY PRODUCTION
assessed into a local
context. It is recognized Off-target effects on Clear-cut and burn
plants Pesticides (humans
that the growing of illicit Effects on humans and non-target
crops such as coca and organisms
poppy as well as the Effects on aquatic
organisms Increased erosion
refining of the cocaine and Effects on terrestrial Fertilizer
heroin involves organisms
considerable impacts on

the environment through
clearing of forests and the
use of a number of

substances for promoting IMPACTS OF REFINING
crop growth and refining of CHEMICALS
the drugs (Figure 2). Humans
Although the identity of the
Non-target organisms
substances is known, the
quantities used, and their Figure 9 Diagrammatic representation of potential impacts of coca
manner of use is largely production, refining, and spraying.

unknown and exposures in
workers cannot be easily estimated. While the hazard of these substances is known
(CICAD/OAS 2004a, 2005), the risks cannot be estimated as the logistics of collecting

the human and environmental exposure data are very difficult and not without other
risks. Because of this and as it was the initial mandate of the Panel, the focus of this
risk assessment is on the use of glyphosate and adjuvants for control of the illicit crops.

In this case, the locations and amounts of application are known with accuracy and
environmental risk can be estimated.
In humans, there are no specific biomarkers for exposure to glyphosate that can

be used to estimate historical exposures. For logistical reasons, it was not possible to
measure exposures resulting from eradication spraying directly in the field. For that
reason, in epidemiology studies, indirect measures of exposures such as ecological

studies, where the indicator variable or exposure is a defined by eradication spraying
and crops production patterns, must be used.

2.1 STRESSOR CHARACTERIZATION
The potential stressors in this risk assessment are glyphosate, its formulants,

and adjuvants, such as surfactants, that are added to the spray formulation to modify its
efficacy. The properties of glyphosate and these substances are described in the
following sections.

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2.1.1 Glyphosate

Glyphosate is one of the most widely used pesticides on a global basis. Uses
include agricultural, industrial, ornamental garden and residential weed management.
In agriculture, the use of glyphosate is increasing and use in soybeans is probably

greater since the introduction of glyphosate-tolerant crops (Wolfenbarger and Phifer
2000). Other agricultural uses for glyphosate-based products include its use by farmers
as a routine step in pre-plant field preparation. Non-agricultural users include public

utilities, municipalities, and regional transportation departments where glyphosate is
used for the control of weeds or noxious plants. The environmental and human-health
properties of glyphosate have been extensively reviewed in the literature (Giesy et al.

2000, Solomon and Thompson 2003, Williams et al. 2000) and by regulatory agencies
(NRA 1996, USEPA 1993a, 1997, 1999, World Health Organization International
Program on Chemical Safety 1994). The following sections highlight key issues with

regard to those properties of glyphosate that are fundamental to the assessment of risks
associated with the coca and poppy eradication programs in Colombia.

2.1.1.1 Structure and chemical properties

The chemical name of glyphosate (acid) is N-(phosphonomethyl) glycine (MW =
167.09) and that of the most common technical form, the isopropylamine salt (IPA) is N-

(phosphonomethyl) glycine isopropylamine salt (MW = 226.16). The Chemical
Abstracts Registry (CAS) number of the acid is 114370-14-8 and for the IPA salt is
1071-83-6. The chemistry of

glyphosate is important in Glyphosate
determining its fate in the O O
environment. Glyphosate (Figure 3)
is a weak organic acid comprising a HO P CH 2 CH 2 OH

glycine moiety and a O H H
phosphonomethyl moiety.
Chemically and physically, AMPA Sarcosine

glyphosate closely resembles O H H O
naturally occurring substances and itHO P CH N H+ CH N + CH C OH
is not chemically reactive, not mobile 2 3 2
OH H H
in air or soils, does not have great Glycine
biological persistence, and does not
bioaccumulate or biomagnify through H O
+ H N+ CH C OH
the food chain (CWQG 1999, Giesy Pi + CH 3H 3 2
et al. 2000, USEPA 1993a, Williams H
et al. 2000, World Health
Organization International Program
Figure 10 The structure of glyphosate and its
on Chemical Safety 1994). major metabolic and breakdown products. From
Glyphosate is readily ionized (Liu et al. 1991)

and, as the anion, will be strongly
adsorbed to organic matter in soils of normal pH (Figure 4). It thus has low mobility in
soils and is rapidly removed from water by adsorption to sediments and suspended

particulate matter.

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45Annex 116

2.1.1.2 Mechanism of action of glyphosate
O O
The mechanism of action of glyphosate is via HO C CH 2 CH P 2P OH
the inhibition of the enzyme 5-enolpyruvyl shikimate- H OH

3-P synthetase, an essential enzyme on the pathway +
to the synthesis of the aromatic amino acids in plants O O H +
(Devine et al. 1993). This inhibition results in - H
HO C CH N2CH P 2P O
decreases in the synthesis of the aromatic amino H O-
acids, tryptophan, phenylalanine, and tyrosine, as well + +
+ + +
as decreased rates of synthesis of protein, indole
acetic acid (a plant hormone), and chlorophyll. The Particle of organic
death of the plant is slow and is first seen as a matter

cessation of growth, followed by chlorosis and then
necrosis of plant tissues. Inhibition of 5-enolpyruvyl Figure 11 Binding of glyphosate
to soil particles
shikimate-3-P synthetase is specific to plants. Many
animals obtain their aromatic amino acids from plants
and other sources and do not possess this pathway of synthesis. For this reason,

glyphosate is relatively non-toxic to animals but is an effective herbicide in plants.

2.1.1.3 Global and local registration and use
Glyphosate has been registered since 1971 and is currently widely used as a

broad-spectrum, non-selective, post-emergence herbicide in a number of countries
around the world (World Health Organization International Program on Chemical Safety
1994). It is rapidly translocated from the leaves of treated plants to other parts of the

plant, including the growing tips of stems and roots, and to underground storage organs,
such as rhizomes and tubers. It is very effective for the control of perennial weeds and

is more efficacious than many other non-selective herbicides that only affect the above-
ground parts of the plant. Applied to soil, glyphosate shows low activity because the

strong binding to soil organic matter makes the substance biologically unavailable to
plants. Glyphosate has been used extensively in Colombia and many other countries
for agricultural and other purposes for many years. Use of glyphosate in the coca and

poppy spray program is shown in Table 3 and represents a relatively small fraction of
the total use in Colombia.

Table 3. Use glyphosate in eradication spraying in Colombia 2000 to 2004
Year Amount sold in Amount used in Percent of total
Colombia (L) a the eradication of amount sold
b
illicit crops (L)
2000 7,037,500 603,970 8.6%

2001 9,473,570 984,848 10.4%
2002 NA 1,061,538 11% c
c
2003 1,381,296 14%
2004 1,420,130 14% c
aData from (ICA 2003).Data from (Direccion Nacional de Estupefacientes 2002, Policia
c
Nacional Direccion Antinarcoticos 2005).mated from total used in 2001 but likely less than
this value.

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46 Annex 116

2.1.1.4 Environmental fate
The environmental fate of glyphosate has been extensively reviewed (CWQG
1999, Giesy et al. 2000, NRA 1996, World Health Organization International Program

on Chemical Safety 1994); only key issues relevant to water and soil/sediment are
summarized below.
As a result of its specific physicochemical properties, glyphosate is immobile or

only slightly mobile in soil. The metabolite of glyphosate, aminomethyl phosphoric acid
(AMPA, Figure 3), is somewhat more mobile in soil but is rapidly broken down, resulting
in minimal amounts leaching in normal agricultural soils. The strong binding of
glyphosate to soil results in almost immediate loss of biological activity, however, the
bound residues do break down sufficiently rapidly that accumulation will not occur, even

over many years of regular use. Contamination of groundwater from the normal use of
glyphosate is unlikely except in the event of a substantial spill or other accidental and
uncontrolled release of large amounts into the environment.

The great water solubility of glyphosate and its salts suggests that it would be
mobile in water, however, strong and rapid binding to sediments and soil particles,
especially in shallow, turbulent waters, or those carrying large loads of particulates,
removes glyphosate from the water column (Tooby 1985). In normal agricultural uses, it

is not expected to run-off or leach into surface waters.
In water, the two major pathways of dissipation are microbiological breakdown
and binding to sediments (Giesy et al. 2000, World Health Organization International

Program on Chemical Safety 1994). Glyphosate does not degrade rapidly in sterile
water, but in the presence of microflora (bacteria and fungi) in water, glyphosate is
broken down to AMPA (Figure 3) and eventually to carbon dioxide (Rueppel et al.
1977). Other metabolic pathways have been reported (Liu et al. 1991), including further
degradation of AMPA to inorganic phosphate and CH -NH , and via sarcosine to glycine
3 3
(Figure 3). None of these products are considered herbicidal and they would not be
expected to be highly toxic to aquatic organisms at concentrations that would result
from field use of glyphosate in aquatic systems. Photodegradation also may take place
under field conditions where sufficient penetration of UV light occurs.

The dissipation of glyphosate from treated foliage and from leaf litter has also
been characterized. As would be expected, most of the glyphosate sprayed on the
plants penetrates into plant tissues after application, but some is available for washoff

for several days after application (World Health Organization International Program on
Chemical Safety 1994). If the plant dies as a result of this exposure, glyphosate would
be present in the dead and decaying plant tissues. Glyphosate residues in leaf litter
dissipate rapidly with a time to 50% disappearance (DT50) of 8-9 days under temperate
forestry conditions (Feng and Thompson 1990). Similar rapid dissipation from fruits and

lichen has also been observed (Stiltanen et al. 1981).
Dissipation under tropical conditions such as in Colombia will likely be more rapid
than in temperate regions because of higher temperatures and moisture content which

promote microbiological activity as well as chemical degradation of many pesticides.
Large areas of Brazil, Colombia, Central America, most of Africa between the Sahara
and Kalahari deserts, India, inland Indochina, and portions of Northern Australia share
similar tropical conditions and some of those countries depend heavily on herbicides

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47Annex 116

such as glyphosate (Racke et al. 1997). Glyphosate has been used in large areas of

Brazil on no-tillage crops in general and, more recently, on transgenic soybeans.
Comparing the fate of pesticides in tropical and temperate conditions, Racke et al.
(1997) found no evidence of particular behavior of the pesticides in the tropics, they
even concluded a greater rate of degradation under tropical conditions. The authors

stated:
“Since soil microbial activities are strongly modulated by temperature,
pesticide degradation would be expected to be greater in tropical soils,
which experience higher year-round temperatures, than in temperate soils.

This explanation would be consistent with observations of the elevated
rates of soil organic matter turnover that characterize udic and ustic (rainy
season) tropical environments. The few available studies which have
directly compared pesticide fate in temperate and tropical soils held under

identical conditions (i.e., laboratory) reveal no significant differences in
either the kinetics or pathway of degradation. It appears that there are no
inherent differences in pesticide fate due to soil properties uniquely
possessed by tropical soils. Tropical soils themselves defy easy
categorization, and their properties are as varied in nature as those from

temperate zones. Pesticides appear to dissipate significantly more rapidly
from soil under tropical conditions than under temperate conditions. The
most prominent mechanisms for this acceleration in pesticide dissipation
appear to be related to the effect of tropical climates, and would include
increased volatility and enhanced chemical and microbial degradation

rates on an annualized basis.

2.1.2 Formulants and adjuvants
Formulants are substances that are added to a pesticide active ingredient at the

time of manufacture to improve its efficacy and ease of use. These formulants serve
many purposes and comprise a large range of substances, ranging from solvents to
surfactants to modifiers of pH. The glyphosate formulation used in Colombia includes
several formulants. Adjuvants are added to formulated pesticides at the time of
application and, like formulants, increase efficacy, or ease of use in special situations

where pests are difficult to control or where non-target effects need to be minimized. In
the control program in Colombia, an adjuvant, Cosmo-Flux®, is added at the time of
spraying.

The relatively great water solubility and the ionic nature of glyphosate retard
penetration through plant cuticular waxes (Figure 5). For this reason, glyphosate is
commonly formulated with surfactants which decrease the surface tension of the
solution and increase penetration into the tissues of the plants (Giesy et al. 2000, World
Health Organization International Program on Chemical Safety 1994).

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48 Annex 116

Droplet Droplet
2.1.2.1 Surfactants in the without with
glyphosate surfactant surfactant
formulation

The glyphosate
formulation as used in Wax layer
Cuticle
eradication spraying in proper
Colombia contains several
formulants which are Pectin
layer
common to the commercial
product as used in Primary wall

agricultural. Secondary
wall
2.1.2.2 Cosmoflux 411F

As mentioned above, Plasma
an adjuvant, Cosmo-Flux®, is membrane
added to the glyphosate at
Cytoplasm
the time of spraying. Cosmo-
Flux® is an agricultural Figure 12 Penetration of an herbicide such as glyphosate
through plant cuticular waxes in the absence (left) and
adjuvant containing non-ionic presence of surfactants (right).
surfactants (a mixture of
linear and aryl polyethoxylates – 17% w/v) and isoparaffins (83% v/v) (Cosmoagro

2004). Adjuvants such as these are commonly added to pesticide formulations to
improve efficacy through several mechanisms (Reeves 1992, Tadros 1994).

For example, surfactants such as the polyethoxylates in Cosmo-Flux®, increase
efficacy through increasing target surface adherence, promoting better droplet spread,
better dispersion, prevention of aggregation, and enhanced penetration of herbicides

into target plant tissues through the reduction of surface tension on plants. Surfactants
can also disrupt the water insoluble wax cuticle, thus increasing the penetration of
herbicide active ingredient.

Base oils, such as the isoparaffins in Cosmo-Flux®, are another class of
adjuvants used in pesticide formulations. They are used primarily to aid foliar

absorption of the pesticide by disrupting the waxy cuticle on the outer surface of foliage
which increases cell membrane permeability (Manthey and Nalewaja 1992).

2.1.3 Coca and poppy control programs
As discussed briefly above, the coca and poppy control programs make use of

several procedures to identify, locate, map coca and poppy fields. The initial step in this
process is the use of satellite images to locate the coca and poppy fields. These
images are provided by North American and European governments to the Government

of Colombia. The images are used to locate potential areas of coca and poppy
production. Further visual observations are made using overflights with observers

and/or photographs from a low-altitude aerial-photography plane, such as a Cessna
Caravan, to verify the presence of the coca and poppy fields. The camera used for this
purpose is multi spectral high-resolution. Maps are generated in a Geographic

Information System (GIS) and are used to produce updated co-ordinates for the spray
pilots as well as information for downloading into the aircraft navigation systems

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49Annex 116

(Figures 6 and 7). The field operation offices for the control program have computers

and a satellite uplink for data transfer. The spray-planes, such as AT 65s, AT 802s, or
OV 10s, are equipped with high resolution tracking equipment and Del Norte positional
data recorders that display position, provide directional guidance, and store positional
data on data cards for later analysis. Thus the locations of the fields, the flight-paths of

the spray-planes, and the areas where spray is released are known to within a
resolution of several meters.
Since 1994, the coca and, more recently, poppy fields have been identified and
sprayed during the eradication program. Total areas of identified fields, and area

sprayed in Colombia are shown in Figure 8. With increasing areas sprayed, the total
area planted to coca has generally decreased since 2000.

2.1.3.1 Receiving environment

Colombia is located between about 4ºS and 12°N of the equator. The country
presents very varied topography ranging from snow-capped peaks through high
mountain plateaus to low-lying tropical regions. In general, coca tends to be grown at
altitudes below 1,500 m and poppy at greater altitudes, usual 2,200 m. The biodiversity
hotspot for the tropical Andean region includes significant areas of Colombia (Figure 9).

The tropical Andes biodiversity region is estimated to contain 15-17 percent of the
world’s plant life in only 0.8 percent of its area. It has a area of 1,258,000 square
kilometers, and extends from Western Venezuela to Northern Chile and Argentina and
includes large portions of Colombia, Ecuador, Peru, and Bolivia (Centre for Biodiversity

2004).
Because the diversity hotspots are mainly associated with the Andean highlands
and coca is mostly grown in lower altitudes, there is only some overlap between the
areas of coca production and regions of high biodiversity. Poppy is grown at greater

altitude and this overlaps with the biodiversity hotspot; however, the total areas grown at
this time are small (Figure 8). E xact areas used for coca and poppy production within
the diversity hotspot are not known, however, this information would be useful for
assessing total impacts of production, especially for rare and endangered species of

plants.

2.1.3.2 Method of application
All coca and poppy fields are sprayed by aerial application from fixed-wing
aircraft. The procedure described below is based on observations recorded for the AT

65, AT 802, and OV 10 aircraft.

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50 Annex 116

Figure 13 Map showing production of coca in Colombia in 2005. Bright green shows coca
production. Blue boundaries indicate indigenous areas, red boundaries indicate national parks (Policia
Nacional Direccion Antinarcoticos 2005).

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51Annex 116

Figure 14 Map showing areas of poppy production in 2005. Bright red circles show poppy
production. Blue boundaries indicate indigenous areas, red boundaries indicate national parks (Policia
Nacional Direccion Antinarcoticos 2005).

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52 Annex 116

The spray- 200

planes are loaded in a Areaofcoca panted

special area of the Areaofcoca spayed
tarmac at one of a 150 Areaofpoppy panted
Areaofpoppy spayed
number of bases

throughout Colombia
100
(Figure 10).
Glyphosate and

Cosmo-Flux® are
50
stored in plastic
containers in a tarp- Heecaress 10000s))
10
lined area protected 8

by a berm to contain 6
accidental spills. The 4

areas may be in the 2
0.20
open or covered. The
glyphosate is Coca
0.15 Poppy
transferred from 200-

L plastic barrels to a 0.10

larger plastic storage
tank (Figure 10-A). 0.05
% ooffoallandd aeaa
Cosmo-Flux® is 0.00

transferred from 20-L 1994 1996 1998 2000 2002 2004
plastic containers to a

mixing tank. The Yeaarr

required amounts of
the components of Figure 15 Areas planted with coca and poppy in Colombia from 1994
to 2002 as ha (above) and as a percent of the total land area of
the application Colombia (below). From (Direccion Nacional de Estupefacientes 2002,

mixture (glyphosate, Policia Nacional Direccion Antinarcoticos 2005)
Cosmo-Flux®, and

water from a local source) are pumped through a metering pump (Figure 10-B) into the

aircraft using a Table of Mixing Proportions to ensure the correct ratio of amounts are
loaded. Appropriate protective equipment is used by the mixer-loaders who are trained

in the loading procedures (Figure 10-C).

The spray boom (Figure 10-D) on the aircraft is equipped with rain-drop nozzles

(Figure 10-E). These nozzles produce droplets with a volume mean diameter (VMD)

between 300-1,500 µm and are similar to those used in forestry spraying for site
preparation (Payne 1993). The aircraft spray systems are electronically calibrated to

disperse a specified quantity of spray mix per hectare, compensating for variances in

ground speed. These electronic spray controls are checked each day by technicians
and also during the pilot’s preflight inspection. During actual spray operations, the pilot

monitors the spray system by observing the readings of the spray pressure and the

spray flow rate gauges (U.S. Department of State 2002).

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53Annex 116

Figure 16 Map showing the region of Colombia identified as part of the Andean Biodiversity Region.
(From Centre for Biodiversity 2004).

The same nozzles are used for both coca and poppy applications but twice as
many are used for the poppy applications and different boom pressures are used. As a

result, coca and poppy applications are done at separate times. The currently-used
application rates are shown in Table 4.

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54 Annex 116

Table 4. Application rates of glyphosate and Cosmo-Flux® for control of coca and

poppy
Litres/ha Kg/ha
Coca Poppy Coca Poppy
Glyphosate 10.4 2.5 4.992 1.2
Cosmo-Flux® 0.24 0.51

From (Direccion Nacional de Estupefacientes 2002)

Each spray operation (Figure 10-F and G), which may consist of 2 or more spray-

planes, is escorted by search-and-rescue (SAR) helicopter(s) in case of an accident or
incident. Spraying is only conducted in daylight hours before mid-afternoon to ensure
that conditions are appropriate for application. If rain is imminent, visibility is poor, or
the wind speed is in excess of 7.5 km/h (4 knots), spraying is not carried out. Wind
speed is checked during the operation by the SAR and other helicopters with the aid of

smoke generated by the spray-planes. The spraying is done at about 30 m above
ground and, although the flight path is determined from the GIS information and the Del
Norte guidance system (Figure 10-H), the actual spraying is controlled by the pilots. In
personal communications with five of the pilots, it was stated that, according to spraying
guidelines, fields are not sprayed if people are seen to be present.

After a spray operation, the flight path of the spray-planes and the areas sprayed
is downloaded from the Del Norte system (Figure 10-I) and processed by GIS to show
the spray patterns and calculate the areas spayed (Figure 10-J). This information is
transmitted to the DIRAN where records of the spray operations are retained and used

for compilation of annual reports and statistics (Direccion Nacional de Estupefacientes
2002).

2.1.3.3 Frequency of application
The frequency of application varies with the local conditions and the actions

taken by the growers after the coca or poppy is sprayed. When coca is sprayed, some
growers will prune the bushes down to about 10 cm above ground in an attempt to
prevent translocation of the herbicide to the roots. Sometimes, these plants will recover
and resprout; however, they will not yield large amounts of coca leaves for several
months. If the field is replanted to coca from seedlings, reasonable productivity may not

be achieved 4-6 months. If the field is replanted from cuttings, productivity may be
achieved sooner. Thus, spraying of a particular coca field may have a return frequency
of about 6 to 12 months.
Being an annual, poppy is grown from seed. In the climatic conditions under

which it is grown in Colombia, poppy fields would be harvested twice a year. If sprayed
before reaching maturity and replanted immediately after spraying, they may be sprayed
four times a year.

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55Annex 116

A)Mixing area or gyphhosateannd aduvvansts B) Mixer orglyphossae and
adjuvantt

C)Miixe-loader D))Spray boom E))Noozzele

F)ATT-65sppayyplane G)OVV110Sppray pane being oaaded

I)Poosionaaldataa J)Sppray ocatonns
H)DeelNoore GPSS system

Figure 17 Photographs of aspects of the spray operation (photographs K R
Solomon).

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56 Annex 116

2.1.3.4 Exposure pathways in soil, air, water, and other media
In terms of the application, there are several pathways through which the

glyphosate and adjuvants may come into contact with the environment (Figure 11).

Deposition on
nontarget
organisms in
the field

Spray drift Deposition on the target

Direct
deposition or
spray drift
Deposition onto water
on soil
Runoff with
soil

Figure 18 Diagram showing exposure routes for various environmental compartments when
glyphosate is used for the control of illicit crops.

Deposition on the target crop (field) is the desired outcome of the operation;
however, from the purposes of assessing risks in humans and the environment,

exposures that result in movement and deposition off the field are important. Spray drift
would result in movement off the target field and could result in adverse effects in
nontarget plants and animals. Given the strong adsorption of glyphosate to soil,

deposition on soil in the field will likely not result in significant effects on nontarget
organisms, however, runoff of residues bound to soil particles may result in
contamination of surface waters with sediment-bound residues. Direct deposition and

spray drift may result in contamination of local surface waters with glyphosate if these
are in the spray-swath or drift envelope of the application. Depending on the depth of
the water, turbulence, flow, and suspended particles, this would result in exposures of
aquatic organisms to both glyphosate and any adjuvants present in the spray mixture.

Organisms present in the field during spraying would be exposed to the spray droplets
and would receive a theoretical dose, depending on surface area exposed and body
mass. Exposures that may occur via these routes are discussed in Section 3.1.4.

2.1.3.5 Off-target deposition
There are two types of off-target deposition. The first is related to incorrect

application where the spray pilot initiates application too soon or turns off the spray too
late, or the spray swath includes a non-target area on one or both sides of the target
field. The second type of off-target deposition that may occur is spray drift. Experience

with spray equipment of the type used in Colombia suggests that spray drift will be

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57Annex 116

minimal (Payne et al. 1990). Estimates of accidental overspray have been made during
assessments of the efficacy of the spray program (Helling 2003). Based on site-visits to

86 fields sprayed in 2002, and on observations of damaged plants beyond the boundary
of the area cleared and planted with coca, 22 fields showed evidence of off-field
deposition. Using the size of these areas, it was estimated that between 0.25 and

0.48% of the areas cleared for coca production were damaged by offsite spray
deposition (Helling 2003). Applying this to the total area of coca sprayed (Figure 8) and
calculating upper and lower intervals, the areas potentially affected are small when

compared to the total area of Colombia (Table 5).

Table 5. Estimates of areas affected by off-target deposition of glyphosate in the

spraying of coca in Colombia
Year Ha Area affected by off-target deposits (ha)Upper interval as a % of the
sprayed Lower interval Upper interval total area of Colombia

0.25% 0.48%
1994 3,871 9.7 18.6 0.0000002
1995 23,915 59.8 114.8 0.0000010
1997 41,861 104.7 200.9 0.0000018

1998 66,029 165.1 316.9 0.0000028
1999 43,111 107.8 206.9 0.0000018
2000 58,074 145.2 278.8 0.0000024
2001 94,152 235.4 451.9 0.0000040

2002 130,364 325.9 625.7 0.0000055
2003 132,817 332.0 637.5 0.0000056
2004 136,551 341.4 655.4 0.0000057

While the areas affected by off-target are estimated to be small, this estimate is

based on visual observations of a relatively small number of fields. These data were
only available for coca, not poppy, however, the total areas planted to poppy at this time
are not large, and similar off-target deposition would be proportionately smaller than that

associated with coca production. This is thus a source of uncertainty in the
assessment. It is not logistically possible to visually inspect all sprayed fields, however,

the routine monitoring of the areas planted to coca and poppy that is undertaken by
satellite and low altitude imagery could be used to assess any off-target deposition
which results in damage to plants. Changes in the size of sprayed fields over time

could be used to extend these estimates over larger areas and increase their accuracy,
although extension of the fields by growers may confound the data. The lower

resolution of satellite imagery may preclude its use for this purpose; however, greater
coverage by low-altitude images could facilitate this process.

2.2 Framework for risk assessment
The following sections outline the conceptual model and hypotheses for the

assessment of the human health and environmental impact of coca and poppy
production in Colombia. Although this document is focused on the risks associated with

the coca and poppy eradication program, it is recognized that the eradication program is
not conducted in isolation. There are a number of other activities associated with the

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58 Annex 116

process that result in risks to human health and the environment. While data are not
available to quantify all these risks, some of them may be estimated on the basis of

other knowledge and expert judgment. This was done using an adaptation of a risk

prioritization scheme that has been used in ecological risk assessment (Harwell et al.
1992).

2.2.1 Context of the risks

2.2.1.1 Human health risks

Risks of the cycle of coca and poppy production were estimated as discussed
above and are shown

in Figure 12. For the Coca or poppy
purposes of this field developed in

ranking process, the a natural area

intensity score
ranged from 0 to 5,

with 5 being a severe

effect such as a
physical injury or MPPACCTSS INTENSSTYT RECCOVERYY FREEQUENCCY IMPACCT % IMPACTT
SCORRE SCORRE % SCORE
toxicity. The recovery
Cleaarr
score also ranged cuutng and
from 0 to 5 and was buuning 5 3 3 55 16.77

based on the Planttng heh
potential for complete
coocaorr
recovery from the pooppy 0 1 100 0 0.00

adverse effect. Feerrtzerrliiz
Frequency was
inputss 0 05.5 10 0 0.00
based on an estimate
Peestcdee
of the proportion (%) inputss 5 3 10 100 55.66
of the total number of
Erradcatonno
persons involved in sppaya ? ? ? ? ?

coca and poppy Prrocessngn
cultivation,
annd efning 5 3 5 55 27.88
production, and the
refinement of cocaine
Figure 19 Potential human health impacts of the cycle of coca or poppy
and heroin. The production Scores for eradication spraying are specifically omitted.

score for impact was
the product of the individual scores and the percent impact is based on the sum of the

impact scores. The scores for the risks associated with the eradication program were

omitted from the ranking in this diagram but are discussed below in the conclusions to
the risk assessment.

2.2.1.2 Ecological risks

A similar procedure to that described above was used for ranking ecological risks

associated with the cycle of coca and poppy production (Figure 13). The intensity score

was ranked from 0 to 5, with 5 being most intense, such as the total destruction of the

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59Annex 116

habitat by clear-cutting and burning when clearing a natural area. Intensity of effects in

this case also included off-field effects such as on non-target animals and plants.

Recovery time in this scheme is the
estimated time for the impacted area Coca or poppy
field developed in
to recover to a state similar to the
a natural area
initial condition. In the case of the
clear cutting and burning, it is

recognized that succession will begin

immediately; however, full recovery to
a mature and diverse tropical forest IMPAACTTS NTTENSTYY REECOVERRY IMPAACT %
SCOORE TMEE Y)) SCOREE IMPAACT
may take considerably more than the
Clearcuttngiin
60 years estimated here. Similarly, in and burnngg 5 60 300 976.6
the absence of cultivation, it was
Plantng heh
estimated that invasive and coca orr
poppy 1 4 4 13.3
competitive species will displace coca
Ferrtzerrliiz
and poppy in several years and an
estimate of four years was used in this npuusts 1 05.5 05.5 02.2

case. Given the need to apply Pestcide
npuusts 2 05.5 1 03.3
fertilizer and pesticides frequently
because of utilization of nutrients and Eradicatonio
spray ? ? ? ?
resurgence of pests, the recovery time
Processing
for these ecological impacts was and effnngn 2 1 2 07.7
judged to be small. The scores were

multiplied to give the impact score and Figure 20 Potential environmental impacts of the cycle
of coca or poppy production. Scores for eradication
the percent impact was based on the
sum of the impact scores. spraying are specifically omitted.

2.2.2 Conceptual model

For the purposes of the risk assessment of the use of glyphosate and adjuvants
in the eradication of poppy and coca, the conceptual model applied was that normally

applied to the agricultural application of pesticides where hazard and risk and directly

related to the toxicity and exposure to the pesticide. Thus, for human health, toxicity
data were compared to exposures estimated from worst-case data and also from more

realistic data obtained in other uses of glyphosate, such as agriculture and forestry.

Because of the low frequency of application of the sprays, exposure from this source is
acute and resulting risks were compared to acute toxicity data. Toxicity data for the

active ingredient, glyphosate, were obtained from the literature and from the results of

acute laboratory-animal tests conducted with the mixture of glyphosate and Cosmo-

Flux® as used in the spray program. It is recognized that glyphosate used in the
eradication program may contribute to exposures via the food chain and drinking water;

these were estimated and compared to toxicity data and exposure guidelines based on

chronic toxicity for glyphosate. In addition, specific human health responses were
assessed in epidemiological studies conducted specifically to address this issue in

Colombia.

In assessing ecological risks, a similar agriculture-based approach was used.

Similar to the above, exposures were estimated from worst-case models, from

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measurements made in other locations, and from measurements based on samples

collected from the environment in Colombia. Because of the frequency of application in
the eradication program (long periods between applications), ecological exposures
resulting from the eradication spray operations were acute and were compared to acute
toxicity data. Toxicity data were obtained from the literature and from laboratory-based

tests on standard test organisms that were specifically conducted on the spray mixture
as used in Colombia. The risk hypotheses are discussed below and the remainder of
the document is focused on tests of these hypotheses.

2.2.3 Risk hypotheses

A large number of hypotheses were actually tested in this risk assessment;
however, they were basically the same hypothesis with minor differences in the
exposure and toxicity parameters. As is normal in the scientific method (Popper 1979),
these hypotheses are stated as the null or negative hypothesis. Again, following the

scientific method, we attempted to falsify or disprove these hypotheses through the use
of appropriate data.
For human health, two main hypotheses were used:

• Exposures to glyphosate and adjuvants as used in the poppy and coca
eradication programs do not cause acute adverse effects to humans exposed
via a number of routes.

• The use of glyphosate and adjuvants in those locations where eradication of
poppy and coca are conducted does not result in acute and chronic health
outcomes that are different from other locations where glyphosate is not used
or is used in other agricultural practices.

For ecological effects, one main hypothesis was used:
• Exposures to glyphosate and adjuvants as used in the poppy and coca

eradication programs do not cause acute or chronic adverse outcomes on
non-target organisms exposed via a number of routes

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3 EXPOSURE CHARACTERIZATION
Exposure characterization is one of the key components to any risk assessment

(NRC 1993, USEPA 1992, 1998). No measurements of farmer or pesticide applicator
exposures have been made in Colombia. An assessment of pesticide use among
farmers in the Amazon Basin of Ecuador has shown that paraquat and glyphosate are

widely used. Risk behaviors were identified as frequent pesticide use, washing
pesticide equipment in water sources used by humans, inadequate disposal of empty
pesticide containers, eating and drinking during pesticide application, and using

inadequate protective clothing (Hurtig et al. 2003). However, agricultural uses such as
these are quite different from the aerial applications of glyphosate for eradication of
coca and poppy in Colombia. In the following sections, the potential for exposures in

humans and the environment to glyphosate as used in the eradication program of
humans is discussed and characterized.

3.1.1 Human exposure groups

In the case of human exposures to pesticides in the agricultural setting there are
usually two groups that are considered – applicators and bystanders. The group that

experiences the greatest probability of exposure is the applicator group, which, in this
case, includes the mixer-loaders, the spray-plane pilots, and the technicians who work
on and service the aircraft. The second group is the made up of bystanders who may

come into contact with the herbicide during application via direct deposition if they are
within the spray swath, are directly exposed to spray drift, are exposed to deposits of
spray when they reenter treated fields, or are exposed to the herbicide through the

consumption of food items that have been sprayed, or drinking water that has been
contaminated.

3.1.2 Applicator exposure

Risk to applicators was not a specific target of this assessment; however,
exposure can be characterized for this group. Based on observations of the spray

operations in several locations in Colombia, a number of measures are taken to reduce
the potential for exposure of applicators (Table 6).

Table 6. Protective measures used to reduce exposure of applicators to glyphosate

and formulants as used in poppy and coca eradication programs in Colombia.
Applicator Mixer-loader Spray pilot Aircraft technician
subgroup

Technology for Use of closed-loading Not involved in mixing Not normally involved in
handling of the systems and pumps to and loading. mixing and loading.
formulation and mix and transfer Aircraft are washed down
spray mix. glyphosate and Cosmo- regularly so that exposure
Flux® to the aircraft. via contaminated
surfaces in reduced.

Protective Long pants, long sleeves, None other than normal Short or long sleeves,
equipment worn. full rubber apron, rubber clothing, long sleeves, shorts or long pants,
gloves, cloth hat or cap, long pants, jacket, and boots or sneakers, cloth
particulate air filter andboots. cap or none.

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Table 6. Protective measures used to reduce exposure of applicators to glyphosate
and formulants as used in poppy and coca eradication programs in Colombia.

Applicator Mixer-loader Spray pilot Aircraft technician
subgroup
dark glasses, leather
military-style boots.

Equipment used Eye-wash station at all Same as is available to Same as is available to
to remove locations, clean water fothe mixer loader. the mixer loader.
contamination, washing hand and any
should it occur. contaminated surfaces, a
shower in some
locations.

No measures of exposure were available for mixer loaders in Colombia;

however, they are likely to be similar to those of applicators in other situations. Based
on observations on forestry and agricultural applicators (Acquavella et al. 2004, and
summarized in Williams et al. 2000), exposures are generally small. From several

studies, peak estimated exposure in applicators from all routes was 0.056 mg/kg body
weight. The estimate of chronic exposure from all routes was 0.0085 mg/kg/day based
on an 8 hour day and a 5 day work week. In the results of the recently published Farm
Family Exposure Study, the greatest estimated systemic dose in a sample of 48

applicators was 0.004 mg/kg (Acquavella et al. 2004). In the spray program in
Colombia, mixing and loading is done by one or two individuals who wear appropriate
protective equipment. Pilots have limited opportunity for exposure and, as has been

observed in other studies (Frank et al. 1985), will likely experience less exposure.
Exposures of mixer-loaders under the conditions of use in Colombia are likely to
be similar to those observed in agricultural applications. Exposures for spray pilots and

technicians will likely also be less than an agricultural applicator.
While most of the protective clothing worn by the mixer loaders is appropriate,

the need for a respirator is questionable and the use of dark glasses in place of a full
face shield is judged inappropriate. Dark glasses will not protect the eyes from a splash
to the forehead that runs into the eyes, a vulnerable area in terms of glyphosate

exposure during mixing and loading (Acquavella et al. 1999). A full face shield would
offer better protection. As glyphosate is not volatile, nor atomized during mixing and
loading, use of a respirator offers little reduction in potential exposure and complicates
the use of a full face shield. The usefulness of a respirator is judged to be small.

3.1.3 Bystander exposure
Bystanders are the second group that can be exposed to glyphosate during

application. Bystanders can be classified into several classes, depending on their route
of exposure. These are discussed in the following sections.

3.1.3.1 Bystanders directly over-sprayed

Although it is unusual for people to be present in a coca field during application, it
is possible that a person could be standing directly in the spray swath and would

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receive a direct application of the spray solution to the body. There are several
scenarios that could occur (Figure 14 and Table 7).

The most likely scenario is the partially clothed human with a cross-sectional
area of 0.25 m² exposed to the spray (bold text in Table 7). Given that glyphosate
penetrates poorly through the skin with maximum penetration of about 2% (Williams et

al. 2000), the body dose under a reasonable worst-case exposure will be approximately
0.08 mg/kg body weight.

Extreme worst case Worst case Most likely case

Figure 21 Illustration of human exposure scenarios

Table 7. Estimates of human exposure to glyphosate during a spray application
Scenario Exposure in mg/kg body weight
Coca sprayed at Poppy sprayed at 1.2

4.992 kg/ha kg/ha
Naked human, total coverage of
body, and complete penetration

through skin. 14.2 3.4
Partially clothed human with cross
sectional area of 0.25 m , complete
penetration. 1.8 0.4

Partially clothed human with 2
cross sectional area of 0.25 m ,
2% penetration – most likely. 0.04 0.01
2
Assumptions: (human weighs 70 kg and has a body surface area of 2 m

Bystander exposure to glyphosate was estimated as 0.0044 mg/kg/day for a

child, 1-6 years of age (Williams et al. 2000). Exposures to glyphosate were measured

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in bystanders to farm applications (Acquavella et al. 2004). These studies were
conducted in spouses and children who were not involved in applications and frequency
of measurable exposure was small with 4 and 12% of the spouses and children
respectively with detectable exposures based on urinary monitoring. The maximum

systemic dose estimates for spouses and children were 0.00004 mg/kg and 0.0008
mg/kg, respectively (Acquavella et al. 2004). If bystanders are not directly sprayed nor
reenter the field immediately after spraying, their exposures will likely be within a factor
of 10 of farm bystanders. All of these measured exposures are considerably less than

those estimated in Table 7. The values in Table 7 were thus considered to be
reasonable worst-case values.

3.1.3.2 Re-entry

If a person was to reenter the sprayed field immediately after spraying and come
into close contact with the treated foliage, such as when attempting to pick leaves from
spayed coca plants, exposure to glyphosate could occur through the hands and arms.

Given the area exposed, the small penetration, and the saturation of the transfer that
would result once the hands were wet, total body dose is likely to be less than the
reasonable worst-case scenario described in Table 7. The potential for re-entry
exposure has been summarized by Williams et al. (2000). Re-entry exposures

decreased with time after application and, on day-7 after application, were 3% of those
estimated for day 1. Re-entry into areas of tall weeds (1.5 m) resulted in 10-fold greater
exposures than in areas of short grass. Based on measurements in farm workers,
estimates of re-entry exposure to glyphosate in adults ranged from 0.0000039 to 0.0026

mg/kg/h of reentry time. Maximum re-entry exposure for a 1-6 year-old child was
estimated at 0.026 mg/kg for a 5 hour contact period. As these estimates are based on
a spray application rate of 1 kg/ha, re-entry exposures under Colombian conditions are
estimated to be somewhat greater (Table 8). These numbers are also greater than the
direct overspray as the people involved may have repeated exposures if they reenter a

field immediately after spraying.

Table 8. Estimates of human exposure to glyphosate during re-entry to treated
fields
Scenario Exposure in mg/kg body weight

Coca sprayed at Poppy sprayed
4.992 kg/ha at 1.2 kg/ha

Maximum re-entry exposure estimated for
an adult human with a 10 hour day. 0.013 0.003

Maximum re-entry exposure estimated for
a 1-6 year-old child with a 10 hour day. 0.259 0.062

3.1.3.3 Inhalation

-3 Because the vapor pressure of glyphosate (isopropylamonium) is small-10.1 x 3
10 -1Pa at 25°C) and it also has a small Henry’s Law Constant (4.6 x 10 Pa m
mol ) (BCPC 2002-2003), it will not be present in air as a vapor at biologically relevant

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concentrations. The droplet sizes resulting from the spray application of glyphosate in

Colombia are large with a mean droplet diameter of about 1000 µm and with very few
droplets <500 µm. As such, they are unlikely to be inhaled and penetrate into the lungs.
Based on measurements of glyphosate concentrations in air during applications, the
maximum estimated daily dose (8 h) resulting from inhalation of spray droplets by
applicators was 0.0062 mg/kg (Williams et al. 2000), a value that is judged to be

applicable as a maximum exposure for bystanders to eradication spraying in Colombia.

3.1.3.4 Dietary and drinking water
As shown in Table 9, dietary and drinking water exposures to glyphosate have
been estimated to be relatively small under conditions of use in N. America (Williams et

al. 2000).

Table 9. Worst-case daily human exposure estimates for glyphosate
(mg/kg/day)
Sources Female adult Female child (1-6 years)
Acute Chronic Acute Chronic

Drinking water 0.000036 0.000002 0.000110 0.000004
Diet 0.024 0.024 0.052 0.052
Wild foods 0.045 0.045

Total from diet and 0.069 0.024 0.097 0.052
water
Values extrapolated from the above (Williams et al. 2000) to the greater

application rate of 4.992 kg/ha used in control of coca
Drinking water 0.000179 0.00001 0.00055 0.000018
Diet 0.119 0.119 0.259 0.259
Wild foods 0.224 0.224 0.224 0.489

Total from diet and 0.343 0.293 0.483 0.747
water

The results of monitoring programs conducted by the Danish Veterinary and
Food Administration from 1997 to 1999, reported on the content of glyphosate and
several other pesticides in cereals produced in Denmark (Granby and Vahl 2001).
Based on the residues of glyphosate in cereals, intake of glyphosate for a 60 kg adult
was estimated at 0.007 mg/day.

Based on a study of 51 streams in nine Midwestern US States, the U.S.
Geological Survey (USGS) reported the presence of glyphosate and a number of other
herbicides in surface waters (Scribner et al. 2003). Of a total of 154 water samples
collected during 2002, glyphosate was detected in 36 percent of the samples, and its

degradation product, aminomethylphosphonic acid (AMPA) was detected in 69 percent
of the samples. The highest measured concentration of glyphosate in any sample was
8.7 µg/L. The highest concentration of AMPA detected in the USGS study was 3.6
μg/L. Concentrations of glyphosate detected in surface waters in Colombia (see below)
were, for the most part, less than 25 μg/L, the method detection limit. Exposures from

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drinking of untreated surface waters in areas where eradication spraying takes place
are judged to be small and infrequent.

3.1.4 Environmental exposures

3.1.4.1 Air
As discussed above, the presence of glyphosate in air is unlikely as it, and the
salt forms commonly used in glyphosate formulations, have essentially negligible vapor

pressure. Spray droplets may, however, be present in air and are the likely reason for
the detection of glyphosate, along with other pesticides, in rainwater in the European
Union (EU) (Quaghebeur et al. 2004). During the period from 1997 to 2001, glyphosate
was only detected in rainwater in Belgium in 2001 and then with a frequency of 10%

and a maximum concentration of 6.2 µg/L.

3.1.4.2 Water
If water is directly over-sprayed during a spray operation, contamination of

surface waters will result. Some coca fields are located near to ponds and lakes and
some are near to streams and rivers (Helling 2003). While surface waters are not
deliberately sprayed by the pilots, some over-spray of small watercourses and the
edges of ponds, reservoirs, and lakes may occur. In the absence of measured

concentrations immediately after spraying in surface waters located close to the fields,
estimates of exposure were made using worst-case assumptions (Table 10) based on
water depth assumptions used by the US EPA (Urban and Cook 1986) and the EU
(Riley et al. 1991).

Table 10. Estimates of concentrations of glyphosate in surface water after a spray
application
Scenario Exposure in μg/L (glyphosate ) a
Coca sprayed at Poppy sprayed

4.992 kg/ha (3.69 at 1.2 kg/ha (0.89
kg AE/ha) kg AE/ha)

Surface water, 2 m deep, rapid mixing
and no absorption to sediments, no flow. 185 44

Surface water, 0.3 m deep, rapid mixing
and no absorption to sediments, no flow. 1,229 296

Surface water, 0.15 m deep, rapid mixing
and no absorption to sediments, no flow. 2,473 595

Surface water, 0.15 m deep, rapid mixing
and 50% absorption to sediments, no
flow. 1,237 297
aNote that the concentration is expressed as glyphosate acid to allow comparison to exposures
used in environmental toxicity testing. In both these exposures and in the toxicity testing Cosmo-
Flux®, proportional amounts are present and the exposure and toxicity values are thus directly
comparable and can be used to assess the hazard of the mixture as applied in Colombia.

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Glyphosate has been detected in surface waters (see above discussion on

human exposures through drinking water) in a number of locations. Glyphosate
residues have been reported in surface waters in Denmark as result of agricultural
activities. These residues were observed as part of the Pesticide Leaching Assessment
Program (PLAP), a project that was intended to study the leaching potential of

pesticides to the groundwater (Kjaer et al. 2005, Kjaer et al. 2003). PLAP was focused
on pesticides used in farming and monitored leaching at six agricultural test sites
representative of Danish conditions. Water from special drilled wells and from normal
tile drains was analyzed for glyphosate and aminomethylphosphonic acid (AMPA, a
major degradate of glyphosate). It is not clear from the report if the samples were

filtered prior to analysis. This is important as glyphosate binds strongly to organic
matter in soils and can be transported in this form. The presence of macropores in the
soil would facilitate transport to the tile drains.

In the samples from PLAP collected following glyphosate applications, there were
no detections of glyphosate or its metabolite, AMPA, that exceeded 0.1 µg/L in any of
the groundwater samples taken from the suction cells (1 and 2 m below ground
surface), the vertical wells (about 1.5 – 5.5 m below ground surface), and the horizontal
wells (about 3.5 m below ground surface).

Glyphosate residues were detected in water from tiles draining the field and were
observed primarily in the autumn. The highest measured concentrations were 5.1 µg/L
for glyphosate and 5.4 µg/L for AMPA. The calculated average annual concentrations
of glyphosate and AMPA in drainage water were 0.54 and 0.17 µg/L, respectively, at

one location, and 0.12 µg/L and 0.06 µg/L, respectively, at a second location. At a third
location, glyphosate and AMPA were detected but average concentrations of both were
below 0.1 µg/L. In other studies in Danish soils, degradation of glyphosate was shown
to be slower in sandy soils than gravel but leaching was observed only in rounded

gravel soils (Strange-Hansen et al. 2004) and leachate concentrations were less than
0.1 µg/L (Fomsgaard et al. 2003). Similarly, a recent study on fate of glyphosate in soils
showed rapid dissipation with almost total dissipation one month after application (Veiga
et al. 2001). Given the small organic content of gravel and the presence of macropores
between the grains of gravel, movement through this matrix is not surprising. Complete

degradation in other types of soil is as would be expected.
Other authors have reported glyphosate residues in surface waters in Europe
(Skark et al. 1998, Skark et al. 2004) the frequency of detection was not large. The

authors of these papers suggested that the contamination was from application to
railroad beds, environments where gravel is used and where adsorption would be
expected to be minimal. This conclusion is supported by other studies on the
dissipation of herbicides applied to railroad beds (Ramwell et al. 2004) and highways
(Huang et al. 2004, Ramwell et al. 2002). Application of glyphosate to hard surfaces in

an urban context (road edges) can give peak run-off concentrations of 650 μg/L
(Ramwell et al. 2002), but only 15 μg/L from a railway trackbed (Ramwell et al. 2004).
In Germany, a study of two catchments found that non-agricultural pesticide use
contributed more than two-thirds of the whole observed pesticide load in the tributaries
and at least one-third in the River Ruhr (Skark et al. 2004). Most of the non-agricultural

pesticides were derived from run-off from domestic, industrial and railway areas.
Nevertheless, in Argentina, where glyphosate-tolerant soybean is now extensively

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grown and regularly treated, no residues have been observed in soil or water, either of

glyphosate or its metabolite, AMPA (aminomethylphosphonic acid) (Arregui et al. 2004).
The USGS study on Midwestern US streams (Scribner et al. 2003), analyzed
samples of water that were filtered through a 0.7 µm filter, thus the concentrations

represent dissolved glyphosate and AMPA. Summary data from this study are shown in
Table 11.

Table 11. Summary data on glyphosate concentration in Midwestern US streams
Herbicide Number of Concentration in μg/L
th
samples 95 centile Maximum
Pre-emergence runoff samples

Glyphosate 51 0.58 1.0
AMPA 51 0.55 1.8

Post-emergence runoff samples
Glyphosate 52 1.5 4.5

AMPA 52 0.94 2.0
Harvest-season runoff samples

Glyphosate 51 0.45 8.7
AMPA 51 1.3 3.6

Data from (Scribner et al. 2003)

Although the concentrations of glyphosate detected in surface waters in other
areas where glyphosate is used in agricultural and other activities are relatively small,
concentrations have not been measured in Colombia. To address this uncertainty, we
conducted a monitoring study to measure concentrations of glyphosate, AMPA and

other pesticides in surface waters.
The surface water monitoring study was conducted in five locations in Colombia
representing areas where spraying of coca was planned to take place or where other

agricultural activities were undertaken and were also close to where human health
studies were being conducted. The sites were selected for safe access as well as ease
of repeated sampling. These locations are summarized in Table 12 and further details
as to temperatures, rainfall, and soil characteristics are provided in separate reports
(PTG 2005a, b, c, d, e)

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Table 12. Characteristics of sampling sites for glyphosate, AMPA and other pesticides

in surface waters and sediments in regions of Colombia
Site name Location Altitude Major crop Known pesticide use
(m) types

Valle del N 03º27.642' 1002 Sugar cane Glyphosate and other
Cauca, Río W 076º19.860' pesticides
Bolo

Boyacá, N 05º40.369' 557 Coca Manual eradication,
Quebrada W 074º00.986' no aerial spraying of
Paunera glyphosate
Sierra Nevada, N 11º13.991' 407 Organic coffee None

Quebrada La W 074º01.588'
Otra
Putumayo, Río N 00º43.259' 329 Coca Aerial eradication

Mansoya W 076º05.634 spraying
Nariño, Rio N 01º27.915' 15 Coca Aerial eradication
Sabaletas spraying
W 078º38.975'

To characterize concentrations of glyphosate and AMPA in surface waters,
samples were taken at weekly intervals for a period or 24 weeks (CICAD/OAS 2004c).
Samples, in plastic bottles, were frozen and held at -17C until shipped to Canada for
analysis using standard methods (Thompson et al. 2004). The Method Detection Limit
(MDL) for the analysis was 25 μg/L. Duplicate samples were taken and one sample

held in Colombia until the duplicate had been analyzed. In addition, field-spiked
samples and blanks were taken at bi-weekly intervals. In addition to water, sediment
samples were taken at monthly intervals for analysis of glyphosate and AMPA if
significant concentrations were detected in surface waters. Appropriate field spikes and
blanks of sediment were also taken bi-monthly. Analytical quality control samples

showed excellent recovery efficiency and precision of the analytical method with 98%
recovery for glyphosate and 8.8% coefficient of variation (CV); 110% recovery efficiency
for AMPA with 20% coefficient of variation. Blank field sample analyses show, on
average, that no co-extractive interferences above the MDL for either glyphosate or
AMPA at any of the sample sites. Field spike samples generally showed no significant

degradation of glyphosate during sample handling and transport with overall average
value of 90% of expected concentrations.
Results of these analyses are summarized in Table 13 (raw data are presented
in Appendix 1). In all locations and on most occasions, residues of glyphosate and

AMPA were present at concentrations below the MDL of 25 μg/L. On one occasion
each in Valle del Cauca and Boyacá, concentrations of 30.1 and 25.5 µg/L, respectively,
were found. These are sites where eradication spraying was not carried out and where
the only use of glyphosate, if any, was in agriculture. These data suggest that little or
no contamination of surface waters with glyphosate at significant concentrations has

resulted from the use of glyphosate in either agricultural or eradication spraying in

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Colombia. As concentrations in surface waters were mostly below the MDL, sediment
analyses were not performed.

Table 13. Concentrations of glyphosate (AE) and AMPA in samples of surface water
collected in Colombia between October 2004 and March 2005

Site name Total number Frequency of detection (n and
of samples %) for site

Glyphosate AMPA
Valle del Cauca, Río Bolo 17 1 (5.9%) 0 (0%)

Boyacá, Quebrada Paunera 18 1 (5.5%) 0 (0%)
Sierra Nevada, Quebrada La Otra 18 0 (0%) 0 (0%)
a
Putumayo, Río Mansoya 16 0 (0%) 0 (0%)
Nariño, Rio Sabaletas a, b 17 0 (0%) 0 (0%)
a
Locations where eradication operations were planned.
bLocation where eradication spraying was carried out during the sampling period.

To characterize concentrations of other pesticides in surface waters and
sediments, samples of water were taken in glass bottles every two weeks for a period of
22 weeks (CICAD/OAS 2004b). Samples were held at 4ºC until shipment to Canada for

analysis. Analyses were conducted at the Laboratory Services Division of the
University of Guelph using standard methods (LSD 2005). Duplicate samples were held
in Colombia until analyses were completed. Field spikes and blanks were taken at 5-
week intervals as were sediment samples. Sediment blanks and spikes were taken

once during the study period.
The results of the analyses for other pesticides are summarized in Table 14 (raw
data are presented in Appendix 2A-G). Blanks showed no contamination of samples

during storage and shipping. Spiked samples showed variable recovery, particularly for
carbaryl. Several pesticides were detected in surface waters. This is not unexpected
as pesticides are widely used in agriculture in Colombia and, based on similar
experience in other locations, some contamination of surface waters will occur. Of

interest is the detection of endosulfan (I and II) and its breakdown product, endosulfan
sulfate, in the samples taken at the Nariño site. Endosulfan is not registered for use in
Colombia and its detection here likely is the result of illegal use. Whether this
contamination resulted from regular agricultural activity or from use in the production of

coca is unknown.

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Table 14. Concentrations of other pesticides in samples of surface water and

sediments taken in Colombia between October 2004 and March 2005
Site name Number of Frequency of detection in
samples surface water

Number Pesticides
detected
Valle del Cauca, Río Bolo 10 3 2,4-D

Boyacá, Quebrada Paunera 8 0 0
Sierra Nevada, Quebrada La Otra 9 0 0

Putumayo, Río Mansoya 9 0 0
Nariño, Rio Sabaletas 8 1 endosulfan I,
endosulfan II,

endosulfan
sulfate
Number of Frequency of detection in

samples sediment
Number Pesticides
detected

Valle del Cauca, Río Bolo 3 0 0
Boyacá, Quebrada Paunera 3 0 0

Sierra Nevada, Quebrada La Otra 3 0 0
Putumayo, Río Mansoya 3 0 0

Nariño, Rio Sabaletas 2 0 0

3.1.4.3 Soil

Concentrations of glyphosate and AMPA in soils can be estimated from the
application rates used in the eradication program (Table 15) and measurements could
be made through the use of residue analysis, however, the more important question is
the biological availability of the glyphosate, as this would determine its potential for

biological effects.

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Table 15. Estimates of glyphosate concentration in the top 25 mm of soil following a
spray application
Scenario Exposure in mg/kg

Coca sprayed at Poppy sprayed
4.992 kg/ha at 1.2 kg/ha
Direct deposition on bare soil with a

density of 1.5 kg/L. 13.3 3.2
Deposition on soil with a density of 1.5

kg/L under a canopy of foliage with an
assumed interception of 50%. 6.7 1.6

While there are no direct measurements of glyphosate and AMPA concentrations
available from treated

coca and poppy fields in
Colombia, the biological
activity of any residues
that may be present is

judged to be small as the
sprayed fields rapidly
become colonized with
invasive plants or are

replanted to coca soon
after spraying. From
visual observations (Figure
15), from observation in
other uses and other

locations (Section 4.3.1),
and from other reports
(Helling 2003), this
recolonization is rapid and
Figure 22 Photograph of coca plants near Caucasia, Colombia,
there have been no replanted from cuttings in a field sprayed with glyphosate 56 days
adverse effects observed previously (Photo, K Solomon, 2004 06 09).
in terms of recolonization
or replanting of the sprayed fields.

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4 EFFECTS CHARACTERIZATION

4.1 GLYPHOSATE

The human-health and environmental effects of glyphosate have been
extensively reviewed in the literature (Giesy et al. 2000, Solomon and Thompson 2003,

Williams et al. 2000) and by regulatory agencies (NRA 1996, USEPA 1993a, 1997,
1999, World Health Organization International Program on Chemical Safety 1994) 1.

The following sections are primarily directed to a critical analysis of original articles
published since 1999 or that were not included in the earlier reviews (Giesy et al. 2000,
Solomon and Thompson 2003, Williams et al. 2000).

4.1.1 Effects of glyphosate on mammals

4.1.1.1 Laboratory toxicity studies
The toxicity of glyphosate and the formulation Roundup® were reviewed recently

(Williams et al. 2000). Glyphosate and its isopropylamine salt have low acute toxicity by
the oral, dermal, and subcutaneous routes of exposure (Table 16).

Table 16. Acute toxicity of glyphosate in selected mammals
Species Route Compound administered a LD50 (mg/kg bw)

Mouse Oral Glyphosate >10,000.
Glyphosate 1,538.
Subcutaneous Glyphosate saline 6,250.(M)
Glyphosate saline 7,810.(F)
Intraperitoneal Glyphosate saline 545.(M)
Glyphosate saline 740.(F)

Glyphosate 134.
Rat Oral Glyphosate, Roundup, Glyphosate >5,000.
isopropylamine salt
Dermal Roundup >17,000.
Inhalation Roundup, Glyphosate saline LC50=3.18 mg/L (4 hours)

Subcutaneous Glyphosate saline 17,500.
Glyphosate saline 281.(M)
467.(F)
Intraperitoneal Glyphosate 238.
Rabbit Oral Glyphosate 3,800.

Dermal Glyphosate, Roundup, Glyphosate >5,000.
isopropylamine salt.
Goat Oral Glyphosate, Roundup, Glyphosate >3,500.
isopropylamine salt.
Data from (Smith and Oehme 1992).

1
It should be noted that several publications on glyphosate have appeared in the literature which
focus on the adverse effects of glyphosate. A pamphlet/brochure by Post (1999) was produced on behalf
of an activist organization. The pamphlet was very brief and was not peer-reviewed. In addition, an
article purporting to be a scientific review was published in 1998 (Cox , 1998) in the “Journal of Pesticide
Reform”. It should be noted that the Journal of Pesticide Reform does not publish original articles, is not
peer-reviewed, is produced by an activist group, and that the editor is often the author of the articles.

Because of this, these articles were not used in this report.

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Toxicity was greatest by intraperitoneal administration. When rats and mice

were given glyphosate orally or intraperitoneally, several stress symptoms, increased
respiration, elevated rectal temperatures, and occasional asphyxial convulsions were
noted. Median lethal doses of 4,704 mg/kg to the rat and 1,581 mg/kg to the mouse
orally were significantly higher than 235 and 130 mg/kg, respectively, median lethal

doses obtained when glyphosate was given intraperitoneally. Lung hyperemia was the
major lesion noted in the glyphosate poisoned animal (Bababurmi et al. 1978).
There is limited information on acute toxicity in dogs. However, there is a
retrospective study conducted of 482 glyphosate calls recorded at the CNITV of France

between 1991 and 1994. Only 31 cases were assessed as certain or highly probable
and were linked with direct ingestion of glyphosate concentrates or spray in 25 dogs.
The symptoms were most frequently described as vomiting, hypersalivation and
diarrhea; prostration and paresis were not common. Symptomatic treatment resulted in

rapid recovery without sequelae (Burgat et al. 1998). Campbell and Chapman (2000)
described the onset of clinical effects in dogs observed in several cases of poisoning as
usually between 30 minutes and 2 hours. Recovery usually occurs over 1-2 days.
Salivation, vomiting, diarrhea, irritation, and swelling of lips are common early features.
Tachycardia and excitability are often present in the early stages, with the animals

subsequently becoming ataxic, depressed, and bradycardic. Inappetence, pharyngitis,
pyrexia, twitching, shaking, and dilated pupils is noted occasionally. Rarely, jaundice,
hepatic damage, and haematuria have been reported. Eye and skin irritation are also
possible. Tachypnoea occurs in glyphosate poisoning in other animals but does not
appear to be a feature of glyphosate toxicity in dogs.

Some recent studies have examined effects of chronic feeding of glyphosate to
Wistar rats. A study was performed to measure the activity of some enzymes with a
function in the pathways of NADPH generation, isocitrate dehydrogenase, glucose-6-

phosphate dehydrogenase and malate dehydrogenase in liver, heart and brain of
pregnant Wistar rats and their fetuses which were exposed to glyphosate solutions
0.5% and 1% at a dose of 0.2 and 0.4 ml/ml water during 21 days of pregnancy.
Glyphosate affects these enzymes in the studied organs of the pregnant rats and their
fetuses (Daruich et al. 2001).

Feeding Glyphosate-Biocarbo® formulation at rates of 4.87 mg/kg every two
days for 75 days resulted in the leakage of hepatic intracellular enzymes, alanine
aminotransferase (ALT) and aspartate aminotransferase (AST), suggesting irreversible

damage in hepatocytes (Benedetti et al. 2004). The formulation used in this study was
from Brazil and the nature of the formulants is unknown. In addition, the exposures
extended over a long period of time and are inappropriate for assessing risks from acute
and infrequent exposures such as may occur in eradication spraying.

The effect of glyphosate on several enzymes was studied in vitro. The enzymes
were: serum acetylcholinesterase (AChE), lactate dehydrogenase (LDH), aspartate
amino-transferase (AST), alanine aminotransferase (ALT, alkaline phosphatase (AP)
and acid phosphatase (AcP). Results revealed that glyphosate inhibited all enzymes

except AcP. IC50 values were 714.3, 750, 54.2, 270.8 and 71.4 mM for ACHE, LDH,
AST, ALT, and AP, respectively (El-Demerdash et al. 2001). The most sensitive
response, that of aspartate amino-transferase was observed at a concentration of 54.2
mM, which is equivalent to a concentration of 9,056 mg/L, a concentration that would

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not occur in vivo. These results of the studies discussed above do not suggest that

glyphosate would have effects at concentrations lower than those previously observed.
Glyphosate has not been found to be genotoxic, mutagenic or carcinogenic.
Glyphosate was not teratogenic or developmentally toxic (Williams et al. 2000) except at

large exposures. Some studies were not reviewed by Williams et al. (2000) or were
published after 2000. These are reviewed below.
In a study on Charles River CD-1 rats, test animals were given oral gavage

doses (direct intubation into the stomach) of 0, 300, 1000 and 3,500 mg/kg body weight
(bw)/day of glyphosate from day 6-19 of gestation. Control animals received 0.5%
methocel. No internal or skeletal anomalies were seen at 300 and 1000 mg/kg bw/day,
although maternal toxicity was apparent at 3,500 mg/kg bw/day with soft stools,
diarrhea, red nasal discharge, reduced body weight, and death by gestation day 17

(6/25). In addition, mean fetal body weights were significantly reduced and early fetal
resorption were significantly increased at this dose level (Rodwell 1980a). Female
Dutch belted rabbits were given oral gavage doses of 0, 75, 175, and 350 mg/kg bw/day
glyphosate from day 6-27 of gestation. Control animals received 0.5% methocel. No
internal or skeletal abnormalities were seen (Rodwell 1980b). In a study from Brazil,

examination of pregnant Wistar rats dosed orally with Roundup® from day 6 to 15 of
pregnancy with rates of 0, 500, 750, or 1000 mg/kg of glyphosate showed skeletal
alteration in fetuses (15.4, 33.1, 42.0, and 57.3%, respectively). There was 50%
mortality of dams at 1000 mg/kg only (Dallegrave et al. 2003). The doses used in this
study were large and considerably greater than those used in an earlier study (reviewed

by Williams et al. 2000). In the earlier study, a No-Observed-Effect-Level (NOEL) of 15
mg/kg/day was described for fetal effects and 300 mg/kg/day for maternal effects.
Given the very large doses used in the Dallegrave et al. study (2003), their results are
not surprising and do not change the assessment of teratogenic potential. The Rodwell

studies discussed above also showed responses at concentrations greater than those
reviewed in Williams et al. (2000) and do not change the assessment of teratogenic
potential.
A number of recent studies have been carried out in tissue culture systems. One

of these assessed the affect of several formulated pesticides on the steroidogenesis
pathway (StAR protein synthesis) in tissue cultures of mouse testicular Leydig tumor
cells (Walsh et al. 2000). Exposure to the formulation at 25 mg/L in the cell culture
medium did cause a reduction in steroidogenesis, but only for a period less than 24

hour during which there was recovery. In another study on tissue cultures, Lin and
Garry reported results of bioassays carried out in cultures of the MCF-7 breast cancer
cell (Lin and Garry 2000). The results presented by the authors indicated that, while
some pesticides caused estrogen-like receptor mediated effects at high exposure
concentrations, both glyphosate and the Roundup® formulation of glyphosate induced

non-estrogen like proliferation, thereby supporting the view expressed by others
(Williams et al. 2000) that neither glyphosate nor Roundup® are endocrine disruptors.
The results of studies on cells in vitro are difficult to interpret as they exclude the normal
pharmacokinetic and metabolic functions that would be present in whole animals. They
should be compared to the multigenerational study used by regulatory agencies

worldwide to assess reproductive/developmental toxicity, which is the most definitive
study design for the evaluation of potential endocrine modulating substances in humans

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and other mammals. Comprehensive reproductive and developmental toxicology

studies carried out in accordance with internationally accepted protocols have
demonstrated that glyphosate is not a developmental or reproductive toxicant and is not
an endocrine disruptor (Williams et al. 2000) (USEPA 1993a) (World Health
Organization International Program on Chemical Safety 1994).

There was no evidence of neurotoxicity in a number of studies on glyphosate
reviewed in Williams et al. (2000). Neurotoxicity was not observed in the large number
of acute, subchronic, and chronic studies conducted in rodents nor was it observed in
two specific neurotoxicity studies conducted in dogs. However, these studies did no

assess potential effects on neurotransmitters and their metabolites in the brain and
other parts of the nervous system –- measures of response used in current testing
protocols for neurotoxicity.

Some reports on the immunotoxicity of glyphosate have appeared in the
literature. Mice exposed to Roundup® at concentrations up to 1.05% in drinking water
for 21 days showed no change in immune function (T-lymphocyte and macrophage-
dependent antibody response) when, on day 21 of the herbicide exposure period, they
were inoculated with sheep erythrocytes (Blakley 1997). In an in vitro study on

cytokine production by human peripheral blood mononuclear cells, glyphosate had only
a slight effect at the greatest concentration tested (1000 μM = 226,000 µg/L)
(Nakashima et al. 2002). Results of both of these studies suggest that glyphosate does
not affect immune response in mammals at realistic exposure concentrations.
However, studies in fish suggest that that there may be some immunotoxic effects.

Short exposures to Roundup® (10 minutes in a concentration of 100,000 µg/L) in carp
(Cyprinus carpio) and European catfish (Silurus glanis) caused a decrease in metabolic
and phagocytic activity as well as proliferative response (Terech-Majewska et al. 2004).
In contrast to these effects at large concentrations, responses on splenic antibody

plaq-2 forming cells in the fish, Tilapia nilotica, were reported at concentrations of 1.65
x 10 µM (= 4.4 μg/L). As responses of the immune system are difficult to interpret in
terms of survival of individuals or the population, they not formally used in assessment
of pestcides by regulatory agencies.

The toxicokinetics of glyphosate were reviewed by Williams et al. (2000).
Between 15 and 36% of ingested glyphosate is absorbed through the intestinal tract and
only about 2% via the skin. Excretion of unabsorbed glyphosate is via the feces but the
absorbed glyphosate is excreted via the urine with only a small amount of metabolism.

Whole-body half-lives were biphasic, with an initial half-life of 6 hours and a terminal
half-life of 79 to 337 hours in rats (Williams et al. 2000). Clearance from most tissues
was rapid but it was cleared more slowly from the bone, possibly because of ionic
binding to the calcium in the bones (Williams et al. 2000). Glyphosate is clearly not
bioaccumulated and any absorbed dose is excreted in the urine relatively rapidly.

4.1.1.2 Cases of human poisoning
A number of anecdotal reports of human poisoning with glyphosate and
formulations have been published in the literature. In some cases, these are reports of

a single event and an observed response. In one such case toxic pneumonitis was
observed after exposure to a glyphosate formulation (Pushnoy et al. 1998). However,
no information was provided to demonstrate how airborne exposure could have

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occurred and the results are at odds with the known inhalation toxicity of the formulation

(Williams et al. 2000) and tests done on the product as used in Colombia (Section
4.2.2).
In another case, a man accidentally sprayed himself with an unidentified

formulation of glyphosate (Barbosa et al. 2001). He developed skin lesions 6 hours
after the accident but these responded to routine treatment. However, one month later,
the patient presented with a case of symmetrical Parkinsonism syndrome. This is an
isolated case and it is impossible to conclude anything about causality as the disease
may have already been present but asymptomatic. In a similar case, a woman of 78

years old presented with extensive chemical burns in legs and trunk caused by an
accidental contact with a glyphosate formulation. These lesions disappeared, without
consequences a month later (Amerio et al. 2004).

Acute intoxication information has been documented in two case-series studies,
from Taiwan, China where glyphosate formulations were apparently used for attempted
suicide (Chang et al. 1999, Lee et al. 2000). The first paper analyzed 15 intentional
intoxications with glyphosate formulation and found that 68% of the patients presented
esophageal, 72% gastric and 16% duodenal injuries. Esophageal injury was the most

serious injury but was minor in comparison with strong acids. Lee et al. (2000)
analyzed 131 suicide attempts in southern Taiwan. The most common symptoms were
sore throat and nausea. Fatality rate was 8.4%. In this study 20.5% presented
respiratory symptoms and more than half of them needed intubations. The authors
propose that direct damage to the airway passage and mention that surfactant (POEA

MON 0818) may be responsible for the toxicity. In many cases, the exact doses
consumed by persons attempting suicide are not known and it is difficult to interpret
these findings in the context of bystander and other accidental exposures which are
usually many orders of magnitude less. It is, however, interesting to note the low fatality

rate compared to what has been reported from other pesticides such as paraquat and
the organophosphorus pesticides (Krieger 2001).
It is well known that the older formulations of glyphosate that contained the
surfactant POEA (MON 0818) were eye irritants. Goldstein et al. (2002) analyzed 815

glyphosate related “calls” to the Pesticide Illness Surveillance Program (PISP), most of
them involving eye irritation (399), skin (250), upper airway (7) and combinations of
these. Of the 187 systemic cases, 22 (12%) had symptoms definitely related to
exposure to formulations of glyphosate. Again, this is not surprising as the formulation

of glyphosate is acidic (similar to strong vinegar) and the surfactant is an eye irritant. In
other studies on eye and skin irritation reviewed in Williams et al. (2000), none of the
reported exposures resulted in permanent change to the structure or function of the eye.
Based on these findings, it was concluded that the potential for severe ocular effects in
users of Roundup herbicides is extremely low. This observation is consistent with the

minimal ocular and dermal effects observed with the formulation of glyphosate used in
Colombia (Section 4.2.2).

4.1.1.3 Human epidemiology studies

A number of studies in the recent epidemiology literature have attempted to
address the issue of glyphosate exposure and disease incidence in humans.
Epidemiology studies on pesticides commonly suffer from two sources of error.

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Possibly the most important of these is the error in assigning exposures. Exposures in

the studied population are never measured directly and it is common to use surrogates
for exposures such as areas treated with pesticides, number of applications made,
and/or number of years of application. Recent studies have shown that these
surrogates are susceptible to significant errors (Arbuckle et al. 2004), leading to the

following quote from the authors of the paper:
“As the present analysis has shown, the consequences of this assumption could
be a high false-positive rate in classification of exposure. The impact of this kind
of error can be profound and has rarely been quantified. Until improvements are

made in classifying pesticide exposure in epidemiologic studies, results on health
effects will be subject to misclassification bias….”
Similar conclusions have been put forward in other papers (Arbuckle et al. 2005,

Harris et al. 2002, Solomon et al. 2005). A second possible source of error in these
studies is the fact that the populations that are studied (farmers and professional
applicators) typically use many pesticides. Thus, any substance-specific responses and
causality are difficult to ascertain.

Cancer Studies. The work of Hardwell et al. (Hardell et al. 2002) presented a
pooled analysis of two case-control studies, one on non-Hodgkin’s Lymphoma (NHL)
(Hardell and Eriksson 1999) and another one related to a Hairy Cell Leukemia (HCL), a
rare subtype of NHL. In the 1999 study, the authors employed a case control type of

study design for their investigation. Case control studies can suffer from poor exposure
histories and recall bias in that study subjects will be required to recall exposures to a
putative agent which may have occurred decades prior to the onset of the disease
under study. In some cases, study subjects may be deceased (in this study, 192 of the
442 study subjects were deceased) requiring exposure information to be provided by

next of kin, thereby further eroding confidence in data related to exposure histories.
The study reported their results in terms of odds ratio (OR). An OR of >1.0 implies a
greater disease rate for exposed individuals than for the unexposed, while an OR <1.0
suggests a decreased rate of disease in the exposed population. The data for the study
were based on small numbers; only four cases and three controls, or less than 1% of

the overall study subjects, reported the use of glyphosate. Furthermore, the confidence
interval (CI) reported by the authors for exposure to glyphosate was 0.4-13, implying a
lack of statistical confidence. In their pooled analysis (Hardell et al. 2002), they reported
a positive association with use of glyphosate (OR 3.04, 95%CI of 1.08-8.52) when

analyzed using univariate statistics with the highest risk for exposure during the latest
decade before diagnosis. However, the OR was reduced when using multivariate
statistics (OR 1.85, 95%CI of 0.55-6.20). In addition, the study was based on a small
number of cases and controls (8/8) and lacked power to differentiate linkages.

De Roos et al. (2005) evaluated associations between glyphosate exposure and
cancer incidence in the Agricultural Health Study (AHS), a prospective cohort study of
57,311 licensed pesticide applicators in Iowa and North Carolina. Among private and
commercial applicators, 75.5% reported having ever used glyphosate, of which > 97%

were men. In their analysis, glyphosate exposure was defined as a) ever personally
mixed or applied products containing glyphosate; b) cumulative lifetime days of use, and
c) intensity-weighted cumulative exposure. Glyphosate exposure was not associated
with incidence of 12 common cancer types (the relative risk, RR, included 1 in all

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cases), however, the RR for multiple myeloma incidence was 2.6 (95% CI of 0.7–9.4

based on 32 cases in the total of 2,088 cancers), prompting the authors to suggest that
this should be followed up in future studies.
Overall, there is no strong evidence to link glyphosate exposure to increased risk

of cancer. Taken with the lack of any evidence of genotoxicity or carcinogenicity of
glyphosate in laboratory studies (Williams et al. 2000), it is highly unlikely that
glyphosate is carcinogenic in humans.

Neurological effects. A recent study on farmers in the Red River Valley in MN,
USA, reported on the link between glyphosate and Attention Deficit Disorder and
Attention Deficit Hyperactivity Disorder (ADD/ADHD) in children of farmers who applied
it (Garry et al. 2002). They reported OR of 3.6 (95% CI, 1.3–9.6), however, the study
suffered from several potential sources of error. The authors noted the lack of uniform

diagnostic neurobehavioral information related to (ADD/ADHD) and that their study
identified 14 cases of ADD/ADHD among 1,532 live births, a frequency that was actually
considerably lower than background rates of ADD/ADHD that had previously been
reported by researchers in Canada and the US. Notwithstanding, while Garry et al.
(2002) concluded that their study showed a tentative association between ADD/ADHD

and the use of glyphosate, they also noted that other experimental evidence did not
support this conclusion, including that glyphosate was not genotoxic and that little, if
any, evidence of neurotoxicity has been associated with exposure to glyphosate, except
in cases of intentional oral overdose. Finally, the authors did express concern that their
tentative conclusions could be explained by random chance alone, and stated the need

for further detailed neurodevelopmental studies to resolve these outstanding issues.
Overall, there appears to be little evidence to support a link between glyphosate
exposure and neurobehavioral problems in children of exposed applicators.

Reproductive outcomes. Several papers have reported on the relation
between adverse reproductive outcomes and the use of glyphosate. In a study in
Ontario, Canada, Arbuckle et al. (2001) observed a moderate increase in the risk of late
abortions associated with preconception exposure to glyphosate (OR = 1.7 95%CI,1.0-
2.9). Another study in Ontario (part of the Ontario Farm Family Health Study) reported

a positive association (decrease in fecundability of 20%, ratio range = 0.51-0.80) when
both spouses participated in activities where they could be exposed to pesticides. This
was observed for 6 of 13 pesticides categories, one of which was glyphosate (Curtis et
al. 1999). The study was based on 2,012 planned pregnancies. There was no strong

or consistent pattern of associations of pesticide exposure with time to pregnancy. For
exposure intervals in which only the men participated in pesticide activities or in which
neither men nor women participated in pesticide activities but pesticides had been used
on the farm, conditional fecundability ratios ranged from 0.75 to 1.50, with no apparent
consistency among pesticide classes, chemical families, or active ingredients. Again,

while this study did suggest a linkage between pesticide exposure and fecundability,
there is no evidence from laboratory studies that glyphosate is a reproductive toxicant at
exposures that would be expected in humans (Williams et al. 2000).

Overall, there is little epidemiological evidence to link glyphosate to any specific
diseases in humans. This conclusion is supported by laboratory toxicity studies.
However, responses related to reproductive outcomes such as fecundability measured
through time to pregnancy offer a useful measure of possible effects that can be applied

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in situations such as Colombia where other health data are difficult to gather. With this

in mind, we designed a preliminary study to gather human epidemiological data in
several regions in Colombia. These regions were the same as those selected for the
surface-water sampling (Table 12). The design and results of the study are
summarized in the following section. A detailed report is given in a separate document
(Sanin 2005).

4.1.2 Human health epidemiology study in Colombia
The question that this study addressed was: Is glyphosate exposure associated
with adverse reproductive effects? The specific objective was thus to elucidate possible
effects on reproductive health from exposure to glyphosate by assessing

fertility/fecundability among women resident in different areas of the country with
different pesticide use patterns. The design was cross-sectional with retrospective
collection of data and is equivalent to a retrospective cohort. The study population
consisted of 600 women of reproductive age in each of five different areas (Table 17)

Table 17. Characteristics of the areas used in the epidemiology study

Site name Focal crop Known pesticide use
Valle del Cauca Sugar cane Glyphosate and other pesticides. Glyphosate
applied by air.

Boyacá Coca Manual eradication, no aerial spraying of
glyphosate. Use of other pesticides unknown.
Sierra Nevada Organic No pesticide use and no coca known to be

coffee grown. Use of other pesticides unknown.
Putumayo Coca Aerial eradication spraying with lower
intensity. Use of other pesticides unknown.

Nariño Coca Aerial eradication spraying with higher
intensity. Use of other pesticides unknown.

The study protocol and questionnaire were approved through the Ethics Review
Board of the Fundación Clínica Santa Fé de Bogotá, Colombia. All females of
reproductive age in each area were informed about the objectives of the study and
invited to participate if their first pregnancy (independent of the result of it) had occurred

during the last 5 years, they had lived in the region at least for the same period, and
they had not visited a physician for treatment of infertility nor used contraceptives during
the year prior to getting pregnant. First pregnancies were the focus of the
questionnaire. This reduced recall bias and other potential biases that are associated
with subsequent pregnancies. Only one pregnancy was used to maintain outcome

independence and minimize the effect of previous reproductive history.
Reproductive health was characterized through the following dependent
variables (retrospectively) assessed by questionnaire:

Time to pregnancy (TTP): Number of months that it takes a couple to
achieve a clinically detectable pregnancy without the use of
contraceptives. A modified version of the key question from the

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questionnaire of Baird et al. (1991) was used to elicit TTP. Valid data on

TTP can be derived retrospectively, with a recall time of 14 years or more
(Joffe et al. 1995).
Fertility: Percentage of women who achieved pregnancy during the first

year after intent.
The independent variable in the study was exposure to glyphosate for eradication

of illicit crops. This was measured through use information from the region as indicated
in Table 12. There were a number of possible confounders or independent predictors of
the reproductive variables in study. These are listed below:

General Health and Nutrition Status
Women and their partner

Age Complete years
Education Highest grade achieved

Active smoking Smoke or not; number of years number of cigarettes per
day
Alcohol consumption Number of drinks per month

Coffee consumption Number of cups per day
Type of family Nuclear or extended

Socioeconomic (Almost all all participants were stratum 1 – rural)
stratum
Only from Women:

Body Mass Index Weight (Kg) / (Height - m)
Reproductive history Information on the father was also available

Techniques and procedures were as follows: In the five areas we started at the
closest household to the location where water and sediment samples were taken from.

Interviewers visited house by house to identify women who met inclusion criteria until
the sample size (600 women in each zone) was completed. Those who met the
inclusion criteria were informed about the project in a general way and were informed
that there would no be reprisals for participation or non-participation and that the
investigators guaranteed the privacy of the information collected. Each participant

provided written informed consent.
Interviewers and supervisors were trained on the objectives of the project and the
questionnaire for two days. All interviewers lived in the study area and were supervised
by local epidemiologists who knew the study area and who were well known by the

population. These local epidemiologists were supervised by PTG team. All the
information collected was submitted to a quality control procedure. The data were
captured in Microsoft Excel (Microsoft Corporation 2003) and processed with the
STATA 7.0. (Stata Corporation, College Station, Texas) with macros developed by
Dinno (2002). The modified version of the key question from the questionnaire of Baird

et al. (1991) was used to elicit TTP was, “How many months were you having sexual

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intercourse before you became pregnant for the first time?” TTP was defined as

duration in months, not divided by menstrual cycle duration in days, because women
are more able to recall time in months than in cycles (Joffe 1997). For analysis
purposes, if TTP was reported as zero months, the answer was interpreted as one
month. Cutoff points for categorization of continuous variables were set as follows:

Age at time of interview - 25;

Age when started to try to get pregnant and age when first got pregnant -
20.

Of a total of 3005 women interviewed, 413 exclusions were made. These
included: 233 women without TTP data and 21 with TTP values greater than 60 months
and 159 women who consulted to physician about infertility. Hence, 2592 (86.3%) were
included in the analyses reported here.

For each exposure and potential determinant variable, non-parametric ANOVAs
of TTP were conducted. In the fecundability predictor models, censoring of TTP was
introduced, in order to reduce the effect of other medical causes on TTP. If a woman
took more than 12 months to conceive, a value of “null” for a separate censor variable
was included with a value equal to 0 if TTP was 12 months or less and 1 if TTP was

greater than 12 months.
Each month was classified according to the ecological exposure and determinant
variables and an indicator variable was generated for every month giving information on

whether the cycle under this exposure resulted in a pregnancy or not. Fecundability
odds ratios (fOR) were calculated with 95% confidence intervals (95% CI) using a
discrete time analogue of Cox’s proportional hazard model (Baird et al. 1986, Curtis et
al. 1999, Zhou and Weinberg 1999). This process generate a fOR for which values
below unity indicate sub-fertility.

The initial saturated multivariate model included all variables significant on
bivariate analysis (p<0.10) and variables of prime biological importance (age at time of
trying to get pregnant). Variables were eliminated one by one according to the p values

(>0.05) and effects of elimination on the coefficients of other variables in the model
assessed. Several goodness of fit statistics for logistic regression were checked
(Hosmer and Lemeshow 1989). The final model consisted of only those variables that
contributed to the explanatory value of the model (coefficient of determination). Co-
linearity was tested with VIF (Variance Inflation Factor). The assumption that the fOR

was constant across time (Weinberg and Wilcox 1998) was tested graphically and by
including an interaction term between cycle (time) and exposure or determinant
variables in the final model. The latter were not significant, implying that the
proportional assumption was not violated. Finally, to evaluate a possible selection bias
based on willingness to participate, the analyses were repeated excluding the

pregnancies occurring by the first month (Weinberg et al. 1994). No significant changes
in the final model were observed.
The distribution of pregnancies in relation TTP (Figure 16) was different between

the five regions. In previous work in Colombia (Idrovo et al. 2005), the percentage for
first month was about 30% - low compared with data from developed countries. In this
case, Valle del Cauca had very low initial percentage and Boyacá had high values for

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the first and twelfth months

(Figure 1). The mean for 100

12 months in developed
countries is 85-90%.
80
Participating women

were generally young
(mean and median age 21
60
years old) and had

completed at least some
secondary education. The 40

vast majority had regular Boyacá
menstrual cycles (96.7%); Percenttofpregnaancies Narñoiñ
20
a substantial proportion SerraNevada
had irregular partner Putumayo
ValedelCauca
relationships. Most
0
experienced their first 2 4 6 8 10 12
pregnancy at young ages
Tmee to fstpreggnancyy moonths))
(73.6% at < 20 years).
During the year before first Figure 23 Summary of the results of the time to pregnancy study

pregnancy, most were free

of illness (84%), had not had x-rays (95.4%), and did not smoke tobacco (95.1%).
Alcohol and coffee consumption were 51.8% and 80.3% respectively. The majority of

women were housekeepers at the time of first pregnancy.

In the crude analyses, longer TTP was associated with a number of factors such
as, region, older maternal age, ethnic group, irregular menstrual cycles, and irregular

partner relationship. Previously visit to physician for problems related with fertility, x-
rays taken in the year before pregnancy (YBP), and coffee consumption in the YBP also

were associated with longer TTP. Coffee consumption had a significant test for trend.

Maternal overweight was associated with a longer TTP. A tendency to longer TTP was
observed among those engaged in some waged work and with higher education.

Paternal unemployment or self work, were associated with longer TTP. No other
paternal data were related with the TTP.

After adjustment of the model for region, several associations were identified

(Table 18). Although non-significant in the adjusted model (p< 0.1), coffee intake and

self perception about bad quality of water was associated with longer TTP and all
sources of water presented greater risk of longer TTP when they were compared with

pure water (“nacimiento”), except for a few cases which use carried water (“carro-
tanque”).

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a
Table 18. Causes of fecundability adjusted for the relationship between time to
preganacy (TTP) and region b based on an alternative model.
c d e
Variable fRMa SE 95% CI p
f
Region

Nariño 0.56 0.048 0.47, 0.66 <0.01
Sierra Nevada 0.36 0.031 0.31, 0.43 <0.01

Putumayo 0.35 0.029 0.29, 0.41 <0.01

Valle del Cauca 0.15 0.014 0.13, 0.18 <0.01

Age at first preganacy > 20 0.81 0.048 0.73, 0.91 <0.01
years g
h
Irregular relationship 0.76 0.041 0.68, 0.84 <0.01
i
Consumption of coffee

Medium (1-3 cups per day) 0.91 0.059 0.81, 1.04 0.15
High ( 4 and more cups per day) 0.84 0.083 0.69, 1.02 0.08

Perception of contamination 0.91 0.51 0.81, 1.01 0.08
j
of water
n = 2592 mothers 11,270 cycles.
aProportional risk model of Cox, modified after Dinno (2002) Modelo de Riesgos proporcionales de
b
Cox, modificado (Dinno, 2000).Rcstricted to those mothers who did dot consult a phyeician
regarding problemsfin conceiving.RMa Adjusted causegof fecundabilityStandard Error. 95%
confidence interval. Compared to Boyacá as referenceCompared to ≤20 years as reference.
hCompared to regular relationship as reference. Compared to no consumption as reference.
Compared to no contamination as reference and based on self-perception and source of water
normally consumed.

In the final multivariate model, the main predictor of TTP was the region adjusted
by irregular relationship with partner and maternal age at first pregnancy. Boyacá had
the minimal risk and was the reference region. Nariño, Sierra Nevada, and Putumayo,

had slightly higher risk. The greatest risk was in the Valle del Cauca region. There was
no association between TTP and use of herbicides in the eradication of illicit crops in
the regions studied. The reason(s) for the increased risk for longer TTP in the Valle del

Cauca region where sugar cane is grown is not known. In this study, the increased risk
in Valle del Cauca cannot be attributed to exposure to pesticides alone since Sierra
Nevada, where organic crops are grown, also showed a statistically significant

difference from the reference location (Boyacá). This study was designed to test
hypotheses related to the use of glyphosate in eradication spraying and the data cannot
be used to identify causality associated with other risk factors. To test this question in

Valle del Cauca or any other region, a new study would have to be designed and
conducted. Some of the factors associated with higher TTP that were identified in our
study should be included in any future studies that may be conducted.

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4.1.3 Effects of glyphosate in non-target organisms in the environment.

The mechanism of action of glyphosate is via the disruption of the shikimate
metabolic pathway that leads to the synthesis of aromatic compounds in numerous
microorganisms and plants. Glyphosate inhibits the shikimate pathway by blocking 5-

enolpyruvyl-shikimate-3-phosphate synthase (EPSPS). This reduces the synthesis of
aromatic amino acids and causes accumulation of high concentrations of shikimic acid
and its derivatives. Glyphosate translocates to active growing tissues, particularly
effective in most plants because its degradation is slow. Thus, the herbicide moves
throughout the plant before symptoms are noticed. The shikimate pathway is absent

from mammals (Eschenburg et al. 2003, Roberts et al. 2002, Roberts et al. 1998).
However, toxic effects of the compound on, for example, non-mammalian aquatic
organisms, have been observed at large concentrations. These effects are discussed in
more detail below.

A common question in conducting risk assessments in tropical regions and other
non-temperate regions is the paucity of toxicity data for “tropical species”. It is true that
most of the test species used in toxicity testing, particularly of pesticides, are “temperate
species” largely because of the location of testing laboratories that are able to conduct

guideline toxicity tests under Good Laboratory Practice (GLP). Except for a few
substances with defined mechanisms of action, there is no reason to believe that
organisms from tropical regions are inherently more or less sensitive than organisms
from temperate regions. It is well known that DDT and some related pesticides become
more toxic at lower temperatures (Dyer et al. 1997); however the mechanisms here are

well understood. Comparison of responses of tropical and temperate organisms to a
number of pesticides other than DDT has shown that there are not significant
differences in sensitivity (Maltby et al. 2005). With this in mind, we used the rich data
set of toxicity values that has accumulated in the literature for glyphosate and its

formulations.

4.1.3.1 Effects in non-target terrestrial animals
The potential environmental effects of glyphosate and Roundup® were

extensively reviewed in 1999 (Giesy et al. 2000). Some additional papers have
appeared since that time. Glyphosate is not considered directly toxic to terrestrial
organisms.
Soil invertebrates. The effects of glyphosate on earthworms have been

reviewed (Giesy et al. 2000) and risks were judged to be essentially negligible. A recent
study on the earthworm Eisenia fetida reported that, although a commercial formulation
of glyphosate was not directly toxic to the earthworms, it did cause effects on
locomotory activity that may be detrimental to the earthworms (Verrell and Van Buskirk
2004). The formulation used in the study was Ortho Groundclear Total Vegetation Killer

which contains 5% by volume of glyphosate as the isopropylamine salt (IPA). In this
study, the authors applied 82 ml of a 1:4 solution of Groundclear to 2 L of soil in a
plastic box. This amount of glyphosate is much greater than would be applied under
normal agricultural uses or in the control of illicit crops. Assuming that the boxes of soil
2
were cubes, the area of the surface would be 12.6 x 12.6 cm or 159 cm . This being so,
the application rate used by the authors was equivalent to 518 kg glyphosate/ha, a
totally unrealistic application rate and 100 times more than that used in the control of

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coca. This study was obviously seriously flawed and the results are not applicable to

any use of glyphosate. This study has no relevance to the use of glyphosate for the
control of illicit crops in Colombia.
Soil microorganisms. Glyphosate has little effect on soil microorganisms

(Giesy et al. 2000). Since the symbiotic soil and root-associated microorganisms may
be partially dependent on the plant for nutrients, the death or injury of the plant will
result in effects on the organisms associated with it. Similarly, death of the plants would
release organic matter and nutrients into the soil and this would affect soil
microorganisms in a similar way to the application of compost or fertilizer. This effect

was reported for glyphosate and its effects on grass (Tenuta and Beuchamp 1995).
This would also occur with other herbicides and with mechanical control of plants.
Effects have been demonstrated in hydroponically grown plants exposed through the
watering solution, however, this route of exposure is not relevant to field conditions

where glyphosate would bind strongly to soil particles and not be biologically available.
Effects on symbiotic microbiota have also been demonstrated in glyphosate tolerant
plants treated at 10 times the normal field application rates but these are not relevant
exposures as the studies were done in vitro and in the absence of soil (Mårtensson
1992). Some effects on metabolism of phenolic substances in symbiotic bacteria in

glyphosate-tolerant soybeans have been shown; however, these changes did not alter
nitrogenase activity (Hernandez et al. 1999). Microbial systems in soil are complex and
considerable variation can be expected among tests and among soil types. More recent
studies on the effects of glyphosate on microbiological activity in soils have shown an
increase in microbiological activity, mainly in fungi, which are likely using the glyphosate

as a source of carbon, nitrogen, and phosphorus (Araujo et al. 2003, Haney et al. 2002,
Laatikainen and Heinonen-Tanski 2002). These changes in microbiological activity are
not judged to be deleterious.

The effects of several fungicides and herbicides on the growth of the
ectomycorrhizal fungi Lactarius deliciosus, strain LDF5, and Pisolithus tinctorius, strains
30AM, 3SR and Mx, in pure culture have been studied. Glyphosate at concentrations of
0, 1, 10, 100, and 1000 mg/Kg had no effect (Diaz et al. 2003). Some 64 strains of
ectomycorhizal fungi were tested against the most common pesticides used in forestry

in Finland. Glyphosate did not produce strong inhibition in any of the strains, most were
unaffected, and some were stimulated by 1 mg/L Roundup Bio® in agar (Laatikainen
and Heinonen-Tanski 2002). Laboratory tests on four species of entomopathogenic
fungi have shown that technical glyphosate has no effect, but a range of formulated
products did have fungicidal properties, especially RoundUp Ready-To-Use® (Morjan

and Pedigo 2002). In fact, as fungi and bacteria have the shikimate pathway, this
suggests the potential use of shikimate pathway inhibitors for the beneficial control of
fungal pathogens and apicomplexan parasites, such as Toxoplasma gondii,
Plasmodium falciparum, and Cryptosporidium parvum (Roberts et al. 2002, Roberts et

al. 1998).
Analysis of all lines of evidence for effects of glyphosate on soil microorganisms
indicates that adverse effects would be unlikely as a result of application at normal field
rates. Any minor effects to communities, such as described above, would be expected

to disappear rapidly (Giesy et al. 2000, World Health Organization International
Program on Chemical Safety 1994). After reviewing several studies conducted in many

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climates, different soils over the past 10 years and under various cropping systems,

Motavalli et al. (2004) have concluded that so far no conclusive evidence shows that
glyphosate has any relevant effect on nutrient transformations by microbes. However,
they point out that this topic needs further study, as not every situation has been
adequately researched. Further, because of lack of bioavailability on soils, adverse

effects on beneficial soil fungi and bacteria are unlikely to occur under field conditions of
use. Glyphosate binds strongly to soil particles and would not be available for uptake
by these microorganisms, many of which are actually inside the tissues of the plants.
The fact that seeds will readily germinate in soils soon after treatment with glyphosate
and that nitrogen-fixing Roundup Ready® soybeans grow and develop high yields

despite treatment with glyphosate demonstrates the practical insignificance of these
effects under actual conditions of use.
Terrestrial invertebrates. As glyphosate is a non-selective herbicide, it will

cause habitat alteration. Habitat alteration also results from a number of human
activities in the production of food and fiber. The most important of these is the clearing
of land for agricultural production. Whether this is through slash and burn processes
such as are used in the initial preparation of coca and poppy fields in Colombia or the
application of a herbicides such as glyphosate and paraquat, also used in coca

production, the effects on non-target species are the same. Use of cultural, mechanical
controls, or herbicides, to alter habitat (remove plants) will have effects on organisms
that normally use these plants for food or shelter.
After applying glyphosate at double the recommended application rates, no

effects were observed in microarthropods in soil (Gomez and Sagardoy 1985). As
weed species compositions and densities are directly affected by the glyphosate,
indirect effects are more likely to occur. Jackson and Pitre (2004a) found that
populations of adult Cerotoma trifurcata, adult Spissistilus festinus, larvae of Plathypena

scabra, and the caterpillar of Anticarsia gemmatalis were unaffected by glyphosate but,
populations of adult Geocoris punctipes, a Hompoteran insect predator, were decreased
by the herbicide. The authors concluded that this effect was due to reduced weed
densities after glyphosate treatment. Populations of green cloverworm (Hypena scabra)
were evaluated on soybean glyphosate-resistant varieties, with and without exposure to

glyphosate and no differences among treatments were detected on developmental time
and survivorship (Morjan and Pedigo 2002). Weed management systems, more than
glyphosate, that allowed more weeds to grow generally had higher insect population
densities (Buckelew et al. 2000).

Effects of glyphosate and associated cultural practices can affect arthopods
indirectly. In studies conducted in the United Kingdom, indirect effects of glyphosate
were observed in the spider Lepthyphantes tenuis. These were a result of habitat
alteration and were related to death of plants and decreasing height of vegetation.

Glyphosate applications only had a within-season indirect habitat effect on L. tenuis as
field margins sampled 16 months after an application of 360 g glyphosate/ha showed no
detrimental effects (Bell et al. 2002, Haughton et al. 2001). Tests of the fecundity and
mortality of Geocoris punctipes (Say), exposed to glyphosate as Roundup® on soybean
found no effects over a 10-d post-treatment period. Exposure of G. punctipes eggs to

glyphosate spray had no effect on egg hatch (Jackson and Pitre 2004b). Some

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reductions in numbers of this species 3 weeks after treatment probably reflect weed

removal, i.e. habitat alteration (Jackson and Pitre 2004a).
Similarly, studies on populations of leaf litter invertebrates in areas of Australia
where glyphosate was spayed at 1 to 1.4 kg/ha for the control of an invasive weed,

showed no significant effects four months after spraying (Lindsay and French 2004).
The authors pointed out that variability in treated and untreated areas was large and
suggested that the nature of the vegetative community and its structure and the post-
spray weather may also be important. In agriculture, these effects are part of the risk
assessment related to integrated pest management (IPM) and potential effects on

beneficial organisms are weighed in the risk benefit equation. In conclusion, there is
little evidence of any direct effect of glyphosate on insects in the field or in natural
environments.

Terrestrial vertebrates. Technical glyphosate, formulated glyphosate, and
glyphosate mixed with Cosmo-Flux® are not acutely toxic to mammals via several
routes of exposures (reviewed in this report). Although wild mammals have not been
specifically tested with the mixture as used in Colombia, the data from these laboratory
studies suggest that they would be insensitive and not directly affected by a direct

overspray.
Birds are not susceptible to glyphosate. In studies on Bobwhite quail, Colinus
virginianus and Mallard duck, Anas platyrhynchos, acute oral LD50 values of >4,640

and >4,640 mg/kg bw have been reported (USEPA 2001). Again, direct effects of
formulated glyphosate or glyphosate plus Cosmo-Flux® are judged to very unlikely.
Indirect effects on terrestrial wildlife have been reported to be associated with the
use of glyphosate in agriculture and forestry uses. Alteration of habitat is more of an

issue in semi-wild areas such as forests where herbicides may be used to control
competing vegetation and allows conifers to grow and mature more rapidly. In these
cases, short-term effects on birds and other wildlife do occur, however, these
populations usually recover in 2-3 years (Kimball and Hunter 1990, Santillo et al. 1989a,

Santillo et al. 1989b) and even the vegetation will recover in less than ten years (BC
Ministry of Forests 2000, Boateng et al. 2000). Normally, in these uses, the actual
areas treated with herbicides are relatively small and are surrounded by or adjacent to
untreated areas that can act as refugia or sites for repopulation by animals that have
moved away because of the changes in habitat. As new vegetation develops to replace

that controlled by the herbicide, the habitat will again become usable to these animals
(Giesy et al. 2000, World Health Organization International Program on Chemical Safety
1994).
Glyphosate is widely used for vegetation management, including in the

restoration of native plant communities where exotic or invasive species are controlled,
(e.g. Hartman and McCarthy 2004). The use of glyphosate for “conifer release” from
competition has minimal effects on wildlife and can be used to enhance biodiversity if
used for spot and patch treatments, (e.g. Sullivan and Sullivan 2003). A review of

management of northern US forests, including the use of herbicides including
glyphosate, indicated no adverse ecological effects (Lautenschlager and Sullivan 2002).
However, the impacts of vegetation removal by manual clearance and glyphosate
application in conifer plantations has effects on bird communities in British Colombia,

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mediated by the removal of deciduous plants. Where the herbicide was used, number

of bird species declined, total number of individuals increased, and common species
dominated. Populations of residents, short-distance migrants, ground gleaners, and
conifer nesters increased significantly after herbicide treatment. Deciduous nesters and
foliage gleaners increased in abundance (nonsignificantly) in control and manually

thinned areas. Warbling Vireos (Vireo gilvus), which are deciduous specialists, declined
in areas treated with herbicide and may be particularly susceptible to the indirect effects
of glyphosate application on plant removal (Easton and Martin 1998, Easton and Martin
2002).

Nevertheless, control of Cirsium arvense (Canada thistle) using wick application
of glyphosate in wildfowl areas can enhance plant diversity that is of benefit to water
birds (Krueger-Mangold et al. 2002). However, the broad spectrum activity of
glyphosate means that accidental overspray of rare non-target plant species during

control of invasive plants will cause damage (Matarczyk et al. 2002).
Beneficial insects. Glyphosate is not considered toxic to honeybees, with a
reported LD50 of >100 μg/bee (USEPA 2001), however, the formulation, with the
adjuvant Cosmo-Flux®, as used in Colombia may have different toxicity because of the

surfactants added to the mixture. To test this hypothesis, toxicity testing of a mixture of
a commercial formulation of glyphosate and the surfactant Cosmo-Flux® 411F, was
conducted to determine the acute contact toxicity to honey bees (Apis mellifera L.)
(Stantec 2005a). This was done in accordance with the testing methods and guidelines
developed by the Organization for Economic Cooperation and Development (OECD)

Method #214, “Honeybees, Acute Contact Toxicity Test” (OECD 1998a) and the United
States Environmental Protection Agency (U.S. EPA) Office of Prevention, Pesticides
and Toxic Substances (OPPTS) Ecological Effects Test Guideline 850.3020, “Honey
Bee Acute Contact Toxicity” (USEPA 1996a). The results of this study showed that the

mixture of glyphosate and Cosmo-Flux® 411F is acutely nontoxic via contact exposure
to honey bees (i.e., did not cause mortality or stress effects in bees within 48-hours of
treatment) at concentrations equal to or less than 56.8 mg AE/bee. These results are
similar to those for glyphosate and formulations from the US EPA ECOTOX data base
(USEPA 2001) and show that the formulated product as used in Colombia is not

hazardous to bees and, by extrapolation, to other beneficial insects.

4.1.3.2 Effects in aquatic animals
Several extensive reviews of the effects of glyphosate on aquatic organisms

have concluded that glyphosate presents an essentially negligible risk to aquatic
organisms (Giesy et al. 2000, Solomon and Thompson 2003, World Health Organization
International Program on Chemical Safety 1994). Several recent publications have
reported on the effects of glyphosate and several of its formulations in frogs. The acute
toxicity of technical-grade glyphosate acid, glyphosate isopropylamine and three

glyphosate formulations to Australian frogs was measured (Mann and Bidwell 1999).
The authors reported the acute toxicity for adults of one species and tadpoles of four
species of southwestern Australian frogs in 48-h static/renewal tests. The 48-h LC50
values for Roundup® Herbicide (MON 2139) tested against tadpoles of Crinia

insignifera, Heleioporus eyrei, Limnodynastes dorsalis, and Litoria moorei ranged
between 8,100 and 32,200 μg/L (2,900 and 11,600 μg/L glyphosate acid equivalent

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[AE], while the 48-h LC50 values for Roundup® Herbicide tested against adult and

newly metamorphosed C. insignifera ranged from 137,000-144,000 μg/L (49,400-51,800
μg/L AE). These values were different, depending on the type of dilution water (lake or
tap water). For the purposes of this risk assessment, the most sensitive stage was
used.

Touchdown® Herbicide (4 LC-E) tested against tadpoles of C. insignifera, H.
eyrei, L. dorsalis, and L. moorei was slightly less toxic than Roundup® with 48-h LC50
values ranging between 27,300 and 48,700 μg/L (9,000 and 16,100 μg/L AE).
Roundup® Biactive (MON 77920) was practically nontoxic to tadpoles of the same four

species producing 48-h LC50 values of 911,000 μg/L (328,000 μg/L AE) for L. moorei
and >1,000,000 μg/L (>360,000 μg/L AE) for C. insignifera, H. eyrei, and L. dorsalis.
Technical glyphosate isopropylamine salt was practically nontoxic, producing no
mortality among tadpoles of any of the four species over 48 h, at concentrations

between 503,000 and 684,000 μg/L (343,000 and 466,000 μg/L AE). The toxicity of
technical-grade glyphosate acid (48-h LC50, 81,200 -121,000 μg/L) is likely to be due to
acid intolerance. Slight differences in species sensitivity were evident, with L. moorei
tadpoles showing greater sensitivity than tadpoles of the other four species. Adult and
newly emergent metamorphs were less sensitive than tadpoles.

A series of studies on frogs were conducted with several formulations of
glyphosate in relation to its use in forestry in Canada (Chen et al. 2004, Edginton et al.
2004, Thompson et al. 2004, Wojtaszek et al. 2004). Using a formulation of glyphosate
(Vision®) containing glyphosate and ethoxylated tallowamine surfactant - POEA, LC50

values as low as 880 µg/L (as glyphosate) were reported for tadpoles of Xenopus
laevis, Bufo americanus, Rana clamitans, Rana pipiens (Edginton et al. 2004). Embryo
stages were less sensitive than older larvae and toxicity was affected by the pH of the
exposure medium, although not in a consistent manner. For the purposes of this

assessment, values obtained at the most sensitive pH and for the most sensitive stage
were used.
In a related study on the toxicity of the Vision® formulation of glyphosate to the
zooplankton organism, Simocephalus vetulus, and tadpoles (Gosner stage 25) of Rana

pipiens, interactions between pH and food availability were reported (Chen et al. 2004).
Both high pH (7.5 vs. 6.5) and food deprivation increased the toxicity of this formulation.
As only two concentrations were tested (750 and 1,500 µg/L), LC50 values could not be
determined.

Field studies conducted on larvae of Rana clamitans and Rana pipiens with the
Vision® formulation of glyphosate showed that, in the presence of natural factors such
as sediment and environmentally relevant pH, the toxicity of the formulation was
reduced as compared to laboratory observations (Wojtaszek et al. 2004). The authors

reported 96-h median lethal concentration (LC50) values ranging from 2,700 to 11,500
μg/L (as glyphosate) (Wojtaszek et al. 2004). Although the authors used a formulation
of glyphosate containing the more toxic surfactant POEA, the results confirm that, in the
presence of sediments, reduction in the bioavailability of glyphosate (and formulants)

occurs and this further reduces risks, a conclusion reached for this forestry use
(Thompson et al. 2004) but which is equally relevant to the use of glyphosate in
Colombia.

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In another study on amphibians, the toxicity of a number of glyphosate

formulations to frogs (Rana clamitans, R. pipiens, R. sylvatica, and Bufo americanus)
was reported (Howe et al. 2004). The formulations included Roundup Original®,
glyphosate technical, the POEA surfactan used in some glyphosate-based herbicides,
and five newer glyphosate formulations of glyphosate. As expected, the most toxic of

the materials was the POEA surfactant, followed by Roundup Original®, Roundup
Transorb®, and Glyfos AU®. No significant acute toxicity was observed with glyphosate
technical material (96-h LC50 >17,900 µg/L). LC50 values for Roundup Original® in R.
clamitans, R. pipiens, and R. sylvatica were 2,200, 2,900, and 5,100 μg/L (AE),
respectively. These values were used in this risk assessment. Several other

formulations of glyphosate were also tested in R. clamitans and these (Roundup
Biactive®, Touchdown®, and Glyfos BIO®) were essentially non-toxic with LC50 values
of >57,000 μg/L.

In a study carried out with several commercial pesticide formulations in leopard
frogs (Rana pipiens), green frogs (R. clamitans), bullfrogs (R. catesbeiana), the
American toad (Bufo americanus), and gray tree frogs (Hyla versicolor), effects of
Roundup® and interactions with other pesticides were reported (Relyea 2004). The
formulation of Roundup® used in this study contained the more toxic POEA surfactant.

Survival and growth over a 16 day period were not significantly affected by the
glyphosate formulation at 1,000 µg/L (glyphosate AE) but some species were affected
at 2,000 µg/L. Some interactions were observed between the glyphosate formulation
and other pesticides such as the insecticides diazinon, carbaryl, and malathion. A
recent paper reported that a glyphosate formulation containing POEA was highly toxic

to tadpoles of several species of frogs exposed under realistic conditions in small (1000-
L) field microcosms (Relyea 2005). The tadpoles (Wood frog, Rana sylvatica; leopard
frog, Rana pipiens; American toad, Bufo americanus; gray tree frog, Hyla versicolor;
and the spring peeper, Pseudacris crucifer) were exposed to a concentration of 3,800

μg/L (AE) of glyphosate formulation applied as a commercial formulation (unspecified)
directly to the surface of the water. The rate of application was equivalent to 16 kg/ha, a
value that is unrealistic and probably the result of an error in the methods. At this
concentration, glyphosate formulated with POEA would be expected to be lethal to
tadpoles. The discussion in the paper that suggests that use of glyphosate may be

having adverse effects on frogs thus based on a flawed study design and is not
supported by other data, much of which is discussed above.
Effects on other non-target aquatic organisms have also been recently reported
in the literature. In studies on the toxicity of glyphosate to several aquatic algae and

zooplankton, Tsui and Chu (2003) showed that technical glyphosate was considerably
less toxic than the product Roundup®, which is formulated with the POEA surfactant.
LC and EC50 values for technical glyphosate ranged from 5,890 to 415,000 µg/L. In
tests conducted in the presence of sediment (Tsui and Chu 2004), these same authors

showed that biological availability of glyphosate was significantly reduced by binding to
sediment. The reduction in porewater concentration that resulted from the presence of
sediments was proportional to the amount of organic carbon in the sediments.
Tests on the fish Oreochromis niloticus (Nile tilapia) exposed for 3 months to

sublethal concentrations (5,000 and 15,000 μg/L) of glyphosate as Roundup® caused
significant damage to gill, liver, and kidney tissue. The structural damages could be

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correlated to the significant increase (p ≤0.05) in aspartate aminotransferase, alanine
aminotransferase, and alkaline phosphatase activities in the second and third months of
exposure. The results indicated that long-term exposure to Roundup® at large,
although sublethal concentrations had caused histopathological and biochemical

alterations of the fish (Jiraungkoorskul et al. 2003). Because technical glyphosate was
not tested and the contribution of the surfactants to this response cannot be judged.
In studies on the freshwater mussel Utterbackia imbecillis, a commercial
formulation of Roundup® was reported to have low toxicity (24-h LC50 of 18,300 µg/L

and a No Observed Effect Concentration (NOEC) of 10,040 µg/L – 7,442 μg/L AE) to
larval mussels (Conners and Black 2004). In studies on genotoxicity in these mussels,
there was no significant difference in response between the control and mussel larvae
treated at ¼ the NOEC, ≈ 2,500 µg/L (1,850 AE).

Response of total free amino acids profiles of snails to glyphosate exposures has
been studied (Tate et al. 2000). These authors showed that exposure of the aquatic
snails (Pseudosuccinae columella) to technical glyphosate at nominal concentrations of

1000-10,000 μg/L lead to increased egg-laying and increased amino acid
concentrations in the tissues. Technical glyphosate was not particularly toxic with a
24-LC50 of 98,900 μg/L. The effect on egg-laying and amino acid concentrations was
stimulative rather than adverse but the authors speculate that it could lead to increases

in incidence of diseases for which the snails are intermediate hosts. Increases in
parasites may affect organisms in the environment. Similar stimulation was observed in
the rotifer Brachionus calyciflorus where growth rates and sexual and asexual
reproduction were stimulated in the presence of glyphosate (formulated, but formulation
unknown) at concentration of ≥4,000 μg/L (growth) and ≥2,000 μg/L for reproduction

and resting egg production (Xi and Feng 2004). Again, although stimulatory and not
“adverse” the authors point out that the increases in one species may affect other
species indirectly.

In a study on grazing of the alga, Scenedesmus spp. by the aquatic crustacean,
Daphnia pulex, technical glyphosate was shown to have no adverse effect, although it
appeared to stimulate the growth of the algae (Bengtsson et al. 2004). Stimulation of
algal growth was suggested to be due to release of nitrogen and phosphorus from the

metabolism of glyphosate by the Daphnia. Similar stimulation was also seen in the
effects of glyphosate (Rodeo®, a formulation without any surfactants) on the primary
productivity of a natural phytoplankton algal assemblage dominated by species of
diatoms and a dinoflagelate (Schaffer and Sebetich 2004). A 60% increase in
productivity as measured by assimilation of 14CO was observed at concentrations of
2
glyphosate of 125, 1,250, and 12,500 µg/L, with no apparent concentration-response.
The authors speculate that the increase was caused by the release of nitrogen and
phosphorus from the breakdown of glyphosate.

The effects of glyphosate on fish and other aquatic organisms are clearly related
to the surfactant in the formulation rather than the glyphosate itself. Surfactants can
disrupt cell membranes and this type of response would be expected. For this reason,
the glyphosate formulation and the surfactant (Cosmo-Flux®-411) as used in Colombia

for the eradication of coca and poppy were tested for toxicity to the aquatic organisms,
algae, crustacea, and fish (Section 4.2.2). The protocols used are described below and
results are summarized in Table 19.

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Algal tests. The testing of a mixture of a commercial formulation of glyphosate
and the surfactant Cosmo-Flux® 411F, was conducted to determine growth inhibition of
the freshwater green alga, Selenastrum capricornutum Printz, according to the

Organization for Economic Co-operation and Development (OECD) Method # 201,
“Alga, Growth Inhibition Test” (OECD 1984b) and in general accordance with OPPTS
Method 850.5400, “Algal Toxicity, Tiers I and II” (USEPA 1996b).

Water Flea. Tests were conducted to determine the acute toxicity of a
commercial formulation of glyphosate and the surfactant Cosmo-Flux® 411F to Daphnia
magna according to OECD Method #202, “Daphnia sp., Acute Immobilization Test and

Reproduction Test” (OECD 1984a), however, the reproduction component of the test
was not conducted.
Rainbow Trout and Fathead Minnow. Tests were conducted to determine the

acute toxicity of a commercial formulation of glyphosate and the surfactant Cosmo-
Flux® 411F to Oncorhynchus mykiss and Pimephales promelas according to OECD
Method #203, “Fish, Acute Toxicity Test” (OECD 1992). In all of these tests, OECD
Principles of GLP (OECD 1998b) were followed.

Table 19. Toxicity values obtained from toxicity tests conducted on a mixture of

glyphosate and Cosmo-Flux®.
Test species Common name 96 hour LC/EC50 Reference

in μg/L (as
glyphosate AE)
Selenastrum Algae 2,278-5,727 a (Stantec 2005b)

Daphnia magna Water flea 4,240 (3,230- (Stantec 2005e)
5,720)b

Onchorynchus Rainbow trout 1,850 (1,410- (Stantec 2005c)
mykiss 2,420)b

Pimephales Fathead minnow 4,600 (1,810b (Stantec 2005d)
promelas 1,173)
a Lowest and highest effect measures in the study
b
LC/EC50 and 95% Confidence Interval

The acute toxicity data for formulated glyphosate in aquatic animals from
Solomon and Thompson (2003) were combined with some of the new data for
amphibians described above and are displayed graphically as a point of reference for

characterizing the toxicity of glyphosate plus Cosmo-Flux® as used in Colombia (Figure
17). The graph is presented as a cumulative frequency distribution in a manner similar
to that used in probabilistic risk assessments for pesticides (Solomon and Takacs
2002). These data show that the combination of formulated glyphosate and Cosmo-

Flux®, as used in Colombia, is more toxic to the aquatic organisms tested than
formulations without the addition of surfactants and adjuvants. This is not altogether
surprising. It has been shown that the toxicity of glyphosate itself to aquatic organisms
is very small (Solomon and Thompson 2003) but, when mixed with some surfactants

and adjuvants, this toxicity can be increased. The toxicity of Cosmo-Flux® was not
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tested on its own; however, from experience with other adjuvants, it is clearly the cause
of the increased toxicity of the mixture.

90
Fathead
minnow

70 Waaerreafffllle

Green agalg
50

Peercenttrankk Formuuated LC500sampphibanns
Rainbow
touttto Formuuated LC500sarthropods
Formuuated LC500s shiis
10
Glyphosate echnncalLC550s

Glyphosate pus Cossmo-Flux animaasls
Glyphosate pus Cossmo-Flux -agaae

1
1 2 3 4 5 6 7 8
10 100 10 100 10 10 10 10

Coonccentration (µg/L ggypphossate AEE))

Figure 24 Distribution of toxicity values for glyphosate technical, formulated glyphosate (Roundup®) in
all aquatic organisms and in fish and the glyphosate and Cosmo-Flux® 411 mixture as used in
Colombia

It is interesting to note that larval amphibians appear to be more susceptible to

glyphosate formulation than are other aquatic animals. The reason for this is likely the
surfactants in the formulation of Roundup®; as discussed above, other formulations of
glyphosate are less toxic to amphibians (Howe et al. 2004).

4.1.3.3 Effects of glyphosate on plants

There are differences in glyphosate uptake between different coca species and
between young and mature plants of Erythroxylum coca and E. novogranatense
(Ferreira and Reddy 2000). Leaf absorption is greater in young plants of both species

and greater in E. novogranatense. Earlier studies showed that control of regrowth was
better in E. novogranatense for equivalent dose of glyphosate (Ferreira et al. 1997).

This study also indicated that defoliation of E. coca 24 hours prior to application resulted
in no significant effect of glyphosate (applied up to 6.7 Kg/ha) on regrowth. This
confirms that, as for other plants, uptake via the leaves is the major route of penetration

into the plant.
A study on the control of the perennial weed pepperweed (Lepidium latifolium)

has shown better control with glyphosate following mowing. The mechanism is via the

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better movement of glyphosate to roots from leaves lower in the canopy. Following

mowing, the leaf distribution and the spray deposition is closer to the ground, giving
better basipetal translocation to roots and better subsequent control (Renz and
DiTomaso 2004). In forestry situations with an aerial application, spray deposition is
typically much greater higher in the canopy, (e.g. Thompson et al. 1997). Studies of
glyphosate efficacy on annual weeds have indicated that application during the day

(09:00 and 18:00h) gives best control (Martinson et al. 2002, Miller et al. 2003).
Resistance to glyphosate is known for an increasing number of species, including
Conyza canadensis (Mueller et al. 2003), Illinois waterhemp (Amaranthus rudis and A.
tuberculatus) (Patzoldt et al. 2002), Eleusine indica (Baerson et al. 2002), Lolium

multiflorum (Perez and Kogan 2003) and Lolium rigidum (Neve et al. 2003a, b). Rates
of evolution of resistance in the latter species are dependent on herbicide use patterns
as part of crop production.
Non-target impacts of glyphosate on seed germination and growth characteristics

of the F1 generation of treated wild plant species have been reported. Blackburn and
Boutin (2003) noted effects on seven out of 11 species tested with 1%, 10% or 100% of
a 0.89 Kg a.i./ha label rate of glyphosate formulated as Roundup® solution sprayed
near seed maturity. Effects of glyphosate drift on rice seed germination were reported
by (Ellis et al. 2003) and (May et al. 2003) noted reduced seed production in alfalfa in

the year following applications of glyphosate at 1.760 Kg a.i./ha for Cirsium arvense
control. Nevertheless, applications of glyphosate at 0.420 kg AE/ha on susceptible
soybean had adverse effects on sprayed plants, but not on progeny (Norsworthy 2004).
Subtle adverse effects of glyphosate on pollen viability and seed set in glyphosate-
resistant cotton were noted by (Pline et al. 2003). Pollen viability of glyphosate-resistant

corn was also significantly reduced by glyphosate applied at 1.12 kg AI/ha, but yield and
seed set is not significantly affected (Thomas et al. 2004). These data indicate that drift
might cause subtle ecological changes to plant communities associated with changes in
plant recruitment. However, this would be significant only for communities largely made
up of monocarpic plant species (that flower once and die, especially annuals)

dependent on seeds for recruitment.

4.2 SURFACTANTS
There are a number of formulations of glyphosate on the market and these may

contain a number of surfactants (Giesy et al. 2000, Solomon and Thompson 2003,
Williams et al. 2000). Normally, this would not be an issue in the risk assessment of a
pesticide, however, in the case of glyphosate; the active ingredient is of very low toxicity
to non-target organisms, thus making the surfactant toxicity more important in the risk
assessment process. For example, tests on Ca 2+-activated ATPase and cholinesterase

(ChE) activities in the nervous system of the slug Phyllocaulis soleiformis showed no
effects of pure glyphosate. An effect noted with the formulation Gliz 480CS® was
caused by non-glyphosate components of the formulation (da Silva et al. 2003).
Technical grade glyphosate at concentrations of 52 mM (870 mg/L) did not affect the
protozoans Tetrahymena thermophila or the parasite Ichthyophthirius multifiliis.

However, the commercial formulation Glyphosate® was up to 100 times more toxic,
reflecting data for fish species and other aquatic invertebrates and caused by
surfactants in the formulation (Everett and Dickerson 2003).

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Because the spray solution as used in the eradication of coca and poppy in

Colombia contains surfactants as part of the formulation as well as additional
surfactants (Cosmo-Flux®) added to the spray mix, the toxicity of the formulants and the
adjuvants may interact to change the toxicity of the mixture. This was the reason why
standardized toxicity tests for mammals and environmental non-target organisms were

conducted with the spray mixture itself. These are discussed below.

4.2.1 Effects on glyphosate and Cosmo-Flux® on non-target aquatic organisms
A base set of toxicity data is required for all pesticide registrations. For

freshwater environments, the set normally makes use of a coldwater fish such as
rainbow trout fingerlings (Onchorynchus mykiss), a warmwater fish such as fathead
minnows (Pimephales promelas), an invertebrate such as the water flea (Daphnia
magna), and an alga such as Selanastrum capricornutum. These are standard test
organisms, have been used for testing glyphosate itself and several other formulations,

and thus are useful for comparison purposes. To reduce the requirement for animals in
the testing, one combination of glyphosate and Cosmo-Flux®, the combination for
poppy (Table 4), was selected. This mixture contains more Cosmo-Flux® than used for
coca and thus represents a worst-case exposure. These data are summarized in Table
19 and Figure 17, above.

4.2.2 Effects of glyphosate and Cosmo-Flux® on mammals
Two series of mammalian toxicity tests on the formulation of glyphosate and
Cosmo-Flux® as used for eradication of coca in Colombia were conducted. One set of

these studies was conducted in the USA under good laboratory practices (GLP) and
using the quality control assurance as appropriate for regulatory decision making. The
other studies were conducted in Colombia, also in compliance with GLP and according
to US EPA guidelines.

4.2.2.1 Analysis of the formulation
The objective of this study was to assess the concentration(s) of glyphosate
(active ingredient) in the formulation (Springborn 2003a). Three 500 mL samples of
each mixture were collected from the top/middle/bottom of Air Tractor N8513Q PNC

4003 (Test Article Mixtures 1 and 3), Air Tractor N8514G PNC 4005 (Test Article
Mixtures 2 and 4), and Air Tractor N8513V PNC 4004 (Test Article Mixture 5). Test
Article Mixtures 1 and 2 were prepared as follows:

Ingredient Amount Added (gallons)

Herbicide: glyphosate 131.7
Surfactant: Cosmo Flux-411F 3.0

Lake Water 165.3
Mixing Time: Test Article Mixture 1 - 13 minutes; Test Article Mixture 2 - 12
minutes.

Test Article Mixtures 3, 4 and 5 were prepared as follows:

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Ingredient Amount Added (gallons)

Herbicide: glyphosate 110.0
Surfactant: Cosmo Flux-411F 2.5

Lake Water 137.5
Mixing Time: Test Article Mixture 3 - 12 minutes; Test Article Mixture 4 - 11
minutes;

Test Article Mixture 5 - 13 minutes.

The test article mixtures were prepared on December 5, 2002. The overall
concentration of the formulation was 16.53 [in terms of % glyphosate (AE)] before use
at SLI and 15.20 [in terms of % glyphosate (AE)] after use at testing laboratory,
indicating that the test material was stable during the period of testing. The overall

result (16.53% glyphosate AE) was higher than the anticipated 14.80% glyphosate (AE)
value but within acceptable error of mixing conditions in the field. Since the results of
the analysis were appropriate and would provide conservative results for toxicity,
irritation and sensitization because they were slightly higher than expected, the five test
article mixes were pooled into a single container for use in the remaining studies.

4.2.2.2 Acute oral toxicity
The single-dose oral toxicity of glyphosate and Cosmo-Flux® was carried out in
Sprague Dawley rats (Springborn 2003b). A limit test was carried out in which one

group of 10 young adult rats (5 male and 5 female) weighed 325-356 g and 190-208 g
respectively and received the test article at a single dose of 5,000 mg/kg body weight
(bw). Following dosing, the rats were observed daily and weighed weekly. All animals
were humanely killed 14-days post-exposure and subjected to a gross pathology

examination. No mortality occurred during the study. Clinical abnormalities observed
during the study included transient incidences of soft stools, fecal staining, rough coat,
congested breathing, rales (wet, crackly lung noises heard on inspiration which indicate
fluid in the air sacs of the lungs), and dark material around the facial area. Body weight
gain was noted for all animals during the test period. No significant macroscopic

findings were observed at necropsy on study day 14. The oral LD50 for test article in
rats was estimated to be greater than 5,000 mg/kg.
Other rat oral acute studies were performed on a mixture of glyphosate (44%),
Cosmo-Flux® (1%), and water (55%) (Immunopharmos 2002a) and a mixture of

glyphosate (5%), Cosmo-Flux® (1%), and water (95%) (Immunopharmos 2002b).
Both studies were performed according to using EPA guidelines 870-1100. In
the first, groups of 5 male and 5 female Wistar rats, approx. 135 g bw, were treated with

the test substance by gavage at concentrations of 1,250, 2,500 and 5,000 mg/kg bw
(Immunopharmos 2002a). The test substance was dissolved in distilled water. The
animals were observed for 5 hours during the first day and later on all days during the
14 day post-dosing period. During the study, the animals did not show any adverse
effects. The Reed and Muench test was used for the calculation of LD50. The LD50

value of test substance was greater than 5,000 mg/kg bw for males and females.

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In the second study (Immunopharmos 2002b), groups of 10 Wistar rats (5 male

and 5 female), ranging from 116 to 138 g bw, were treated with the test substance by
gavage at concentrations of 1,250, 2,500 and 5,000 mg/kg bw. The test substance was
dissolved in distilled water. The animals were observed as above. During the study, the
animals showed no adverse effects. The Reed and Muench test was used for the

calculation of LD50. The LD50 value of test substance was greater than 5,000 mg/kg
bw for males and females.

4.2.2.3 Acute Inhalation toxicity

A limit test was performed in 10 young adult Sprague Dawley rats (5 male and 5
female) weighing 248-275 g and 201-212 g respectively received a 4-hour nose-only
inhalation exposure at an aerosol concentration of 2.60 mg/L (Springborn 2003c). The
mass median aerodynamic diameter and geometric standard deviation of the sampled
particles were 2.9 µm ± 2.17. The percentage of particles ≤ 4.0 µm was determined to

be 66%. After exposure, the rats were observed daily and weighed weekly. All animals
were humanely killed at 14-days post-exposure and subjected to a gross pathology
examination on day 14. There was no mortality during the study. The clinical
abnormalities observed during the study included breathing abnormalities,

no/decreased defecation, urine staining, rough hair coat, dark material around the facial
area and decreased food consumption. Body weight loss was noted in 2 males and 1
female during days 0 to 7. Body weight gain was noted for all other animals during the
test period. At study termination, the animals had exceeded/maintained their initial body
weight. No macroscopic findings were observed at necropsy (day 14). The inhalation

LC50 of test material was estimated to be greater than 2.60 mg/L but exposures greater
than or equal to this value may be harmful.
Other rat acute inhalation toxicity studies were performed on a mixture of
glyphosate (44%), Cosmo-Flux® (1%), and water (55%) (Immunopharmos 2002a) and

a mixture of glyphosate (5%), Cosmo-Flux® (1%), and water (95%) (Immunopharmos
2002b).
Both studies were performed under EPA guideline 870-1300. In the first, ten

Wistar rats (5 male and 5 female) were used for each concentration (Immunopharmos
2002c). The test substance was dissolved in sterile water to achieve concentrations of
5, 10, and 20 mg/L air/hour during 4 hours of exposure. After the exposure period, the
animals were kept for a 14-day observation period. The mass median aerodynamic
diameter and geometric standard deviation of the sampled particles were not indicated.

There were no deaths during exposure period and no signs of systemic toxicity were
observed at the three concentrations tested. All animals were humanely killed at 14
days post-exposure and subjected to a gross pathology and histopathology
examinations and no abnormalities were observed. The LC50 value of the test

substance was higher than 20 mg/L of air. Therefore, the test substance is not
considered as harmful at concentrations less than 20 mg/L.
In the second study (Immunopharmos 2002d), ten Wistar rats (5 male and 5
female) were used for each concentration. The test substance was dissolved in sterile

water to achieve concentrations of 5, 10, and 20 mg/L air/hour during 4 hours of
exposure. After the exposure period, the animals were kept for a 14-day observation
period. The mass median aerodynamic diameter and geometric standard deviation of

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the sampled particles were not indicated. There were no deaths during the exposure

period and no signs of systemic toxicity at the three concentrations tested. All animals
were humanely killed 14 days post-exposure and subjected to a gross pathology and
histopathology examinations. At necropsy the surviving animals showed petechial lung
(3/10) while the remaining organs were normal. The LC50 value of the test substance

was higher than 20 mg/L of air.

4.2.2.4 Acute dermal toxicity
A limit test was performed in 10 Sprague Dawley rats (5 male and 5 female)

receiving a single dermal administration of the test article at a dose of 5,000 mg/kg bw
(Springborn 2003d). Following dosing, the rats were observed daily and weighed
weekly. All animals were humanely killed after 14-days exposure and subjected to a
gross pathology examination. No mortality occurred during the study. Clinical
abnormalities observed during the study included transient incidences of dark material

around the facial area and decreased defecation. Dermal irritation was noted at the site
of test article application. Body weight loss was noted in 1 male and 2 females during
the study (day 7 to 14). Body weight gain was noted for all other animals during the test
period. At necropsy (day 14), no significant macroscopic findings were observed. The
acute dermal LD50 of test article was estimated to be greater than 5,000 mg/kg in the

rat.

4.2.2.5 Skin irritation
A potential irritation of the test material was evaluated on the skin of New

Zealand White rabbits (Springborn 2003e). Each of 3 rabbits (13 weeks of age and
weighed 2.5-2.8 kg prior to dosing) received a 0.5 ml dose of the test article as a single
dermal application. The dose was held in contact with the skin under a semi-occlusive
binder for an exposure period of 4 hours. Following the exposure period, the binder
was removed and the remaining test article was wiped from the skin using gauze

moistened with deionized water followed by dry gauze. Test sites were subsequently
examined and scored for dermal irritation for up to 72 hours following patch application.
Exposure to the test article produced very slight erythema on 3/3 test sites at the 1-hour
scoring interval. The dermal irritation resolved completely on all test sites by 24-hour.

The test article was considered to be a slight irritant to the skin of the rabbit. The
calculated Primary Irritation Index for the test article was 0.25.
Other skin irritation studies were performed on a mixture of glyphosate (44%),
Cosmo-Flux® (1%), and water (55%) (Immunopharmos 2002g) and a mixture of

glyphosate (5%), Cosmo-Flux® (1%), and water (95%) (Immunopharmos 2002h). Both
studies were performed using EPA guidance 870-2500.
In the first, 0.5 ml of test substance was applied to the clipped and abraded skin

of 3 male and 3 female New Zealand White rabbits (2.3-2.4 kg bw) (Immunopharmos
2002g). The application site of the test substance was covered with three occlusive
dressings for 15 minutes, 1 hour, and 4 hours, after which the site was washed. Skin
reactions were measured for erythema and edema using a modified Draize test. The
readings were made at 24, 48, and 72 hours after treatment. Body weight was not

measured. There were no signs of irritation at the application site or systemic toxicity.
In the second study, 0.5 ml of test substance was applied to the clipped and abraded

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skin of 3 male and 3 female New Zealand White rabbits (2.3-2.4 kg bw)

(Immunopharmos 2002h). The application site of the test substance was covered with
three occlusive dressings for 15 minutes, 1 hour, and 4 hours, after which the site was
washed. Skin reactions were measured for erythema and edema using a modified
Draize test. The readings were made at 24, 48, and 72 hours after treatment.

Bodyweight was not measured. There were no signs of irritation and/or edema on the
shaved skin.

4.2.2.6 Eye irritation

The eye irritation for the test article was evaluated in rabbits (Springborn 2003f).
Each of 3 New Zealand White rabbits received a 0.1 mL dose of the test article in the
conjunctival sac of the right eye. The left eye of each untreated animal served as a
negative control. Test and control eyes were examined for signs of irritation for up to 7
days after dosing. Exposure to the test article produced iritis (3/3 test eyes) at the 1-

hour scoring interval which resolved completely in all eyes by 24-hour. Conjunctivitis
(redness, swelling and discharge) was noted in 3/3 test eyes at the 1-hour. The
conjunctival irritation resolved completely in all treated eyes by day 7. An additional
ocular finding of slight dulling of normal luster of the cornea was noted in 1/3 test eyes.
Based on these results, the test material is considered to be a moderate irritant to the

eye.
Other eye irritation studies were performed on a mixture of glyphosate (44%),
Cosmo-Flux® (1%), and water (55%) (Immunopharmos 2002e) and a mixture of

glyphosate (5%), Cosmo-Flux® (1%), and water (95%) (Immunopharmos 2002f). Both
studies were performed using EPA guidance 870-2400.
In the first, 18 New Zealand White rabbits were used (Immunopharmos 2002e).
The test substance (0.1 ml) was placed into the conjunctival sacs of rabbits. The left

eye of each untreated animal served as negative control. The eyes of 3 rabbits of each
sex were rinsed for 30 second after the test substance application. A further 6 rabbits
were left with unrinsed eyes. The eyes were examined for irritation at 1, 24, 48, 72, 96
hours, and 7 days after instillation. The animals showed the following signs: opacity

(5/12, from grade 1 to 3); corneal damage (4/12 neovascularization on cornea); iritis
(5/12 grade 1, disappearing 4 days latter); conjunctivitis (12/12 from grade 1 to 3);
chemosis (10/12 from grade 1 to 3); discharge (4/12 animals presented discharge the
first days of the study).

The eyes of the 6 animals rinsed 30 seconds after application of the test
substance presented as follows: opacity (6/6 did not present corneal opacity); corneal
damage (6/6, with no damage); iritis (6/6 with no iritis); conjunctivitis (6/6 animals
presented from grade 1 to 3, which was diminishing which disappeared at the end of the
study, 7 days); chemosis (3/6 animals presented grade 1 which disappeared in 24

hours); discharge (6/6 animals presented discharge the first two days of the study). In
conclusion, the test substance caused slight to moderate irritation in the eyes from
animal that were treated and then not rinsed. This irritation was observable between
days 1 and 7. In contrast, the test substance did not produce irritation in animals, the

eyes of which were treated and then rinsed for 30 seconds after the application of test
substance.

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In the second study, 18 New Zealand White rabbits were used (Immunopharmos

2002). Again, 0.1 ml of the test substance was placed into the conjunctival sacs of
rabbits. The left eye of each untreated animal served as negative control. The eyes of
3 rabbits of each sex were rinsed for 30 seconds after the test substance application. A
further 6 rabbits were left with unrinsed eyes. The eyes were examined for irritation at

1, 24, 48, 72, 96 hours after instillation. The test substance did not cause irritation in
the eyes from animals treated and not rinsed (observed between days 1 and 4). The
test substance did not produce irritation in the eyes of animals treated and rinsed 30
seconds after the application of test substance and then observed for 4 days.

4.2.2.7 Skin sensitization

The dermal sensitization potential of test substance was evaluated in guinea pigs
(Springborn 2003g). Twenty Hartley albino guinea pigs (10 male and 10 female) were
topically treated with 100% test substance, once per week, during three weeks.

Following a 2-week rest period, a challenge was performed [20 animals treated and 10
animals untreated (challenge control)] were topically treated with 100% test substance.
A positive control group was given hexylcinnamaldehyde (HCA). Based on the results
of this study, test substance was not considered to be a sensitizer.

Other skin sensitization studies were performed on a mixture of glyphosate
(44%), Cosmo-Flux® (1%), and water (55%) (Immunopharmos 2002j) and a mixture of
glyphosate (5%), Cosmo-Flux® (1%), and water (95%) (Immunopharmos 2002i). Both
studies were performed according to EPA guideline 870-2600. In the first, 30 Hartley

guinea-pigs (300-350 g bw), were divided into 6 groups; 2 groups of males with 5
animals and 2 groups of females with 5 animals for the study, and 2 groups of 5 animals
of both sexes that serves as control. The test substance (0.5 ml) was applied to the
skin of albino guinea-pigs three times with an interval between each exposure of 1 week
(0, 7, and 14 days) and for a duration of 6 hours in each application. The animals were

inspected at 24, 48, and 72 hours after applications. The control group (5 male and 5
female) received sterile distilled water. A positive sensitization study was conducted
every 6 month using a sensitizing agent (data not given). The test material caused no
dermal adverse reactions even after several applications (Buehler test). It was noted
that the test material was not a sensitizer for the skin

In the second study (Immunopharmos 2002i), 30 Hartley guinea-pigs (300-350 g
of weight), were divided in 6 groups; 2 groups of males with 5 animals and 2 groups of
females with 5 animals for the study, and 2 groups of 5 animals of both sexes that

served as a control. The test substance (0.5 ml) was applied to the skin of albino
guinea-pigs, three times with an interval for each exposure of 1 week (0, 7, and 14
days) and 6 hours for each application (Buehler test). A total of 0.5 ml was applied over
the exposed skin. The animals were inspected at 24, 48, and 72 hours after application.
The control group (5 male and 5 female) received sterile distilled water. The positive

sensitization study was conducted in the laboratory every 6 months using a sensitizing
agent (data not given). The test material caused no adverse dermal reactions even
after several applications (Buehler test). It was concluded that the test material was not
a sensitizer for the skin.

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4.2.2.8 General conclusions on the mammalian acute toxicity of glyphosate and

Cosmo-Flux®
Based on the results of these studies undertaken with the mixture glyphosate
and Cosmo-Flux®, the following conclusions can be drawn:

• The acute oral and dermal LD50 value was estimated to be greater than
5,000 mg/kg bw in the rat. Therefore, this formulation is considered as
practically non-toxic by the oral route.

• The acute inhalation LC50 value was estimated to be greater than 2.60 mg/L
in the rat. In one study the rats showed breathing abnormalities after
exposures at 2.6 mg/L for 4 hours. This value for the test substance is
considered as potentially harmful for durations of exposure of the order of 4
hours. In two other studies, the mixture was shown to not be harmful at

exposures up to 20 mg/L for 4 hours. Exposures via the inhalation route in
these animal studies were via small droplets. Exposures via inhalation under
field conditions will be smaller as the droplets are larger and less easily
inhaled.

• The formulation is considered to be a slight and moderate irritant to the skin
and eyes of the rabbit. The calculated Primary Irritation Index for the test
article was 0.25.

Based on these observations, the hazard to the humans via application or
bystander exposures are considered small and are limited to slight to moderate skin and
eye irritation. These responses will be reduced if the affected areas are rinsed shortly
after exposure to remove contamination. It was also concluded that the addition of the
adjuvant Cosmo-Flux® to the glyphosate did not change its toxicological properties to

mammals.

4.3 EFFECTS IN THE FIELD

4.3.1 Duration of effects in the field

In tropical forest situations, similar to some of the locations of the coca
eradication programs, there are limited data on vegetation recovery following
glyphosate application. Nevertheless, there are a number of studies of successional
patterns following land clearance and for tree gaps. Forest clearance has been a

historical feature of the development of agriculture from across the globe, (e.g. Boahene
1998, Matlack 1997). In Central America, agricultural intensification and forest
clearance in Mayan and other cultures has been determined from the pollen record,
(e.g. Clement and Horn 2001, Curtis et al. 1998, Goman and Byrne 1998). Patterns of
successional change (recovery) in Neotropical forests have been reviewed by

(Gauriguata and Ostertag 2001). The authors note:
“the consensus of these analyses is that the regenerative power of
Neotropical forest vegetation is high, if propagule sources are close by and
land use intensity before abandonment has not been severe.

Nevertheless, the recovery of biophysical properties and vegetation is
heavily dependent on the interactions between site-specific factors and

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land use, which makes it extremely difficult to predict successional

trajectories in anthropogenic settings.”
In relation to the eradication program, patterns of vegetation recovery will be
dependent on size of plot, location of plot in relation to surrounding vegetation types and

local anthropogenic management, i.e., subsequent cultivation activities.
A study of tree regeneration in dry and humid selectively-logged Bolivian tropical
forests indicated that tree release with glyphosate in logging gaps had no significant

impact on target tree species growth (Pariona et al. 2003). While glyphosate controlled
vegetation for a limited period, there were problems with the recruitment of commercial
trees in logging gaps, suggesting a silvicultural need for site preparation treatments and
more judicious seed tree retention.

Glyphosate has been widely used for controlling deciduous understorey
vegetation in managed northern forests, so-called conifer-release treatments, (e.g.
Lautenschlager and Sullivan 2002). Recovery of the deciduous herb and shrub layers
occurs over a period of 2-3 years in general and the tree layer over 10 years (See
Section 4.3.2.3). Often, total structural diversity is unaffected by glyphosate treatments

after one year.

4.3.1.1 Forest clearance and soils
The impacts of forest clearance on soil fertility are generally well-understood.

Typically, tropical forest soils are fragile, being nutrient-poor and subject to leaching.
Tree clearance can quickly result in loss of nutrients, change in pH, and therefore
change in element availability to plants (McAlister et al. 1998). Such conditions often
allow only shifting cultivation under subsistence production, so-called slash-and-burn

agriculture. Studies in Jamaican forests have shown that cultivations result in large
amounts of soil erosion compared with secondary forest. An agroforestry treatment with
Calliandra calothyrsus contour hedges reduced erosion and increased rainfall infiltration
within the hedges (McDonald et al. 2002). As coca is a shrub, typically grown in rows, it
might be argued that soil and water changes associated with forest clearance may be

less than for annual crops such as maize, but clearly both have significant adverse
effects on primary forest sites.
Whilst vegetation recovery may be rapid, in eastern North America, research has
led to the surprising conclusion that 19 century agricultural practices decreased forest

floor nutrient content and C:N and C:P ratios and increased nitrifier populations and net
nitrate production, for approximately a century after abandonment (Compton and Boone
2000). The level of agricultural intensity, in terms of cultivation and fertilizer use, may
have significant long-term impact on soils.

4.3.1.2 Effects on associated fauna
In an area of highly disturbed tropical dry forest in Cordoba Department, northern
Colombia, small mammals were censused by live-trapping, running from secondary
growth forest into agricultural areas (Adler et al. 1997). The results suggest that the

disturbed habitat supports a small mammal fauna of low diversity. However, several of
the species appear to have benefited from forest clearance and agricultural activities
and may occasionally reach extremely high numbers, though populations were not

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stable. A similar effect on reduced diversity of termites with increasing disturbance has

been shown in dry forest in Uganda (Okwakol 2000). Changes in bird populations of a
eucalypt forest in Australia following clear-felling indicate that full recovery may take up
to 70 years (Williams et al. 2001).

Whilst some species are adapted to disturbed conditions and can utilize
agricultural land and secondary forest, there are many species associated with primary
forest only, for example the Great Argus pheasant in Indonesian tropical forests (Nijman
1998). With much of Colombia associated with extremely high biodiversity, there are
very many endemic plant and animal species associated with National Parks and

indeed with eradication areas.
Studies on the impacts of vegetation change caused by glyphosate use on
associated fauna in northern environments are available for some species. For

example, following the application of glyphosate in clear-cut forest areas in Maine, USA,
the use by moose (Alces alces) of treated and untreated areas was compared 1-2 years
and 7-11 years post application (Eschenburg et al. 2003, Eschholz et al. 1996). At 1
and 2 years post-treatment, tracks of foraging moose were 57 and 75% less abundant
on treated than untreated clear-cuts (P = 0.013). However, at 7-11 years post-

treatment, tracks of foraging moose (P = 0.05) and moose beds (P = 0.06) were greater
on treated than untreated clear-cuts. Less foraging activity at 1-2 years post-treatment
appeared to be the result of reduced browse availability, because conifer cover for
bedding was similar on treated and untreated clear-cuts. The authors hypothesized that
greater counts of tracks of foraging moose on older treated clear-cuts was due to

increased foraging activity on sites with more abundant conifer cover (Eschholz et al.
1996, Raymond et al. 1996), i.e. tree cover had returned sufficiently after 10 years.
Studies of small mammal responses to glyphosate vegetation control in similar
environments (Sullivan et al. 1998) have indicated that vegetation recovery 2-3 years

after treatment was sufficient to return population dynamics to expected ranges.
Spot applications of glyphosate to reduce invasive ground flora in forests can
have the beneficial effect of opening up the ground layer and encouraging spring
ephemeral species to establish larger populations. Carlson (2004) reported this effect

when controlling Alliaria peteolata, an invasive biennial plant. The impact of glyphosate
on the target species was only for a single season.

4.3.1.3 Interactions with surfactants

Surfactants significantly improve coca control with glyphosate (Collins and
Helling 2002) and control of Salvinia molesta, an aquatic fern (Fairchild et al. 2002).
Nevertheless, the behavior of surfactants is complex (Liu 2004). Spray droplet size
affects retention on the target plant, but also the absorption into the plant. Smaller
droplets are better retained on the plant, but absorption through the leaf is better from

larger “coarse” droplets (Feng et al. 2003). A study of volume rate effects of glyphosate
on grasses has shown that reduced application volumes give better control, partly
affected by the concentration of surfactants in formulated products (Ramsdale et al.
2003).

Studies of biodegradable non-phytotoxic rapeseed oil derivatives (triglyceride
ethoxylates; Agnique RSO(R) series containing an average of 5, 10, 30 and 60 units of
ethylene oxide) indicate that these adjuvants gave similar or better control of Phaseolus

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vulgaris L. compared with 0.36 Kg AE/L SL Roundup Ultra®. In these studies Agnique

RSO 60 generally was most effective (Haefs et al. 2002). Tests with a range of
surfactants and different herbicides on several plant species indicate that the optimum
surfactant structure is both herbicide and plant species dependent (Johnson et al.
2002).

Studies of synergism between amino acid biosynthesis-inhibiting herbicides
indicate that, in most cases associated with glyphosate, the lack of effects with technical
herbicide confirm that surfactants are important components of formulated products
(Kudsk and Mathiassen 2004).

4.3.2 Recovery from effects

4.3.2.1 Principles
Glyphosate, as a well-translocated herbicide, affects most plant species, if

sufficient herbicide can penetrate plant tissues, particularly leaves. Effects typically
result in plant death over a period of 2 to 3 weeks, though species with extensive
storage organs, e.g. long rhizomes, large size, or particularly impenetrable leaf
surfaces, may survive. A low dose of glyphosate can result in growth abnormalities in

plants, most typically localized accelerated branching. If the dose of herbicide is
insufficient to cause death, it has been proposed that plant fitness may also be reduced,
such that if there is competition with other plants, death may result indirectly, though
there is little published evidence for this.

The effect of glyphosate is limited to the plants that receive spray at the time of
application, as the herbicide is rapidly adsorbed onto soil and root uptake does not
occur. The broad spectrum of plant species controlled and the pattern of foliar uptake,
together with the safety of the compound, have led to widespread use of the herbicide

for total vegetation control, pre-harvest weed control in annual crops and for the
eradication of perennial plants.
Recovery of treated areas is dependent on the initial level of control, the
quantities of material (and the methods used) for plant regeneration and the

environmental conditions of the site. Plants have a variety of adaptations for
regenerating, with some life forms showing a range of methods, while others have only
a single strategy. Monocarpic species, typically annuals, have seeds for recruitment of
the next generation. Polycarpic species may also produce seeds, but many also have a
variety of vegetative means of regenerating, such as rhizomes, bulbs, corms and

runners. Patterns of secondary succession, the resultant plant communities over time,
reflect the plant-environment interactions and the opportunities for regeneration
provided by the local species pool. Seeds in the soil or that can reach a site from the
surroundings, together with vegetative fragments, will establish initially. Continued

agricultural operations, such as cutting or soil disturbance, will have a major influence
on the species that survive. In most situations, vegetation recovery is rapid, with ruderal
and pioneer plant species establishing within weeks of application.

4.3.2.2 Tropical situations

In tropical forests, similar to some of the locations of the coca eradication
programs, there is limited published data on vegetation recovery following glyphosate

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application. Nevertheless, there are a number of studies of successional patterns

following land clearance and for tree gaps. Secondary succession (forest recovery) has
become more common in some forest areas, for example in Puerto Rico (Chinea 2002).
Forest recovery is generally fairly rapid, but recovery of the full complement of forest
species can take many years (>30 y) and the effects of bulldozing for initial clearance

can reduce diversity of native species and enhance establishment of non-native
species. Comparisons of different aged plots (2-40 y) in the Bolivian Amazon forests
have contributed to the knowledge of secondary succession (Pena-Claros 2003). Not
surprisingly, it takes longer for the forest canopy to achieve similar diversity to mature
forest, compared with the understory and subcanopy communities.

In relation to the eradication program, patterns of vegetation recovery will be
dependent on size of plot, location of plot in relation to surrounding vegetation types and
local anthropogenic management, i.e., subsequent cultivation activities. Nevertheless, it
2
should be noted that naturally occurring tree gaps (20-460 m ) are an important
component of overall forest diversity, providing opportunities for understory and
subcanopy species and regeneration of canopy species in the modified light climate
(Martins et al. 2004, Martins and Rodrigues 2002). In Brazilian varzea (white-water)
forests, natural patterns of succession are affected by both light and local flooding

(Wittmann et al. 2004). The patch scale of eradication applications of glyphosate may
or may not be at the scale of natural gap dynamics; this deserves further scientific
study.
In the high Andes alpine paramo habitats, patterns of succession were described

(Sarmiento et al. 2003). Following cultivation, usually for potato, patterns of secondary
succession were such that after 12 years, the species diversity of the undisturbed
paramo had still not been attained. The characteristic paramo life forms, sclerophilous
shrubs (e.g. Baccharis prunifolia, Hypericum laricifolium) and giant rosettes (e.g.,

Espeletia schultzii), appear very early and gradually increase in abundance during
succession (Sarmiento et al. 2003).
In situations of agricultural expansion over large areas in Europe and North
America, there is evidence that, where the proportion of remaining ancient habitat is

low, subsequent forest recovery on abandoned agricultural land can be extended over
long time periods (Vellend 2003). It is unlikely that habitat fragmentation and intensity
of agriculture will combine to provide such a scenario in the coca eradication program
areas.

4.3.2.3 Temperate situations
Glyphosate has been widely used for controlling deciduous understorey
vegetation in managed northern forests, so-called conifer-release treatments, (e.g.
Lautenschlager and Sullivan 2002). Effects on the successional patterns of vegetation

in such temperate and boreal situations are that woody and herbaceous species are
most reduced by glyphosate, (e.g. Bell et al. 1997). In a study in British Colombia,
species richness, diversity, and turnover of the herb, shrub, and tree layers were not
significantly (p>0.10) different between mechanical and glyphosate spray cut stump

treatments and a control. Similarly, the structural diversity of herb, shrub, and tree
layers were also not significantly (p>0.10) different between treatments and control. By
opening the canopy and decreasing the dominance of the deciduous tree layer, both

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manual and cut-stump treatments showed greater total structural diversity (herb, shrub,

and tree layers combined) relative to the control. However, differences in total structural
diversity between treatments and control were, for the most part, not significant
(p>0.10). Therefore, these vegetation management treatments affected only the
volume of the targeted deciduous tree layer and did not adversely affect the species

richness, diversity, turnover, or structural diversity of the plant community. The authors
note that the results may be applicable to other temperate forest ecosystems where
conifer release is practiced in young plantations (Lindgren and Sullivan 2001). Herb
biomass and cover usually recover to untreated values within 2-3 years of conifer
release treatment (Sullivan 1994). Meanwhile, the reduced competition on target

conifers allows enhanced growth with little adverse effect on plant diversity (Sullivan et
al. 1996, Sullivan et al. 1998). Nevertheless, some plant groups may take longer to
recover from glyphosate application. For example, cryptogams (ferns) may take longer
than 5 years to recover in boreal forest situations (Newmaster and Bell 2002), probably
reflecting longer generation times and poor dispersal. Spot applications of glyphosate

to reduce invasive ground flora in forests can have the beneficial effect of opening up
the ground layer and encouraging spring ephemeral species to establish larger
populations. Carlson and Gorchov (2004) reported this effect when controlling Alliaria
peteolata, an invasive biennial plant. The impact of glyphosate on the target species

was only for a single season. Reviewing the effects of glyphosate use in forestry,
(Sullivan and Sullivan 2003) noted that:
“…the magnitude of observed changes in mean species richness and
diversity of vascular plants, birds, and small mammals, from the effects of

herbicide treatment, were within the mean values of natural fluctuations of
these variables. The biological significance of this impact is limited to shifts
in species composition based on changes in floral composition and
structure of habitats. Management for a mosaic of habitats within forest

and agricultural landscapes, which provide a range of conditions for plant
and animal species, should help ameliorate the short-term changes in
species composition accompanying vegetation management with
glyphosate”.

Single applications of glyphosate control much of the vegetation that receives
spray, but recovery is generally rapid and within the range of natural disturbances.

4.3.2.4 Conclusions

The experience of glyphosate use in northern temperate forests is such that
vegetation and fauna recover over a period of 2 to 3 years, following a single conifer-
release treatment. With generally rapid plant growth under tropical conditions, available
data confirm this scenario for Colombian conditions. In comparison, land clearance for
agriculture (or coca production) is a much more environmentally damaging operation,

impacting adversely on soils in particular. Land clearance for illicit crops is already a
threat to the conservation of bird species diversity in Colombia (Álvarez 2002). Whilst
there are legitimate scientific questions as to the effects of a) the spatial scale of
individual glyphosate applications and b) the return frequency of eradication treatments,

field operational factors set these parameters. Spray areas reflect the patch scale of
coca and poppy growing, averaging 1-2 ha each in a total of ~150,000 ha. Re-

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application frequencies are generally greater than 6 months for coca and greater than 3
months for poppy and, bearing in mind the molecule is biologically unavailable in the
soil and soil-bound residues have a half life of between 14 and 32 days, the
environmental impacts are no greater than single applications.

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5 RISK ASSESSMENT

The risk assessment was conducted by comparing estimated exposures to effect
values for glyphosate from specific toxicity studies, from the literature, and from
regulatory guidelines such as those established by the US EPA (1993b). The estimated
exposures used were those calculated for the use of glyphosate for eradication spraying
in Colombia. This was done for human and environmental risks and is outlined above.

5.1 HUMAN HEALTH
From an assessment of the results of toxicity testing of the formulation of
glyphosate and Cosmo-Flux® as used in Colombia (Section 4.2.2), it was concluded
that the addition of Cosmo-Flux® to the spray mixture did not affect the toxicity of the

glyphosate to mammals. For this reason, it was possible to compare the toxicity of
glyphosate and its formulations to exposures estimated under conditions of use in
Colombia.
Exposures for the assessment were taken from Tables 7-9. The greatest values

were taken as reasonable worst-case for a hazard assessment. These results are
shown in Table 20 and illustrated graphically in Figure 18. In comparing the exposure
and effect concentrations a margin of exposure approach was used. Thus a number
greater than 1 (Table 20) means that the exposure was less than the exposure or dose
that caused the response in the toxicology study.

From the data in Table 20, it is clear that potential exposures to glyphosate and
Cosmo-Flux® as used for the eradication of coca and poppy in Colombia do not present
a risk to human bystanders. In all cases, the margin of exposure for the most sensitive
endpoint in laboratory animal studies with glyphosate was greater than 100 – a
conservative value often used to account for uncertainty in risk assessments of this
type. As well, estimated worst- case exposures were below the Reference Dose (RfD)

established for glyphosate by the US EPA. The toxicity values used in both of these
approaches were derived from chronic exposures where the animals were dosed over
extended time periods. They are thus additionally protective of short and infrequent
exposures that would occur during the use of glyphosate in the eradication spray
program. Some exposure values were close to the inhalation toxicity value but, but as
discussed above, droplet size is large and inhalation will be less than in the laboratory

animal studies as well as the droplet size used in agricultural uses, from which the
potential inhalation exposure was derived.

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110 Annex 116

Table 20. Summary of reasonable worst-case estimated exposures of humans to

glyphosate resulting from use in the eradication of coca and poppy in Colombia
and margins of exposure.
Source of exposure Exposure value in Margin of exposure
mg/kg compared to the most
sensitive NOEL(175 mg/kg)

Coca Poppy Coca Poppy
Direct overspray 0.04 0.01 4,918 20,417

Reentry 0.26 0.06 676 2,804
Inhalation 0.01 0.01 28,226 28,226
Diet and water 0.75 0.18 234 972
Worst case total exposure

from all sources 1.05 0.26 167 680
Source of exposure Exposure value in Margin of exposure for the US
mg/kg EPA RfD (2 mg/kg/day)

Coca Poppy Coca Poppy

Direct overspray 0.04 0.01 56 233
Reentry 0.26 0.06 8 32
Inhalation 0.01 0.01 323 323

Diet and water 0.75 0.18 2.7 11.1
Worst case total exposure
from all sources 1.05 0.26 1.9 7.8

5.2 ENVIRONMENT

Assessment of the environmental risks of glyphosate and Cosmo-Flux® to
aquatic organisms was based on data from the literature and from studies conducted on
the mixture of formulated glyphosate and Cosmo-Flux® as used in Colombia. As
discussed in Section 4.1.2, the toxicity of the mixture of glyphosate and Cosmo-Flux®

was greater than that reported for formulated glyphosate itself. When the toxicity values
for the mixture as used in Colombia are compared to the range of estimated exposures
that would result from a direct overspray of surface waters (Table 10), it is clear that
aquatic animals and algae in some shallow water bodies may be at risk (Figure 19).

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111Annex 116

Formulated plus Cosmo-Flux®

All sources Inhalation

Toxicity
Diet and water Coca Dermal
Poppy

Inhalation Oral

Acute exposures
Glyphosate chronic
Reentry NOEL

Direct overspray Margin of exposure RfD

0.001 0.01 0.1 1 10 100 1000 10000

Glyphosate (mg/kg)

Figure 25 Illustration of acute toxicity values in laboratory mammals for glyphosate plus Cosmo-

Flux®, the NOEL from the most sensitive chronic study in laboratory animals, and the RfD
(glyphosate) and the estimated worst-case acute exposures that may be experiences under
conditions of use in Colombia.

While the overlap of the range of estimated exposure concentrations with the

toxicity values for the green alga and rainbow trout suggests that there may be
increased risk in situations where an accidental overspray will occur, this would have to

be in a location where a shallow water body is in close enough proximity to the coca
field that it is accidentally over-sprayed, that it is less than 30 cm deep, and that it is not

flowing. Because flow of the water would likely result in rapid hydraulic dilution to
concentrations to below the threshold of biological activity, organisms in flowing water

would not be at risk. It was not possible to determine the actual frequency of these risks
as data on proximity of surface water to coca fields is not available at this time. Based

on the toxicity data with formulated Roundup® in amphibians, this group of organisms
may be at risk, however, specific testing in amphibians has not yet been conducted on

the mixture of glyphosate plus Cosmo-Flux® as used in Colombia.

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112 Annex 116

99 Estimated concentration in
surface water (cm deep)

MDL surface
water 200 30 15

90
Fathead
minnow

70 Waaerr ealle

50 Green agag

Percent rank Formulated LC50s amphibians
Raanbow
toutt Formulated LC50s arthropods

10 Formulated LC50s fish

Glyphosate technical LC50s
Glyphosate plus Cosmo-Flux -animals

Glyphosate plus Cosmo-Flux - algae

1
1 2 3 4 5 6 7 8
10 10 10 10 10 10 10 10

Concentration (µg/L glyphosate AE)

Figure 26 Distribution of toxicity values for glyphosate technical, formulated glyphosate (Roundup®) in
all aquatic organisms and in fish and the toxicity values in four aquatic species for glyphosate and
Cosmo-Flux® 411 mixture as used in Colombia. The yellow rectangle shows the range of predicted
worst-case exposures resulting from direct overspray of surface waters ranging from 15 to >200 cm in

depth. Lines are the regressions through the log-probability transformed data.

Based on the toxicity data for honeybees (Section 4.1.2.1), the mixture of
glyphosate and Cosmo-Flux® 411F is not acutely toxic via contact exposure to honey

bees. It did not cause mortality or stress effects in bees in the normal 48 hour period
after treatment at concentrations equal to or less than 56.8 mg AE/bee. These results

show that the formulated product is not directly hazardous to bees and, by
extrapolation, to other beneficial insects.

Although no acute or chronic data are available on wild animals, extrapolation of

the mammalian data discussed above (Sections 4.1.2 and 4.2.2) and from reports in the
literature support the conclusion that glyphosate and Cosmo-Flux®, as used in the

eradication program in Colombia, will not have adverse direct effects on wild mammals
or birds. Indirect effects through habitat alteration are possible. However, it is unlikely

that the coca and poppy fields are significant habitats for wildlife. Human activities
related to cultivation and harvesting the crop will be more disruptive to wildlife and death

of the coca bushes or the poppy plants as a result of spraying with glyphosate will not
add an additional stressor. In fact, if the sprayed area is not replanted and allowed to

naturalize, this new successional habitat may be more attractive to birds and mammals
than an old-growth forest. Given that coca and poppy fields are usually located in

remote areas and are often surrounded by natural habitats, sources for recolonization or

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113Annex 116

alternate habitats will be close by. Some habitat alteration will result from accidental
over-sprays that affect non-target vegetation, however, as discussed above (Section
2.1.3.5), these areas are small in relation to the sprayed fields < 0.48%), represent a
very small proportion of the total habitat available << 0.001%, and will undergo rapid

recolonization and succession to habitats suitable for wildlife.

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114 Annex 116

6 CONCLUSIONS

Because of differences in the approaches to human and ecological risk

assessment, the conclusions of this report are discussed separately in the following

sections. In these discussions, the risks associated with the use of glyphosate and
Cosmo-Flux® in the coca and poppy eradication program in Colombia are related to the

total impacts of coca and poppy production as discussed in the Problem Formulation

(Section 2.2.1).

6.1 HUMAN HEALTH RELEVANCE

Based on all of the evidence and information presented above, the Panel

concluded that the risks to humans and human health from the use of glyphosate and

Cosmo-Flux® in the
eradication of coca and
Coca or poppy
poppy in Colombia were field developed in

minimal (Figure 20). The a natural area
acute toxicity of the

formulated product and

Cosmo-Flux® to laboratory

animals was very low, the NTTENSITY REECOVERYY FREEQUENCYY MPPACT % IMPACCT
likely exposures were low, IMPPACTTS SCOORE SCORRE % SCORRE

and the frequency of Clearr

exposures was low. When cuttng and
these risks are compared burning 5 3 3 45 167.7

to other risks associated Plantng the
coca orr
with clearing of land, the popppy 0 1 100 0 0.00
uncontrolled and
Ferrtzerrliiz
unmonitored use of other
inputss 0 05.5 10 0 0.00
pesticides to protect the
Peestcded
coca and poppy, and inputss 5 3 10 150 555.5
exposures to substances
Eradicattono
used in the refining of the sprayy <01.1 0 1 <01.1 0.11

raw product into cocaine Processing
and heroin, they are and refnngg 5 3 5 75 278.8

essentially negligible.
Figure 27 Potential human health impacts of the cycle of coca or

poppy production and the spray eradication program.

6.2 ECOLOGICAL RELEVANCE

Based on the evidence and data discussed above and the results of a number of
specific studies conducted specifically for this assessment, the Panel concluded that the

risks to the environment from the use of glyphosate and Cosmo-Flux® in the eradication

of coca and poppy in Colombia were small in most circumstances (Figure 21). Risks of

direct effects in terrestrial wildlife such as mammals and birds were judged to be
negligible as were those to beneficial insects such as bees. Moderate risks to some

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115Annex 116

aquatic wildlife may exist in some locations where shallow and static water bodies are
located in close proximity to

coca fields and are accidentally Coca or poppy

over-sprayed. However, when field developed in
taken in the context of the a natural area

environmental risks from other
activities associated with the

production of coca and poppy,

in particular, the uncontrolled MPPAACTTS NTTENSITY RECOVVERY IMPACCT %
and unplanned clearing of SCCORE TIME Y)) SCOORE IMPACCT

pristine lands in ecologically Clearcuttngiin

important areas for the and burnngn 5 60 300 7.66
purposes of planting the crop,
Panntng heh
the added risks associated with coca orr
the spray program are small. poppy 1 4 4 1.33

Ferrtzerrliiz
nputss 1 0.55 0.55 0.22

Pestcide

nputss 2 0.55 1 0.33

Eradicatonno
spray 1 0.55 0.55 0.22

Processing
and effnngn 2 1 2 0.77

Figure 28 Potential environmental impacts of the cycle of coca
or poppy production and the spray eradication program.

6.3 STRENGTHS AND UNCERTAINTIES IN THE ASSESSMENT

This assessment has both strengths and uncertainties. These are discussed in

the following sections. These strengths and uncertainties lie in the exposure and effects

characterizations and, because these are used in the risk characterization, are also
reflected in the risk assessment. Uncertainties are inherent in all risk assessments and,

in some cases, can be easily addressed though additional data collection or specific

studies. Recommendations for additional studies and data collection are addressed in
the final section of this report.

6.3.1 Exposures

6.3.1.1 Environmental exposures

Applications of glyphosate are well characterized. State of the art equipment is

used. The locations of application and the areas sprayed are well documented and
measured with resolutions only equaled in some applications in forestry in other

jurisdictions. The mixing and application rates are well characterized and the probability

of application of amounts of glyphosate and Cosmo-Flux® greater than those specified

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are judged to be small. The resultant concentrations in soil and water that may result

from an accidental overspray also have high certainty. The environmental behavior of
glyphosate is well characterized and, under the conditions of use in the eradication
program in Colombia, will not persist, accumulate, or biomagnify in the environment.
Analyses of surface waters and sediments in one watershed where eradication spraying

was carried out did not reveal the presence of significant concentrations of glyphosate,
confirming the conclusion based on its properties that it is not mobile in the
environments where it is applied. Residues of glyphosate were not frequently detected
in areas where eradication spraying was not conducted but where glyphosate use was
known to occur in agriculture. Given that considerably more glyphosate is used in

agriculture and other non-eradication uses (~85%), this further confirms that glyphosate
is not sufficiently mobile to result in significant contamination of surface waters in
Colombia, regardless of the use pattern.

Uncertainties in the exposure characterization lie in lack of precise
measurements of the proximity of sprayed fields to surface waters and the proportion of
treated areas that are in close proximity to these surface waters. The sampling of the
surface waters only took place for a period of 24 weeks and only 5 locations were
sampled in this way. Although two of these were scheduled to be sprayed, only one

location was treated during the sampling period. For logistical reasons, it was also not
possible to sample close to the application sites. If sampling had been conducted at
more sites closer to the sprayed fields and over a longer time period, residues may
have been detected more frequently.

6.3.1.2 Human exposures
Human exposures to glyphosate were estimated from extensive and well
documented studies in other jurisdictions and are judged to be accurate with respect to
bystanders who are directly over-sprayed. Exposures were judged to be small and, in

all cases, considerably below thresholds of concern.
Application rates of glyphosate used for coca eradication are greater than those
used in conventional agriculture suggesting that experience and exposures measured

under these conditions may not be applicable to bystander exposures in eradication
spraying in Colombia. While this may be true, the margins between exposures doses at
which chronic effects may occur are great enough to provide a wide margin of safety to
bystanders. Less information is available regarding the likelihood of exposure upon
reentry to coca fields immediately after spraying. This relates to the anecdotal evidence

that picking of leaves or pruning of plants immediately after they are sprayed with
glyphosate will “save” the plants. Exposures under these conditions are unmeasured,
but are estimated to be considerably below the US EPA reference dose.

6.3.2 Effects

6.3.2.1 Environmental effects
The environmental toxicology database for glyphosate is relatively large and its
effects in non-target organisms are well known or can be extrapolated. Glyphosate
itself is of low toxicity to non-target organisms, however, there are a number of

formulations of glyphosate that exist in the marketplace and these may contain many

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different surfactants and/or adjuvants. It is also known that it is the surfactants that

determine the toxicity of the formulation as many are more toxic than technical
glyphosate itself. Because of this, the Panel had several toxicity tests conducted with
the formulated product of glyphosate plus Cosmo-Flux® as used in the eradication
program in Colombia. This reduced uncertainty with respect to toxicity to beneficial

insects such as the honeybee and to aquatic organisms. Recent studies have reported
that amphibians, such as frogs, are amongst the more sensitive aquatic organisms with
respect to formulations of glyphosate such as Roundup® and Vision®. We did not
conduct toxicity studies in amphibians with the mixture of glyphosate plus Cosmo-Flux®
and this is a source of some uncertainty for ecological risks for frogs.

6.3.2.2 Effects in humans
The database of effect data for glyphosate is large and its risks to humans and
the environment have extensively reviewed and assessed in a number of national and

international jurisdictions as well as in the open scientific literature. In all cases,
glyphosate has been judged to be of low risk. However, some of the studies on which
these assessments are based were conducted prior to the refinement of testing
guidelines and the availability of new and more sensitive methods of analysis and effect
characterization, such as those based on alteration in the concentrations of

neurotransmitters and their metabolites in the central nervous system. In the process of
reassessment and re-registration, older studies will be replaced with newer tests
conducted according to current guidelines. Given the large and expanding use of
glyphosate in agriculture, the priorities for updating the database will likely be high.

Changes in the regulatory status of glyphosate should be monitored and any newly
identified risks included in an updated risk assessment.
There is considerable literature on the epidemiology of pesticides and possible
effects on human health. As a result of recent work, it is clear that many epidemiology

studies are confounded by the use of poor and inaccurate surrogates for exposures to
pesticides. The Panel also conducted a preliminary epidemiological study to assess
possible linkages between the use of glyphosate and adverse human-health outcomes
and recognizes that, for clear logistical reasons, no measures of exposure were
available for the various groups enrolled in the study other than the use of glyphosate

for eradication spraying in the region. The results of this study do not suggest that there
is an association between the use of glyphosate in the eradication program and time to
pregnancy (TTP) as a reproductive outcome. A somewhat greater risk for longer TTP
was observed in one region (Valle del Cauca) where eradication spraying is not

conducted but it was not possible to identify any specific factors that may have been
responsible for this observation.

6.3.3 Confounding risks
Through the Tier-1 and Tier-2 hazard assessments of the other substances used

in the production and refining of cocaine and heroin, the Panel recognizes that some of
these substances present a significantly greater hazard to both humans and the
environment than does the mixture of glyphosate and Cosmo-Flux® used in the
eradication program in Colombia. Exacerbating these hazards is the lack of information

about the conditions of use of these substances. Because of the lack of specific data

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118 Annex 116

on use and exposure, it was not possible to conduct detailed risk assessments for these

substances. From anecdotal evidence and observations in other locations, it is clear
that, in most cases, these substances are used without adequate safety training, without
adequate protective equipment, without suitable disposal methods, and without
supervision. This represents a significant and serious potential risk to humans and the
environment.

6.4 RECOMMENDATIONS
The Panel has identified a number of uncertainties in its review of the data and
from these makes the following recommendations. These recommendations are

grouped into two classes, recommendations to retain current practices that were judged
to be essential or useful (Table 21) and recommendations related to new activities or
data collection that will address key uncertainties identified in our study (Table 22).

Table 21. Recommendations for the continuance of current practices in the coca and
poppy eradication program in Colombia

Practice Benefit of continuance Ranking of
importance
(5 = most
important)

Mixer-loader, worker, and Protection of the humans and the 5
environmental protection in the environment from excessive

storage, mixing, and loading exposures.
operations.
Use of state of art application Accurate records of location and 5

technology. areas sprayed.
Replace the respirator worn by This recommendation is 5
the mixer-loader with a full face modification of current procedures
shield to reduce the potential for that will reduce the risk of

splashed material to run down the splashes of concentrated
face into the eyes. formulation into the eyes.
Use of glyphosate in the The risk of this product to humans 4
eradication program. and the environment is judged to
be lower than any currently-

available alternatives. However, if
new candidate products become
available, their use should only be
considered after an appropriate
risk assessment has been

conducted.

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119Annex 116

Table 22. Recommendations for the collection of new data and information in the

coca and poppy eradication program in Colombia
Recommendation Benefit of new data Ranking of
importance
(5 = most
important)

Conduct a study to identify risk This is a recommendation 3

factors associated with time to resulting from the observation of
pregnancy (TTP). increased risk of longer TTP in
one region of Colombia (Valle del
Cauca) where eradication
spraying was not carried out. The

study should be considered for
prioritization in the general human
health research programs
conducted in Colombia.
Including proximity to surface Better indication of likely 2

waters in (geographic Information frequency of contamination of
System (GIS) analysis of locations these habitats. This would help to
and areas of coca and poppy better quantify the risks to aquatic
fields. organisms in shallow-water non-
flowing habitats.

Identify mixtures of glyphosate Reduction in possible 2
and adjuvants that are less toxic environmental impacts to non-
to aquatic organisms than the target organisms in shallow
currently used mixture. The surface water environments.
priority of this recommendation

would depend on the results of
the GIS analysis.
Testing of the glyphosate-Cosmo- Decrease in uncertainty regarding 2
Flux® formulation for toxicity to the toxicity to amphibians.
amphibians.

Use of GIS to quantify areas of Better quantification of proportion 2
coca and poppy production in of regions identified as important
biodiversity hotspots. sources of biodiversity that are
being adversely impacted
because of clear-cutting and

planting of coca and poppy.
Use of GIS to quantify size of Allow more accurate quantification 2
fields planted to coca and poppy of potentially impacted areas as
and track these over time to judge well as recovery when these fields
extent of environmental impact as are abandoned.

well as recovery.
Review the regulatory status of Ensure that new testing and 2
glyphosate on a regular basis. toxicity data on glyphosate are
included in the risk assessment of

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120 Annex 116

Table 22. Recommendations for the collection of new data and information in the

coca and poppy eradication program in Colombia
Recommendation Benefit of new data Ranking of
importance
(5 = most

important)

its use in eradication spraying in
Colombia.
Measurement of exposures to Better characterization of 1
glyphosate in bystanders to exposures under conditions of use

sprays and reentry into sprayed in Colombia.
fields. This recommendation
would follow selection of new
formulations and mixtures of
adjuvants that have lower

environmental toxicity.

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121Annex 116

7 REFERENCES

Acquavella JF, Weber JA, Cullen MR, Cruz OA, Martens MA, Holden LR, Riordan S,
Thompson M, Farmer D. 1999. Human ocular effects from self-reported
exposures to Roundup® herbicides. Human and Experimental Toxicology
18:479-486.

Acquavella JF, Alexander BH, Mandel JS, Gustin C, Baker B, Chapman P, Bleeke M.
2004. Glyphosate biomonitoring for farmers and their families: Results from the
farm family exposure study. Environmental Health Perspectives 112:321-326.

Adler GH, Arboledo JJ, Travi BL. 1997. Diversity and abundance of small mammals in

degraded tropical dry forest of northern Colombia. Mammalia 61:361-370.

Álvarez MD. 2002. Illicit crops and bird conservation priorities in Colombia.
Conservation Biology 16:1086-1096.

Amerio P, Motta A, Toto P, Pour SM, Pajand R, Feliciani C, Tulli A. 2004. Skin toxicity
from glyphosate-surfactant formulation. Journal of Toxicology-Clinical Toxicology
42:317-319.

Araujo AS, Monteiro RT, Abarkeli RB. 2003. Effect of glyphosate on the microbial
activity of two Brazilian soils. Chemosphere 52:799-804.

Arbuckle T, Cole DC, Ritter L, Ripley BD. 2005. Biomonitoring of herbicides in Ontario
farm applicators. Scandanavian Journal of Work and Environmental Health 31
Suppl 1:In Press.

Arbuckle TE, Lin Z, Mery LS. 2001. An exploratory analysis of the effect of pesticide
exposure on the risk of spontaneous abortion in an Ontario farm population.
Environmental Health Perspectives 109:851-857.

Arbuckle TE, Cole DC, Ritter L, Ripley BD. 2004. Farm children's exposure to
herbicides: Comparison of biomonitoring and questionnaire data. Epidemiology

15:187–194.

Arregui MC, Lenardon A, Sanchez D, Maitre MI, Scotta R, Enrique S. 2004. Monitoring
glyphosate residues in transgenic glyphosate- resistant soybean. Pest
Management Science 60:163-166.

Bababurmi EA, Olorunsogo OO, Bassir O. 1978. Toxicology of glyphosate in rats and
mice. Toxicology and Applied Pharmacology 45:319-320.

Baerson SR, Rodriguez DJ, Tran M, Feng Y, Biest NA, Dill GM. 2002. Glyphosate-
resistant goosegrass. Identification of a mutation in the target enzyme 5-

enolpyruvylshikimate-3-phosphate synthase. Plant Physiology 129:1265-1275.

Page 97 of 121

122 Annex 116

Baird DD, Wilcox AJ, Weinberg CR. 1986. Use of time to pregnancy to study
environmental exposures. American Journal of Epidemiology 124:470-480.

Baird DD, Weinberg CR, Rowland AS. 1991. Reporting errors in time-to-pregnancy data

collected with a short questionnaire. Impact on power and estimation of
fecundability ratios. American Journal of Epidemiology 133:1282-1290.

Barbosa ER, Leiros da Costa MD, Bacheschi LA, Scaff M, Leite CC. 2001.
Parkinsonism after glycine-derivate exposure. Movement Disorders 16:565-567.

BC Ministry of Forests. 2000. Boreal plant community diversity 10 years after
glyphosate treatment: A report summary. Victoria, BC, Canada: Silviculture
Practices Section. 4 p.

BCPC. 2002-2003. The e-Pesticide Manual. Tomlin CDS, editor. Farnham, Surrey, UK:
British Crop Protection Council.

Bell FW, Lautenschlager RA, Wagner RG, Pitt DG, Hawkins JW, Ride KR. 1997. Motor-

manual, mechanical, and herbicide release affect early successional vegetation
in northwestern Ontario. Forestry Chronicle 73:61-68.

Bell JR, Johnson PJ, Hambler C, Haughton AJ, Smith H, Feber RE, Tattersall FH, Hart
BH, Manley W, Macdonald DW. 2002. Manipulating the abundance of

Lepthyphantes tenuis (Araneae: Linyphiidae) by field margin management.
Agriculture Ecosystems & Environment 93:295-304.

Benedetti AL, Vituri CD, Trentin AG, Domingues MAC, Alvarez-Silva M. 2004. The
effects of sub-chronic exposure of Wistar rats to the herbicide Glyphosate-

Biocarb (R). Toxicology Letters 153:227-232.

Bengtsson G, Hansson LA, Montenegro K. 2004. Reduced grazing rates in Daphnia
pulex caused by contaminants: Implications for trophic cascades. Environmental

Toxicology and Chemistry 23:2641-2648.

Blackburn LG, Boutin C. 2003. Subtle effects of herbicide use in the context of
genetically modified crops: A case study with glyphosate (Roundup (R)).
Ecotoxicology 12:271-285.

Blakley BR. 1997. Effect of Roundup and Tordon 202C herbicides on antibody
production in mice. Veterinary and Human Toxicology 39:204-206.

Boahene K. 1998. The challenge of deforestation in tropical Africa: Reflections on its

principal causes, consequences and solutions. Land Degradation & Development
9:247-258.

Page 98 of 121

123Annex 116

Boateng JO, Haeussler S, Bedford L. 2000. Boreal plant community diversity 10 years

after glyphosate treatment. Western Journal of Applied Forestry 15:15-26.

Bray W, Dallery C. 1983. Coca chewing and high altitude stress: A spurious correlation.
Current Anthropology 24:269-274.

Buckelew LD, Pedigo LP, Mero HM, Owen MDK, Tylka GL. 2000. Effects of weed
management systems on canopy insects in herbicide-resistant soybeans. Journal
of Economic Entomology 93:1437-1443.

Burgat V, Keck G, Guerre P, Pineau X. 1998. Glyphosate toxicosis in domestic animals:
A survey from the data of the Centre National d'Informations Toxicologiques
Veterinaires (CNITV). Veterinary and Human Toxicology 40:363-367.

Campbell A, Chapman M. 2000. Handbook of poisoning in dogs and cats. Oxford, UK:

Blackwell Science.

Carlson AM, Gorchov DL. 2004. Effects of herbicide on the invasive biennial Alliaria
petiolata (garlic mustard) and initial responses of native plants in a southwestern

Ohio forest. Restoration Ecology 12:559-567.

[Centre for Biodiversity] Centre for Biodiversity. 2004. Andean Biodiversity Region.
Centre for Biodiversity website
http://www.biodiversityhotspots.org/xp/Hotspots/andes/, Accessed December 30

2004.

Chang CY, Peng YC, Hung DZ, Hu WH, Yang DY, Lin TJ. 1999. Clinical impact of
upper gastrointestinal tract injuries in glyphosate-surfactant oral intoxication.
Human and Experimental Toxicology 18:475-478.

Chen CY, Hathaway KM, Folt CL. 2004. Multiple stress effects of Vision herbicide, pH,
and food on zooplankton and larval amphibian species from forest wetlands.
Environmental Toxicology and Chemistry 23:823-831.

Chinea JD. 2002. Tropical forest succession on abandoned farms in the Humacao
Municipality of eastern Puerto Rico. Forest Ecology and Management 167:195-
207.

CICAD/OAS. 2004a. The Toxicology of Chemicals Used in the Production and Refining
of Cocaine and Heroin: A Tier-one Assessment. Washington, DC, USA: CICAD,
Organization of American States. OAS/CICAD 2004-01. 536 p.

CICAD/OAS. 2004b. STANDARD OPERATING PROCEDURE. Stream Water and

Sediment Sampling for Other Pesticides. Washington DC, USA: CICAD. 14 p.

Page 99 of 121

124 Annex 116

CICAD/OAS. 2004c. STANDARD OPERATING PROCEDURE. Stream Water and

Sediment Sampling for Glyphosate/AMPA. Washington DC, USA: CICAD. 14 p.

CICAD/OAS. 2005. The Toxicology of Selected Chemicals Used in the Production and
Refining of Cocaine and Heroin: A Tier-two Assessment. Washington, DC, USA:

CICAD, Organization of American States. OAS/CICAD 2005-01. ??? p.

Clement RM, Horn SP. 2001. Pre-Columbian land-use history in Costa Rica: a 3000-
year record of forest clearance, agriculture and fires from Laguna Zoncho.
Holocene 11:419-426.

Collins RT, Helling CS. 2002. Surfactant-enhanced control of two Erythroxylum species
by glyphosate. Weed Technology 16:851–859.

Compton JE, Boone RD. 2000. Long-term impacts of agriculture on soil carbon and

nitrogen in New England forests. Ecology 81:2314-2330.

Conners DE, Black MC. 2004. Evaluation of lethality and genotoxicity in the freshwater
mussel Utterbackia imbecillis (Bivalvia: Unionidae) exposed singly and in

combination to chemicals used in lawn care. Archives of Environmental
Contamination and Toxicology 46:362-371.

[Cosmoagro] Cosmoagro S.A. 2004. Cosmo-Flux® 411F Label. Cosmoagro website
www.cosmoagro.com, Accessed October 20, 2004

Cox C. 1998. Herbicide Factsheet: Glyphosate (Roundup). Journal of Pesticide Reform
18:3-17.

Curtis JH, Brenner M, Hodell DA, Balser RA, Islebe GA, Hooghiemstra H. 1998. A multi-

proxy study of Holocene environmental change in the Maya lowlands of Peten,
Guatemala. Journal Of Paleolimnology 19:139-159.

Curtis KM, Savitz DA, Weinberg CR, Arbuckle TE. 1999. The effect of pesticide

exposure on time to pregnancy. Epidemiology 10:112-117.

CWQG. 1999. Canadian Water Quality Guidelines (and updates). Ottawa, ON: Task
Force on Water Quality Guidelines of the Canadian Council of Resource and
Environment Ministers.

da Silva RS, Cognato GD, Vuaden FC, Rezende MFS, Thiesen FV, Fauth MD, Bogo
MR, Bonan CD, Dias RD. 2003. Different sensitivity of Ca2+-ATPase and
cholinesterase to pure and commercial pesticides in nervous ganglia of
Phyllocaulis soleiformis (Mollusca). Comparative Biochemistry And Physiology C

-Toxicology & Pharmacology 135:215-220.

Page 100 of 121

125Annex 116

Dallegrave E, Mantese FD, Coelho RS, Pereira JD, Dalsenter PR, Langeloh A. 2003.

The teratogenic potential of the herbicide glyphosate-Roundup (R) in Wistar rats.
Toxicology Letters 142:45-52.

Daruich J, Zirulnik F, Gimenez MS. 2001. Effect of the herbicide glyphosate on enzyme

activity in pregnant rats and their fetuses. Environmental Research A 85:226-
231.

De Roos AJ, Blair A, Rusiecki JA, Hoppin JA, Svec M, Dosemeci M, Sandler DP,
Alavanja MC. 2005. Cancer incidence among glyphosate-exposed pesticide

applicators in the agricultural health study. Environmental Health Perspectives
113:49-54.

Devine MD, Duke SO, Fedtke C. 1993. Physiology of Herbicide Action. Englewood
Cliffs, NJ: PTR Prentice Hall.

Diaz G, Carrillo C, Honrubia M. 2003. Differential responses of ectomycorrhizal fungi to
pesticides in pure culture. Cryptogamie Mycologie 24:199-211.

[Dinno A] 2002. DTHAZ: Stata module to compute discrete-time hazard and survival
probability. Dinno A website http://ideas.repec.org/c/boc/bocode/s420701.html,
Accessed March 15 2005.

Direccion Nacional de Estupefacientes. 2002. La lucha de Colombia contra las drogas

ilicitas: Acciones y resultados 2002. Colombia's war against drugs: Actions and
results 2002. Bogota, D.C. Colombia: Direccion Nacional de Estupefacientes.
374 p.

Dyer SD, Belanger SE, Carr GJ. 1997. An initial evaluation of the use of Euro/North

America fish species for tropical effects assessments. Chemosphere 35:2767-
2781.

Easton WE, Martin K. 1998. The effect of vegetation management on breeding bird

communities in British Columbia. Ecological Applications 8:1092-1103.

Easton WE, Martin K. 2002. Effects of thinning and herbicide treatments on nestsite
selection by songbirds in young, managed forests. Auk 119:685-694.

Edginton AN, Sheridan PM, Stephenson GR, Thompson DG, Boermans HJ. 2004.
Comparative effects of pH and Vision herbicide on two life stages of four anuran
amphibian species. Environmental Toxicology and Chemistry 23:815-822.

El-Demerdash FM, Yousef MI, Elagamy EI. 2001. Influence of paraquat, glyphosate,

and cadmium on the activity of some serum enzymes and protein electrophoretic
behaviour (in vitro). Journal of Environmental Science and Health B 36:29-42.

Page 101 of 121

126 Annex 116

Ellis JM, Griffin JL, Linscombe SD, Webster ER. 2003. Rice (Oryza sativa) and corn

(Zea mays) response to simulated drift of glyphosate and glufosinate. Weed
Technology 17:452-460.

Eschenburg S, Kabsch W, Healy ML, Schonbrunn E. 2003. A new view of the

mechanisms of UDP-N-acetylglucosamine enolpyruvyl transferase (MurA) and 5-
enolpyruvylshikimate-3- phosphate synthase (AroA) derived from X-ray
structures of their tetrahedral reaction intermediate states. Journal of Biological
Chemistry 278:49215-49222.

Eschholz WE, Servello FA, Griffith B, Raymond KS, Krohn WB. 1996. Winter use of
glyphosate-treated clearcuts by moose in Maine. Journal of Wildlife Management
60:764-769.

Everett KDE, Dickerson HW. 2003. Ichthyophthirius multifiliis and Tetrahymena

thermophila tolerate glyphosate but not a commercial herbicidal formulation.
Bulletin of Environmental Contamination and Toxicology 70:731-738.

Fairchild JF, Allert AL, Riddle JS, Gladwin DR. 2002. Efficacy of glyphosate and five

surfactants for controlling giant Salvinia. Journal Of Aquatic Plant Management
40:53-58.

Feng JC, Thompson DG. 1990. Fate of glyphosate in in a Canadian forest watershed. 2
Persistence in foliage and soils. Journal of Agricultural and Food Chemistry

38:1118-1125.

Feng PCC, Chiu T, Sammons RD, Ryerse JS. 2003. Droplet size affects glyphosate
retention, absorption, and translocation in corn. Weed Science 51:443-448.

Ferreira JFS, Smeda RJ, Duke SO. 1997. Control of coca plants (Erythroxylum coca
and E. novogranatense) with glyphosate. Weed Science 45:551-556.

Ferreira JFS, Reddy KN. 2000. Absorption and translocation of glyphosate in

Erythroxylum coca and E. novogranatense. Weed Science 48:193-199.

Fomsgaard IS, Spliid NH, Felding G. 2003. Leaching of pesticides through normal-
tillage and low-tillage soil--a lysimeter study. II. Glyphosate. Journal of
Environmental Sciences and Health B 38:19-35.

Frank R, Campbell RA, Sirons GJ. 1985. Forestry workers involved in aerial application
of 2,4-dichlorophenoxyacetic acid (2,4-D): Exposure and urinary excretion.
Archives of Environmental Contamination and Toxicology 14:427-435.

Garry VF, Harkins ME, Erickson LL, Long-Simpson LK, Holland SE, Burroughs BL.
2002. Birth defects, season of conception, and sex of children born to pesticide

Page 102 of 121

127Annex 116

applicators living in the Red River Valley of Minnesota, USA. Environmental

Health Perspectives 110 Suppl 3:441-449.

Gauriguata MR, Ostertag R. 2001. Neotropical secondary forest succession: changes in
structural and functional characteristics. Forest Ecology and Management

148:185-206.

Giesy JP, Dobson S, Solomon KR. 2000. Ecotoxicological risk assessment for
Roundup® herbicide. Reviews of Environmental Contamination and Toxicology
167:35-120.

Goldstein DA, Acquavella JF, Mannion RM, Farmer DR. 2002. An analysis of
glyphosate data from the California Environmental Protection Agency Pesticide
Illness Surveillance Program. Journal of Clinical Toxicology 40:885-892.

Goman M, Byrne R. 1998. A 5000-year record of agriculture and tropical forest
clearance in the Tuxtlas, Veracruz, Mexico. Holocene 8:83-89.

Gomez MA, Sagardoy MA. 1985. Effect of glyphosate herbicide on the microflora and

mesofauna of a sandy soil in a semiarid region. Revista Latinoamericana de
Microbiologia 27:351-357.

Gottlieb A. 1976. The pleasures of cocaine. Berkeley, CA, USA: And/Or Press.

Granby K, Vahl M. 2001. Investigation of the herbicide glyphosate and the plant growth
regulators chlormequat and mepiquat in cereals produced in Denmark. Food
Additives and Contamination 18:898-905.

Haefs R, Schmitz-Eiberger M, Mainx HG, Mittelstaedt W, Noga G. 2002. Studies on a

new group of biodegradable surfactants for glyphosate. Pest Management
Science 58:825-833.

Haney RL, Senseman SA, Hons FM. 2002. Effect of Roundup ultra on microbial activity

and biomass from selected soils. Journal of Environmental Quality 31:730-735.

Hardell L, Eriksson M. 1999. A case-control study of non-hodgkin lymphoma and
exposure to pesticides. Cancer 85:1353-1360.

Hardell L, Eriksson M, Nordstrom M. 2002. Exposure to pesticides as risk factor for non-
Hodgkin's lymphoma and hairy cell leukemia: pooled analysis of two Swedish
case-control studies. Leukemia and Lymphoma 43:1043-1049.

Harris SA, Sass-Kortsak AM, Corey PN, Purdham J. 2002. Development of models to

predict dose of pesticides in professional turf applicators. Journal of Exposure
Analysis and Environmental Epidemiology 12:130-144.

Page 103 of 121

128 Annex 116

Hartman KM, McCarthy BC. 2004. Restoration of a forest understory after the removal

of an invasive shrub, Amur honeysuckle (Lonicera maackii). Restoration Ecology
12:154-165.

Harwell MA, Cooper W, Flaak R. 1992. Prioritizing ecological and human welfare risks

from environmental stresses. Environmental Management 16:451-464.

Haughton AJ, Bell JR, Wilcox A, Boatman ND. 2001. The effect of the herbicide
glyphosate on non-target spiders: Part I. Direct effects on Lepthyphantes tenuis
under laboratory conditions. Pest Management Science 57:1033-1036.

Helling CS. 2003. Eradication of coca in Colombia -2002. Results of the Colombia coca
verification mission #9, December 2002. Beltsville, MD, USA: US Department of
Agriculture. 289 p.

Hernandez A, Garcia-Plazaola JI, Becerril JM. 1999. Glyphosate effects on phenolic
metabolism of nodulated soybean (Glycine max L. merr.). Journal of Agricultural
and Food Chemistry 47:2920-2925.

Hosmer DW, Lemeshow S. 1989. Applied Logistic Regression. New York, NY, USA:
Wiley. 307 p.

Howe CM, Berrill M, Pauli BD, Helbing CC, Werry K, Veldhoen N. 2004. Toxicity of
glyphosate-based pesticides to four North American frog species. Environmental

Toxicology and Chemistry 23:1928-1938.

Huang X, Pedersen T, Fischer M, White R, Young TM. 2004. Herbicide runoff along
highways. 1. Field observations. Environmental Science and Technology 38.

Hurtig AK, Sebastian MS, Soto A, Shingre A, Zambrano D, Guerrero W. 2003. Pesticide
use among farmers in the Amazon Basin of Ecuador. Archives of Environmental
Health 58:223-228.

ICA. 2003. Comercialización de Plaguicidas 2000-2001. Bogota, Colombia: Instituto
Colombiano Agropecuario. 124 p.

Idrovo J, Sanin LH, Cole D, Chavarro J, Caceres H, Narváez J, Restrepo M. 2005. Time
to first pregnancy among women working in agricultural production. International

Archives of Occupational and Environmental Health In Press.

Immunopharmos. 2002a. Toxicidad oral aguda-DL50 con Glifosato 44% + Cosmoflux
1% + agua 55%. Cota, Cundinamarca, Colombia: Immunopharmos Ltda.
Laboratorios.

Page 104 of 121

129Annex 116

Immunopharmos. 2002b. Toxicidad oral aguda-DL50 con Glifosato 5% + Cosmoflux 1%

+ agua 94%. Cota, Cundinamarca, Colombia: Immunopharmos Ltda.
Laboratorios.

Immunopharmos. 2002c. Toxicidad inhalatoria aguda-CL50 con Glifosato 44% +

Cosmoflux 1% + agua 55%. Cota, Cundinamarca, Colombia: Immunopharmos
Ltda. Laboratorios.

Immunopharmos. 2002d. Toxicidad inhalatoria aguda-CL50 con Glifosato 5% +
Cosmoflux 1% + agua 94%. Cota, Cundinamarca, Colombia: Immunopharmos

Ltda. Laboratorios.

Immunopharmos. 2002e. Irritación ocular primaria con Glifosato 44% + Cosmoflux 1% +
agua 55%. Cota, Cundinamarca, Colombia: Immunopharmos Ltda. Laboratorios.

Immunopharmos. 2002f. Irritación ocular primaria con Glifosato 5% + Cosmoflux 1% +
agua 94%. Cota, Cundinamarca, Colombia: Immunopharmos Ltda. Laboratorios.

Immunopharmos. 2002g. Irritación dérmica primaria con Glifosato 44% + Cosmoflux 1%

+ agua 55%. Cota, Cundinamarca, Colombia: Immunopharmos Ltda.
Laboratorios.

Immunopharmos. 2002h. Irritación dérmica primaria con Glifosato 5% + Cosmoflux 1%
+ agua 94%. Cota, Cundinamarca, Colombia: Immunopharmos Ltda.

Laboratorios.

Immunopharmos. 2002i. Sensibilidad cutánea en cobayos con Glifosato 5% +
Cosmoflux 1% + agua 94%. Cota, Cundinamarca, Colombia: Immunopharmos
Ltda. Laboratorios.

Immunopharmos. 2002j. Sensibilidad cutánea en cobayos con Glifosato 44% +
Cosmoflux 1% + agua 55%. Cota, Cundinamarca, Colombia: Immunopharmos
Ltda. Laboratorios.

Jackson RE, Pitre HN. 2004a. Influence of Roundup Ready soybean production
systems and glyphosate application on pest and beneficial insects in narrow-row
soybean. Journal of Entomological Science 39:62-70.

Jackson RE, Pitre HN. 2004b. Influence of Roundup Ready soybean and roundup ultra
herbicide on Geocoris punctipes (Say) (Heteroptera: Lygaeidae) in the
laboratory. Journal of Entomological Science 39:56-61.

Jiraungkoorskul W, Upatham ES, Kruatrachue M, Sahaphong S, Vichasri-Grams S,

Pokethitiyook P. 2003. Biochemical and histopathological effects of glyphosate
herbicide on Nile tilapia (Oreochromis niloticus). Environmental Toxicology
18:260-267.

Page 105 of 121

130 Annex 116

Joffe M, Villard L, Li Z, Plowman R, Vessey M. 1995. A time to pregnancy questionnaire
designed for long term recall: validity in Oxford, England. Journal of
Epidemiology and Community Health 49:314-319.

Joffe M. 1997. Time to pregnancy: A measure of reproductive function in either sex.
Occupational and Environmental Medicine 54:289-295.

Johnson HE, Hazen JL, Penner D. 2002. Citric ester surfactants as adjuvants with
herbicides. Weed Technology 16:867-872.

Kimball AJ, Hunter ML. 1990. Intensive Silviculture. In: Hunter M, L, editor. Wildlife,
Forests and Forestry. Englewood Cliffs, NJ, USA: Prentice Hall. p 200-234.

Kjaer J, Ullum M, Olsen P, Sjelborg P, Helweg A, Bügel Mogensen B, Plauborg F, Grant

R, Fomsgaard IS, Brüsch W. 2003. The Danish Pesticide Leaching Assessment
Programme. Monitoring Results May 1999 – June 2002. 3rd Report.
Copenhagen, Denmark: Geological Survey of Denmark and Greenland. 158 p.

Kjaer J, Olsen P, Ullum M, Grant R. 2005. Leaching of glyphosate and amino-
methylphosphonic acid from Danish agricultural field sites. Journal of
Environmental Quality 34:609-620.

Krieger RI, editor. 2001. Handbook of Pesticide Toxicology. 2nd ed. New York, NY,

USA: Academic Press. 1908 p.

Krueger-Mangold J, Sheley RL, Roos BD. 2002. Maintaining plant community diversity
in a waterfowl production area by controlling Canada thistle (Cirsium arvense)
using glyphosate. Weed Technology 16:457-463.

Kudsk P, Mathiassen SK. 2004. Joint action of amino acid biosynthesis-inhibiting
herbicides. Weed Research 44:313-322.

Laatikainen T, Heinonen-Tanski H. 2002. Mycorrhizal growth in pure cultures in the
presence of pesticides. Microbiological Research 157:127-137.

Lautenschlager RA, Sullivan TP. 2002. Effects of herbicide treatments on biotic
components in regenerating northern forests. Forestry Chronicle 78:695-731.

Lee HL, Chen KW, Chi CH, Huang JJ, Tsai LM. 2000. Clinical presentations and
prognostic factors of a glyphosate-surfactant herbicide intoxication: a review of
131 cases. Academic Emergency Medicine 7:906-910.

Lin N, Garry VF. 2000. In vitro studies of cellular and molecular developmental toxicity
of adjuvants, herbicides, and fungicides commonly used in Red River Valley,
Minnesota. Journal of Toxicology and Environmental Health A 60:423-439.

Page 106 of 121

131Annex 116

Lindgren PMF, Sullivan TP. 2001. Influence of alternative vegetation management
treatments on conifer plantation attributes: abundance, species diversity, and
structural diversity. Forest Ecology and Management 142:163-182.

Lindsay EA, French K. 2004. The impact of the herbicide glyphosate on leaf litter
invertebrates within Bitou bush, Chrysanthemoides monilifera ssp rotundata,
infestations. Pest Management Science 60:1205-1212.

Liu C-M, McLean PA, Sookdeo CC, Cannon FC. 1991. Degradation of the herbicide

glyphosate by memebrs of family Rhizobiaceae. Applied Environmental
Microbiology 57:1799-1840.

Liu ZQ. 2004. Effects of surfactants on foliar uptake of herbicides - a complex scenario.
Colloids and Surfaces B-Biointerfaces 35:149-153.

[LSD] 2005. Trace Organic Analyses. LSD website
http://www.uoguelph.ca/labserv/Services/services.html, Accessed January 10
2005.

Maltby L, Blake NN, Brock TCM, Van den Brink PJ. 2005. Insecticide species sensitivity
distributions: the importance of test species selection and relevance to aquatic
ecosystems. Environmental Toxicology and Chemistry Accepted.

Mann RM, Bidwell JR. 1999. The toxicity of glyphosate and several glyphosate
formulations to four species of Southwestern Australian frogs. Archives of
Environmental Contamination and Toxicology 36:193-199.

Manthey FA, Nalewaja JD. 1992. Relative wax solubility and phytotoxicity of oil to green

foxtail [Setaria viridis (L.) Beauv.]. In: Foy CL, editor. Adjuvants for
Agrichemicals. Boca Raton, FL, USA: CRC Press. p 463-471.

Mårtensson AM. 1992. Effects of agrochemicals and heavy metals on fast-growing

Rhizobia and their symbiosis in small-seeded legumes. Soil Biology and
Biochemistry 25:435-445.

Martins SV, Rodrigues RR. 2002. Gap-phase regeneration in a semideciduous
mesophytic forest, south-eastern Brazil. Plant Ecology 163:51-62.

Martins SV, Junior RC, Rodrigues RR, Gandolfi S. 2004. Colonization of gaps produced
by death of bamboo clumps in a semideciduous mesophytic forest in south-
eastern Brazil. Plant Ecology 172:121-131.

Martinson KB, Sothern RB, Koukkari WL, Durgan BR, Gunsolus JL. 2002. Circadian
response of annual weeds to glyphosate and glufosinate. Chronobiology
International 19:405-422.

Page 107 of 121

132 Annex 116

Matarczyk JA, Willis AJ, Vranjic JA, Ash JE. 2002. Herbicides, weeds and endangered
species: management of bitou bush (Chrysanthemoides monilifera ssp
rotundata) with glyphosate and impacts on the endangered shrub, Pimelea
spicata. Biological Conservation 108:133-141.

Matlack GR. 1997. Four centuries of forest clearance and regeneration in the hinterland
of a large city. Journal of Biogeography 24:281-295.

May WE, Loeppky HA, Murrell DC, Myhre CD, Soroka JJ. 2003. Preharvest glyphosate

in alfalfa for seed production: Effect on alfalfa seed yield and quality. Canadian
Journal of Plant Science 83:189-197.

McAlister JJ, Smith BJ, Sanchez B. 1998. Forest clearance: Impact of landuse change
on fertility status of soils from the Sao Francisco area of Niteroi, Brazil. Land

Degradation & Development 9:425-440.

McDonald MA, Healey JR, Stevens PA. 2002. The effects of secondary forest clearance
and subsequent land- use on erosion losses and soil properties in the Blue

Mountains of Jamaica. Agriculture Ecosystems & Environment 92:1-19.

[Microsoft Corporation] Microsoft Corporation. 2003. Microsoft Office Excel 2003®
[computer program]. Redmond, WA, USA

Miller RP, Martinson KB, Sothern RB, Durgan BR, Gunsolus JL. 2003. Circadian
response of annual weeds in a natural setting to high and low application rates of
four herbicides with different modes of action. Chronobiology International
20:299-324.

Morjan WE, Pedigo LP. 2002. Suitability of transgenic glyphosate-resistant soybeans to
green cloverworm (Lepidoptera: Noctuidae). Journal of Economic Entomology
95:1275-1280.

Motavalli PP, Kremer RJ, Fang M, Means NE. 2004. Impact of genetically modified
crops and their management on soil microbially mediated plant nutrient
transformations. Journal of Environmental Quality 33:816-824.

Mueller TC, Massey JH, Hayes RM, Main CL, Stewart CN. 2003. Shikimate

accumulates in both glyphosate-sensitive and glyphosate-resistant horseweed
(Conyza canadensis L. Cronq.). Journal of Agricultural and Food Chemistry
51:680-684.

Nakashima K, Yoshimura T, Mori H, Kawaguchi M, Adachi S, Nakao T, Yamazaki F.

2002. Effects of pesticides on cytokines production by human peripheral blood
mononuclear cells - fenitrothion and glyphosate. Chudoku Kenkyu 15:159-165
(Abstract only, original in Japanese).

Page 108 of 121

133Annex 116

Neve P, Diggle AJ, Smith FP, Powles SB. 2003a. Simulating evolution of glyphosate
resistance in Lolium rigidum II: past, present and future glyphosate use in
Australian cropping. Weed Research 43:418-427.

Neve P, Diggle AJ, Smith FP, Powles SB. 2003b. Simulating evolution of glyphosate
resistance in Lolium rigidum I: population biology of a rare resistance trait. Weed
Research 43:404-417.

Newmaster SG, Bell FW. 2002. The effects of silvicultural disturbances on cryptogam

diversity in the boreal-mixedwood forest. Canadian Journal Of Forest Research-
Revue Canadienne De Recherche Forestiere 32:38-51.

Nijman V. 1998. Habitat preference of Great Argus pheasant (Argusianus argus) in
Kayan Mentarang National Park, east Kalimantan, Indonesia. Journal Fur

Ornithologie 139:313-323.

Norsworthy JK. 2004. Conventional soybean plant and progeny response to glyphosate.
Weed Technology 18:527-531.

[NRA] National Registration Authority for Agricultural and Veterinary Chemicals. 1996.
NRA Special Review of Glyphosate. Canberra, Australia: NRA.

NRC. 1986. Ecological Knowledge and Environmental Problem-Solving: Concepts and

Case Studies. Committee on the Applications of Ecological Theory to
Environmental Problems, editor. Washington, DC, USA: National Academy
Press,. 388 p.

NRC. 1993. Issues in Risk Assessment. Washington, DC, USA: National Academy

Press.

OECD. 1984a. OECD Guidelines for testing of chemicals. Method 202. Daphnia
species, acute immobilisation test and reproduction test. Paris, France:

Organisation for Economic Co-Operation and Development.

OECD. 1984b. OECD Guidelines for testing of chemicals. Method 201. Alga, growth
inhibition test. Paris, France: Organisation for Economic Co-Operation and
Development.

OECD. 1992. OECD Guidelines for testing of chemicals. Method 203. Fish acute toxicity
test. Paris, France: Organisation for Economic Co-Operation and Development.

OECD. 1998a. OECD Guidelines for testing of chemicals. Method #214. Honeybees,

acute contact toxicity test. Paris, France: Organisation for Economic Co-
Operation and Development.

Page 109 of 121

134 Annex 116

OECD. 1998b. The OECD Principles of Good Laboratory Practice. Paris, France:

Organisation for Economic Co-Operation and Development. Environment
Monograph No. 45.

Okwakol MJN. 2000. Changes in termite (Isoptera) communities due to the clearance

and cultivation of tropical forest in Uganda. African Journal Of Ecology 38:1-7.

Pariona W, Fredericksen TS, Licona JC. 2003. Natural regeneration and liberation of
timber species in logging gaps in two Bolivian tropical forests. Forest Ecology
and Management 181:313-322.

Patzoldt WL, Tranel PJ, Hager AG. 2002. Variable herbicide responses among Illinois
waterhemp (Amaranthus rudis and A. tuberculatus) populations. Crop Protection
21:707-712.

Payne NJ, Feng JC, Reynolds PE. 1990. Off-target deposits and buffer zones required
around water for aerial glyphosate applications. Pesticide Science 30:183-198.

Payne NJ. 1993. Spray dispersal from aerial silvicultural glyphosate applications. Crop

Production 12:463-469.

Pena-Claros M. 2003. Changes in forest structure and species composition during
secondary forest succession in the Bolivian Amazon. Biotropica 35:450-461.

Perez A, Kogan M. 2003. Glyphosate-resistant Lolium multiflorum in Chilean orchard.
Weed Research 43:12-19.

Pline WA, Edmisten KL, Wilcut JW, Wells R, Thomas J. 2003. Glyphosate-induced
reductions in pollen viability and seed set in glyphosate-resistant cotton and

attempted remediation by gibberellic acid (GA(3)). Weed Science 51:19-27.

2005. Maps showing areas in Colombia where coca and poppy are grown. Bogota, D.C.
Colombia: Direccion Nacional de Estupefacientes.

Popper KR. 1979. Truth, rationality and the growth of scientific knowledge. Frankfurt am
Main: V. Klostermann. 61 p.

Post D. 1999. Glyphosate - Actions and Reactions, July 29, 1999. Chevy Chase,

Maryland, USA: Rachel Carson Council, Inc. Brochure. 4 p.

PTG. 2005a. Informe Valle de Cauca. Bogota, Colombia: PGT.

PTG. 2005b. Informe Boyacá. Bogota, Colombia: PGT.

PTG. 2005c. Informe Sierra Nevada. Bogota, Colombia: PGT.

Page 110 of 121

135Annex 116

PTG. 2005d. Informe Nariño. Bogota, Colombia: PGT.

PTG. 2005e. Informe Putumayo. Bogota, Colombia: PGT.

Pushnoy LA, Avnon LS, Carel RS. 1998. Herbicide (Roundup) pneumonitis. Chest

114:1769-1771.

Quaghebeur D, De Smet B, De Wulf E, Steurbaut W. 2004. Pesticides in rainwater in
Flanders, Belgium: results from the monitoring program 1997-2001. Journal of
Environmental Montitoring 6:182-190.

Racke KD, Skidmore MW, Hamilton DJ, Unsworth JB, Miyamoto J, Cohen SZ. 1997.
Pesticide fate in tropical soils. Pure and Applied Chemistry 69:1349-1371.

Ramsdale BK, Messersmith CG, Nalewaja JD. 2003. Spray volume, formulation,

ammonium sulfate, and nozzle effects on glyphosate efficacy. Weed Technology
17:589-598.

Ramwell CT, Heather AI, Shepherd AJ. 2002. Herbicide loss following application to a

roadside. Pest Management Science 58:695-701.

Ramwell CT, Heather AI, Shepherd AJ. 2004. Herbicide loss following application to a
railway. Pest Management Science 60:556-564.

Raymond KS, Servello FA, Griffith B, Eschholz WE. 1996. Winter foraging ecology of
moose on glyphosate-treated clearcuts in Maine. Journal of Wildlife Management
60:753-763.

Reeves BG. 1992. The rationale for adjuvant use with agrichemicals. In: Foy CL, editor.

Adjuvants for Agrichemicals. Boca Raton, FL, USA: CRC Press. p 487-488.

Relyea RA. 2004. Growth and survival of five amphibian species exposed to
combinations of pesticides. Environmental Toxicology and Chemistry 23:1737-

1742.

Relyea RA. 2005. The impact of insecticides and herbicides on the biodiversity and
productivity of aquatic communities. Ecological Applications 15:618-627.

Renz MJ, DiTomaso JM. 2004. Mechanism for the enhanced effect of mowing followed
by glyphosate application to resprouts of perennial pepperweed (Lepidium
latifolium). Weed Science 52:14-23.

Riley CM, Weisner CJ, Sexsmith WA. 1991. Estimating off-target spray deposition on

the ground following aerial application of glyphosate for conifer release in New
Brunswick. Journal of Environmental Science and Health B 26:185-208.

Page 111 of 121

136 Annex 116

Roberts CW, Roberts F, Lyons RE, Kirisits MJ, Mui EJ, Finnerty J, Johnson JJ,

Ferguson DJ, Coggins JR, Krell T, Coombs GH, Milhous WK, Kyle DE, Tzipori S,
Barnwell J, Dame JB, Carlton J, McLeod R. 2002. The shikimate pathway and its
branches in apicomplexan parasites. Journal of Infectious Diseases 185 Suppl
1:S25-36.

Roberts F, Roberts CW, Johnson JJ, Kyle DE, Krell T, Coggins JR, Coombs GH,
Milhous WK, Tzipori S, Ferguson DJ, Chakrabarti D, McLeod R. 1998. Evidence
for the shikimate pathway in apicomplexan parasites. Nature 393:801-805.

Rodwell DE. 1980a. Teratology study in rats with technical glyphosate. Mattawan, MI,
USA: International Research and Development Corporation. 79/016. Cited in
Pesticide Residues in Food 1986, Evaluations 1986 Part II Toxicology:
Sponsored jointly by FAO and WHO, 1980.

Rodwell DE. 1980b. Teratology study in rabbits with technical glyphosate. Mattawan,
MI, USA: International Research and Development Corporation. 79/018. Cited in
Pesticide Residues in Food 1986, Evaluations 1986 Part II Toxicology:
Sponsored jointly by FAO and WHO, 1980.

Rueppel ML, Brightwell BB, Schaefer J, Marvel JT. 1977. Metabolism and degradation
of glyphosate in soil and water. Journal of Agricultural and Food Chemistry
25:517-528.

Sanin L-H. 2005. Analysis of Time to Preganancy in Five Regions of Colombia, 2005.
Chihuahua, Mexico: Autonomous University of Chihuahua and National Institute
of Public Health, Mexico.

Santillo DJ, Brown PW, Leslie DM. 1989a. Response of songbirds to glyphosate-

induced habitat changes on clearcuts. Journal of Wildlife Management 53:64-71.

Santillo DJ, Leslie DM, Brown PW. 1989b. Responses of small mammals and habitat to
glyphosate application on clearcuts. Journal of Wildlife Management 53:164-172.

Sarmiento L, Llambi LD, Escalona A, Marquez N. 2003. Vegetation patterns,
regeneration rates and divergence in an old-field succession of the high tropical
Andes. Plant Ecology 166:63-74.

Schaffer JD, Sebetich MJ. 2004. Effects of aquatic herbicides on primary productivity of
phytoplankton in the laboratory. Bulletin of Environmental Contamination and
Toxicology 72:1032-1037.

Scribner EA, Battaglin WA, Dietze JE, Thurman EM. 2003. Reconnaissance Data for

Glyphosate, Other Selected Herbicides, Their Degradation Products, and
Antibiotics in 51 Streams in Nine Midwestern States, 2002. Lawrence, Kansas,
USA: US Geological Survey. Open-File Report. 03–217. 109 p.

Page 112 of 121

137Annex 116

Skark C, Zullei-Seibert N, Schöttler U, Schlett C. 1998. The occurrence of glyphosate in
surface water. International Journal of Environmental and Analytical Chemistry
70:93–104.

Skark C, Zullei-Seibert N, Willme U, Gatzemann U, Schlett C. 2004. Contribution of
non-agricultural pesticides to pesticide load in surface water. Pest Management
Science 60:525-530.

Smith EA, Oehme FW. 1992. The biological activity of glyphosate to plants and animals:

A literature review. Veterinary and Human Toxicology 34:531-543.

Solomon KR, Takacs P. 2002. Probabilistic risk assessment using species sensitivity
distributions. In: Posthuma L, Traas T, Suter GW, editors. Species Sensitivity
Distributions in Risk Assessment. Boca Raton, FL, USA: CRC Press. p 285-313.

Solomon KR, Thompson DG. 2003. Ecological risk assessment for aquatic organisms
from over-water uses of glyphosate. Journal of Toxicology and Environmental
Health B 6:211-246.

Solomon KR, Houghton D, Harris SA. 2005. Nonagricultural and residential exposures
to pesticides. Scandinavian Journal of Work and Environmental Health 31 suppl
1:In Press.

Springborn. 2003a. Purity analysis for glyphosate of Spray-Charlie (active ingredient).
Spencerville, Ohio, USA: Springborn Laboratories. 3596.18. 52 p.

Springborn. 2003b. An acute oral toxicity study in rats with Spray--Charlie. Spencerville,
Ohio, USA: Springborn Laboratories. 3596.16. 29 p.

Springborn. 2003c. An acute nose-only inhalation toxicity study in rats with Spray-
Charlie. Spencerville, Ohio, USA: Springborn Laboratories. 3596.18. 52 p.

Springborn. 2003d. An acute dermal toxicity study in rats with Spray-Charlie.
Spencerville, Ohio, USA: Springborn Laboratories. 3596.17. 29 p.

Springborn. 2003e. An primary skin irritant study in rabbits with Spray-Charlie.
Spencerville, Ohio, USA: Springborn Laboratories. 3596.20. 23 p.

Springborn. 2003f. A primary eye irritation study in rabbits with Spray-Charlie3596.19.
Spencerville, Ohio, USA: Springborn Laboratories. 3596.19. 24 p.

Springborn. 2003g. A dermal sensitization study in guinea-pigs with Spray-Charlie.

Modified Buheler design. Spencerville, Ohio, USA: Springborn Laboratories.
3596.21. 40 p.

Page 113 of 121

138 Annex 116

Stantec. 2005a. Mixture of Glyphosate and Cosmo-Flux® 411F: Acute Contact Toxicity

to Honey Bee (Apis mellifera). Guelph, ON, Canada: Stantec. 132 p.

Stantec. 2005b. Mixture of Glyphosate and Cosmo-Flux® 411F: Growth Inhibition Test
With the Freshwater Green Algae Selanastrum capricornutum. Guelph, ON,

Canada: Stantec. 186 p.

Stantec. 2005c. Mixture of Glyphosate and Cosmo-Flux® 411F: Acute Toxicity to
Rainbow Trout OECD 203. Guelph, ON, Canada: Stantec. 167 p.

Stantec. 2005d. Mixture of Glyphosate and Cosmo-Flux® 411F: Acute Toxicity to
Fathead Minnow OECD 203. Guelph, ON, Canada: Stantec. 186 p.

Stantec. 2005e. Mixture of Glyphosate and Cosmo-Flux® 411F: Acute Toxicity to
(Daphnia magna). Guelph, ON, Canada: Stantec. 174 p.

Stiltanen H, Rosenberg C, Raatikainen M, Raatikainen T. 1981. Triclopyr, glyphosate
and phenoxyherbicide residues in cowberries, bilberries and lichen. Bulletin of
Environmental Contamination and Toxicology 27:731-737.

Strange-Hansen R, Holm PE, Jacobsen OS, Jacobsen CS. 2004. Sorption,
mineralization and mobility of N-(phosphonomethyl)glycine (glyphosate) in five
different types of gravel. Pest Management Science 60:570-578.

Sullivan TP. 1994. Influence of herbicide-induced habitat alteration on vegetation and
snowshoe hare populations in sub-boreal spruce forest. Journal of Applied
Ecology 31:717-730.

Sullivan TP, Lautenschlager RA, Wagner RG. 1996. Influence of glyphosate on

vegetation dynamics in different successional stages of sub-boreal spruce forest.
Weed Technology 10:439-446.

Sullivan TP, Nowotny C, Lautenschlager RA, Wagner RG. 1998. Silvicultural use of

herbicide in sub-boreal spruce forest: Implications for small mammal population
dynamics. Journal of Wildlife Management 62:1196-1206.

Sullivan TP, Sullivan DS. 2003. Vegetation management and ecosystem disturbance:
impact of glyphosate herbicide on plant and animal diversity in terrestrial

systems. Environmental Reviews 11:37-59.

Tadros TF. 1994. Surfactants in Agrochemicals. Schrick MJ, Fowkes FM, editors. New
York, NY: Marcel Dekker, Incorporated. 264 p.

Tate TM, Jackson RN, Christian FA. 2000. Effects of glyphosate and dalpon on total
free amino acids profiles of Pseudosuccinae columella snails. Bulletin of
Environmental Contamination and Toxicology 64:258-262.

Page 114 of 121

139Annex 116

Tenuta M, Beuchamp EG. 1995. Denitrification following herbicide application to a grass
sward. Canadian Journal of Soil Science 76:15-22.

Terech-Majewska E, Siwicki AK, Szweda W. 2004. Modulative influence of lysozyme

dimer on defence mechanisms in the carp (Cyprinus carpio) and European
sheatfish (Silurus glanis) after suppression induced by herbicide Roundup. Polish
Journal of Veterinary Science 7:123-128.

Thomas WE, Pline-Srnic WA, Thomas JF, Edmisten KL, Wells R, Wilcut JW. 2004.

Glyphosate negatively affects pollen viability but not pollination and seed set in
glyphosate-resistant corn. Weed Science 52:725-734.

Thompson DG, Pitt DG, Staznik B, Payne NJ, Jaipersaid D, Lautenschlager RA, Bell
FW. 1997. On-target deposit and vertical distribution of aerially released

herbicides. Forestry Chronicle 73:47-59.

Thompson DG, Wojtaszek BF, Staznik B, Chartrand DT, Stephenson GR. 2004.
Chemical and biomonitoring to assess potential acute effects of Vision herbicide

on native amphibian larvae in forest wetlands. Environmental Toxicology and
Chemistry 23:843-849.

Tooby TE. 1985. Fate and biological consequences of glyphosate in the aquatic
environment. In: Gossbard E, Atkinson D, editors. The Herbicide Glyphosa
te.

London, GB: Butterworth and Co. p 206-217.

Tsui MTK, Chu LM. 2003. Aquatic toxicity of glyphosate-based formulations:
Comparison between different organisms and the effects of environmental
factors. Chemosphere 52:1189-1197.

Tsui MTK, Chu LM. 2004. Comparative toxicity of glyphosate-ased herbicides: Aqueous
and sediment porewater exposures. Archives of Environmental Contamination
and Toxicology 46:316-323.

[U.S. Department of State] United States Department of State. 2002. Aerial Eradication
of Illicit Coca in Colombia Report on Issues Related to the Aerial Eradication of
Illicit Coca in Colombia. U.S. Department of State website
http://www.state.gov/g/inl/rls/rpt/aeicc/c7470.htm, Accessed April 2005.

United Nations. 2002. Global Illicit Drug Trends. New York, NY, USA: Drug Control and
Crime Prevention Office. ??? p.

Urban DJ, Cook NJ. 1986. Standard Evaluation Procedure for Ecological Risk

Assessment. Washington, DC.: Hazard Evaluation Division, Office of Pesticide
Programs, U.S. Environmental Protection Agency. EPA/540/09-86/167.

Page 115 of 121

140 Annex 116

[USEPA] US Environmental Protection Agency. 1992. Framework for ecological risk

assessment. Washington, DC, USA: USEPA.

USEPA. 1993a. R.E.D. Facts Glyphosate. Washington, DC, USA: U.S. Environmental
Protection Agency. EPA 738-R-93-014.

USEPA. 1993b. Reregistration Eligibility Decision (RED). Washington, DC, USA: U.S.
Environmental Protection Agency. EPA 738-R-93-014. 74 p.

[USEPA] U.S. Environmental Protection Agency. 1996a. Ecological effects test

guidelines. OPPTS 850.3020 Honey bee acute contact toxicity. Washington, DC,
USA: USEPA.

[USEPA] U.S. Environmental Protection Agency. 1996b. Ecological Effects Test
Guidelines. OPPTS 850.5400 Algal Toxicity, Tiers I and II. Washington, DC,

USA: USEPA.

USEPA. 1997. Glyphosate: pesticide tolerances for emergency exemptions. Federal
Register 62:42921-42928.

[USEPA] U.S. Environmental Protection Agency. 1998. Guidelines for Ecological Risk
Assessment. Washington, DC, USA: USEPA.

USEPA. 1999. Glyphosate; Pesticide Tolerance. Federal Register 64:18360-18367.

[USEPA] U.S. Environmental Protection Agency, Office of Pesticide Programs,
Environmental Fate and Effects Division, U.S. EPA, Washington, D.C. 2001.
Environmental Effects Database (EEDB). ECOTOX Database System. USEPA
website http://www.epa.gov/ecotox/, Accessed February 23 2005.

Veiga F, Zapata JM, Fernandez Marcos ML, Alvarez E. 2001. Dynamics of glyphosate
and aminomethylphosphonic acid in a forest soil in Galicia, north-west Spain.
Science of the Total Environment 271:135-144.

Vellend M. 2003. Habitat loss inhibits recovery of plant diversity as forests regrow.
Ecology 84:1158-1164.

Verrell P, Van Buskirk E. 2004. As the worm turns: Eisenia fetida avoids soil

contaminated by a glyphosate-based herbicide. Bulletin of Environmental
Contamination and Toxicology 72:219-224.

Walsh LP, McCormick C, Martin C, Stocco DM. 2000. Roundup inhibits steroidogenesis
by disrupting steroidogenic acute regulatory (StAR) protein expression.

Environmental Health Perspectives 108:769-776.

Page 116 of 121

141Annex 116

Weinberg CR, Baird DD, Wilcox AJ. 1994. Sources of bias in studies of time to

pregnancy. Statistical Medicine 13:671-681.

Weinberg CR, Wilcox AJ. 1998. Reproductive epidemiology. In: Rothman KJ,
Greenland S, editors. Modern Epidemiology2 ed. Philadelphia, PA, USA:

Lippincott-Raven. p 585-608.

Williams GM, Kroes R, Munro IC. 2000. Safety evaluation and risk assessment of the
herbicide Roundup® and its active ingredient, glyphosate, for humans.
Regulatory Toxicology and Pharmacology 31:117-165.

Williams MR, Abbott I, Liddelow GL, Vellios C, Wheeler IB, Mellican AE. 2001.
Recovery of bird populations after clearfelling of tall open eucalypt forest in
Western Australia. Journal of Applied Ecology 38:910-920.

Wittmann F, Junk WJ, Piedade MTF. 2004. The varzea forests in Amazonia: flooding
and the highly dynamic geomorphology interact with natural forest succession.
Forest Ecology and Management 196:199-212.

Wojtaszek BF, Staznik B, Chartrand DT, Stephenson GR, Thompson DG. 2004. Effects
of Vision herbicide on mortality, avoidance response, and growth of amphibian
larvae in two forest wetlands. Environmental Toxicology and Chemistry 23:832-
842.

Wolfenbarger LL, Phifer PR. 2000. The ecological risks and benefits of genetically
engineered plants. Science 290:2088-2093.

World Health Organization International Program on Chemical Safety. 1994.
Glyphosate. Geneva. 161 p.

Xi YL, Feng LK. 2004. Effects of thiophanate-methyl and glyphosate on asexual and
sexual reproduction in the rotifer Brachionus calyciflorus Pallas. Bulletin of
Environmental Contamination and Toxicology 73:644-651.

Zhou H, Weinberg CR. 1999. Potential for bias in estimating human fecundability
parameters: a comparison of statistical models. Statistical Medicine 18:411-422.

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142 Annex 116

8 GLOSSARY
Absorption: The movement of a substance across an exposed surface (e.g., skin,
respiratory / digestive mucous) and into the circulation to be distributed
throughout the body. This will vary depending on a compound’s inherent ability

to cross a particular barrier.
AE - Acid Equivalent: The concentration of a substance (glyphosate) expressed in
terms of the amount of glyphosate acid, rather than the salt.

A.I. - Active Ingredient: The component of a mixture / formulation which is ultimately
responsible for the physiological effects.
Acute toxicity: The potential of a compound to cause injury or illness when given in a
single dose or in multiple doses over a short period of time (e.g. 24 h). These
effects are based on mechanisms of chemical action where perceptible

physiological alterations can be appreciated shortly after administration (e.g.
death).
ADI - Acceptable daily intake: This is an estimate of the maximum amount of a
compound (often in food) which can be ingested daily over a lifetime without any
appreciable detrimental health effects. This parameter has been developed

primarily by the WHO and FAO.
Adjuvant: Ingredient added to a particular formulation in order to enhance the
availability and efficacy of the active ingredient. These often act by increasing
the spreading or uptake of the active ingredient(s).

Adsorption: The process by which a compound is held or bound to a surface by
chemical or physical attraction.
Anthropogenic: Chemicals artificially developed by man.

Aromatic: Organic compound in which constituent atoms form a ring (s). These ring
structures may grant a compound its characteristic properties such as solubility in
lipids.
Bioaccumulation: The accumulation of a particular compound in certain body tissues.

This occurs when rate of uptake exceeds that of metabolism and/or excretion.
Over time this results in a higher concentration of the substance in the organism
than in its environment. Important factors governing the extent of this process
include the lipid solubility of the compound as well as how readily it is
metabolized.

Bioactivation: The process by which a chemical becomes more reactive due to
alterations in its structure and hence chemical properties. This can occur in the
environment or within a biological system.
Bioconcentration Factor (BCF): Measure of the tendency of a substance in water to
accumulate within the tissues of fish or other organisms. The concentration in

the organism can be roughly calculated by multiplying the concentration in the
water by the bioconcentration factor. The value determined is useful in helping to
determine the possible human consumption level.
CAS No.: Chemical Abstract System registry number. Pertains to a database providing

chemical substance information.

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Carcinogenic: Any chemical that can cause the formation of cancerous lesions. Often

this is achieved through the formation of genetic mutations within a cell(s)
resulting in the loss in ability to regulate proliferation.
Chlorosis: A disease in plants, causing the flowers to turn green or the leaves to lose

their normal green color.
Chronic toxicity: The nature of adverse effects over a prolonged period of chemical
exposure. Such effect measures can include the development of cancer or
decrease in growth.

Dermatitis: Inflammation of the skin.
Dose-response: The change in the intensity of physiological effect with dosage. The
relationship of response to dose will vary depending on the mechanism through

which the compound is acting.
EC50: Median effective concentration. The concentration of a substance in a medium
(such as water) which produces an defined effect in 50% of test organisms.

Ecosystem: A collection of populations (microorganisms, plants, and animals) that
occur in the same place at the same time and that can therefore potentially
interact with each other as well as their physical and chemical environment and
thus form a functional entity.

Emulsification: The mixture of two immiscible (non-mixable) liquids by the dispersion
of one into the other in the form of tiny droplets.
Environmental fate: The movement, accumulation, and disappearance of chemicals in
the environment after their release.

EPA: Environmental Protection Agency (and in the U.S. EPA).
Epidemiological study: The study of the distribution and determinants of health-
related states and events within populations. The prevalence of a particular

disease as well as various risk factors for its development are studied.
Exposure: Amount of a chemical which comes into contact with a body surface (skin,
respiratory tract, digestive tract) from which it can be absorbed into the body.
Exposure does not include any chemical that is nearby but not in contact or

which is intercepted by clothing or protective equipment.
Exposure route: The means by which a compound comes into contact an absorptive
interface such as dermal or inhalation.

Formulant: A substance normally added to a pesticide to increase its ease of use,
penetration into the target organism, or to facilitate its application.
Genotoxic: Describes any substance capable of damaging DNA resulting in mutations
or the development of cancer.

Half-life: The time for the concentration of a particular chemical or drug to decrease by
half of its initial concentration. This will vary depending on its rate of degradation,
metabolism, and/or elimination.

Hazard quotient: The ratio of exposure concentration to a reference (threshold) value.
If this value is above acceptable concentration, an adverse effect is possible.
Inert ingredients: All components of a mixture not classified as the primary active
ingredient. See, Formulant.

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144 Annex 116

Intraperitoneal: Within the peritoneal cavity, the area that contains the abdominal
organs.
Intravenous: The injection or entry of a substance directly into a vein and hence into

general circulation.
K OW (Log): The octanol-water partition coefficient OW ) is a ratio of the concentration of
the chemical in n-octanol and water at equilibrium. Chemicals with a K OW greater

than 1 preferentially partition into octanol. May be expressed as a l10. The
value obtained from this determination gives an indication of the potential for the
substance to bioconcentrate into organisms.

LC50 - Lethal Concentration 50: The concentration that is lethal to 50% of test
organisms. This value is usually used when referring to the toxicity of a
substance to organisms exposed via a matrix such as water.
LD50 – Lethal Dose 50: The dose that is lethal to 50% of test animals. This value is

used when referring to the toxicity of a substance to organisms that exposed to a
specific amount of substance such as via the oral or the injection route.
Leaching: The movement of a substance through the soil.

LOAEL - Lowest Observed Adverse Effect Level: The lowest dose of a toxin at which
an adverse effect can be noted in a particular test species. This value will vary
depending on the species being utilized.

Matrix: The medium through which an organism may be exposed to a substance.
Water for aquatic organisms, soil for soil organisms, air, etc.
Mechanism of action: The process by which a substance produces its characteristic

effects. It is often used interchangeably with “toxic mode of action” however it is
usually a more specific term. This is a description of the physiological processes
that are altered and the consequences of such changes.

Metabolite: A product of natural metabolic processes.
MRL-Maximum Residue Limit: The maximum amount of a substance permissible on
food products as well as animal feeds. This value is recommended by the Codex
Alimentarius Commission. This takes into account various safety factors as does

the ADI.
MTD-Maximum Tolerated Dose): The dose at which significant toxic effects occur
without resulting in death.

Mutagen: Any substance or agent that is capable of creating changes in DNA that are
subsequently passed on to future cells. These changes may sometimes lead to
the development of cancer or changes in organism characteristics.

NOAEL-No Observable Adverse Effects Level: The highest dose that results in no
adverse effects being noted in test organisms.
Oxidation: An alteration of chemical structure by the removal of an electron. This is

accomplished by any compound that is capable of achieving this (oxidant).
Percutaneous: Pertaining to any agent than can traverse or is administered through
the skin.

Persistence: The resistance of a substance to metabolism or environmental
degradation. A chemical deemed as persistent will have a long half-life and will
remain in the environment for an extended period of time.

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PPB-Parts Per Billion: A measure of concentration where the proportion is such that

one part of solute exists per one billion parts of solvent or matrix.
PPM-Parts Per Million: A measure of concentration where the proportion is such that
one part of solute exists per one million parts of solvent or matrix.

RfD-Reference Dose: A numerical estimate of a daily oral exposure to humans of a
substance. This dose level considered unlikely to cause harmful effects during a
lifetime. This value takes into account sensitive subgroups whom can be
exposed to this agent.

Safety factor: The difference between the NOAEL and the dose allowed in routine
exposure. This value is calculated by using the NOAEL for the most sensitive
species and dividing it by various uncertainty factors depending on the readily
available scientific data. For example if a value is being extrapolated to man

from animals, the NOAEL will be divided by a factor of 10. Such numerical
factors will vary depending on the size of the uncertainty (i.e. more related
species extrapolation will utilize a smaller safety factor).
Sensitizer: A chemical that is capable of causing the development of an allergic

response upon subsequent exposure.
Solubility: The relative ability of a certain substance to be dissolved in a particular
solvent. For example, compounds that are very readily dissolved in water may
be only minimally dissolved in a more lipid-like solvent such as organic solvents

(e.g. octanol).
Sub-chronic: Refers to a period of repeated exposure which is usually about 10% of an
organism's expected life-span.

Synergism: The process by which two or more substances interact via a biological
mechanism to produce a greater than additive response.
Teratogenesis: The development of a deformed offspring after exposure of the fetus to
a certain chemical insult. The various developmental stages at which this

exposure occurs will result in different abnormalities.
Toxicity test: The determination of the toxic potential of a particular substance on a
group of selected organisms under defined conditions.

Toxicodynamics: The mechanism through which a toxic compound exerts its
physiological effect. This includes the relationship between the structure of a
compound and the means by which it acts.
Toxicokinetics: The movement of chemicals through the body. This includes

rate/extent of absorption, distribution, metabolism and elimination.
TWA-Time Weighted Average: The average exposure concentration over an 8-hour
work shift.

Volatility: The ability of a compound to evaporate and partition into the air.
Xenobiotic: Any substance to which an organism is exposed which is not produced
internally in that organism at that time.

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37THR EGULAR SESSION OCICAD,S ANTO DOMINGO ,DOMINICANR EPUBLI, 26-29
APRIL2005,DOCUMENT 1421

(Available at: http://www.cicad.oas.org/apps/Document.aspx?Id=265 (last visited 7 March 2010))

147Annex 117

ENVIRONMENTAL AND
HUMAN HEALTH

ASSESSMENT OF THE
AERIAL SPRAY

PROGRAM FOR COCA
AND POPPY CONTROL

(PECIG) IN COLOMBIA

Rpara el Control del Abuso de Drogas (CICAD)ana
Organization of American States (OAS)
37 session
Santo Domingo, April 26, 2005

148 Annex 117

• Dr. Keith R Solomon
• University of Guelph, Canada

• Dr. Arturo Anadón

• Universidad Complutense de Madrid, Spain
• Dr. Antonio Luiz Cerdeira

• EMBRAPA, Brazil
• Dr. Jon Marshall

• Marshall Agroecology, Somerset, UK

• Dr. Luz-Helena Sanin
• University of Toronto, Canada and
Autonomous University of Chihuahua, Mexico

APPROACH

•Scientific team with expertise in several areas

•Followed the standard approach to risk assessment
•Problem formulation and stressor characteristics

•Exposure characterization

•Effects characterization
•Risk assessment

•Reviewed the open scientific literature and government
reports

•Conducted special studies in Colombia and elsewhere to
characterize effects and exposures
•Science-based assessment for publication in the scientific
literature

149Annex 117

IMPACTS OF
PRODUCTION

Clear-cut and burn
Pesticides (humans

and non-target
organisms
Increased erosion

Fertilizer

150 Annex 117

IMPACTS OF
PRODUCTION

Clear-cut and burn
Pesticides (humans

and non-target
organisms
Increased erosion

Fertilizer

IMPACTS OF REFINING
CHEMICALS

Humans

Non-target organisms

151Annex 117

IMPACTS OF IMPACTS OF
SPRAY PRODUCTION

Off-target effects Clear-cut and burn
on plants
Pesticides (humans
Effects on humans and non-target
organisms
Effects on aquatic
organisms Increased erosion
Effects on
Fertilizer
terrestrial
organisms

IMPACTS OF REFINING
CHEMICALS

Humans

Non-target organisms

152 Annex 117

CHARACTERISTICS

Glyphosate
O O
HO P CH N CH C OH
2 2
OH H

AMPA Sarcosine

O H H O
HO P CH N H+ CH N+ CH C OH
2 3 2
OH H H
Glycine

H O
Pi + CH3 NH3 H N+ CH2C OH

GLYPHOSATE

• Glyphosate is not highly mobile in the

environment
• Rapidly and tightly bound on contact with

soil and aquatic sediments

• Very short biological activity in soils and
water

• Does not biomagnify or move through the

food chain

• Does not leach into groundwater from soil.

153Annex 117

USE
2,000,000

Total use on Colombia (L)
Percent use in spray eradication
1,500,000

1,000,000

15%

Glyphosate use

0%
2000 2001 2002 2003 2004

Year

Droplet Droplet with
without surfactant

surfactant

Wax layer

Cuticle proper

Pectin layer

Primary wall

Secondary wall

Plasma membrane

Cytoplasm

154 Annex 117

AREAS SPRAYED

200

Area of coca planted
Area of coca sprayed
Area of poppy planted
150
Area of poppy sprayed

100

Hec10res (1000s)
8

6
4

0.20
Coca
0.15 Poppy

0.10

0.05
% of total land area
0.00

1994 1996 1998 2000 2002 2004

Year

155Annex 117

BIODIVERSTY HOTSPOT

156 Annex 117

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158 Annex 117

PATHWAYS OF EXPOSURE

Deposition on
nontarget
fieldisms in the

Spray drift Deposition on the target

or spray drift onto
Deposition water
on soil Runoff with
soil

OFF-TARGET DEPOSITION

Photo C Helling

159Annex 117

OFF-TARGET DEPOSITION

140000
120000 Total ha sprayed
100000 Estimated range of off-target drift
80000
60000

Ha sprayed
20000
max
1000
800

600

400 min
Ha200f-target

0
1994 1996 1998 2000 2002 2004
Year

EXPOSURES

• Applicators

• Mixer-loader

• Pilots

• Technicians

• Bystanders

160 Annex 117

DIRECT OVERSPRAY
Extreme Worst case Most likely
worst case case

Whole body (2 m )0.25 m 0.25 m

100% absorption 100% absorption 2% absorption
14.2 mg/kg bw 1.8 mg/kg bw 0.04 mg/kg bw

TOTAL ESTIMATED EXPOSURES

Source of Exposure value in mg/kg

exposure bw
Coca Poppy

Direct overspray 0.04 0.01

Reentry 0.26 0.06

Inhalation 0.01 0.01

Diet and water 0.75 0.18
Worst case total

exposure from all
sources 1.05 0.26

161Annex 117

ENVIRONMENTAL EXPOSURES

Surface water scenario Exposure in µg/L

Coca sprayed Poppy sprayed
at 4.982 kg/ha at 1.2 kg/ha

(3.69 kg AE/ha) (0.89 kg AE/ha)

2 m deep, rapid mixing and no
absorption to sediments, no flow. 185 44

0.3 m deep, rapid mixing and no
absorption to sediments, no flow. 1,229 296

0.15 m deep, rapid mixing and no

absorption to sediments, no flow. 2,473 595
0.15 m deep, rapid mixing and

50% absorption to sediments, no
flow. 1,237 297

SAMPLING IN COLOMBIA

Site name Location Altitude Major crop Known pesticide
(m) types use
Valle del Cauca, N 03º27.642' 1002 Sugar cane Glyphosate and
Río Bolo other pesticides
W 076º19.860'

Boyacá, N 05º40.369' 557 Coca Manual eradication,
Quebrada W 074º00.986' no aerial spraying of
Paunera glyphosate

Sierra Nevada, N 11º13.991' 407 Organic None
Quebrada La Otra W 074º01.588' coffee

Putumayo, Río N 00º43.259' 329 Coca Aerial eradication
Mansoya spraying
W 076º05.634
Nariño, Rio N 01º27.915' 15 Coca Aerial eradication
Sabaletas spraying
W 078º38.975'

162 Annex 117

Sierra Nevada

Boyacá

Valle del Cauca

Nariño

Putumayo

DETECTION OF GLYPHOSATE

Surface water Total number of Frequency of detection (n and %)
collection site samples for site

Glyphosate AMPA

Valle del Cauca, Río 17 1 (5.9%) 0 (0%)
Bolo

Boyacá, Quebrada 18 1 (5.5%) 0 (0%)
Paunera

Sierra Nevada, 18 0 (0%) 0 (0%)
Quebrada la Otra

Putumayo, Río 16 0 (0%) 0 (0%)
Mansoya

Nariño, Rio 17 0 (0%) 0 (0%)
Sabaletas

Other pesticides detected at Nariño - 2,4-D, endosulfan I,
MDL = 25 µg/L endosulfan II, endosulfan sulfate

163Annex 117

EFFECTS IN MAMMALS

• Glyphosate
• Very low acute and chronic toxicity
• Not teratogenic
• Not mutagenic

• Not carcinogenic
• Not immunotoxic in mammals
• Cancer epidemiology
• No strong association with cancer
• Neurological epidemiology
• No strong association
• Reproductive epidemiology
• Association with reproductive responses – Time to
Pregnancy

GLYPHOSATE AND COSMOFLUX®

• ACUTE STUDIES (GLP guideline
studies)

• Very low acute oral toxicity
• Very low acute dermal toxicity
• Low to moderate inhalation toxicity

• Low to moderate skin irritant
• Moderate eye irritant (recovery)

• Not a skin sensitizer
• Addition of the adjuvant Cosmo-Flux® to

the glyphosate did not change its
toxicological properties to mammals

164 Annex 117

EPIDEMIOLOGY STUDY IN

COLOMBIA

• Questionnaire study in 5 regions in
Colombia

• Time to (1 s) Pregnancy (TTP)

• 600 women in each location (3000 total)

• Ecologic study based on region – use of

glyphosate for eradication

• Other factors were also assessed

EPIDEMIOLOGY REGIONS

Site name Focal Known pesticide use
crop
Valle del Sugar Glyphosate and other pesticides. Glyphosate
Cauca cane applied by air.

Boyacá Coca Manual eradication, no aerial spraying of
glyphosate. Use of other pesticides unknown.
Sierra Organic No pesticide use and no coca known to be
Nevada coffee grown. Use of other pesticides unknown.

Putumayo Coca Aerial eradication spraying with lower intensity.
Use of other pesticides unknown.
Nariño Coca Aerial eradication spraying with higher intensity.
Use of other pesticides unknown.

165Annex 117

TIME TO PREGNANCY

100

Percent of pregnancies Boyacá
Nariño
20 Sierra Nevada
Putumayo
Valle del Cauca
0

2 4 6 8 10 12
Time to first pregnancy (months)

ALTERNATIVE MODEL

Variable OR 95% CI

Region

Boyacá 1.00 --
Nariño 0.56 0,47; 0,66

Sierra Nevada 0.36 0,31; 0,43

Putumayo 0.35 0,29; 0,41
Valle del Cauca 0.15 0,13; 0,18

Age at first pregnancy > 20 years 0.81 0,73; 0,91
Irregular relationship 0.76 0,68; 0,84

Consumption of coffee

Medium (1-3 cups per day) 0.91 0,81; 1,04
High ( 4 and more cups per day) 0.84 0,69; 1,02

Perception of contamination of water 0.91 0,81; 1,01

166 Annex 117

RESULTS OF EPIDEMIOLOGY

• The greatest risk (TTP) was in the Valle del Cauca
region
• No association between TTP and eradication of illicit
crops
• Reason(s) for the increased risk for longer TTP in the
Valle del Cauca region not known
• crops) also showed a significant difference from referencerganic
(Boyacá)
• Study designed to test hypotheses related to the use of
glyphosate in eradication spraying - data cannot be used
to identify causality associated with other risk factors
• To test this question in Valle del Cauca or any other
region, a new study would have to be designed and
conducted

EFFECTS IN THE ENVIRONMENT

• GLYPHOSATE AND ROUNDUP®
• Published papers

• Government documents (U.S.EPA, EU, etc)

• GLYPHOSATE AND COSMOFLUX®
• Special studies on the mixture

• Honey bee
• Daphnia magna (aquatic invertebrate)

• Aquatic alga
• Two fish (fathead minnow, rainbow trout)

167Annex 117

ECOTOXICOLOGY DATA

90
Fathead
minnow

70 Water flea

Green alga
50

30
Percent Rainbow Formulated LC50s amphibians
trout Formulated LC50s arthropods

Formulated LC50s fish
10
Glyphosate technical LC50s
Glyphosate plus Cosmo-Flux -animals

Glyphosate plus Cosmo-Flux - algae
1
1 2 3 4 5 6 7 8
10 10 10 10 10 10 10 10

Concentration (µg/L glyphosate AE)

HUMAN RISK ASSESSMENT

Formulated plus
All sources InhalatCosmo-Flux®

Diet and water Toxicity Dermal
Coca
Poppy

Inhalation Oral

Acute exposures
Glyphosate
Reentry NOEL chronic

Direct overspray Margin of exposure RfD

0.001 0.01 0.1 1 10 100 1000 10000
Glyphosate (mg/kg)

Margin of exposure is protective for all sources of contamination and is even

lower because acute exposures are compared to chronic effect doses

168 Annex 117

ENVIRONMENTAL RISK

99 Estimated concentration in
surface water (cm deep)
MDL surface
water 200 30 15

90
Fathead
minnow

70
Water flea

50 Green alga

Percent rank Formulated LC50s amphibians
Rainbow
trout Formulated LC50s arthropods
Formulated LC50s fish
10
Glyphosate technical LC50s
Glyphosate plus Cosmo-Flux -animals

Glyphosate plus Cosmo-Flux - algae
1
1 2 3 4 5 6 7 8
10 10 10 10 10 10 10 10

Concentration (µg/L glyphosate AE)

HUMAN HEALTH

CONCLUSIONS

IMPACTS INTENSITY RECOVERY FREQUENCY IMPACT % IMPACT

SCORE SCORE % SCORE

Clear cutting
and burning 5 3 3 45 16.7

Planting the

coca or poppy 0 1 100 0 0.0

Fertilizer inputs 0 0.5 10 0 0.0

Pesticide

inputs 5 3 10 150 55.5

Eradication
spray <0.1 0 1 <0.1 <0.1

Processing

and refining 5 3 5 75 27.8

169Annex 117

ENVIRONMENTAL

CONCLUSIONS

INTENSITY RECOVERY IMPACT %
IMPACTS
SCORE TIME (Y) SCORE IMPACT
Clear cutting

and burning 5 60 300 97.6
Planting the

coca or poppy 1 4 4 1.3
Fertilizer inputs 1 0.5 0.5 0.2

Pesticide inputs 2 0.5 1 0.3

Eradication
spray 1 0.5 0.5 0.2

Processing and
refining 2 1 2 0.7

RECOMMENDATIONS FOR

CURRENT PRACTICES

Practice Benefit of continuance Rank
Mixer-loader, worker, anProtection of the humans and the environme5t
environmental protectionfrom excessive exposures.

storage, mixing, and
loading operations.
Use of state of art Accurate records of location and areas spr5yed
application technology.

Replace the respirator Reduce the risk of splashes of concentrate5
worn by the mixer-loaderformulation into the eyes.
with a full face shield to
reduce the potential
exposure of the eyes.

Use of glyphosate in theRisk to humans and the environment is judg4d
eradication program. to be lower than any currently-available
alternatives. New candidate products should
only be considered after an appropriate risk
assessment has been conducted.

170 Annex 117

NEW RECOMMENDATIONS (1)

Recommendation Benefit of new data Rank
Conduct a study to identify other Better understand and manage 3
factors associated with time to human health risks.

pregnancy (TTP).
Including proximity to surface watBetter indication of likely 2
in Geographic Information System frequency of contamination of
(GIS) analysis of locations and areas these habitats.
of coca and poppy fields.

Identify mixtures of glyphosate anReduction in possible 2
adjuvants that are less toxic to environmental impacts to non-
aquatic organisms than the currenttarget organisms in shallow
used mixture. The priority of thissurface water environments.
recommendation would depend on
the results of the GIS analysis.

Testing of the glyphosate-Cosmo- Decrease in uncertainty 2
Flux® formulation for toxicity to regarding the toxicity to
amphibians. amphibians.

PROXIMITY TO WATER

Photo C Helling

171Annex 117

NEW RECOMMENDATIONS (1)

Recommendation Benefit of new data Rank

Conduct a study to identify risk Better understand and manage 3
factors associated with time to human health risks
pregnancy (TTP).

Including proximity to surface waterBetter indication of likely 2
in Geographic Information System frequency of contamination of
(GIS) analysis of locations and areas these habitats.
of coca and poppy fields.

Identify mixtures of glyphosate and Reduction in possible 2
adjuvants that are less toxic to environmental impacts to non-
aquatic organisms than the currentlytarget organisms in shallow
used mixture. The priority of this surface water environments.
recommendation would depend on

the results of the GIS analysis.
Testing of the glyphosate-Cosmo- Decrease in uncertainty 2
Flux® formulation for toxicity to regarding the toxicity to
amphibians. amphibians.

NEW RECOMMENDATIONS (2)

Recommendation Benefit of new data Rank
Use of GIS to quantify areas of Better understand potential effects on2
coca and poppy production in important sources of biodiversity from
biodiversity hotspots. clear-cutting and planting of coca and

poppy.
Use of GIS to quantify size of Allow more accurate quantification of 2
fields planted to coca and poppy potentially impacted areas as well as
and track these over time. recovery.

Review the regulatory status of Ensure that new testing and toxicity 2

glyphosate on a regular basis. data on glyphosate are included in the
risk assessment of its use in
eradication spraying in Colombia.
If new mixtures are used, Better characterization of human 1

measurement of exposures to exposures under conditions of use in
glyphosate in bystanders to sprays Colombia.
and reentry into sprayed fields.

172 Annex 117

ACKNOWLEDGEMENTS

SAT members
Jorge Rios and Adriana Henao of the

CICAD office and the Executive
Secretariat of CICAD
PTG Team members

Staff of the Ministry of Foreign Affairs
and Ministry of Justice of Colombia

Staff of the National Police
(Antinarcoticos)

Captain James Roa

THANK YOU

173174 Annex 118

CICAD,R.A.B RAIN ET A., HE TOXICOLOGY OF SUBSTANCESUSED IN THE
PRODUCTION ANDR EFINING OCOCAINE ANDH EROIN: ATIE-TWO H AZARD
ASSESSMENT”, OAS,WASHINGTON , D.C.,3ULY 2005,(CICADI)

(Complete Report without appendices. Full Report available at:
http://www.cicad.oas.org/Desarrollo_Alternativo/ENG/Projects%20By%20Cou…
AD_Tier_2_Hazard_Assessment_July_2005%5B1%5D.pdf (last visited 7 March 2010))

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219220 Annex 119

2006 MEMORANDUM OF U NDERSTANDING BETWEEN THG ENERALS ECRETARIAT OF

THEO RGANIZATION OAMERICAN STATES(SG/OAS) AND THEGOVERNMENT OF
C OLOMBIA FOR THE EXECUTION OF THE STUDY ON THE EFFECTS PROGRAM
FOR THEERADICATION OFLLICITCROPS BY AERIAL SPRAYING WIGLYPHOSATE
H ERBICID(PECIG) ON HUMAN HEALTH AND THE ENVIRONMEN,23M AY2006

(Archives of the Ministry of Foreign Affairs of Colombia)

MEMORANDUM OF UNDERSTANDING BETWEEN THE GENERAL
SECRETARIAT OF THE ORGANIZATION OF AMERICAN STATES (SG/OAS)
AND THE GOVERNMENT OF COLOMBIA FOR THE EXECUTION OF THE
STUDY ON THE EFFECTS OF THE PROGRAM FOR THE ERADICATION OF

ILLICIT CROPS BY AERIAL SPRAYING WITH GLYPHOSATE HERBICIDE
(PECIG) ON HUMAN HEALTH AND THE ENVIRONMENT

[PAGE 2]

MEMORANDUM OF UNDERSTANDING BETWEEN THE GENERAL
SECRETARIAT OF THE ORGANIZATION OF AMERICAN STATES (SG/OAS)
AND THE GOVERNMENT OF COLOMBIA FOR THE EXECUTION OF THE

STUDY ON THE EFFECTS OF THE PROGRAM FOR THE ERADICATION OF
ILLICIT CROPS BY AERIAL SPRAYING WITH GLYPHOSATE HERBICIDE
(PECIG) ON HUMAN HEALTH AND THE ENVIRONMENT

The Parties to this Memorandum of Understanding, the General Secretariat of the
Organization of American States (hereinafter SG/OAS), through the Executive
Secretariat of the Inter-American Drug Abuse Control Commission (hereinafter,
SE/CICAD), represented by its Assistant Executive Secretary, Abraham Stein, and the

Government of Colombia through the Ministry of Foreign Affairs, represented by the
Minister of Foreign Affairs, Carolina Barco:

CONSIDERING

That the SG/OAS, is the main and permanent organ of the Organization of
American States (hereafter OAS), and is authorized to establish and promote relations

221Annex 119

of cooperation with member States pursuant to Article 112(h) of the OAS Charter and
with its General Assembly resolution AG/RES. 57 (l-O/71).

That the Inter-American Drug Abuse Control Commission (hereafter CICAD or

the Commission) is an agency of the OAS, established by Article 52 of the Charter of
the Organization. This agency is technically autonomous and carries out its duties
within the context and scope of the Rio de Janeiro Action Plan against Consumption,
Production, and Illicit Trafficking on Drugs and Psychotropic Substances, the mandates
of the General Assembly, and the decisions internally adopted by the Commission.

That the purpose of CICAD is to contribute to eliminate illicit trafficking and

That within the framework of its Hemispheric Strategy, CICAD promotes
actions against the illicit crops of raw materials destined for the production of illicit

[PAGE 3]

That the Colombian State implemented the Program for the Eradication of Illicit
Crops by Aerial Spraying with Glyphosate Herbicide (PECIG) in Colombia, in

accordance with paragraph g) of Article 91 of Law 20 of 1986, whereby Colombia
adopted the National Anti-Narcotics Statute that assigns to the National Narcotics
Council the duty to “provide for the destruction of marihuana, coca and other crops
from which substances causing dependency may be extracted, using the most adequate
means, following a favourable opinion of the agencies entrusted with protecting the
health of the population and the preservation and balance of the ecosystem in the
country”. [The Program] is regulated through resolution 0013 of 2003 and operates in

all the regions in the country the presence of illicit crops is evidenced.

That for the Colombian State, the adoption and implementation of the Program
for the Eradication of Illicit Crops by Aerial Spraying with Glyphosate Herbicide
(PECIG) has become an inexorable necessity in view of the fact of the extended
presence of illicit crops in the national territory and the security problems that, in many
cases, preclude resort to other eradication methods.

222 Annex 119

That the Government of Colombia understands the PECIG as the plan of the
State for the mitigation of the adverse environmental impact caused by illicit crops and
the subsequent processing of illicit drugs.

That, in accordance with Colombian law and abiding by the provisions of the
1961 Single Convention on Narcotic Drugs as amended by its 1972 Protocol and the
1988 United Nations Convention Against Trafficking of Illicit Drugs and Psychotropic
Substances as regards the obligation to adopt the necessary measures to eradicate the
poppy crops, coca bushes and cannabis plants that are illicitly grown, and in light of the
unusual increase of illicit crops in the national territory, the Government of Colombia

set out to strengthen its strategy to confront the problem of illicit drugs production and
trafficking through forced eradication by aerial spraying with glyphosate herbicide.

That in view of the growing domestic and international concern as to the alleged
effects of the Program for the Eradication of Illicit Crops by Aerial Spraying with
Glyphosate Herbicide (PECIG), the Governments of Colombia, the United States and

the Untied Kingdom requested CICAD to conduct a study in order to document such
effects in a scientific and independent manner.

That the panel of scientists contracted by SE/CICAD to carry out that study,
conducted under the Memorandum of Understanding between the OAS and the
Government of Colombia for the execution of the study on the effects of the PECIG on

human health and ththenvironment, concluded in the city of Bogotá, D.C., Republic of
Colombia, on the 4 day of February 2004, identified certain queries in their reviews of
the data and on the basis thereof makes certain recommendations aimed at resolving the
queries identified in the study.

[PAGE 4]

That having learned of the results of the Initial Phase, the Government of
Colombia and the Government of the United States request the cooperation of CICAD
in order to be able to conduct a complementary phase.

That scientific team identified the strengths and queries as a result of the

assessment and recommended to maintain the current practices of the PECIG, additional
data collection for a longer period with the purposes of gathering a between
characterization of the impacts of coca and poppy crops in areas of the Andean region
characterized by their biodiversity and the definition of the alleged effects on superficial
waters adjacent to the crops. It is also recommended that other adjuvants be tested, that
represent a higher efficacy and, at the same time, eliminate or minimize any risk that
could affect aquatic organisms. Although no relation was observed between aerial

sprayings with glyphosate herbicide and the results in human reproduction, it is

223Annex 119

recommended to conduct additional studies in order to identify possible risk factors
associated to other human activities or environmental factors.

STATING the importance of coordinating their efforts with the purpose of
fulfilling their objectives,

AGREE to conclude the present Memorandum of Understanding that will be
governed by the following provisions:

FIRST CLAUSE: Object and purpose

The object and purpose of the present Memorandum is to conclude an agreement
that serves as framework for the development of an independent scientific study on the
alleged effects of the PECIG on human health and the environment.

The description of the study is set out in detail in Annex I that is an integral part

of the present Memorandum, that was vetted by the Government of Colombia and the
SE/CICAD.

SECOND CLAUSE: Framework for cooperation

Cooperation and assistance provided in pursuance of the present project will be

carried out in observance of the respect for national sovereignty, confidentiality,
transparency and veracity of conclusions.

THIRD CLAUSE: Study areas

For comparative purposes and of statistic and methodological precision, the

study will focus both on the areas where the Program for the Eradication of Illicit Crops
is implemented, as well as on areas where glyphosate herbicide is used for the
cultivation of lawful produce,

[PAGE 5]

on areas where manual eradication programs are carried out and in areas of
organic production. By mutual agreement between the Parties, other areas the
assessment of which is considered relevant may be included.

FOURTH CLAUSE: Responsibilities of the Parties

A. SE/CICAD undertakes to:

224 Annex 119

1. Supervise and follow-up on the works carried out by the Scientific
Assessment Team (SAT) and the Permanent Technical Group for Mobile
Monitoring (PTGMM [shortened form PTG]).

2. Contract and supervise the required personnel and laboratories for
conducting the study that is the object of the present Memorandum.
3. Conduct and follow-up on the study that is the object of the present
Memorandum.
4. Coordinate and supervise the adequate progress of the activities foreseen in
Annex I, “Monitoring of the Aerial Spraying Program for the Control of
Illicit Coca and Poppy Crops on the Environment and Human Health in

Colombia.”
5. Take all actions required for the effective and timely execution of the
project’s activities mentioned in Table 6 of Annex I.
6. Review and approve periodical reports on the progress of the established
work plan.
7. Periodically inform the Government of Colombia on the progress of the

completion of the study that is the object of the present Memorandum.
8. In accordance with the provisions of the Fifth Clause, publicly present the
results of the study and widely publicize the corresponding final report that
will have been previously presented to the Government of Colombia for its
information. The results of the study and the final report to which this
paragraph refers will be presented in Spanish and English.

B. The Government of Colombia undertakes to:
1. Facilitate the compliance with and implementation of the present
Memorandum.
2. Provide any information required for the formulation and implementation of
the project as requested by SE/CICAD, including, among other, information
relating to the areas considered within the aerial spraying program.

3. Appoint an agency that, in direct coordination with the Ministry of Foreign
Affairs (Division of Multilateral Political Affairs, Sub-division for Drug
Affairs), will be entrusted with the following responsibilities:
a. To facilitate communication between SE/CICAD, implementing
personnel and the Government of Colombia.
b. To arrange the required logistics required for the mobilization of the

personnel to and within the areas under study described in Annex I.

[PAGE 6]

c. To ensure the timely and coordinated action of the different
authorities in charge of providing security to the personnel
implementing the study.

225Annex 119

4. To provide the required elements for the correct execution of the
components of the study.

5. To provide a security detail for the mobilization of the personnel involved in
the study to and within its areas, in accordance with the resources allocated
for these purposes in the project budget. All field visits to the areas under
study shall be conducted by mutual agreement with the Colombian
authorities in charge of providing security, and under the terms
recommended by such authorities according to the security situation.
Pursuant to these same reasons, any scheduled visit may be suspended prior

to the agreed date.

FIFTH CLAUSE: Confidentiality

The Parties to this Memorandum undertake to preserve the strictest

confidentiality while the study is being developed. Neither Party may, without the
express prior consent of the other, publish partial results of the study under way.

Once the Parties have learned, under reserve, the results of the study, the final
report will be made public and will be widely publicized.

SIXTH CLAUSE: Termination

The present Memorandum may be terminated by mutual agreement or by either
Party, through written advance notice of at least three months to the other.

SEVENTH CLAUSE: Settlement of disputes

The Parties undertake to settle controversies that may arise of the interpretation
or application of the present Memorandum of Understanding, preferably by mutual
agreement. In case a satisfactory solution is not reached, recourse will be had to the
arbitration procedure mutually agreed by the CICAD and the Government of Colombia.
If there is no agreement on the procedure, arbitration will be conducted pursuant to the

arbitral procedures in force of the United Nations Commission on International Trade
Law (UNCITRAL). The arbitral tribunal constituted in accordance with those Rules
will rule as amiable mediator or ex aequo et bono and its decision will be final and
binding.

None of the provisions in this Memorandum signifies or shall be construed as a
relinquishment of the privileges and immunities

226 Annex 119

[PAGE 7]

enjoyed by the Parties in accordance with the relevant agreements and laws on

the matter and the general principles of international law.

EIGHTH CLAUSE: Entry into Force, Duration and Amendments

The present Memorandum shall enter into force on the date of its signature by
the last of the Parties, and shall be in force until the completion of the study and the
publication of its results.

Addition or amendment to this Memorandum will be made by mutual agreement
between the Parties, following compliance with legal requirements. The instruments
registering those modifications will be appended as annexes to the present
Memorandum and shall become integral parts thereof.

In witness whereof, the present Memorandum between the General Secretariat
of the Organization of American States (SG/OAS) and the Government of Colombia for
the Execution of a Study on the Effects of the Program for the Eradication of Illicit
Crops by Aerial Spraying with Glyphosate Herbicide (PECIG) on Human Health and

the Environment, is signed by the duly authorized representatives of the Parties, in two
copies in Spanish, both equally authentic.

For the General Secretariat of the For the Government of Colombia

Organization of American States

[signed illegibly] [signed illegibly]
ABRAHAM STEIN CAROLINA BARCO
Assistant Executive Secretary of the Minister of Foreign Affairs
Inter-American Drug Abuse Control

Commission

Date: 23 May 2006 Date:

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228 Annex 119

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230 Annex 119

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232 Annex 119

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234 Annex 119

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236 Annex 119

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238 Annex 119

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240 Annex 119

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242 Annex 119

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244 Annex 119

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246 Annex 119

247248 Annex 120

A NNUALREPORT OF THENTER-AMERICAN DRUG ABUSEC ONTROLC OMMISSION
(CICAD)TO THEG ENERALA SSEMBLY OF THORGANIZATION OFAMERICAN
STATES AT I36TH REGULAR SESSIO,SANTOD OMINGO, OMINICAN REPUBLI,4-
6 UNE2006

(Available at: http://www.cicad.oas.org/AnnualReports/2005/1474-rev2-
AnnualReport_CICAD2005_en.pdf (last visited 7 March 2010), p. 13)

249Annex 120

ORGANIZATION OF AMERICAN STATES

INTER-AMERICAN DRUG ABUSE CONTROL COMMISSION

THIRTY-EIGHTH REGULAR SESSION OEA/Ser.L/XIV.2.38
December 6-9, 2005 CICAD/doc.1474/05 rev.2
Washington, D.C. 22 May 2006
Original: English

ANNUAL REPORT OF THE
INTER-AMERICAN DRUG ABUSE CONTROL COMMISSION (CICAD) TO THE
GENERAL ASSEMBLY OF THE ORGANIZATION OF AMERICAN STATES
AT ITS THIRTY-SIXTH REGULAR SESSION

250 Annex 120

local market for unprocessed and processed produce to strengthen those capabilities
that might later be applied to exporting processed products.

Colombia

Study of the Effects of Aerial Glyphosate Spraying and Illicit Crop Cultivation on Human
Health and the Environment
In April, the results of the study were presented to the Government of Colombia and
made available on the CICAD web site. The scientific team held a news conference in
Bogotá, reported the findings to several scientific fora, and prepared responses to

commentaries made by several organizations.

Dominica

Organic Banana Production and Pest Management
In 2005, this project expanded its showcase plots to demonstrate the use of organic
farming and pest management techniques. This change also brought in new project

personnel.

Peru

Tropical Crops Institute (ICT) – Training farmers in the Apurimac and Ene River Valley
(VRAE)
The Tropical Crops Institute (ICT) renewed agreements with both CICAD and the
Narcotics Affairs Section of the US Embassy in Peru to conduct agricultural extension

training in more than 700 locations in Tingo María, Tocache, Juanjui and Tarapoto for
3,000 beneficiaries. Training was provided on topics such as using more technical
methods to increase cacao yield, propagation systems, fertilization, pruning, and pest
management. Leadership scholarships also enabled farmers to live and study for five
days at the ICT-NAS/CICAD Experimental Station in Tarapoto.

Publication

Comité Andino para le Desarrollo Alternativo (CADA), Estrategia andina de desarrollo
alternativo integral y sostenible. Bogotá, Colombia: 2005

D. LEGAL DEVELOPMENT

Overview

The Legal Development unit provides counseling and legal assistance to the different
units of CICAD, giving legal advice on topics and procedures in the areas of drugs and
related subjects. One of its most important tasks is to review model regulations. This unit
is also in charge of administrative and legal tasks, advising the CICAD on OAS policies

and procedures as well as funding obligations. As such, it is responsible for formulating
and drafting of Memoranda of Understanding and other agreements.

Control of Arms and Explosives
The Legal Development unit is concentrating on prevention and control of trafficking in

firearms through application of the Inter-American Convention Against the Illicit
Manufacturing of and Trafficking in Firearms, Ammunition, Explosives, and Other

251252 Annex 121

ANNUAL REPORT OF THPERMANENTC OUNCIL TO TGEENERALASSEMBLY OF THE
O RGANIZATION AFMERICANSTATES2006-2007

(OAS, AG/DOC.4698/07, pp. 33, 34)

253Annex 121

GENERAL ASSEMBLY

THIRTY-SEVENTH REGULAR SESSION OEA/Ser.P
June 3 to 5, 2007 AG/doc.4698/07 corr. 1
Panama City, Panama 4 June 2007
Original: Spanish

Item 7 on the agenda

ANNUAL REPORT OF THE PERMANENT COUNCIL

TO THE GENERAL ASSEMBLY
2006-2007

254 Annex 121

- 33 -

7. Other business

The Permanent Mission of Brazil requested the distribution of a CD-ROM of the
Special Session of Freedom of Thought and Expression that took place on October

26 and 27, 2006. The Alternative Representative of Brazil also informed the Council
that his government ratified the Inter-American Convention on Transparency in
Conventional Weapons Acquisitions.

The Permanent Council bid farewell to Ambassador John Maisto, Permanent

Representative of the United States.

15. Record of the regular meeting held on January 9, 2007
CP/ACTA 1576/07 24/

1. Adoption of the order of business

The Council adopted the order of business, document CP/OD-1576/07.

Remarks by the Chair of the Permanent Council

Ambassador María del Luján Flores, Permanent Representative of Uruguay, made
some remarks on the occasion of her first meeting as Chair of the Permanent
Council. She said, among other things, that she intended to promote steadfastly the

general principles of international law and, with support from the various diplomatic
missions, some topics such as the environment and regional agreements, protection
and promotion of the rights of children, and in particular the right of identity and of
citizen participation.
Presentation of the gavel

As is customary, the Permanent Council presented the gavel to Ambassador Marina
Annette Valère, Permanent Representative of Trinidad and Tobago, in recognition
for her work as Chair of the Council from October to December 2006.

2. Note from the Permanent Mission of Ecuador requesting inclusion of the topic “Resumption
of glyphosate spraying in an area adjacent to the Ecuadorian border”

At the request of the Permanent Mission of Ecuador, the Permanent Council considered the
topic “Resumption of glyphosate spraying in an area adjacent to the Ecuadorian border”

(CP/INF.5428/07).

Present for consideration of this matter were His Excellency Mr. Francisco Carrión Mena,
Minister of Foreign Affairs of Ecuador, and His Excellency Mr. Camilo Reyes, Vice Minister of
Foreign Affairs of Colombia.

24. Las declaraciones y comentarios formulados por la Delegaciones se encuentran en el acta de la sesión
CP/ACTA 1576/07.

255Annex 121

- 34 -

During the meeting, the Minister of Foreign Affairs of Ecuador presented the OAS Secretary
General, José Miguel Insulza, with three volumes containing “Studies and documents on aerial
spraying of glyphosate and its chemical components.” The three volumes have been placed in the
Columbus Memorial Library. The statements by the Minister of Foreign Affairs of Ecuador and the
Vice Minister of Foreign Affairs of Colombia have been published, respectively, as documents

CP/INF.5433/07 and CP/INF.5432/07.

Lastly, the Council decided to take note of the information presented on the topic.

3. Update on preparations for the thirty-seventh regular session of the General Assembly

Ambassador Albert R. Ramdin, Assistant Secretary General, commented on the preparatory
work for the thirty-seventh regular session of the General Assembly.

In that connection, the Permanent Council decided:

To take note of the information presented by the Assistant Secretary General on the
preparatory work for the next regular session of the General Assembly.
To reiterate to the organs, agencies, and entities of the Organization that they should
transmit, no later than March 1 of this year, their respective annual reports in order to
meet statutory deadlines and as indicated by the Assistant Secretary General.

4. Applications from civil society organizations to participate in OAS activities

Ambassador Marina Annette Valère, Permanent Representative of Trinidad and Tobago and
Chair of the Committee on Inter-American Summits Management and Civil Society Participation in

OAS Activities, presented the applications from the following civil society organizations (CP/CISC-
301/06) to the Permanent Council for consideration:

1. Transparency International Costa Rica (CP/CISC-255/07)

2 Transparência Brasil (CP/CISC-256/07)

3 Caja de Compensación Familiar de Antioquia (COMFAMA) (CP/CISC-257/07)

4. Coalición Regional contra el Tráfico de Mujeres y Niñas en (CP/CISC-258/07)

América Latina y el Caribe, A.C. (CATW-LAC)

5. Centro Latinoamericano de Estudios y Cooperación para el (CP/CISC-259/07)
Desarrollo (CENLAT)

6. Corporación Participación Ciudadana Ecuador (Participación (CP/CISC-260/07)

Ciudadana)

7. Federación Latinoamericana de Ciudades, Municipios y (CP/CISC-261/07)

256 Annex 122

ANNUAL REPORT OF THENTER-AMERICAN DRUG ABUSEC ONTROLC OMMISSION
(CICAD)TO THEG ENERALA SSEMBLY OF THORGANIZATION OFAMERICAN
STATES AT I39 R EGULAR SESSIO, AN PEDROSULA ,HONDURAS, 2-3UNE2009

(OEA/Ser.G, CP/doc.4395/09 corr. 1, 26 May 2009, p. 12)

257Annex 122

PERMANENT COUNCIL

OEA/Ser.G
CP/doc. 4395/09 corr. 1
26 May 2009
Original: Spanish

ANNUAL REPORT OF THE
INTER-AMERICAN DRUG ABUSE CONTROL COMMISSION
TO THE GENERAL ASSEMBLY

258 Annex 122

Andean Countries Cocoa Export Support Opportunity (ACCESO)

The Andean Countries Cocoa Export Support Opportunity (ACCESO) initiative started in June 2005 with
support from the World Cocoa Foundation (private business interests), the U.S. Agency for International
Development (USAID) and the Inter-American Institute for Agricultural Cooperation (IICA). The
ACCESO initiative aims to strengthen the entire supply chain of cacao production, from the field to the

consumer, in Bolivia, Colombia, Ecuador and Peru. CICAD took specific responsibility for developing
technical assistance and training for cacao farmers with the ―farmer field school‖ (FFS) methodology.
This participatory approach allows farmers to learn by doing. The method integrates farmer and trainer
through a two-way channel -- adoption of practical and theoretic knowledge about the cultivation of cacao
and the use of simple methodological tools. The activities of farmer field schools incorporate elements of

organization, observation, analysis, reflection and action that aim towards developing the skills needed to
improve decision-making and problem-solving.
In 2008, CICAD financed the implementation of 30 farm field schools in Peru, which trained 750 growers

and certified 60 of them as FFS instructors. The FFS extension methodology is also being used in Bolivia.
In addition, CICAD underwrote the monitoring and evaluation of the impact achieved by the farm field
schools program in Peru, which reached 5,840 farmers through 234 farm field schools over the three years
of the project. The project’s first phase finished in late 2008, and the ACCESO team is planning a follow-
on stage of activities.

Colombia: Study on the Effects of Aerial Glyphosate Spraying
In 2005, CICAD’s scientific evaluation team presented the results of an independent study, undertaken at

the request of the governments of Colombia, the United States and the United Kingdom, to measure the
impact of aerial spraying of coca fields in Colombia on human health and the environment. Although no
association between spraying and human reproduction was found, the team proposed to carry out
additional studies to identify possible risk factors associated with other human activities or the
environment. The independent scientific evaluation team that CICAD hired in 2006 presented most of its

findings of the follow-up study on the human heath and environmental evaluation of the aerial spraying to
control coca and poppy crops in Colombia in late 2008. The completed study, which consisted of several
technical articles, was submitted for consideration in the peer-reviewed scientific periodical Journal of
Human and Environmental Toxicology, and was also to be published on the CICAD web site. The
findings will also be presented to the public in Washington, DC and in Bogota in 2009.

The components of the study are the following:

Risk to human and environmental health posed by the use of Glyphosate for the control of coca
crops;

Differences in gestation period in fertile women in five Colombian regions;

Bio-monitoring of genotoxic risks for farm workers in five Colombian regions, considering their
work exposure to Glyphosate;

Identification of the geographic distribution of amphibian fauna exposed to the use of pesticides;
Risk posed to amphibians due to the production and eradication of coca;

Analysis of the drift from aerial spraying with Glyphosate and Cosmo-Flux, as employed in the
Colombian eradication program; and

Identification of the mixtures of Glyphosate and additives that might be less toxic for aquatic
organisms than the one currently in use by the Government of Colombia.

259260 Annex 123

SGS (SOCIETÉ G ÉNERALE DE SURVEILLANCE , S.A)COLOMBIA S.A.,“REPORT OF
CONTAMINATION C ONTROL FOR GLYPHOSATE APPLICATION AT THE SIERRA OF
S ANTA M ARTA”,1987

(SGS Colombia S.A. Bogotá, Report of Contamination Control for Glyphosate
Application at the Sierra of Santa Marta, 1987. pp. 2, 5, 6, 7, 8, 12)

[Page 2]

INTRODUCTION

This study has been undertaken for the need of the Colombian National Police to

establish the effects on the jungle as a consequence of the intensive application (by
means of spraying) of glyphosate used to destroy marijuana crops. In the participation
of this effort, SGS has been contracted as an independent, private institution to carry out
control of the current contamination resulting from the use of glyphosate. For this
preliminary study, it was decided to sample some sites recently sprayed by the National

Police and considered by it the most heavily sprayed with Glyphosate.

Soil, foliage, and water samples were taken in seven (7) recently sprayed sites as well as
water samples from the Ciénaga Grande de Santa Marta or Puente de la Parra, and the
Cordoba River downstream the sprayed areas.

[…]

[Page 5]

2. Inspection Period

Inspectors from SGS carried out the inspection on 2 and 3 February 1998. On those
days, an aerial reconnaissance was made on a helicopter of the National Police. During

this reconnaissance of the northern and western area of the Sierra de Santa Marta, a lot
of overflights on previously sprayed sites were made, landing on the sites with the
greatest spraying to take soil, foliage, and water samples.

All of the above mentioned sites (See 2 a.) were widely sprayed during the August-
November 1987 period, according to the Police officers that carried out the task and to
the documentation submitted ) See Annex DOC 9774/5-6-7-8)

261Annex 123

Thus, we can consider that the inspection sites with no exemption were sprayed with
glyphosate in a 2 to 5 month period prior to our sampling process.

[…]

[Page 6]

4. Procedure to collect samples

Taking into account the glyphosate characteristics, a sampling scheme was designed so
that it were reliable and allowed us to get representative result of contamination of the
sites sprayed with glyphosate.

[…]

[Page 7]

Sampling Scheme

In every sampling site; that is, a site intensively sprayed with glyphosate, soil, foliage,
and water samples were taken at random with the purpose of obtaining a representative

compound soil, foliage, and water sample of each site.

The scheme is designed to determine if

-There is or there is not presence of glyphosate in soils

- There is or there is not presence of glyphosate in weeds and/or food plants that grew
again in the sprayed sites.

- There is or there is not contamination in the rivers water, resulting from leaching and
erosion of soils in hilly landscape.

Soil Sampling

Only the superficial horizon was sampled (A) which corresponds to the zone in contact
with the glyphosate that has fallen and that due to the characteristics of the product
(quick absorption) may contain contaminants.

262Thus, we can consider that the inspection sites with no exemption were sprayed with
glyphosate in a 2 to 5 month period prior to our sampling process.

[…]

[Page 6]

4. Procedure to collect samples

[…]

[Page 7]

Sampling Scheme

In every sampling site; that is, a site intensively sprayed with glyphosate, soil, foliage,
and water samples were taken at random with the purpose of obtaining a representative

compound soil, foliage, and water sample of each site.

The scheme is designed to determine if

-There is or there is not presence of glyphosate in soils

- There is or there is not presence of glyphosate in weeds and/or food plants that grew
again in the sprayed sites.

- There is or there is not contamination in the rivers water, resulting from leaching and
erosion of soils in hilly landscape.

Soil Sampling

-in soils

-in weed and/or food plants that grew again
-in river waters

And the results from the analyses made to the 26 samples to detect the presence of
glyphosate in soil, foliage, and water samples duly collected and packed by SGS, we
can certify that (see sampling and quality certificates by SGS Colombia S.A.) there is

no detectable contamination with glyphosate in the sampled sites.

SGS COLOMBIA S.A.
[Signed]

264 Annex 124

J.P.GIESY, S.OBSON S& K.R. S OLOMON ,“ECOTOXICOLOGICAL RISK ASSESSMENT
FOR R OUNDUP HERBICIDE”

(Reviews of Environmental Contamination and Toxicology 167: 35-120, 2000, pp. 69, 74.)

265Annex 124

266 Annex 124

267Annex 124

268 Annex 125

G. M.W ILLIAMS ET AL., “AFETY EVALUATION AND RISK ASSESSMENT OF THE
HERBICIDE R OUNDUP ® AND ITS ACTIVE INGREDIENT,GLYPHOSATE ,FOR HUMANS ”

(Regulatory Toxicology and Pharmacology 31:117–165, 2000, pp. 117, 160.)

269Annex 125

270 Annex 125

271272 Annex 127

R.E. RAMOS C.,J.P.R AMOS B.,ENVIRONMENTAL ASSESSMENT OF THE IMPACT OF
COCA CROPS AND THE PROCESSING OF COCA LEAF ,U NIVERSIDAD DE LOS A NDES ,
BOGOTÁ , COLOMBIA , 2002

(R.E. Ramos C., J.P. Ramos B., Evaluación Ambiental del Impacto de cultivos de coca y el
procesamiento de la hoja de coca, Universidad de los Andes, Bogotá, Colombia, 2002, pp. 1, 5, 10-11)

[Page 1]

ABSTRACT: This document analyzes environmental impacts caused by coca crops and

the processing of coca leaf in the province of Norte de Santander, Tibu municipality. To
determine environmental impacts, forest cover loss was analyzed using multi-temporal
analysis of SPOT satellite images for the years 1999, 2000, and 2001. The cover loss
due to coca crops corresponded to 30% of the total for that period. The environmental

effects generated by the use of pesticides and processing laboratories were established
by determining residues and pouring resulting from the processing of coca paste and
their location. It, combined with the permanence of coca crops, allows to determine
sites of potential chemical substances accumulation, given that coca crops use 10 time
more agrochemicals compared to cocoa crops, the traditional agricultural crop in the

municipality.

[…]

[Page 5]

[…]

Acid solutions are poured directly on the soil or in the nearest water course depending

on the laboratory location. It is likely that the effects of pouring acid solutions alter the
pH in soils and water where they deposit.
Transformation of coca leaf into coca paste and cocaine has negative environmental
effects. Studies of the United States Department of State show that 10 million liters of
sulfuric acid, 16 million liters of Ethyl Ether, 8 million liters of acetone, and 40 to 770

million liters of kerosene are poured every year on the soil by coca processors in the
Andean Region, mainly in Colombia (Scheafer 2002).

5.2 Chemical precursors used in processing

275Annex 127

Processing of coca leaf requires a great deal of chemical precursors and water to extract
the alkaloid (DNE, 2002). According to data obtained from the Antinarcotics Police, it
is estimated that every hectare of coca crops requires the use of approximately 127

kilos/ha of solid precursors, 447 litres/ha of liquid precursors and 400 litres/ha of water
(DIRAN 2002).

[…]

[Page 10 and 11]

8. CONCLUSIONS
[…]

Gramaxone (A.i. paraquat), Faena (A.i. glyphosate), insecticides such as Tamaron (I.a.
Metamidaphos), and fungicides such as Manzate (I.a. Mancozeb). The analysis allowed
to establish that the risk of substances accumulation per environmental component and
the monitoring needs are: In soils, substances that need to be monitored are Gramaxone
and Faena, in underground waters, Anikilamina and Tamaron, and in superficial water
sediments all these compounds must be monitored.

6. Processing of coca is made in order to extract or wash the alkaloid, which represents
only 0,5 to 1,5 % of the total substances in the leaf. This extracting and purification
process requires the use of acids, bases, water, and organic solvents that are added to the
process along the different stages until the pure alkaloid or coca paste is obtained. Of
these substances, organic solvents are recycled and acids (sulfuric acid) and bases

(ammoniac), and water are dumped on the environment without any control. Likewise
vegetation residues are produced. They get contaminated along the process and then are
dumped. Based on analyses made, in the production of 1 kg of coca paste, 1.9 litres of
sulphuric acid, 1.25 litres of ammonia, 193,75 litres of contaminated water, and
625 kg of solid waste are released into the environment.

7. In the study zone, the following amounts of chemical substances and waste were
released in the 1999-2001 analysis period. 91,962 litres of sulphuric acid or 434 tins of
55 gallons each, 60.501 liters or 286 tins of ammonia, and 9.378 m3 of contaminated
water and 30 tons 0f vegetation waste were poured. Therefore, there is a great chance
that of existence of these substances on superficial water and soil, which affects the
populations that depend on these two components.

276Processing of coca leaf requires a great deal of chemical precursors and water to extract
the alkaloid (DNE, 2002). According to data obtained from the Antinarcotics Police, it
is estimated that every hectare of coca crops requires the use of approximately 127
kilos/ha of solid precursors, 447 litres/ha of liquid precursors and 400 litres/ha of water

(DIRAN 2002).

[…]

[Page 10 and 11]

8. CONCLUSIONS

[…]

5. after establishing the types of pesticides used by coca growers, it was possible to
identify 5 pesticides that due to its use intensity and toxicological classification are
classified from extremely to highly toxic. These pesticides are: herbicides such as
Gramaxone (A.i. paraquat), Faena (A.i. glyphosate), insecticides such as Tamaron (I.a.
Metamidaphos), and fungicides such as Manzate (I.a. Mancozeb). The analysis allowed

to establish that the risk of substances accumulation per environmental component and
the monitoring needs are: In soils, substances that need to be monitored are Gramaxone
and Faena, in underground waters, Anikilamina and Tamaron, and in superficial water
sediments all these compounds must be monitored.

6. Processing of coca is made in order to extract or wash the alkaloid, which represents
only 0,5 to 1,5 % of the total substances in the leaf. This extracting and purification

process requires the use of acids, bases, water, and organic solvents that are added to the
process along the different stages until the pure alkaloid or coca paste is obtained. Of
these substances, organic solvents are recycled and acids (sulfuric acid) and bases
(ammoniac), and water are dumped on the environment without any control. Likewise
vegetation residues are produced. They get contaminated along the process and then are
dumped. Based on analyses made, in the production of 1 kg of coca paste, 1.9 litres of
sulphuric acid, 1.25 litres of ammonia, 193,75 litres of contaminated water, and
625 kg of solid waste are released into the environment.

M IGUELSAN SEBASTIÁ& ANNA -KARINHURTIG,“OIL EXPLOITATION IN THE
AMAZON BASIN OFECUADOR:A PUBLIC HEALTH EMERGEN”,PAN AM JPUBLIC
HEALTH, 15(3):205-211,2004

(Pan Am J Public Health, 15(3):205-211, 2004)

289Annex 129

Temas de actualidad / Current topics

Oil is a major source of income for Ecuador and
Oil exploitation in
since the 1970s has been the “engine” of the nation’s
economy. Before the 1970s oil price boom, Ecuador
the Amazon basin was one of the poorest countries in Latin America.
Since then, oil production has been the primary
of Ecuador: a public cause of Ecuador’s economic growth, which has av-

health emergency eraged 7% annually. Per capita income rose from
US$ 290 in 1972 to US$ 1 200 in 2000. Today, oil con-
tinues to account for 40% of the nation’s export
earnings and of the budget of the national Govern-
1 ment (1, 2). Most of this oil comes from the north-
Miguel San Sebastián and
Anna-Karin Hurtig 1 eastern part of the country, the Amazon basin.
The Amazon basin of Ecuador, known as el
Oriente (the provinces of Sucumbios, Orellana,
Napo, Pastaza, Morona Santiago, and Zamora-
2
Chinchipe), consists of more than 100 000 km of
tropical rain forest lying at the headwaters of the
Amazon river network. The region contains one of
the most diverse collections of plant and animal life
in the world (3). The Oriente region is also the home

of some 500 000 people, or about 4.5% of the coun-
try’s population. These half-million persons include
eight groups of indigenous people as well as peas-
ants who, encouraged by land policies of the na-
tional Government, moved to the area from Ec-

uador’s coastal and highland regions in the 1970s
and the 1980s (4).
In 1967 a Texaco-Gulf consortium discovered
a rich field of oil beneath the rain forest, leading to
an oil boom that has permanently reshaped the

region. The Amazon of Ecuador now houses a
vast network of roads, pipelines, and oil facilities.
While the national Government has retained do-
minion over all mineral rights, several private for-
eign companies have built and operated most of the

oil infrastructure.
Current oil production activities in the Oriente
region span nearly one million hectares, with over
300 producing wells and 29 production camps. The

country has 4.6 billion barrels of proven oil reserves,
with crude production of around 390 000 barrels per
day. Of this production, Petroecuador, the Govern-
Key words: petroleum, extraction and processing ment-owned company, accounts for about 55% of
industry, environmental pollution, water pollution, Ecuador’s total output, with private companies ac-
Ecuador.
counting for the remaining 45%. Petroecuador is at-
tempting both to attract foreign investment to the
1 Umeå University, Umeå International School of Public Health, Pub-rgest oil fields and to boost its own pro-
lic Health and Clinical Medicine, Umeå, Sweden, and Iduction from around 215 000 barrels per day today
Epidemiología y Salud Comunitaria “Manuel Amunárriz,”to 600 000 barrels per day by 2005 (5).
bastián, Public Health and Clinical Medicine, Umeå Internationale-
School of Public Health, Umeå University, SE-901 85 Umeå, Sweden;967 many different companies have
telephone: 00-46-90 7851328; e-mail: [email protected] in the oil exploitation process. There
umu.se are currently 16 companies operating in the coun-

Rev Panam Salud Publica/Pan Am J Public Hea1l5th(3), 2004 205

290 Annex 129

try: Petroecuador, 3 private Ecuadorian companies, THE ENVIRONMENTAL EXPOSURE
and 12 foreign companies (6). Figure 1 shows the oil
companies now operating in the country and the Source and extent of pollution
blocks where they are located.

Since the beginning of oil exploitation, foreign Oil development activities include several
oil companies and Petroecuador have extracted contaminating processes. The extent of these pollut-
more than two billion barrels of crude oil from the ing processes depends mainly on the environmen-

Ecuadorian Amazon. However, in this development tal practices and technology used by oil companies.
process, billions of gallons (1 gallon = 3.7853 liters) In Ecuador these practices have repeatedly been
of untreated wastes, gas, and crude oil have been questioned (8–10).
released into the environment (7). Deep below the earth’s surface, oil is usually

This paper examines the environmental and mixed with natural gas and “formation water,”
health impacts brought about by the oil develop- which contains hydrocarbons, heavy metals, and a
ment process in the Amazon region of Ecuador. high concentration of salts. In the Amazon basin of

FIGURE 1. Oil blocks operated by oil companies, Ecuador, 2003

†
COLOMBIA

Galapagos
Tarapoa

PACIFIC OCEAN

Espol

Legend
1 Canada Grande 17 Vintage
3 EDC 18 Ecuador TLC
7 Perenco 21 Perenco
10 Agip 23 CGC
11 Lumbaqui Oil 24 Burlington
14 Vintage 27 City
15 Occidental 28 Tripetrol
PERU 16 Repsol-YPF 31 Perez Companc
Tarapoa block AEC Ecuador

Source:Petroecuador (http://petroecuador.com.ec/donde.htm). Used with permission.

206 Temas de actuali• Current topics

291Annex 129

Ecuador, each exploratory well that is drilled pro- Environmental analysis
duces an average of 4 000 cubic meters of drilling
wastes, including formation water and drilling Numerous reports have indicated that the
muds (which are used as lubricants and sealants). contamination has occurred since the beginning of

These wastes were frequently deposited into open, the oil exploration in the Ecuadorian Amazon (8–
unlined pits called separation ponds, from which 10, 15) even though longitudinal data on the levels
they were either directly discharged into the envi- of population exposure over time do not exist.
ronment or they leached out as the pits degraded or A study in 1987 by the Ecuadorian Govern-
overflowed from rainwater (7, 8). Although some ment found elevated levels of oil and grease in all

companies have modified this practice in the last of the 36 samples taken from rivers and streams
10 years by building protected ponds, these prac- near productions sites. That study also found that
tices still occur. There are currently nearly 200 open a shortage of dissolved oxygen in the majority of
ponds in the Amazon region (11). water samples had seriously harmed the aquatic

If commercial quantities of oil are found, the ecosystem (16). In 1989 another Ecuadorian Gov-
production stage starts. During production, oil is ernment study of 187 wells found that crude oil was
extracted in a mixture with formation water and regularly dumped into the forests and into bodies
gas and then separated in a central facility. At each of water (13).
facility, over 4.3 million gallons (16.3 million li- In 1994 a study carried out by the Ecuadorian

ters) of liquid wastes are generated every day and environmental and human rights organization Cen-
discharged without treatment into pits. Roughly tro de Derechos Económicos y Sociales (the Center
53 million cubic feet (1.5 million cubic meters) of for Economic and Social Rights) also found highly
“waste” gas from the separation process is burned elevated levels of oil pollutants in the streams and

daily without temperature or emissions controls. rivers of the Oriente area. Concentrations of poly-
Air contamination can also be generated at pits and nuclear aromatic hydrocarbons were 10 to 10 000
oil spills by hydrocarbons coming from standing oil times greater than the levels recommended by the
slicks (1, 7). Environmental Protection Agency of the United States
Routine maintenance activities at over 300 of America (9).

producing wells discharge an estimated five mil- In 1998 an independent local laboratory that
lion gallons (18.9 million liters) of untreated toxic is frequently used by the oil companies surveyed
wastes into the environment every year. Leaks from 46 streams in the Oriente region (17). The labora-
wells and spills from tanks have been common (12). tory found contamination by total petroleum hy-

According to a study conducted by the Govern- drocarbons (TPH) in areas of oil activities, while no
ment of Ecuador in 1989, spills from the flowlines water contamination was found in areas without
that connect the wells to the stations were dumping such activities.
an estimated 20 000 gallons (75 800 liters) of oil In 1999 the Instituto de Epidemiología y Salud
every two weeks (13). Comunitaria “Manuel Amunárriz” (“Manuel Amuná-

Spills from the main and secondary pipelines, rriz” Institute of Epidemiology and Community
which connect the separation stations to the re- Health), a local nongovernmental organization con-
finery in the coastal region, are also common. In cerned with health issues, undertook water analyses
1992 the Ecuadorian Government recorded approx- for TPH in communities near oil fields and also in

imately 30 major spills, with an estimated loss of communities far away from the fields. Those analy-
16.8 million gallons (63.6 million liters) of crude oil ses showed high levels of TPH concentrations in
(7). In 1989 a spill of at least 294 000 gallons (1.1 mil-rivers used by the communities that were close to
lion liters) of crude oil caused the Napo River, the oil fields. In some streams, hydrocarbon concen-
which has a width of one km, to run black for a trations exceeded by more than 100 times the limit

week; the same thing happened in 1992, when there permitted by European Community regulation (18).
was a spill of about 275 000 gallons (1.0 million Since 1999 the oil companies have been re-
liters) of crude oil (12). It was estimated in 2002 that quired by law to regularly monitor the level of pol-
two big spills per week were occurring from the lution in the environment and to send reports to the

main oil fields in the Oriente region (14). national Government of Ecuador. This information
Overall, during the period of 1972 through is not open for public scrutiny. However, in 1999,
1993, more than 30 billion gallons (114 billion liters) when one of these reports was presented to a com-
of toxic wastes and crude oil were discharged into munity that had made several complaints to the
the land and waterways of the Oriente (7). This Ministry of Environment, it showed that streams

compares to the 10.8 million gallons (40.9 million in the community had concentrations of TPH that
liters) spilled in the Exxon Valdez tanker disaster in were over 500 times the limit permitted by Euro-
1989 in Alaska, one of the largest sea oil spills that pean Community regulations (19). Nevertheless,
has ever occurred. the oil company and a representative of the Ecua-

Rev Panam Salud Publica/Pan Am J Public Hea1l5th(3), 2004 207

292 Annex 129

dorian Government insisted that the levels that had fects of oil (27). Another study found that the risk
been found were acceptable. of spontaneous abortions was 2.5 times as high in
For the Amazon basin of Ecuador, there is a women living in the proximity of oil fields (28).
lack of data on soil pollution and its possible im- Research done in 1998 found an excess of
pact, and no study has been conducted on the im- cancers among males in a village located in an oil-

pact that oil development has had on fish and fish- producing area in the Oriente region (29). Another
ing. However, studies from the Amazon basin of study, from 2000, examined the differences in can-
Peru found, after an oil spill in the Marañon River, cer incidences over the period of 1985 to 1998 in the
high concentrations of TPH in the stomach and Amazon region of Ecuador. This study found a sig-

muscles of fish (20). nificantly higher overall incidence of cancer in both
men and women in the cantones (“counties,” or
divisions of provinces) where oil exploitation had
THE HEALTH EFFECTS been going on for at least 20 years. Significantly el-
evated levels were observed for cancers of the

Several studies have focused on residents ex- stomach, rectum, skin melanoma, soft tissue, and
posed to major coastal oil spills from tankers (21– kidney in men and for cancers of the cervix and
23). However, there are few epidemiological stud- lymph nodes in women. An increase in hematopoi-
ies concerning persons who live in communities etic cancers was observed in children (30).

that are near oil fields and who are exposed to acute
and/or long-term contamination (24).
For many years residents of the oil-producing GOVERNMENT RESPONSES
areas of the Ecuadorian Amazon have raised con-
cerns over pollution related to oil development. Peasants and indigenous people from the

Both peasants and indigenous people have reported Amazon have presented their complaints to vari-
that many local streams and rivers, once rich in fish, ous administrations of the national Government of
now support little or no aquatic life; further, cattle Ecuador. The inhabitants of the Ecuadorian Ama-
are reported to be dying from drinking from con- zon have asked for a better quality of life and for

taminated streams and rivers. These are typically technical assistance; that electricity, water, health
the same waters that people use for drinking, cook- services, and other basic services be provided; and,
ing, and bathing. Residents have also reported that above all, that the oil pollution be remediated.
bathing in the river waters causes skin rashes, espe- Through their own organizations and with support
cially after heavy rains, which accelerate the flow of from national environmental groups, Oriente resi-

wastes from nearby pits into the streams (25). dents have demanded that the companies clean up
In 1993 a community health workers asso- the environmental pollution and compensate them
ciation in the Ecuadorian Amazon conducted a for damages caused by oil-related contamination.
descriptive study in its communities. The study The measures adopted so far by oil companies and

suggested that, compared to communities free from the various administrations of the national Govern-
oil exploitation, communities in oil-producing areas ment have been described as “patches,” such as cov-
had elevated morbidity rates, with a higher occur- ering some waste pits, building some schools, and
rence of abortion, dermatitis, skin mycosis, and mal- constructing roads, all without facing the root
nutrition, as well as higher mortality rates (26). causes of the problem (10, 31, 32).

In 1994 the Center for Economic and Social Various administrations of the national Gov-
Rights released a study reporting skin problems ernment of Ecuador have declared the essential im-
(dermatosis) in the population in the Ecuadorian portance of oil to Ecuador’s development. However,
Amazon, apparently related to crude oil contami- despite the oil revenues, improvements in socioeco-

nation of local rivers (9). nomic conditions in the country have fallen short of
In recent years the “Manuel Amunárriz” Insti- expectations. Ecuador now has the highest per
tute of Epidemiology and Community Health has capita debt of any country in South America, nearly
been involved in a research process to assess the po- US$ 1 100 per person (1). In the period from 1970
tential health impact of oil pollution in communities to 2002 the unemployment rate rose from 6.0% to

near oil fields. In the first of these studies, women 7.7%, and the percentage of people living in poverty
living in communities near oil fields reported climbed from 47.0% to 61.3% (2, 33). The ratio of the
higher rates of various physical symptoms than did income received by the poorest 5% of the population
women in control areas. These symptoms included and by the richest 5% changed from 1:109 in 1988 to

skin mycosis, tiredness, itchy nose, sore throat, head- 1:206 in 1999 (34). The Amazon region has the worst
ache, red eyes, ear pain, diarrhea, and gastritis. Af- infrastructure and the lowest socioeconomic and
ter adjustment for possible confounding factors, health indicators in the country (35).
the symptoms significantly associated with exposure In response to the nearly $16 billion in exter-
were those expected from known toxicological ef- nal debt that Ecuador has, one of the main eco-

208 Temas de actuali• Current topics

293Annex 129

nomic strategies of the national Government and tion standards and environmental management
the International Monetary Fund has been to ex- plans should be accessible to and appropriately

pand the oil exploitation in the country. The na- discussed with communities and independent en-
tional Government’s proposals include opening vironmental groups. Without such basic informa-
two million hectares of pristine rain forest in the tion, these groups are left unaware of potential
south of the Amazon to oil exploitation and con- risks, they cannot participate meaningfully in for-
structing a new heavy crude oil pipeline in the mulating public policy, and they cannot hold

north of the Amazon, to allow further oil exploita- companies accountable for their actions. In addi-
tion in that area (36, 37). tion, an environmental monitoring system should
be established, with the involvement of the af-
fected communities. As a minimum, this system

WHAT NEEDS TO BE DONE should include regular detailed chemical sam-
pling of the environment and reporting on the
Modern oil and gas development, if compa- emissions and effluent controls.
tible with sustainable development and the well- • Oil development policies have an impact on
being of Amazonian peoples, must be based on health, and the consequences of those policies

comprehensive environmental planning that fully need to be assessed and taken into account. The
considers the cumulative impact of ongoing and Ecuadorian Government should acknowledge the
planned oil exploitation throughout the region. need for health impact assessments as an inte-
Strict environmental controls and careful long-term gral feature of policy development and evalua-

monitoring of oil activities—with both of those tion. Community consultation and participation
firmly grounded in the rule of law and broad par- are essential in assessing impacts on the environ-
ticipation by local communities, local governments, ment and health (42).
and nongovernmental groups—are necessary in • Ecuador enacted a new constitution in 1998. That
order to prevent further negative environmental document acknowledges the right of communi-

and health impacts in the Oriente region (38). Five ties to be consulted by oil companies before the
interrelated actions are urgently needed: companies begin the exploratory stage of oil de-
velopment. To enforce these rights, it is essential
• The Ecuadorian Government should conduct an for community organizations to work with re-

evaluation of the environmental situation in the gional, national, and international environmental
Oriente region. It is also necessary to develop and groups. The Ecuadorian Government has already
oversee the implementation of a plan to repair the given a commitment to develop mechanisms to
damage that has already occurred and to limit enforce the laws protecting the environment and
further destruction. While oil pollution persists, the health of their citizens, but developing those

the health of the population of the Oriente area mechanisms will be difficult. This should be ad-
and other populations in similar situations will dressed within the context of promoting human
remain at risk. Some indigenous and environ- rights, combating corruption, and strengthening
mental groups have called for the application of democratic institutions.

the precautionary principle. (The precautionary • Concern has been raised around the world that
principle has been defined as “when an activity globalization of trade does not bode well for the
raises threats of harm to human health or the en- environment and for people’s health (43–45).
vironment, precautionary measures should be Shifting trade policies in the direction of environ-
taken even if some cause and effect relationships mental sustainability and social justice is urgently

are not fully established scientifically” (39)). That needed if environmental protection, economic se-
principle has been developed by scientists in the curity, and health benefits are to be received by
face of scientific uncertainty, and it is a strong call the majority of the world’s population.
for prevention of potential harm and for caution

in actions taken. Those indigenous and environ- We believe that oil exploitation in the Ama-
mental groups have also asked the national Gov- zon basin of Ecuador has resulted in a public health
ernment for a moratorium on oil and gas de- emergency because of its adverse impact on the en-
velopment in new areas of the Amazon. Such vironment and health. So far, the Ecuadorian Gov-
development alternatives as ecotourism and rain ernment has not designed an adequate strategy to

forest conservation have been proposed, and they prevent further negative environmental and health
should be seriously considered (40, 41). impacts. The oil industry argues that it has a role to
• Oil companies operating in the Ecuadorian Ama- play in the development of the country (46-48), but
zon should change their practices to minimize en- that development should not come with the added

vironmental impacts and to build partnerships cost of pollution and poor health.
with local communities so that local residents At first, it may appear that the oil industry
benefit from development. Environmental protec- and public health are not related. However, we

Rev Panam Salud Publica/Pan Am J Public Hea1l5th(3), 2004 209

294 Annex 129

have shown that they are closely interconnected. her professional affiliation and mailing address, and
Unfortunately, Ecuador is not the only country in should list any potential conflicts of interest related
Latin America to suffer the negative consequences to the subject of the letter. Letters are welcome in

of oil exploitation; Bolivia, Colombia, and Peru are English, Spanish, or Portuguese. Letters can be sent
in a similar situation (49, 50). There are already via electronic mail to: publiper@ paho.org; letters can
public health problems, and these problems may also be sent via postal mail to: Editor,Revista Pan-
grow if unregulated oil exploitation continues to americana de Salud Pública/Pan American Journal of

expand in Latin America. Preventing additional Public Health, 525 Twenty-third Street, N.W., Wash-
health and environmental damage will require ac- ington, D.C. 20037, United States of America.
tion on a local, national, and international level.

Acknowledgments. The studies on the health
impact of oil exploitation in Ecuador carried out by
SINOPSIS
the “Manuel Amunárriz” Institute of Epidemiology
and Community Health have been funded by the
Vicariato de Aguarico (the local Catholic church) La explotación petrolera en la cuenca
amazónica de Ecuador: una emergencia
and Medicus Mundi Gipuzkoa (a Spanish non- para la salud pública
governmental organization).

Note on conflicts of interest. In 1993 a law- Desde la década de 1970, el petróleo ha sido una de las prin-
cipales fuentes de ingresos del Ecuador y ha servido como
suit was filed in the United States of America “motor impulsor” de la economía nacional. La mayor parte
against Texaco, an oil company that had worked in del petróleo ecuatoriano se extrae en la cuenca amazónica del
Ecuador for more than 20 years. The plaintiffs—
nordeste del país. Desde que comenzó la explotación petrolera,
some 30 000 indigenous persons and peasants— compañías extranjeras y la empresa petrolera estatal Petro-
claimed that the oil company had caused irrepara- ecuador han extraído más de dos mil millones de barriles de
ble damage to the rain forest of the Amazon region petróleo crudo de la Amazonía ecuatoriana. A lo largo de este
proceso se han liberado al medio ambiente miles de millones de
of Ecuador. In 2000 the suit was dismissed in the galones de desechos sin tratar, gas y petróleo crudo. Este artí-
United States and sent to Ecuador to be considered.
In October 2003 one of the authors, Miguel San culo analiza el impacto ambiental y sanitario provocado por el
desarrollo petrolero en la región amazónica del Ecuador. Por
Sebastián, presented testimony in an Ecuadorian ejemplo, el análisis del agua de varias corrientes fluviales de
court on behalf of the plaintiffs; he did not receive la localidad ha demostrado la presencia de altas concentracio-
any payment for his testimony. nes de productos químicos derivados del petróleo en las zonas
While the Revista Panamericana de Salud
petrolíferas en explotación. Los estudios epidemiológicos han
Pública/Pan American Journal of Public Health is affil- encontrado un mayor riesgo de sufrir síntomas asociados con
iated with the Pan American Health Organization el petróleo y abortos espontáneos en las mujeres que viven en
(PAHO), the Revista/Journal is an independent sci- las proximidades de los campos petroleros. También se ha en-
contrado una incidencia excesiva de cáncer. Se necesitan in-
entific publication whose articles do not necessarily
reflect the opinions or official positions of PAHO or tervenciones locales, nacionales e internacionales para evitar
of its Member States on specific issues. que se empeoren los efectos negativos que ejerce sobre el medio
ambiente y la salud el desarrollo petrolero. Estas intervencio-
nes deben abarcar un sistema de monitoreo y remediación am-
Note on alternative points of view.The Re- biental, consultas a la comunidad y participación comunita-
vista Panamericana de Salud Pública/Pan American Jour- ria, mecanismos para hacer cumplir las leyes que protegen el
nal of Public Health welcomes letters to the editor that
medio ambiente y la salud de la población, y cambios en las
clarify, discuss, or comment in a constructive man- políticas comerciales dirigidos a lograr la sostenibilidad en
ner on ideas presented in theRevista/Journal.Letters materia ambiental y la justicia social.
should be signed by the author, should specify his or

REFERENCES

1. Centro de Derechos Económicos y So- http://www.ildis.org.ec/estadisticas/4. Fundación José Peralta. Ecuador: su rea-
ciales. El petróleo no es eterno. Quito:estadisticas.htm. Accessed 25 May 2003. lidad. Quito: Fundación de Investiga-
Centro de Derechos Económicos y So- 3. Ecuador rainforest. Available from: ción y Promoción Social José Peralta;
ciales; 1999. http://www.rainforestweb.org/ 2001.
2. Instituto Latinoamericano de Investiga-Rainforest_Regions/South_America/ 5. United States of America, Environ-
ciones Sociales. Economía ecuatoriana Ecuador/?state=more. Accessed 25 May mental Information Agency. Ecuador:
en cifras, 1970–2003. Available from: 2003. country analysis brief. Available from:

210 Temas de actualid• Current topics

295Annex 129

http://www.eia.doe.gov/cabs/ecuador. derrame de petróleo sobre los recursos 35. Terán C. Sucumbios 2000. Lago Agrio,
html. Accessed 28 May 2003. hidrobiológicos entre San José de Sara- Ecuador: Vicariato de Sucumbíos; 2000.
6. Petroecuador. Bloques concesionados. muro y Nauta, Río Marañón. Iquitos, 36. World Bank. Ecuador overview. Avail-

Available from: http://www.petroecuador. Perú: Dirección Regional de Pesquería; able from: http://www.worldbank/
com.ec/donde.htm Accessed 28 May 2000. org Accessed 28 May 2003.
2003. 21. Campbell D, Cox D, Crum J, Foster K, 37. Centro de Derechos Económicos y So-
7. Jochnick C, Normand R, Zaidi S. Rights Christie P, Brewster D. Initial effects ciales. Apertura 2000 . . . la solución al
violations in the Ecuadorian Amazon: of the grounding of the tanker Braer país? Boletín número 2, marzo. Quito:
the human consequences of oil develop- on health in Shetland. BMJ. 1993;307: Centro de Derechos Económicos y So-
ment. Health Human Rights. 1994;1(1): 1251–5. ciales; 2000.
82–100. 22. Palinkas LA, Petterson JS, Russell J, 38. Kimerling J. The human face of petro-

8. Kimerling J. Amazon crude. New York: Downs MA. Community patterns of leum: sustainable development in Ama-
Brickfront Graphics Inc.; 1991. psychiatric disorders after the Exxon zonia? Rev Eur Community Int Environ
9. Centro de Derechos Económicos y So- Valdez oil spill. Am J Psychiatry. 1993; Law. 2001;10(1):65–81.
ciales. Violaciones de derechos en la Ama- 150(10):1517–23. 39. Raffensperger C, Tickner J. Protecting
zonía Ecuatoriana. Quito: Abya-Yala; 23. Lyons RA, Temple MF, Evans D, Fone public health and the environment:
1994. DL, Palmer SR. Acute health effects of implementing the precautionary prin-
10. Varea A, Ortiz P, eds. Marea negra en la the Sea Empress oil spill. J Epidemiol ciple. Washington, D.C.: Island Press;

Amazonía: conflictos socioambientales Community Health. 1999;53(5):306–10. 1999.
vinculados a la actividad petrolera en el4. Falaschi C, ed. Informe socio ambiental40. Centro de Derechos Económicos y So-
Ecuador. Quito: Abya-Yala; 1995. sobre Loma de la Lata. Available from: ciales. Una opción para el país: deuda
11. Frente de Defensa de la Amazonía- http://www.ecoportal.com.ar/articulos/ por conservación de la Amazonía.
Petroecuador. Estudio para conocer el lomalata.htm Accessed 26 May 2003. Quito: Centro de Derechos Económicos
alcance de los efectos de la contamina- 25. Kimerling J. Rights, responsibilities, andy Sociales; 2000.
ción en los pozos y estaciones perfora- realities: environmental protection law 41. Acosta A. ¿Es posible la transición a una
dos antes de 1990 en los campos Lago in Ecuador’s Amazon oil fields. South- economía post petrolera? In: Martínez E,

Agrio, Dureno, Atacapi, Guanta, Shushu- western J Law Trade Americas. 1995; ed. El Ecuador post petrolero. Quito: Ac-
findi, Sacha, Yuca, Auca y Cononaco. 2(2):293–384. ción Ecológica; 2000. Pp. 38–55.
Quito: Frente de Defensa de la Amazo- 26. Unión de Promotores Populares de 42. British Medical Association. Health and
nía-Petroecuador; 2003. Salud de la Amazonía Ecuatoriana. Cul- environmental impact assessment: an
12. Almeida A. Reseña sobre la historia eco- turas bañadas en petróleo: diagnóstico integrated approach. London: Earthscan;
lógica de la Amazonía ecuatoriana. In: de salud realizado por promotores. 1998.
Martínez E, ed. El Ecuador post petro- Quito: Abya-Yala; 1993. 43. Korten D. When corporations rule the
lero. Quito: Acción Ecológica; 2000. Pp.27. San Sebastián M, Armstrong M, Ste- world. London: Earthscan; 1995.

27–38. phens C. La salud de mujeres que viven 44. United Nations Environmental Pro-
13. Dirección General de Medio Ambiente cerca de pozos y estaciones de petróleo gramme. Global environment outlook.
de Ecuador. Estudio de impacto am- en la Amazonía ecuatoriana. [The health Nairobi, Kenya: Earthscan Publications;
biental 42. Quito: Dirección General de of women who live near oil wells and oil 1999.
Medio Ambiente; 1989. production stations in the Amazon re- 45. Stephens C, Lewin S, Leonardi G, San Se-
14. 2 derrames de petróleo al mes en el campo gion of Ecuador] Rev Panam Salud Pu- bastián M, Shaw R. Health, sustainability
Auca. El Comercio, 18 de octubre 2002. blica. 2001;9:375–84. and equity: global trade in the brave new
15. Fabra A. Indigenous peoples, environ- 28. San Sebastián M, Armstrong M, Stephens world. Global Change Human Health.

mental degradation and human rights: C. Outcome of pregnancy among women 2000;1(1):44–58.
a case study. In: Boyle A, Anderson M, living in the proximity of oil fields in theShell. The Shell report 2002. Meeting the
eds. Human rights approaches to envi- Amazon basin of Ecuador. Int J Occup energy challenge. Available from: http://
ronmental protection. Oxford: Claren- Environ Health. 2002;8:312–9. www.shell.com/home/Framework?site
don Press; 1998. Pp. 245–64. 29. San Sebastián M, Armstrong M, Cor- Id=shellreport2002-en Accessed 30 May
16. Corporación Estatal Petrolera Ecuato- doba JA, Stephens C. Exposures and 2003.
riana. Análisis de la contaminación am- cancer incidence near oil fields in the 47. British Petroleum. Environment and so-
biental en los campos petroleros Liberta- Amazon basin of Ecuador. Occup Envi- ciety. Available from: http://www.bp.

dor y Bermejo. Quito: CEPE; 1987. ron Med 2001;58:517–22. com/environ_social/index.asp Accessed
17. Zehner R, Villacreces LA. Estudio de la30. Hurtig AK, San Sebastián M. Geograph- 30 May 2003.
calidad de aguas de río en la zona de ical differences of cancer incidence in 48. Oxy. Social responsibility. Available
amortiguamiento del Parque Nacional the Amazon basin of Ecuador in relation from: http://www.oxy.com/HTML/
Yasuní. Primera fase: monitoreo de aguas to residency near oil fields. Int J Epi- socialrespons.html Accessed 30 May
— screening Octubre de 1997. Coca, demiol 2002;31:1021–7. 2003.
Ecuador: Laboratorio de Aguas y Suelos 31. Frente de Defensa de la Amazonía. La 49. La Torre López L. ¡Sólo queremos vivir

P. Miguel Gamboa-Fepp; 1998. Texacontaminación en el Ecuador. Lago en paz! Experiencias petroleras en terri-
18. San Sebastián M. Informe yana curi: Agrio, Ecuador: Frente de Defensa de la torios indígenas de la Amazonía pe-
impacto de la actividad petrolera en Amazonía; 1999. ruana. Copenhague: Grupo Internacio-
la salud de poblaciones rurales de la 32. Acción Ecológica. Amazonía por la vida. nal de Trabajo sobre Asuntos Indígenas;
Amazonía ecuatoriana. Quito: Cicame & Available from: http://www.accion 1998.
Abya-Yala; 2000. ecologica.org/petroleo.htm Accessed 50. Oilwatch. Fluye el petróleo, sangra la
19. Ministerio de Medio Ambiente de Ecua- 27 May 2003. tierra. Quito: Oilwatch; 1999.
dor. Informe de inspección ambiental al 33. Instituto Nacional de Estadísticas y Cen-

área de las comunidades Flor de Man- sos del Ecuador. VI censo de población y
duro y Centro Manduro ubicadas en el V de vivienda. Quito: INEC; 2001.
bloque siete operado por la compañía 34. Acosta A. El petróleo en el Ecuador: una
Oryx. Quito: Ministerio de Medio Am- evaluación crítica del pasado cuarto de
biente; 1999. siglo. In: Martínez E, ed. El Ecuador post
20. Perú, Dirección Regional de Pesquería petrolero. Quito: Acción Ecológica; 2000Manuscript received 3 July 2003. Accepted for
de Loreto. Monitoreo del impacto post- Pp. 2–19. publication on 17 November 2003.

Rev Panam Salud Publica/Pan Am J Public Hea1l5th(3), 2004 211

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K.R. OLOMON ET AL ., AESPONSE TOTOMÁS LEÓN SICARD ET A.,NTER-
A MERICAN DRUG ABUSE CONTROL C OMMISSION(CICAD), O RGANIZATION OF
A MERICAN STATES,2005

(Available at:
http://www.cicad.oas.org/Desarrollo_Alternativo/ESP/Colombia/Panel%20re…

Sicard%20et%20al.pdf (last visited 7 March 2010))

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309310 Annex 131

JOURNAL OF TOXICOLOGY AND ENVIRONMENTAL HEALTH,PART A: CURRENT
ISSUE, VOLUME 72, UMBERS 15-16,AUGUST1 ANDA UGUST 152009
SPECIALISSUE: RODUCTION OF LLICITDRUGS ,THEENVIRONMENT AND H UMAN
H EALTH

(Taylor & Francis, London, 2009)

311Annex 131

312 Annex 131

313314 Annex 131-A

Annex 131-A

K.R. OLOMON ET AL., “UMANH EALTH ANDENVIRONMENTAL RISKS FROM THE
USE OFGLYPHOSATE FORMULATIONS TOCONTROL THEP RODUCTION OC OCA IN
COLOMBIA: OVERVIEW ANDCONCLUSIONS”

(Journal of Toxicology and Environmental Health, Part A, 72:914-920, 2009)

315 Annex 131-A

Journal of Toxicology and Environmental Health, Part A, 72: 914–920, 2009
ISSN: 1528-7394 print / 1087-2620 online
DOI: 10.1080/15287390902929659

UTEH
Human Health and Environmental Risks from the Use

of Glyphosate Formulations to Control the Production

of Coca in Colombia: Overview and Conclusions

Keith R. Solomon , E. J. P. Marshall , and Gabriel Carrasquilla 3
1
Centre for Toxicology and Department of Environmental Biology, University of Guelph, Guelph,
Ontario, Canada, 2Marshall Agroecology Limited, Barton, Winscombe, Somerset, United Kingdom, and
3
Facultad de Salud, Universidad del Valle, Cali, Colombia

Much of the world’s cocaine originates in Latin America were identified included the influence of spray procedures and
with the majority of the supply coming from Colombia. The
conditions on spray drift and the toxicity of the spray mixture to
control of the coca plants from which cocaine is produced amphibians, with ancillary questions related to alternative prod-
(Erythroxylum coca and E. novogranatense) has been the focus ucts and mixtures that would pose less risk to amphibians and

of considerable effort and expenditure. As part of the supply the distributions of amphibians in relation coca production and
control effort that started in the 1970s, an aerial spraying eradi-the spray program (Solomon et al., 2007b). Other questions have
cation program in Colombia was started in 1997 using the her- also been raised, such as in a recent report in the literature of

bicide glyphosate. The total area planted with coca was 99,000 effects of glyphosate use on humans (Paz-y-Miño et al., 2007),
ha and the cumulative area of coca sprayed with glyphosate was which suggested that drift of spray was affecting humans at dis-

153,134 ha in 2007, 11% less than in 2006 (UNODC 2008). tances of several km from areas of application.
The potential human and environmental risks related to the As a result of these questions, several studies were initiated
use of glyphosate for controlling coca plants have generated to collect data and to test specific hypotheses. These studies

considerable interest and attention in Colombia and other have been completed and are published as a series of articles in
countries. At the request of the Organization of American this issue of Journal of Toxicology and Environmental Health.
States (OAS), an independent scientific review of this issue This article is an overview of the results of these studies which

was completed in 2005 by an expert panel (Solomon et al., places them in the context of other recent publications on the
2005). This review, published in the scientific literature in spray program and the effects of glyphosate on the environment
Downloaded At: 20:07 2 October 2009
2007, noted that, at the time, knowledge of the toxicity of gly- and on human health. As is common for all risk assessments
phosate and its formulated products did not suggest significant (U.S. EPA, 1998), this overview addresses both exposures and
risks to humans or most wildlife (Solomon et al., 2007b). In effects and draws on the data in these articles tointegrate these

addition to assessing the toxicological effects of the eradication observations into a risk assessment and overview.
spray program, the review pointed out that some of the chemicals
used in the production and refining of the final product, cocaine
EXPOSURES IN THE ENVIRONMENT
hydrochloride, presented potential hazards to humans and the
environment (CICAD/OAS 2004, 2005; Solomon et al., 2007a). As exposure is the primary driver of risks, the first article in
the series relates to spray drift (Hewitt et al., 2009). One of the
However, the primary review noted that there were several out-
standing questions and issues. Key environmental issues that primary questions addressed in this article was the effect of
spray conditions on droplet size and the potential for spray

drift. Until recently, the aircraft currently used for eradication
©General Secretariat of the Organization of American States, spraying in Colombia were the OV-10 and the AT 802; however,
2009. This paper was prepared as part of a Study entitled “Productithe only one in current use is the AT 802 (National Police,
of Illicit Drugs, the Environment and Human Health,” financed with
contributions from the Governments of Colombia and the United Colombia, personal communication, September 2008). The
States of America. The conclusions and opinions expressed herein arepeed during spray application is 333 km/h for the OV-10 and
those of the authors and not necessarily those of the Organization 274 km/h for the AT-802, both in excess of the speeds used in

American States and its General Secretariat, which as of the date oconventional agricultural spray applications (approximately
this copyright, have not formulated any opinion with respect to the200 km/h). The greater speed of these aircraft, necessitated by
Address correspondence to Keith R. Solomon, Centre for
Toxicology and Department of Environmental Biology, University of the need to avoid hazards such as gunfire from the growers of
Guelph, Guelph, ON, N1G 2W1, Canada. E-mail: ksolomon@ illicit crops, would be expected to increase velocity of air and
uoguelph.ca shear at the spray nozzles. This, in turn, would increase the

914

316 Annex 131-A

OVERVIEW: HUMAN HEALTH AND GLYPHOSATE FORMULATIONS 915

formation of small droplets with a greater propensity for drift 2007b). This is between 175- and 1750-fold less than the
and offsite damage. Currently, exposure characterization is chronic reference dose of 100 μg/kg/day as determined by the

conducted by using models such as AGDISP (Bilanin et al., U.S. Environmental Protection Agency (EPA) (2008) and
1989) and AgDRIFT (Hewitt et al., 2001) for predicting on- between 3500- and 58,000-fold less than the acceptable operator
and off-target deposition of aerially applied sprays of pesti- exposure level (AOEL) of 200μg/kg/d (IUPAC, 2009).

cides. However, data on droplet size under spray conditions in
Colombia to input into the models were lacking.
EFFECTS IN THE ENVIRONMENT
Data on droplet size spectra were measured in a unique
wind tunnel facility in Australia where the appropriate velocity With regard to effects on organisms in the environment, the
of air could be achieved, and these data were then used to initial review (Solomon et al., 2007b) noted that amphibians

model spray drift in relation to sensitive organisms (Hewitt appeared to be relatively more sensitive to formulations of
et al., 2009). These results showed that the tank mix of Glyphos glyphosate than other aquatic animals, however, there were no
and Cosmo-Flux, as used in the eradication spraying in Colombia, data for the mixture of Glyphos and Cosmo-Flux as used for

produced droplets of median diameter (D v0.5 of 128 to 140μm, coca eradication in Colombia. It was also noted that many
which are classified as very fine to fine sprays. Modeling of other pesticides were used in the production of coca and that
spray drift using AgDRIFT showed that the spray droplets these could present significant risks to humans and non-target

would not evaporate as rapidly as most similarly sized agricul- organisms in the environment (Solomon et al., 2007a). Based
tural sprays because of the large proportion of nonvolatile on a worst-case exposure scenario, and a quotient based on the

components (active and inert adjuvant ingredients). Thus, even reference dose, some of the chemicals used by growers, partic-
under worst-case conditions of a cross-wind of 9.3 km/h, the ularly the organophosphorus insecticides, had hazard quotients
potential for longer range drift was small and most drift that for humans 2000-fold greater than that for the eradication

might occur would deposit relatively close to the application spray (Solomon et al., 2007a). Similarly, hazards to all aquatic
swath. In addition, drift only occurs downwind and with winds organisms were up to 20,000-fold greater (for endosulfan)
of velocity less than the modeled maximum the drift distance than the eradication spray mixture (Solomon et al., 2007a). A

would be less. Based on worst-case spray drift at various dis- refinement of this approach that focused on amphibians (Brain
tances from the application swath, exposures of plants and & Solomon, 2009) is published in this issue and confirmed the

organisms in shallow water (15 cm deep) were estimated and greater hazards to amphibians of the chemicals used to produce
compared to species sensitivity distributions of toxicity values coca. For some species of larval amphibians, sensitivity to sev-
for formulations of glyphosate in plants and amphibians, the eral pesticides (mainly insecticides) was 10- to 1000-fold

most sensitive group of animals. greater than estimated worst-case exposures and hazards were
Based on modeled drift and 5th centile concentrations, much greater than those for the eradication spray mixture. In
which would be protective of 95% of plants, appropriate no- addition, habitat destruction, such as clear-cutting forests for

Downloaded spray buffer zones (distance from the end of the spray boom as production of coca or food crops, was identified as another
recorded electronically ±5%) were 50 m to 120 m for coca major threat to amphibians (Brain & Solomon, 2009; Lynch &
spraying. These buffers are additionally protective of plants, as Arroyo, 2009).

it was shown that, at small rates of application, glyphosate As there were no data on the susceptibility of amphibians to
stimulates plant growth (Velini et al., 2008), which, even in the the mixture of Glyphos and Cosmo-Flux used in the eradica-

long term, does not reduce yields (Cedergreen, 2008). tion sprays, this was also a focus in the collection of additional
The equivalent buffer zone for protection of amphibia in data. An initial lab study with the African clawed frog, Xeno-
shallow water was 5 m, which, as discussed later, is conserva- pus laevis (Wildlife International, 2006a, 2006b), showed that

tive because adsorption of glyphosate and formulants in the the mixture was somewhat less toxic than reported values for
mixture to sediments and particulate matter further reduces other formulations of glyphosate. The LC50 for the mixture as
exposures and therefore, risk. The low toxicity of glyphosate used on coca was the equivalent of 1100 (95% CI; 560-2,300 μ)g

and its formulations to mammals (Williams et al., 2000; glyphosate a.e./L, while the lowest LC50 previously reported
Solomon et al., 2007b) suggests that these aerial applications for formulated glyphosate (Vision) in the same species of frog

are not a concern to bystanders,even those close to the spray was 800 μg a.e./L (Edginton et al., 2004). This then raised two
swath. The assertion that spray drift over long distances was questions; the first was whether there were alternative formula-
adversely affecting humans (Paz-y-Miño et al., 2007) is not sup- tions of glyphosate that were potentially less toxic to frogs but

ported by these observations, asexposures would be extremely as effective as the currently used mixture for the control of
small. For example, at 1 km from the spray swath, deposition coca and, second, were Colombian frogs generally more or less
would be between 1 and 0.1 g glyphosate acid equivalents sensitive to formulated glyphosate than other species tested in

(a.e.)/ha, which is equivalent to between 0.57 and 0.06μg/kg other regions?
body weight (bw), assuming a total exposed skin area for a To investigate the efficacy of other formulations of glypho-
naked 70-kg human of 2 m 2and 2% penetration (Solomon et al., sate on coca, field trials were conducted in Tolima Department,

317 Annex 131-A

916 K. R. SOLOMON ET AL.

Colombia (Marshall et al., 2009). Coca plants (E. coca) were (Maltby et al., 2005) and therefore allows the combination of
grown from seedlings to 75 cm height and then sprayed with a Colombian data with those from other regions for the purposes

range of glyphosate formulations and different adjuvants using of risk assessment.
an experimental ground sprayer. Assessments were made of In contrast to laboratory observations, toxicity studies con-
plant vigor, height, and above-ground standing crop (fresh ducted on Gosner stage 25 tadpoles under field conditions in

weight) 3 wk after application. Resprouting of plants was 15-cm deep microcosms containing a 3-cm layer of sediment
assessed at 9 wk after treatment. Even mixed with adjutants, showed reduced sensitivity (Bernal et al., 2009b). Microcosms

unformulated glyphosate applied as the product Rodeo gave were sprayed with the mixture of Glyphos and Cosmo-Flux as
poorer control of coca than two formulated products, Roundup used in eradication spraying. Mortality >50% was only observed
Biactive (from Europe) and the formulation currently used in in the tested species when the application rates were >2-fold the

eradication spraying, Glyphos. In general, these latter two normal application rate of 3.69 kg glyphosate a.e./ha. LC50 values
products performed well without added adjuvants, giving con- were between 8.9 and 10.9 kg glyphosate a.e./ha (equivalent to
trol similar to that of the mixture of Glyphos and Cosmo-Flux initial nominal concentrations of 5963 to 7303 μg glyphosate

as currently used in Colombia. There was some evidence that a.e./L in the microcosms. These results show that toxicity of
addition of the adjuvant Silwet L-77 and, to a lesser extent, the spray mixture is reduced in the presence of sediments and
Mixture B (from the United Kingdom) resulted in the earlier
particulates in the water column. Although it was not possible
appearance of symptoms of injury. There were also indications to measure concentrations of glyphosate in these systems, the
that glyphosate rates of less than 3.7 kg a.e./ha provided con- reduction in toxicity was similar to that observed by others

trol in the range of 95%. When also considering that glypho- (Tsui & Chu, 2003, 2004, 2008; Tsui et al., 2005) for the
sate appears to inhibit the production of cocaine in coca plants formulated product and also for the POEA surfactant, which
(Casale & Lydon, 2007), effective control of drug production contributes the greatest to the toxicity of the formulation

may be possible with lower rates of application. These results (Wang et al., 2005). In these studies, reductions in toxicity
also illustrate that there are potential alternatives to currently were attributed to reductions in exposure as a result of absorp-
used products, one of which, Roundup Biactive, was shown to tion to sediments and/or breakdown by microbes. Thus, risks to

be less toxic to amphibians (Mann et al., 2003). Before using larval frogs (representing sensitive aquatic organisms) from the
these products, field testing to assess the influence of different eradication sprays as used in Colombia would be reduced by

environmental conditions, varieties of coca, and aerial applica- adsorption to sediments under field conditions and, even with
tion procedures needs to be conducted. Should a different adju- direct overspray, amphibians in shallow water systems (∼15 cm
vant be required, Silwet L-77 and Mixture B would be good deep and theoretically the most vulnerable) would be at low risk.

candidates for further evaluation, including toxicity to nontarget In bioassays where terrestrial stages of frogs (juveniles and
organisms. adults) were exposed to a direct overspray of the Glyphos–
To address the question of sensitivity of Colombian species Cosmo-Flux mixture, LC50 values ranged between 4.5 and

Downloadedof frogs to formulated glyphosate, a series of toxicity bioassays 22.8 kg a.e./ha, all of which were above the application rate of
was conducted on tadpoles under laboratory conditions (Bernal 3.7 kg a.e./ha for eradication spraying. These studies were con-
et al., 2009a). Laboratory studies were conducted in glass con- ducted under realistic conditions with soil and leaf litter

tainers and in the absence of sediments and particulate matter. present in the bottom of the exposure chambers, a different
LC50 values for the 8 species tested (Gosner stage-25 tadpoles exposure system from that used in other studies that claimed

of Scinax ruber, Dendrosophus microcephalus, Hypsiboas high toxicity of formulated glyphosate (Relyea, 2005). The
crepitans, Rhinella granulosa, R. marina, R. typhonius, Centrolene observations of Relyea (2005) on adult frogs may have been
prosoblepon, and Engystomops pustulosus) ranged from 1200 the result of the presence of formulants specific to the product

to 2780 μg glyphosate a.e./L. These values suggest that sensi- used (Dinehart et al. 2009) or incorrect calculation of exposures
tivity to Roundup-type formulations of glyphosate in these spe- as the results reported by Bernal et al. (2009a) are consistent
cies is similar to that observed in other tropical and temperate with those of Mann and Bidwell (1999), who observed that

species of frogs for which data have been published in the liter- adult and juvenile terrestrial stages of the Australian frog,Crinia
ature. The toxicity of the mixture of Glyphos and Cosmo-Flux insignifera were less sensitive to Roundup than tadpoles. The

as used to spray coca was likely driven by the surfactant in the overall conclusions of the studies on Colombian frogs are that,
Glyphos, as the addition of Cosmo-Flux did not increase toxic- under worst-case exposure conditions, the mixture of Glyphos
ity above those values reported in other frogs for studies using and Cosmo-Flux used for control of coca in Colombia is of low

both Vision and Roundup, two similar formulations used in or negligible risk to aquatic and juvenile terrestrial stages of
North America (discussed earlier). Cosmo-Flux is of low toxic- frogs.
ity to fish with an LC50 of 4417 mg formulation/L (Rondon- To provide background information on amphibians and

Barragan et al., 2007). That tropical frog species were of simi- their distribution in relation to coca production and aerial erad-
lar sensitivity to those from temperate regions is also consistent ication spraying in Colombia, data on the more than 53,000
with observations with other pesticides and other organisms records of amphibians in the Instituto de Ciencias Naturales

318 Annex 131-A

OVERVIEW: HUMAN HEALTH AND GLYPHOSATE FORMULATIONS 917

(ICN) (Bogotá) were characterized (Lynch & Arroyo, 2009). interception by foliage and adsorption to soils and sediments
Analyses were based on the proximities of actual museum reduce exposures still further, and risks, even to a direct mix-

records to localities in which illegal crops are being grown and tures of the eradication mixtures, are small to negligible.
the subset of those that have been sprayed with glyphosate.
ARC MAP software was used so that direct distance separating

of collection locations for frogs, known coca fields, and areas EFFECTS IN HUMANS
where aerial spraying was being conducted could be measured In previous reviews of the risk of glyphosate to humans, it

(Lynch & Arroyo, 2009). was concluded that both the active ingredient and the formu-
Based on data for the location of amphibians collected in lated product (Roundup) present low risks to humans whether
Colombia, records existed of 193 species (28% of the national used in agricultural or vegetation management (Williams et al.,

diversity) of frogs and toads from localities within 10 km of 2000) or as used in the eradication of coca in Colombia
areas where coca is grown. Records in or near coca fields (Solomon et al., 2007b). The first article in this series on the
included records for 13 of the 15 families of frogs and toads potential human health effects of the use of Glyphos and

known for Colombia. Only Ceratophryidae and Pipidae were Cosmo-Flux for the eradication of coca addressed the issue of
not reported from these locations and would not be at risk. For possible reproductive effects of the spray program in Colombia
eight species (Dendrobates truncatus, Craugastor raniformis,
(Sanin et al., 2009). This issue was identified as a possible
Pristimantis gaigeae, Smilisca phaeota, Elachistocleis ovale, response by earlier reports of associations between pesticides
Hypsiboas crepitans, Trachycephalus venulosus, and Pseudis and reproductive outcomes. Arbuckle et al. (2001) reported

paradoxa) selected to represent several coca-associated habitat moderate increases in the risk of early abortion when precon-
preferences and lifecycle strategies, large areas of their distriception self-reported exposures to phenoxy acetic acid herbi-
butions lie outside coca production regions and the populations cides were present and for late abortions and self-reported

as a whole are at low risk of exposure. For a limited number of preconception exposure to glyphosate was associated with
species that barely enter Colombian territory, the consequences higher risks. In another study, Curtis et al. (1999) showed a
of coca production may be more serious and may have placed positive association (decrease in fecundability of 20% or more)

several species of frogs at risk. These include Ameerega bilin- measured through time to pregnancy (TTP) when both spouses
gua, Dendropsophus bifurcus, Eleutherodactylus colomai, E. reported exposure to pesticide-related activities, one of which

degener, E. diadematus, E. quaquaversus, E. variablis, and was glyphosate.
Trachycephalus jordani. Other species may be at risk, but The study in Colombia was to test whether there was an
exact numbers are unknown because little investigation association between the use of glyphosate when applied by

occurred in these areas during the past 30 yr. As these species aerial spray for the eradication of illicit crops eradication
are found in Ecuador, it is assumed that healthy populations (cocaine and poppy) and time to pregnancy (TTP) among fer-
persist there. tile women. The study was a retrospective cohort study with an

Downloaded At:Overall, the risks from pesticide used for eradication spray- ecological exposure index related to areas of residence with
ing must be placed in the context of the greater toxicity of othedifferent uses of glyphosate. First pregnancies in 2592 fertile
products used by growers (Brain & Solomon, 2009) and the women from 5 regions were included in the study and the

sensitivity of frogs from Colombia to the mixture of glyphosate women were interviewed regarding potential reproductive, lif-
and Cosmo-Flux as used in the aerial eradication spraying. estyle and work history predictors of TTP. The results showed

Laboratory-based toxicity studies showed that aquatic larval that there were differences in TTP between regions. Boyacá, a
stages of Colombian species are not differently sensitive as region with traditional crops without glyphosate eradication
compared with frogs from other locations (Bernal et al., spraying (manual eradication), had the minimal risk and was

2009b). When tested under realistic conditions—in shallow the reference region. Sierra Nevada, a control area with organic
water (15 cm deep) in the presence of sediment and particu- agriculture and no pesticide use; Putumayo, where illicit crops
lates that will absorb glyphosate and the more toxic surfac- are grown and with an intensive eradication spray program;

tant—toxicity was reduced (Bernal et al., 2009b), resulting in and Valle del Cauca, a sugar cane region where glyphosate and
lower risk. In contrast, some of the products used by growers others chemicals have been used for more than 30 yr, had

may be more bioavailable in the environment and risks to these greater risk of longer TTP, with the highest risk for Valle del
may not be mitigated. Terrestrial stages were less susceptible Cauca (Sanin et al., 2009).
than aquatic stages (Bernal et al., 2009b). Modeling of spray Classification of exposure in the study was by location of

drift from the aerial eradication spraying (Hewitt et al., 2009) residence. Nonexposed participants were those who lived in
showed small downwind exposures to the mixture of glypho- the region where organic crops were produced and who, in
sate and Cosmo-Flux at distances beyond 30 m. Based on addition, did not report any use of pesticides in the interview.

laboratory toxicity data, larval stages of frogs would only be atIn the other four departments, there was exposure not only to
risk if they were in shallow water within 5 m of the spray glyphosate, but also to other herbicides and pesticides.
swath. However, under conditions of exposure in the field, Although place of residence is not an accurate surrogate for

319 Annex 131-A

918 K. R. SOLOMON ET AL.

exposure to pesticides and may generate misclassification history, lifestyle, past and current occupational exposure to
(Arbuckle et al., 2004), this ecological assessment was useful pesticides, and factors known to be associated with increased

to explore, at the population level, whether an association frequency of micronuclei. In regions where glyphosate was
existed between the putative exposure (aerial spraying of gly- being sprayed, blood samples were taken prior to spraying, 5 d
phosate) and outcome (Ritter et al., 2006). Pesticides in generalafter spraying, and 4 mo after spraying. Lymphocytes were

are likely not the cause of observed differences either. Large cultured and MN analysis performed using standardized
differences in TTP were found between two regions of high to techniques on binucleated lymphocytes (BN) with preserved

moderate pesticide use, Valle del Cauca and Boyacá. The cytoplasm.
observed ecological differences remain unexplained, but may The frequency of binucleated lymphocytes with micronuclei
be produced by varying exposures to environmental factors, (BNMN) was smallest in Santa Marta, where organic coffee is

history of contraceptive programs in the region, or psychologi- grown without pesticides. Compared with Santa Marta, the
cal distress. Future studies examining these alternative causes pre-spray baseline frequency of BNMN was significantly
are needed. greater in subjects from the other four regions. The highest fre-

Epidemiological studies have not shown consistent or quency of BNMN was in Boyacá, where no aerial eradication
strong relationships between glyphosate exposures and health spraying of glyphosate was carried out, and Valle del Cauca,
outcomes. Glyphosate and its formulations have been exten-
where glyphosate was used for maturation of sugar cane.
sively investigated for potential adverse effects in humans Boyacá and Valle showed significantly higher frequency on
(Williams et al., 2000). They have been reported to exert a low BNMN than Nariño and Putumayo, where aerial spraying was

acute toxicity to different animal species. Chronic feeding carried out. Region, gender, and older age (≥35 yr) were the
studies have not shown evidence of carcinogenicity or any only variables associated with the frequency of BNMN mea-
other relevant long-term effect (U.S. EPA, 1993; World Health sured before spraying. A significant increase in frequency of

Organization International Program on Chemical Safety, BNMN between first and second sampling was observed in
1994). Glyphosate AI and Roundup were extensively tested for Valle, Putumayo, and Nariño immediately (<5 d) after spraying.
genotoxicity in a wide range of in vitro and in vivo systems Four months after spraying in Nariño, there was a statistically

evaluating different genetic endpoints (gene mutation, chromo- significant decrease in the mean frequency of BNMN com-
some mutation, DNA damage and repair) using bacteria and pared with the second sampling, but in Valle del Cauca the

mammalian somatic cells (Williams et al., 2000). Although decrease was not significant nor was the increase in Putumayo.
effects were reported in some in vitro studies, it was concluded There was no significant association between self-reported
that, in vivo, glyphosate and its formulations were not genotoxicdirect contact with eradication sprays and frequency of

(Williams et al., 2000). Several in vitro and in vivo studies with BNMN. The frequency of BNMN in participants who self-
parallel testing of glyphosate AIand Roundup showed that only reported that they were exposed to glyphosate because they
the commercial formulation was genotoxic (Rank et al., 1993; entered the field immediately after spraying (to pick the coca

DownloadedBolognesi et al., 1997; Gebel et al., 1997; Grisolia 2002), in gen- leaves), felt spray drops in their skin, or they thought they were
eral agreement with the observation that adjutants in the formu- exposed because they had contact with the chemical in the air,
lation may be more toxic to animals than glyphosate itself was not significantly greater than in subjects living in the same

(Giesy et al., 2000; Williams et al., 2000; Richard et al., 2005)areas but who were not present during spraying. Overall, these
Evidence of DNA damage in peripheral lymphocytes from a results suggest that genotoxic damage associated with glypho-

small group of subjects potentially exposed to glyphosate was sate spraying, as evidenced by the MN test, is small and
reported in a recent article (Paz-y-Miño et al., 2007). Problems appears to be transient. The frequencies of BNMN in Nariño
with the study design, such as the small number of subjects and Putumayo during the second and the third sampling fell

(21 control and 24 exposed) and the fact that random selection within the range of values observed in Boyacá, an area where
produced 23 females and 1 male in the exposed group, do not people were exposed to a complex mixture of different pesti-
allow conclusions to be drawn; however, this article did raise cides (including glyphosate). A greater increase in frequency

concerns about possible effects and a study was carried out of BNMN was observed in Valle del Cauca, but it cannot be
using the micronucleus (MN) response in peripheral lympho- attributed only to the glyphosate exposure, because the applica-

cytes as a biomarker (Bolognesi et al., 2009). tion rate of the herbicide in this area was one-third compared
This study was carried out in volunteers from five Colombian with that in Nariño and Putumayo. There was no association
regions, characterized by different exposure to glyphosate and between self-reported direct contact with eradication sprays

other pesticides. The epidemiological design was a prospective and frequency of BNMN. Overall it was concluded that the
cohort study but, for logistical reasons, without exposure genotoxic risk potentially associated with exposure of humans
biomonitoring. A large sample, 274 persons comprising to glyphosate in the areas of Colombia where the herbicide is

137 women of reproductive age (15–49 yr of age) and their applied for coca and poppy eradication is of low biological rel-
spouses (137), were included in the study. Participants were evance. When these conclusions are combined with the lack of
interviewed to obtain relevant details about health status, significant spray drift (Hewitt et al., 2009), there is no support

320 Annex 131-A

OVERVIEW: HUMAN HEALTH AND GLYPHOSATE FORMULATIONS 919

for the earlier conclusion (Paz-y-Miño et al., 2007) that eradi- Overall, the risks to sensitive wildlife and human health

cation spraying is producing adverse effects in humans. from the use of glyphosate in the control of coca (and poppy)
production in Colombia are small to negligible, especially

when compared to the risks to wildlife and humans that result
OVERALL CONCLUSIONS from the entire process of the production of cocaine (and heroin)

The study started out with three questions related to the in Colombia.
risks to the environment and human health of the use of gly-

phosate for eradication of coca (and poppy) in Colombia.
These questions were related to spray drift, effects on sensitive REFERENCES
Arbuckle, T. E., Cole, D. C., Ritter, L., and Ripley, B. D. 2004. Farm
wildlife such as amphibians, and effects on humans. On the children’s exposure to herbicides: Comparison of biomonitoring and ques-
basis of the results of the series of studies reported in this issue
tionnaire data. Epidemiology 15:187–194.
and other observations reported in the literature, several overall Arbuckle, T. E., Lin, Z., and Mery, L. S. 2001. An exploratory analysis of the
conclusions were reached. In terms of spray drift, new data effect of pesticide exposure on the risk of spontaneous abortion in an
Ontario farm population. Environ. Health Perspect. 109:851–857.
showed that drift from eradication spraying is minimal and that Bernal, M. H., Solomon, K. R., and Carrasquilla, G. 2009a. Toxicity of for-
relatively small buffer zones, ranging from 5 to 120 m, are pro- mulated glyphosate (Glyphos®) and Cosmo-Flux ® to larval Colombian

tective of sensitive aquatic animals and, the target organisms, frogs 1. Laboratory acute toxicity. J. Toxicol. Environ. Health A
plants, respectively. 72:961–965.
Bernal, M. H., Solomon, K. R., and Carrasquilla, G. 2009b. Toxicity of
Laboratory and field tests on amphibians showed that formulated glyphosate (Glyphos ) and Cosmo-Flux to larval and juvenile
Colombian species were of similar sensitivity to species tested Colombian frogs 2. Field and laboartory microcosm acute toxicity,J. Toxicol.
Environ. Health A72:966–973.
in other locations and that they were not especially sensitive to
glyphosate formulations. Tests on larvae stages of amphibians Bilanin, A. J., Teske, M. E., Barry, J. W., and Ekblad, R. B. 1989. AGDISP:
The aircraft spray dispersion model, code development and experimental
under realistic conditions showed that toxicity was reduced, validation. Trans. Am. Soc. Ag. Eng. 32:327–334.
most likely because of the rapid absorption of glyphosate and Bolognesi, C., Bonatti, S., Degan, P., Gallerani, E., Peluso, M., Rabboni, R.,
Roggieri, P., and Abbondandolo, A. 1997. Genotoxic activity of glypho-
its adjuvants to sediments and particulate matter. Terrestrial
stages of frogs showed a range of sensitivity, but all had LC50 sate and its technical formulation, Roundup. J. Agric. Food. Chem.
values less than the application rate used for eradication of 45:1957–1962.
Bolognesi, C., Carrasquilla, G., Volpi, S., Solomon, K. R., and Marshall, E. J.
coca. Given interception by foliage, risks to aquatic and terres- P. 2009. Biomonitoring of genotoxic risk in agricultural workers from five
trial stages of frogs from Colombia, even from direct exposure Colombian regions: Association to occupational exposure to glyphosate. J.
Toxicol. Environ. Health A 72:986–997.
to aerial eradication sprays, are judged to be small to negligi-
ble. The study of the large distribution of large diversity of frog Brain, R. A., and Solomon, K. R. 2009. Comparative hazards of glyphosate,
other pesticides, and other human activities to amphibians in the production
species in Colombia in relation to coca production and eradica- of coca. J. Toxicol. Environ. Health A 72:937–948.
tion spraying showed that there were only a few species of Casale, J., and Lydon, J. 2007. Apparent effects of glyphosate on alkaloid pro-
duction in coca plants grown in Colombia. J. Forens. Sci. 52:573–578.
Downloaded Afrogs potentially at risk because of their location in southwest Cedergreen, N. 2008. Is the growth stimulation by low doses of glyphosate
Colombia. As these species are also found in Ecuador, the
sustained over time? Environ Pollut. 156:1099–1104.
likely small risks are to populations in Colombia, not the spe- CICAD/OAS. 2004. The toxicology of chemicals used in the production and
cies as a whole. A much greater risk to frogs in Colombia is refining of cocaine and heroin: A tier-one assessment. Technical report
OAS/CICAD 2004-01, CICAD, Organization of American States.
from the other pesticides used by the growers of coca (and Washington, DC, USA.
poppy) and particularly the deforestation that precedes the CICAD/OAS. 2005.The toxicology of selected chemicals used in the production

planting of these crops. and refining of cocaine and heroin: A tier-two assessmen.tTechnical report
In terms of effects on humans, an epidemiological study OAS/CICAD 2005-01, CICAD, Organization of American States.
Washington, DC, USA.
did not provide evidence of effects on reproductive function in Curtis, K. M., Savitz, D. A., Weinberg, C. R., and Arbuckle, T. E. 1999.
terms of TTP. In a study on potential genotoxicity that com- The effect of pesticide exposure on time to pregnancyEpidemiology

bined epidemiological surveys with biological monitoring of 10:112–117.
the frequency of MN in white blood cells, differences in the Dinehart, S. K., Smith, L. M., McMurry, S. T., Anderson, T. A., Smith, P. N.,
and Haukos, D. A. 2009. Toxicity of a glufosinate-and several glyphosate-
baseline frequency were observed in relation to region sam- based herbicides to juvenile amphibians from the Southern High Plains,
pled. In those regions where spraying of glyphosate was being USA, Sci. Tot. Environ. 407:1065–1071.
Edginton, A. N., Sheridan, P. M., Stephenson, G. R., Thompson, D. G., and
carried out for agricultural and eradication purposes, fre-
quency of MN rose after spraying but these increases were not Boermans, H. J. 2004. Comparative effects of pH and Vision herbicide on
two life stages of four anuran amphibian species. Environ. Toxicol. Chem.
related to the rate of application or to self-reported exposures 23:815–822.
to the spray. In some regions the frequency decreased after Gebel, T., Kevekordes, S., Pav, K., Edenharder, R., and Dunkelberg, H. 1997.
In vivo genotoxicity of selected herbicides in the mouse bone marrow
spraying but in one, it did not. These observations do not ful- micronucleus test. Arch. Toxicol. 71:193–197.
fill all of the criteria for causality, suggesting that if glypho-
Giesy, J. P., Dobson, S., ®nd Solomon, K. R. 2000. Ecotoxicological risk
sate spraying has any influence on MN, this is small and not of assessment for Roundup herbicide. Rev. Environ. Contam. Toxicol.
biological significance. 167:35–120.

321 Annex 131-A

920 K. R. SOLOMON ET AL.

Grisolia, C. K. 2002. A comparison between mouse and fish micronucleus test Series no. 966 (vol. 966), eds. Kennedy, I. R., Solomon, K. R., Gee, S.,
using cyclophosphamide, mitomycin C and various pesticides. Mutat. Res. Crossan, A. N., Wang, S., and Sanchez-Bayo, F., pp. 87–99. Washington,
518:145–150. DC: American Chemical Society.
Hewitt, A. J., Solomon, K. R., and Marshall, E. J. P. 2009. Spray droplet size, Solomon, K. R., Anadón, A., Carrasquilla, G., Cerdeira, A., Marshall, J., and

drift potential, and risksto nontarget organisms from aerially applied glypho- Sanin, L.-H. 2007b. Coca and poppy eradication in Colombia: Environ-
sate for coca control in ColumbiaJ. . Toxicol. Environ. Health A 72:921–929. mental and human health assessment of aerially applied glyphosate. Rev.
Hewitt, A. J., Teske, M. E., and Thistle, H. E. 2001. The development of the Environ. Contam. Toxicol. 190:43–125.
AgDRIFT ® model for aerial application from helicopters and fixed-wing Solomon, K. R., Anadón, A., Cerdeira, A., Marshall, J., and Sanin, L.-H. 2005.

aircraft. Aust. J. Ecotoxicol. 8:3–6. Environmental and human health assessment of the aerial spray program
IUPAC. 2009. IUPAC Agrochemical Information, International Union of Pure for coca and poppy control in Colombia. Technical report. Inter-American
and Applied Chemistry. Accessed January 8, 2009. http://sitem.herts.ac.uk/ Drug Abuse Control Commission (CICAD) section of the Organization of
aeru/iupac/373.htm American States (OAS). Washington, DC, USA.

Lynch, J. D., and Arroyo, S. 2009. Risks to Colombian amphibian fauna from Tsui, M. T., Wang, W. X., and Chu, L. M. 2005. Influence of glyphosate and
cultivation of coca (Erythroxylum coca): A geographical analysis.J. Toxicol. its formulation (Roundup ®) on the toxicity and bioavailability of metals to
Environ. Health A72:974–985. Ceriodaphnia dubia. Environ. Pollut. 138:59–68.

Maltby, L., Blake, N. N., Brock, T. C. M., and van den Brink, P. J. 2005. Insec-Tsui, M. T. K., and Chu, L. M. 2003. Aquatic toxicity of glyphosate-based for-
ticide species sensitivity distributions: The importance of test species mulations: Comparison between different organisms and the effects of
selection and relevance to aquatic ecosystems. Environ. Toxicol. Chem. environmental factors Chemosphere 52:1189–1197.
24:379–388. Tsui, M. T. K., and Chu, L. M. 2004. Comparative toxicity of glyphosate-based

Mann, R. M., and Bidwell, J. R. 1999. The toxicity of glyphosate and several herbicides: Aqueous and sediment porewater exposures. Arch. Environ.
glyphosate formulations to four species of Southwestern Australian frogs. Contam. Toxicol. 46:316–323.
Arch. Environ. Contam. Toxicol. 36:193–199. Tsui, M. T. K., and Chu, L. M. 2008. Environmental fate and nontarget impact
Mann, R. M., Bidwell, J. R., and Tyler, M. J. 2003. Toxicity of herbicide for- of glyphosate in a subtropical wetland. Chemosphere 71:439–446.

mulations to frogs and the implications for product registration: A case UNODC. 2008. Coca cultivation in the Andean Region. A survey of Bolivia,
study from Western Australia. Appl. Herpetol. 1:13–22. Colombia and Peru. Technical report, United Nations Office on Drugs and
Marshall, E. J. P., Solomon, K. R., and Carrasquilla, G. 2009. ar(ythroxylum Crime. Vienna, Austria.

coca) control is affected by glyphosate formultaions and adjuvants.J. ToxicoU.S. Environmental Protection Agency. 1993. R.E.D. Facts Glyphosate. Tech-
Environ. Health A 72:930–936. nical report EPA 738-R-93-014. Washington, DC: U.S. Environmental
Paz-y-Miño, C., Sánchez, M. E., Arév alo, M., Muñoz, M. J., Witte, T., Protection Agency.
De-la-Carrera, G. O., and Paola, L. E. 2007. Evaluation of DNA damage U.S. Environmental Protection Agency. 1998. Guidelines for ecological risk

in an Ecuadorian population exposed to glyphosate. Genet. Mol. Biol. assessment. Technical report. Washington, DC: U.S. Environmental
30:456–460. Protection Agency.
Rank, J., Jensen, A. G., Skov, B., Pedersen, L. H., and Jensen, K. 1993. GenotoxU.S. Environmental Protection Agency. 2008. Integrated Risk Information
icity testing of Roundup and its activeingredient glyphosate isopropylamine System (IRIS) database for risk assessment. United States Environmental

using the mouse bone marrow micronucleus test,Salmonella mutagenicity Protection Agency: Office of Research and Development, National Center
test andAllium anaphase–telophase test.Mutat. Res. 300:29–36. for Environmental Assessment. Accessed September 30, 2008. http://
Relyea, R. A. 2005. The lethal impact of Roundup on aquatic and terrestrial www.epa.gov/iris
amphibians. Ecol. Appl. 15:1118–1124. Velini, E. D., Alves, E., Godoy, M. C., Meschede, D. K., Souza, R. T., and

Richard, S., Moslemi, S., Sipahutar, H., Benachour, N., and Seralini, G.-E. Duke, S. O. 2008. Glyphosate applied at low doses can stimulate plant
2005. Differential effects of glyphosate and Roundup on human placental growth. Pestic. Manage. Sci. 64:489–496.
cells and aromatase. Environ. Health Perspect. 113:716–720. Wang, N., Besser, J. M., Buckler, D. R., Honegger, J. L., Ingersoll, C. G.,
Downloaded At: 20:07 2 October 2009
Ritter, L., Goushleff, N. C. I., Arbuckle, T., Cole, D., and Raizenne, M. 2006. Johnson, B. T., Kurtzweil, M. L., MacGregor, J., and McKee, M. J. 2005.
Addressing the linkage between exposure to pesticides and human health Influence of sediment on the fate and toxicity of a polyethoxylated tallowa-
effects—Research trends and priorities for research 1. J. Toxicol. Environ. mine surfactant system (MON 0818) in aquatic microcosms. Chemosphere
Health B 9:441–456. 59:545–551.
®
Rondon-Barragan, I. S., Ramirez-Duarte, W. F., and Eslava-Mocha, P. R. Wildlife International. 2006a. Glyphosate–Cosmo-Flux–Coca Mix: A 96-hour
2007. Evaluación de los efectos tóxicos y concentración letal 50 del surfac- static-renewal acute toxicity test with the African clawed-frog tadpole
tante Cosmoflux ® 411F sobre juveniles de cachama blanca (Piaractus (Xenopus laevis). Final report, technical report 628A-101, Wildlife
brachypomus). Rev. Colomb. Cienc. Pec. 20:431–446. International. Easton, MD, USA.
®
Sanin, L.-H., Carrasquilla, G., Solomon, K. R., Cole, D. C., and Marshall, E. J.Wildlife International. 2006b. Glyphosate-Cosmo-Flux–Poppy Mix: A 96-
P. 2009. Regional differences in time to pregnancy among fertile women hour static-renewal acute toxicity test with the African clawed-frog tad-
from five Colombian regions with different uses of glyphosate. J. Toxicol. pole (Xenopus laevis). Final report, technical report 628A-102, Wildlife
Environ. Health A 72:949–960. International. Easton, MD, USA.

Solomon, K. R., Anadón, A., Brain, R. A., Cerdeira, A. L., Crossan, A. N., Williams, G. M., Kroes, R., and Munro, I. C. 2000. Safety evaluation and risk
Marshall, A. J., Sanin, L. H., and Smith, L. 2007a. Comparative hazard assessment of the herbicide Roundup ® and its active ingredient, glypho-
assessment of the substances used for production and control of coca and sate, for humans. Regul. Toxicol. Pharmacol. 31:117–165.

poppy in Colombia. In Rational environmental management of agrochemi- World Health Organization International Program on Chemical Safety. 1994.
cals: Risk assessment, monitoring, and remedial action. ACS Symposium Glyphosate (vol. 159), Geneva: WHO IPCS.

322 Annex 131-B

Annex 131-B

H EWITT ET ., “PRAYDROPLET SIZ,DRIFTPOTENTIAL,AND RISKS TO
NONTARGET O RGANISMS FROMAERIALLYAPPLIEDGLYPHOSATE FORC OCA
CONTROL INCOLOMBIA”

(Journal of Toxicology and Environmental Health, Part A, 72:921-929, 2009)

323 Annex 131-B

Journal of Toxicology and Environmental Health, Part A, 72: 921–929, 2009
ISSN: 1528-7394 print / 1087-2620 online
DOI: 10.1080/15287390902929667

UTEH
Spray Droplet Size, Drift Potential, and Risks to Nontarget

Organisms from Aerially Applied Glyphosate for Coca

Control in Colombia

Andrew J. Hewitt , Keith R. Solomon , and E. J. P. Marshall 3
1
The University of Queensland, Centre for Pesticide Application and Safety, Gatton, Queensland,
Australia, Lincoln Ventures/Lincoln University, Lincoln, Christchurch, New Zealand, 2Centre for

Toxicology and Department of Environmental Biology, University of Guelph, Guelph, Ontario,
Canada, and 3Marshall Agroecology Limited, Barton, Winscombe, Somerset, United Kingdom

sate were constructed for plants and amphibians. Based on modeled

A wind tunnel atomization study was conducted to measure drift and 5th centile concentrations, appropriate no-spray buffer
the emission droplet size spectra for water and Glyphos (a glypho- zones (distance from the end of the spray boom as recorded elec-
sate formulation sold in Colombia)+Cosmo-flux sprays for aerial tronically±5%) for protection of sensitive plants were 50–120 m for
application to control coca and poppy crops in Colombia. The coca spray scenarios and considerably lower for poppy spray sce-
narios. The equivalent buffer zone for amphibia was 5 m. The low
droplet size spectra were measured in a wind tunnel for an Accu- toxicity of glyphosate to humanssuggests that these aerial applica-
Flo nozzle (with 16 size 0.085 [2.16 mm] orifices), under appropri-
ate simulated aircraft speeds (up to 333 km/h), using a laser dif- tions are not a concern for human health.
fraction instrument covering a dynamic size range for droplets of
0.5 to 3,500 mm. The spray drift potential of the glyphosate was
modeled using the AGDISP spray application and drift model,

using input parameters representative of those occurring in Aerial applications of glyphosate to control illicit coca and
Colombia for typical aerial application operations. The droplet poppy crops have been made in Colombia since 1997. The area
size spectra for tank mixes containing glyphosate and Cosmo-Flux
were considerably finer than water and became finer with higher of coca sprayed with glyphosate has shown a steady increase
aircraft speeds. The tank mix with 44% glyphosate had a D v0.5f over recent years, reaching approximately 153,000 ha in 2007
Downloaded A128 mm, while the value at the 4.9% glyphosate rate was 140 mm. (personal communication, National Police of Colombia,
These are classified as very fine to fine sprays. Despite being rela-
Bogotá, December, 2007). Concerns have been raised as to the
tively fine, modeling showed that the droplets would not evapo- possible environmental and human health effects of the aerial
rate as rapidly as most similarly sized agricultural sprays because
the nonvolatile proportion of the tank mix (active and inert adju- spray program (International Court of Justice, 2008).
vant ingredients) was large. Thus, longer range drift is small and The aerial application of herbicides involves release of
most drift that does occur will deposit relatively close to the applica- spray liquid over a target area using appropriate equipment
tion area. Drift will only occur downwind and, with winds of velocity
less than the modeled maximum of9 km/h, the drift distance would (aircraft, nozzles, boom setup, etc.) and directing the spray
toward the target, considering any cross-wind, vertical wind, or
be substantially reduced. Spray dritfpotential might be additionallother meteorological effects. Many studies examined the
reduced through various practices such as the selection of nozzles,
tank mix adjuvants, aircraft speeds, and spray pressures that wouldeffects of application, meteorological, chemical, and canopy
produce coarser sprays. Species sensitivity distributinos to glyphoparameters that may influence deposition of the spray and thus

efficacy and potential off-target spray drift. Studies by the
©General Secretariat of the Organization of American States, Spray Drift Task Force (Hewitt et al., 2002) and others have
2009. This paper was prepared as part of a Study entitled “Productiond to the development and validation of accurate models such

of Illicit Drugs, the Environment and Human Health,” financed with as AGDISP (Bilanin et al., 1989) and AgDRIFT (Hewitt et al.,
contributions from the Governments of Colombia and the United 2001) for predicting on- and off-target deposition of aerially
States of America. The conclusions and opinions expressed herein arapplied sprays. Originally developed by the U.S. Forest
those of the authors and not necessarily those of the Organization of
American States and its General Secretariat, which as of the date ofervice, NASA, U.S. Army, and other organizations around
this copyright, have not formulated any opinion with respect to them.e world, this model has been extensively validated for use in

Address correspondence to Dr. Andrew J. Hewitt, Director, Centrespray drift exposure assessments for aerial applications of her-
for Pesticide Application and Safety, University of Queensland, bicides such as glyphosate. The U.S. Environmental Protection
Gatton QLD 4343, Australia. E-mail: [email protected] Agency (EPA) has been a key participant in the development

921

324 Annex 131-B

922 A. J. HEWITT ET AL.

of these modeling tools (Bird et al., 2002), and routinely uses Droplets contained in the spray clouds produced by the noz-
them for risk assessments involving pesticide drift. One of the zles in the present study were not spatially uniform. Droplet size

most important factors affecting spray application and environ- varied across the spray plume (for example,larger droplets typi-
mental fate modeling is the droplet size spectrum applied by cally occurred at the edge of the plume, and smaller droplets in
the aircraft or sprayer (Hewitt 1997). the center). It was therefore important to ensure that a cross-

This article describes a wind tunnel atomization study to sectional average spray sample was obtained at a given axial
measure the emission droplet size spectra for water alone and location that was representativeof the spray plume under evalua-

the herbicide glyphosate (Glyphos) plus the adjuvant Cosmo- tion (Dodge, 1988; Snyder et al., 1989). Cross-section averages
Flux sprays applied under simulated conditions for aerial appli- were obtained in this study by traversing the nozzle vertically
cation to control coca and poppy crops in Colombia. The drop- down across the laser beam while spraying.

let size spectra were measured in a wind tunnel for the same Volumetric and cumulative volu metric droplet size spectra
Accu-Flo nozzles that are used in the field spray applications, as well as the D v0.5value were calculated for water as a
under appropriate simulated aircraft speeds (up to 333 km/h), standard control and two different spray mixtures containing

using a laser diffraction instrument covering a dynamic size Glyphos and Cosmo-Flux. The D v0.5 or volume median diam-
range for droplets of 0.5 to 3500 μm. The nozzle size was eter, is the droplet diameter ( μm) at which 50% of the total
0.085 (2.16 mm) with 16 discharge orifices. The spray pressure spray volume is contained in droplets with larger and smaller

was 2.4 bar at the nozzle. Using the droplet spectrum data from diameter. Droplet parameter size values were calculated by
the wind tunnel study, the spray drift potential of the glypho- the particle size analyzer in compliance with ASTM Standard

sate spray mixture was modeled using the AGDISP model with E799-87 (ASTM, 1987). In this study, there was interest in
application and meteorological input parameters representative the spray volume contained in relatively small droplets, i.e.,
of those occurring in typical aerial application for control of those with diameter below 150 μm. This represents the finer

coca and poppy in Colombia. Based on model predictions, and droplets in the spray, which might present more of an exposure
in combination with data on the potential environmental effects risk for downwind spray drift under unfavora ble conditions.
of glyphosate and the adjuvant Cosmo-Flux (Solomon et al., Experimental runs were conducted using either (a) water

2007; Brain & Solomon, 2009), studies were undertaken to alone, (b) water, 44% Glyphos, 1% Cosmo-Flux as used for
assess the risk to plants and the environment associated with treatment of coca crops, or (c) water, 4.9% Glyphos, 0.5%

the potential for off-target spray drift under the conditions of Cosmo-Flux as used for treatment of poppy crops. Glyphos
use in Colombia. and Cosmo-Flux were the exact same products used in
Colombia for aerial application. Aerial application rates of

glyphosate for coca are 3.69 kg a.e./ha. In order to maximize
MATERIALS AND METHODS penetration and effectiveness of the spray formulation, Gly-
A Sympatec HELOS VARIO laser diffraction particle size phos is tank-mixed with an adjuvant product (Cosmo-Flux

Downloadedanalyzer was used to measure the emission droplet size spectra 411F; Cosmoagro, Bogotá). Cosmo-Flux is an agricultural
for Accu-Flo and solid-stream nozzles in a wind tunnel. All adjuvant containing nonionic surfactants (a mixture of linear
measurements were made using a 2000-mm focal length lens and aryl polyethoxylates: 17% w/v) and isoparaffins (83% v/

which measured droplets in the size range 0.5 to 3,500 μm. v). These tank mixes were prepared by mixing the compo-
Data and results were obtained using model-independent anal- nents on a volumetric basis using graduated measuring cylin-

ysis (MIA) and Windox software. MIA is the usual analysis ders. The mixtures were stirre d thoroughly and placed into
method used by other liquid spray researchers in the Spray 20-L pressurized containers. Compressed air was used to dis-
Drift Task Force (Hewitt, 1994). The wind tunnel, a straight- place the products from the spray tanks through the nozzles

through blower design used for such studies (Foster & French, for sampling the droplet size spectra in the wind tunnel.
1992), was set to deliver airstream velocities up to 333 km/h Applications of each tank mix were made through the Accu-
(92.6 m/s; 207 miles/h), measured using Pitot and hot wire Flo nozzle (0.085 [2.16 mm] with 16 orifices, as used in

anemometer probes. The end fans were also operated to draw Colombia) and an alternative D10 nozzle type at simulated
air through the filter section of the wind tunnel and ensured aircraft speeds of 259, 296, and 333 km/h. The Accu-Flo noz-

uniform air velocities while sampling the sprays. All measure- zle was also tested in a prototype reverse Venturi chamber
ments were made for atomized droplets of spray, i.e., beyond (supplied by Russ Stocker, Woodland) to see if such devices
the initial ligament breakup distance (with the laser 1.5 m used to reduce air shear effects on liquid atomization could

downwind of the nozzle). All measurements were replicated to increase droplet size.
provide two or three measurements per treatment, which were
statistically averaged and characterized. The spray pressure

was measured using a calibrated pressure gauge connected to a Spray Drift Modeling
capillary at the nozzle tip, and set to approximately 2.4 bar (35 psi)ssessments of spray drift and deposition were conducted
for most of the applications. using the AGDISP model. This model was developed and

325 Annex 131-B

NONTARGET ORGANISMS AND AERIALLY APPLIED GLYPHOSATE 923

validated by NASA, the U.S. Forest Service, U.S. Army, Spray was calculated from the intercept of the 5th centile toxicity val-

Drift Task Force, and others over several decades for aerial for- ues and the drift deposition values from the AGDISP model.
estry (Bilanin et al., 1989) and agricultural spray applications Toxicity data were obtained for plants and the most sensi-
(Hewitt et al., 2002). The AGDISP model was run using the tive aquatic species, amphibians. The toxicity data for plants

following inputs: were taken from the U.S. EPA Ecotox database (U.S. EPA,
2001) for effects of glyphosate on plants. The values were the
• Droplet size data measured from the wind tunnel tests,
specified in 32 size classes by spray volume. EC25 from tests for plants sprayed with formulated glyphosate
(mostly Roundup). These values were converted to units of g/ha
• For OV-10 and AT-802, 60 Accu-Flo nozzles (maxi- a.e. to allow for a common basis for comparison. It is recog-
mum allowed by AGDISP model); for Ayres Turbo
nized that Cosmo-Flux may increase potency of the mixture for
Thrush (ATT-65), 35 Accu-Flo nozzles, regularly plants; however, there were no data for sensitivity to the mix-
spaced on the boom, spaced at equivalent of 74 nozzles
at 114 mm for OV-10; and 96 nozzles at 140 mm for ture in plants other than coca, where there was little difference
between Glyphos alone or in combination with Cosmo-Flux
AT-802 and 35 nozzles at 152 mm for ATT-65. (Marshall et al., 2009). The plantdata were fitted to a log-normal
• Aircraft types from the library to include Ayres Turbo
Thrush 65, Bronco OV-10, and Air Tractor 802 with distribution to create a species sensitivity distribution (SSD;
Solomon & Takacs, 2002). In doing this, the test plants were
characteristic configurations, vortex patterns, and
default settings except operational flight speed. used as surrogates for other plants in the environment, a nor-
mal process for assessing risks using SSD. For amphibians, the
• Aircraft flight speeds of 333 km/h for OV-10, 274 toxicity data for formulated glyphosate were taken from litera-
km/h for AT-802, and 226 km/h for ATT-65.
• Tank mix compositions to represent water with 4.992 or ture as summarized elsewhere (Brain & Solomon, 2009).
These data are for formulated glyphosate without the addition
1.2 kg/ha glyphosate isopropylamine salt (as appropriate
for modeling each application scenario), of which it is of Cosmo-Flux, but toxicity tests on the spray mixture with the
most sensitive frog species (Xenopus laevis) (Wildlife Interna-
assumed the acid equivalents (a.e.) rate was 75%. tional, 2006) showed no increased sensitivity over previously
Nonvolatile rate set to assume that all of the glyphosate
and Cosmo-Flux molecules were nonvolatile. Spray published values (Edginton et al., 2004). The 5th centile of the
distribution was calculated using the transformed values for the
volume rate of 10.4 L/ha (1.11 gallon/acre) total tank
mix for coca sprays. slope and intercept of the regression line of the SSD. The 5th
centile of the amphibian SSD was calculated as already
• Aircraft height above ground 30.48 m. described and converted from a concentration to a rate per
• Canopy (tree) height 25.91 m.
• One flight line (i.e., single swath applications as used hectare under the worst-case assumption that the concentration
resulted from direct overspray of a 15-cm-deep pool with no
operationally in Colombia). exposure reduction via adsorption to sediments and organic
• An 18.3-m effective swath width.
Downloaded At: 20:07 2 October 2009 matter, and no interception by surrounding plants.
• 2.57 m/s (9.3 km/h) wind speed at 90º to aircraft flight
direction (i.e., perpendicular cross-wind).
• 35ºC air temperature and 70% relative humidity.

• Weak atmospheric stability (weak solar insolation RESULTS
equivalent to partial or full cloud cover).
2 Droplet Size Spectra
• Evaporation rate 37 μm /ºC/s (representative of gly- The droplet size from the Accu-Flo nozzle became smaller
phosate products).
as the aircraft speed increased, due to higher air shear at the
All other model input values were the model default settings. point of liquid atomization (Table 1). For example, for the coca

spraying tank mix, as aircraft speed increased from 259 to 296
to 333 km/h, the D v0.5(volume median diameter droplet size)
Risk Assessment value of the sprays decreased from 219 to 173 to 128 μm,

Amphibians had been identified as aquatic organisms at respectively, and the fine spray volume in droplets with diame-
greatest risk from a direct overspray in a previous risk assessmentter below 150 μm increased from 35 to 44 to 57%.
(Solomon & Thompson, 2003; Solomon et al., 2007) and, as
The water sprays were much coarser than tank mixes con-
glyphosate is a herbicide for the control of plants, plants would taining glyphosate and Cosmo-Flux. Examples for the two
be potentially at risk from direct overspray (the desired out- active ingredient tank mixes at 333 km/h (180 knots) aircraft

come) as well as from spray drift. As the human health risks of speed are shown in Figures 1 and 2. There was less difference
exposures to a direct overspray of glyphosate–Cosmo-Flux in droplet size between the two active ingredient tank mixes,
mixture were judged to be negligible in previous studies
with both being considerably finer than water sprays. An
(Solomon et al., 2007), any exposures from drift would also be example of the effect of tank mix composition on droplet size
negligible. A deterministic risk value for amphibians and plants is given as follows, for applications through the Accu-Flo

326 Annex 131-B

924 A. J. HEWITT ET AL.

TABLE 1
Volume Median Diameter Droplet Size (D v0.5and Fine Spray Volume in Droplets Below 150 μm, for Water
and the Tank Mixes for Coca and Poppy Control for Accu-Flo and D10 Nozzle Types

Accu-Flo (0.085-16) nozzle D10 nozzle

Speed D v0.5 Percent volume Dv0.5 Percent volume
Spray solution (km/h) (μm) <150 μm (μm) <150 μm

Water 259 253 29 329 17

296 250 31 – –
333 217 37 225 31

Coca spray: 259 219 35 229 30
44% glyphosate 296 173 44 179 41

+1% Cosmo-Flux 333 128 57 147 51
Poppy spray: 259 194 39 – –

4.9% glyphosate 296 178 43 – –
+0.5% Cosmo-Flux 333 139 53 167 45

Note. Data are the means of three runs each.

Downloaded At: 20:07 2 October 2009

FIG. 1.Volumetric droplet size spectra for coca control spray mixture at 333 km/h (180 knots) aircraft speed with the Accu-Flo nozzle.

nozzle at an aircraft speed of 296 km/h. The water spray hadnozzles are effective at increa sing droplet size for spray
D v0.5alue of 250 μm and 31% spray volume in droplets <150 applications by rotary wing (helicopter) aircraft, but with

μm. The tank mixes containing glyphosate and Cosmo-Flux their relatively small orifidiameter are less effective at
were much finer, with Dv0.5values around 175 μm and 44% very high aircraft speeds. Larg e orifice solid stream nozzles
spray volume in droplets <150 μm (Table 1). There was a cleasuch as the D10 (named for its 10/64 in [4 mm] diameter
2
approximately linear relationship (r .996) between droplet orifice) provide a viable altern ative, especially if used at
size and effective aircraft speed within the range tested inhigh spray pressures to increase the spray breakup length.

study for the coca sprays (Figure 3), which allows extrapolaAt the same spray pressure (2.4 bar), this nozzle increased
tion of droplet size for additional aircraft speeds for the the Dv0.5at 333 km/h from 139 to 168 μm, with a propor-
Flo nozzle. tional approximate 15% decrease in droplets <150 μm rela-

The study included assessments of alternative applicationtive to the Accu-Flo nozzle (Table 1). The use of higher
systems to the Accu-Flo nozzle s currently in use. Accu-Flo spray pressures would probably produce even coarser

327 Annex 131-B

NONTARGET ORGANISMS AND AERIALLY APPLIED GLYPHOSATE 925

FIG. 2. Volumetric droplet size spectra for poppy control spray mixture at 333 km/h (180 knots) aircraft speed with the Accu-Flo nozzle.

700 sprays. However, the high proportion of the tank mix contained

Dv0.1 in nonvolatile materials (the active ingredient product and inert
600
Dv0.5 adjuvant) and the high relative humidity (70%) of the air
Dv0.9 during these applications reduced evaporation rates signifi-
500
cantly compared to similar agricultural applications. In addi-
Dv0.9 –6.385 × air speed + 1532.7
400 r = 0.9977 tion, the tree canopy and adjacent vegetation are effective at
intercepting spray and reduce the availability of droplets for

300 off-target drift where these trees and vegetation are present

Dv0.5 –22275 x air speed + 537.9 (AgDRIFT, 2008).
Downloaded At: 22007 2 October 2009 r = 0.9999 The model-predicted off-swath spray deposition rates for

glyphosate (a.e.) using each of the three simulated aircraft
100 Dv0.1 –0.5675 x air speed + 136.23 types is shown in Figure 5. Mean deposition rates for the
r = 0.996
most concentrated application rate, i.e., the coca spray, on the
0
120 130 140 150 160 170 180 190 target spray block were between 3030 and 3260 g/ha, which
is close to the target rate of 3690 g/ha of active product.
Air speed (knots)
Spray drift potential was related to the speed of the aircraft
FIG. 3. Droplet size parameter values (D , D , and D ) at different
v0.1 v0.5 v0.9 and was similar for the AT-802 and OV-10, but less for the
aircraft speeds for the Accu-Flo nozzle with fitted regression lines. slower ATT-65 where lower air shear at the nozzles produced

coarser sprays.

sprays with this nozzle, based on previous research observa- Because the relative humidity (RH) of the air exerts an
influence on spray drift, particularly for smaller droplets, addi-
tions with narrow-angle, la rge-orifice (e.g., large solid
tional modeling was conducted to characterize spray drift at an
stream) nozzles on high-speed aircraft.
Tests with the prototype reverse Venturi chamber showed that RH of 90%, which is more typical of the conditions that occur
in the Nariño and Putamayo areas in the SW part of Colombia.
increases in droplet size, especially for the “fines” in the spray,
The modeling data (Figure 6) demonstrate that the majority of
can be achieved (Figure 4). For example, there was a decrease in
“fines” from 31 to 25% at an aircraft speed of 296 km/h. the active ingredient (>90%) deposits within 100 m of the
swath edge. Under higher humidity conditions, comparatively

lower deposition of active ingredient occurs at downwind dis-

Spray Drift Modeling tance, with the differential being most pronounced at distances
The AGDISP model showed that the spray drift potential beyond 200 m downwind. This pattern reflects the disproportional

for these applications was equivalent to that of fine to very fine influence of relative humidity on smaller droplets (<15μ 0m),

328 Annex 131-B

926 A. J. HEWITT ET AL.

FIG. 4. Droplet size distribution from the Accu-Flo nozzle in the reverse Venturi chamber. VMD for water only at 259, 296, and 333 km/h (140, 160, and 180
knots) were 321, 257, and 187 μm, respectively.

10000
10000
OV-10 OV-10 70% RH
1000 AT-802 1000 AT-802
ATT-65 OV-10 90% RH
AT-802
100
100
10

10
Deposition (g/ha)
Deposition (g/ha)
1 0.1

0.1 0.01
Downloaded At: 20:07 2 October 2009 50 100 150 200 250 300 0 200 400 600 800 1000 1200 1400 1600

Distance from swath (m) Distance from swath (m)

FIG. 5. AGDISP-predicted deposition of glyphosate (g/ha on a log scale) FIG. 6. Influence of relative humidity on spray drift potential of the

for different aircraft types at representative flight speeds. Negative distanglyphosate–Cosmo-Flux mixture as used to spray coca. The RH of 90% is
represent deposition within the target area; 0 m represents downwind edge o more typical of the southwest region of Colombia. Note that the y axis is a log

spray block. scale to better show that the effect of RH on spray drift is most pronounced a
deposition rates of less than 10 g/ha.

which are primarily responsible for drift, but which also carry a
were taken from the U.S. EPA Ecotox database (U.S. EPA,
small proportion of the total amount of active ingredient
2001) for effects of glyphosate on plants. The values were the
released into the air. The effect of evaporation is due to the loss
EC25 from tests for plants sprayed with formulated glypho-
of water, which is important because even the most concen-
trated tank mix included approximately 50% by volume of sate. These values were converted to units of g a.e./ha to allow
for a common basis for comparison (Table 2).
water. This means that at least half of the droplet volume could
The most sensitive plant in the data set waB srassica rapaand
potentially be lost through evaporation.
the least sensitive wasMagnoliaphytaspp. The plant data were fit-
Airborne spray volumes decreased rapidly, with the major-
ity of the spray predicted to deposit within 30 s after release ted to a log-normal distribution to create a species sensitivity dis-
tribution (SSD, Figure 7) using standard methods (Solomon &
from the aircraft.
Takacs, 2002). In doing this, the test plants were used as surro-

gates for other plants in the environment, a normal process for
Toxicity Data
assessing risks using SSDs. The datum foM r agnoliaphytaspp. is
Toxicity data were obtained for plants and the most sensi- plotted on the SSD but was not used in the regression as this

tive aquatic species, amphibians. The toxicity data for plants datum was judged to be an outlier. The 5th centile of the distribution

329 Annex 131-B

NONTARGET ORGANISMS AND AERIALLY APPLIED GLYPHOSATE 927

TABLE 2

Toxicity Data for Formulated Glyphosate in Plants

Species EC25 Exposure
Species scientific name common name (g/ha) Effect duration (days)

Brassica rapa-rapa Turnip 36 Development 28
Raphanus sativus Radish 44 Growth 21
Cucumis sativus Cucumber 60 Development 28

Glycine max Soybean 76 Growth 28
Triticum aestivum Bread wheat 78 Growth 28

Helianthus annuus Sunflower 78 Development 21
Sorghum bicolor Broomcorn 78 Growth 21
Beta vulgaris Beet 87 Growth 28

Abelmoschus esculentus Okra 90 Growth 28
Raphanus sativus Radish 100 Development 28

Beta vulgaris Beet 103 Development 21
Triticum aestivum Bread wheat 108 Growth 21
Lactuca sativa Lettuce 111 Growth 28

Zea mays Corn 117 Development 336
Avena sativa Common oat 120 Development 28

Allium cepa Common onion 137 Development 21
Glycine max Soybean 157 Growth 21
Cucumis sativus Cucumber 220 Development 21

Cyperus rotundus Purple nutsedge 372 Growth 28
Pisum sativum Pea 436 Development 21

Magnoliophyta Angiosperm 1958 Development 28

Note. Data from (U.S. EPA, 2001).

99 TABLE 3
Regression Coefficients and Intercepts for the Toxicity Data
90
Downloaded At: 20:07 2 October 2009 Distributions for Acute Exposures of Plants and Frogs to
70 Formulated Glyphosate
50
Centile intercepts
30
Percent rank y=ax+b (a.e.)
10
Data source n r 2 ab 5% Units
1
10 10 10 10 Plants 21 0.90 2.96 –6.47 43 g/ha
Glyphosate rate (g/ha AE)
Amphibians 11 0.90 3.68 –12.53 917 μg/L (= 1368
FIG. 7. Species sensitivity distribution (SSD) of glyphosate in plants. g/ha in water

15 cm deep)

was calculated using the transformed values for the slope alated as already described (Table 3) and converted from a con-
intercept of the regression line of the SSD (Table 3). centration to a rate per hectare on the assumption that the

For amphibians, the toxicity data for formulated glyphosconcentration resulted from direct overspray of a 15-cm-deep
were taken from literature as summarized in a companion pool with no exposure reduction via adsorption to sediments
paper (Brain & Solomon, 2009). These data are for formulated
and organic matter.
glyphosate without the addition of Cosmo-Flux, but toxicity
tests on the spray mixture with the most sensitive frog species
Environmental Risk
(Xenopus laevis) (Wildlife International, 2006) showed no
change in sensitivity over previously published values (Edgintonterministic risk values for amphibians and plants
et al., 2004). The 5th centile of the amphibian SSD was calcu-e determined from intercept of the 5th centile toxicity

330 Annex 131-B

928 A. J. HEWITT ET AL.

3000
DISCUSSION AND CONCLUSIONS
OV-10 Effective decision making in aerial applications of pesti-
2500 AT-802
ATT-65 cides can be assisted through the use of appropriate risk assess-
ment and modeling information and tools. The present study
2000
5 centile amphibians assessed spray drift exposure risk in aerial field applications of
glyphosate sprays for control of poppy and coca crops in
1500
Colombia. The droplet size spectra generated when the tank
De1000tion (g/ha) mixes in this study were applied through Accu-Flo nozzles

5tcentile plants under the simulated aircraft speeds relative to field applications
500 were classified as very fine to fine sprays. The droplet size,

0 application, herbicide tank mix, and meteorological and can-
0 20 40 60 80 100 120 opy/terrain characteristics for Colombian conditions were
Amphib. Plnt. ATT-Plnt. OV-10
Distance from edge of spray swath (m) input to a spray drift exposure risk model, AGDISP, to assess
off-target spray drift potential and on-target spray perfor-

FIG. 8. Modeled drift deposition values for glyphosate plus Cosmo-Fluxmance. The results predicted that most of the spray safely
overlaid with the 5th centile toxicity values for plants and amphibiansdeposits within the target area or a few hundreds of meters
distances from the edge of the spray swath where sensitive plants and
amphibians may be at risk are shown by the arrows on the x axis. downwind of the application. An appropriate no-spray buffer
to protect sensitive plants from spray drift exposure would be

50 to 120 m for the coca spray. This is proposed to be a direc-
tional buffer because drift only occurs in the downwind direc-

values (given earlier) and the drift deposition values from tion, and not upwind.
Figure 5. These values are shown in Figure 8. These data
The extensive vegetation of the forest canopy and environ-
show that, for the ATT-65 ai rcraft, the worst-case spray ment around the area where the coca and poppy plants are

drift will be such that the 5th centile toxicity value for sprayed in Colombia will afford excellent reductions in spray
amphibians will not be ex ceeded in the off-swath area. drift potential by interception of droplets with leaf and other

This is indicative of negligible risk to amphibians located surfaces (Raupach et al., 2001). This will greatly reduce the
in 15-cm-deep pools located off-field for this type of spray drift exposure risk from the values reported in this study

aircraft. by 50–90% (AgDRIFT, 2008). The present study showed that
For worst-case spray drift from the AT-802 and the OV- the product tank mixes produced up to 50% more small, drift-

10, the model predicts that the 5th centile toxicity value for able droplets than water alone. This is due largely to the rela-
amphibians may be exceeded in 15-cm-deep pools located tively low dynamic surface tension and extensional viscosity of

within 5 m of the edge of the field. Thus amphibians could the tank mix when these active and inert materials are added.
Downloaded be at risk in locations where pools containing larvae of Tests could be conducted to evaluate the effectiveness of alter-

sensitive species were in the coca field or were within 5 m native adjuvants in increasing droplet size, or even alternative
of the downwind edge of the spray swath. Studies in field glyphosate products with lower surfactant loading or more

microcosms show that toxicity to larval amphibians is favorable physical property and atomization characteristics.
reduced in the presence of natural sediments and that Candidate emulsion/ polymer adjuvants for possible screening

amphibian larvae would not be at risk, even from a could include invert suspension agents, esterified seed oils,
direct overspray at twice the normal rate of application polyacrylamide, and/or guar gum with ammonium sulfate and

(Thompson et al., 2004; Wojtaszek et al., 2004; Bernal adjuvants containing lecithin.
et al., 2009). Because a direct overspray of humans with glyphosate plus

For plants, which are the most sensitive to glyphosate, Cosmo-Flux was a negligible health risk to humans (Solomon
the risks from the worst-case modeled spray drift are et al., 2007), exposure to spray drift downwind of the spray

greater. For spray drift from the ATT-65, sensitive plants presents an even smaller risk. Amphibians in shallow pools
within 30 m of the edge of the field may be affected. For within 5 m downwind of the edge of the field may be at risk

spray drift from the AT-802 and OV-10 aircraft, sensitive under worst-case conditions but field microcosm experiments
plants within 50 m of the spray swath may be affected. demonstrated that this is not the case where natural sediments

Adverse effects beyond that distance are unlikely, espe- are present. As a broad-spectrum translocated herbicide, appli-
cially as glyphosate were shown to stimulate the growth of cation of glyphosate may pose a risk to the most sensitive plant

the several species of plants, such as maize, soybean, euca- species in areas within 30–50 m downwind of spray targets.
lyptus ( Eucalyptus grandis ), pine ( Pinus caribea ), and However, drift deposition will only occur downwind and be

tropical spiderwort ( Commelina benghalensis ), at rates reduced at lower wind speeds. Further, droplet capture by adja-
ranging from 1.8 to 36 g glyphosate a.e./ha (Velini et al., cent trees and shrubs may reduce drift from the modeled worst-

2008). case figures.

331 Annex 131-B

NONTARGET ORGANISMS AND AERIALLY APPLIED GLYPHOSATE 929

In sensitive, high-biodiversity environments, risk to nontar- EPA MRID 43485603, Spray Drift Task Force Report. United States Envi-
ronmental Protection Agency, Washington, DC.
get plant species may be reduced by a number of mitigation Hewitt, A. J. 1997. The importance of droplet size in agricultural spraying.
measures. A primary approach would be to use spray nozzles
Atomization Sprays 7:235–244.
that produce larger drops, notably the D10 solid stream nozzle. Hewitt, A. J., Johnson, D., Fish, J. D., Hermansky, C. G., and Valcore, D. L.
This will significantly reduce spray drift. No-spray buffer 2002. The development of the spray drift task force database on pesticide
movements for aerial agricultural spray applications. Environ. Toxicol.
zones or reduced spray target areas, particularly downwind,
can also be implemented to protect sensitive areas. This Chem. 21:648–658.
Hewitt, A. J., Teske, M. E., and Thistle, H. E. 2001. The development of the
approach is regularly used for ground application machinery AgDRIFT model for aerial application from helicopters and fixed-wing
aircraft. Aust. J. Ecotoxicol. 8:3–6.
adjacent to for example watercourses. Other approaches could International Court of Justice. 2008. Ecuador institutes proceedings against
be to use drift control adjuvants, though these are reported to
Colombia with regard to a dispute concerning the alleged aerial spraying
exert little effect on lethal or injury drift distances with glypho- by Colombia of toxic herbicides over Ecuadorian territory. International
sate, when compared with reductions in drift from alternative Court of Justice. Accessed June 12, 2008. http://www.icj-cij.org/docket/
files/138/14470.pdf
nozzle types (Johnson et al., 2006).
Long-distance transport of spray drift particles is small and Johnson, A. K., Roeth, F. W., Martin, A. R., and Klein, R. N. 2006. Glyphosate
spray drift management with drift-reducing nozzles and adjuvants. Weed
not an issue for humans or the environment beyond 50 m Technol. 20:893–897.
downwind at the maximum permitted wind velocity of 9 km/h Marshall, E. J. P., Solomon, K. R., and Carrasquilla, G. 2009ar(ythroxylum
coca) control is affected by glyphosate formulations and adjuvantJs..Toxicol.
for spraying operations. Long-distance movement of glypho-
sate is negligible if appropriate no-spray buffers are used and Environ. Health A72:930–936.
Raupach, M. R., Woods, N., Dorr, G., Leys, J. F., and Cleugh, H. A. 2001. The
nonexistent if the wind direction during spraying is away from entrapment of particles by windbreaks. Atmos. Environ. 35:3373–3383.
Snyder, H. E., Senser, D. W., and Lefebvre, A. H. 1989. Experimental tech-
the areas of concern.
niques for drop-size measurements in fan sprays. Proc. Int. Inst. Liquid
Atomization Spray Systems—Americas, pp. 56–61. Worcester, MA: Inter-
national Institutes for Liquid Atomization and Spray Systems.
REFERENCES Solomon, K.R., and Thompson, D.G. 2003. Ecological risk assessment for

AgDRIFT. 2008. Drift filtration by natural and artificial collectors: A litera- aquatic organisms from over-water uses ofr glyphosate. J. Toxicol. Envi-
ture review. AgDRIFT. Accessed June 17, 2008. http://www.agdrift.com/ ron. Health B 6:289–324.
PDF_FILES/drift_filtration.PDF Solomon, K. R., Anadon, A., Carrasquilla, G., Cerdeira, A., Marshall, E. J. P,
American Society for Testing and Materials. 1987. Standard practice for and Sanin, L. H. 2007. Coca and poppy eradication in Colombia: Environ-
determining data criteria and processing for liquid drop size analysis. mental and human health assessment of aerially applied glyphosate. Rev.

Technical report E799-03. American Society for Testing and Materials. Environ. Contam. Toxicol. 190:43–125.
West Conshohocken, PA, USA. Solomon, K. R., and Takacs, P. 2002. Probabilistic risk assessment using
Bernal, M. H., Solomon, K. R., and Carrasquilla, G. 2009. Tocxiity of glyphosatespecies sensitivity distributions. ISpecies sensitivity distributions in
and Cosmo-Flux to larval and juvenile Colombian frogs. 2. Field and labora- ecotoxicology , ed. L. Posthuma, pp. 285–313. Boca Raton, FL: CRC

tory microcosm acute toxicity.J. Toxicol. Environ. Health A72:961–965. Press.
Bilanin, A. J., Teske, M. E., Barry, J. W., and Ekblad, R. B. 1989. AGDISP: Thompson, D. G., Wojtaszek, B. F., Staznik, B., Chartrand, D. T., and
The aircraft spray dispersion model, code development and experimental Stephenson, G. R. 2004. Chemical and biomonitoring to assess potential
Downloaded At: 20validation. Trans. Am. Soc. Agric. Eng. 32:327–334. acute effects of Vision herbicide on native amphibian larvae in forest wetlands.
Environ. Toxicol. Chem. 23:843–849.
Bird, S. L., Perry, S. G., Ray, S. L., and Teske, M. E. 2002. Evaluation of the
AGDISP aerial spray algorithms in the AgDRIFT model. Environ. Toxicol. U.S. Envirnomental Protection Agency. 2001. Environmental Effects Database
Chem. 21:672–681. (EEDB). ECOTOX Database System, U.S. Environmental Protection
Brain, R. A., and Solomon, K. R. 2009. Comparative hazards of glyphosate, Agency, Office of Pesticide Programs, Environmental Fate and Effects Divi-
other pesticides, and other human activities to amphibians in the productionsion, Washington, DC. Accessed June 10, 2006. http://www.epa.gov/ecotox

of coca. J. Toxicol. Environ. Health A 72:937–948. Velini, E. D., Alves, E., Godoy, M.C., meschede, D.K., Souza, R.T., and Duke,
Dodge, L. G. 1988. Representation of average drop sizes in spray. J. Prop. S.O.2008. Glyphosate applied at low doses can stimulate plant growth.
Power 4:490–496. Pestic. Manage. Sci. 64:489–496.
Edginton, A. N., Sheridan, P. M., Stephenson, G. R., Thompson, D. G., and Boer-dlife International. 2006. Glyphosate–Cosmo-Flux–coca mix: A 96-hour

mans, H. J. 2004. Comparative effects of pH and Vision rhbeicide on two lifestatic-renewal acute toxicity test with the African clawed-frog tadpoXleen(opus
stages of four anuran amphibian species. Environ. Toxicol.23. :815–822. laevis). Final report. Technical report 628A-101, Wildlife International.
Foster, M. P., and French, H. M. 1992. A new wind tunnel facility for atomiza- Easton, MD, USA.
tion studies. Conference of Engineering in Agriculture, 4–7 October, 1992,jtaszek, B. F., Staznik, B., Chartrand, D. T., Stephenson, G. R., and
Albury, New South Wales, Australia. Thompson, D. G. 2004. Effects of Vision herbicide on mortality, avoidance

Hewitt, A. J. 1994. Measurement techniques for atomization droplet size spec- response, and growth of amphibian larvae in two forest wetlands. Environ.
tra using particle size analyzers in wind tunnels. Technical report T94-001.Toxicol. Chem. 23:832–842.

332 Annex 131-C

Annex 131-C

E.J.P.MARSHALL ET AL, “OCA (ERYTHROXYLUM COCA ) ONTROL ISA FFECTED BY
G LYPHOSATE FORMULATIONS AND A DJUVANTS”

(Journal of Toxicology and Environmental Health, Part A, 72:930-936, 2009)

333 Annex 131-C

Journal of Toxicology and Environmental Health, Part A, 72: 930–936, 2009
ISSN: 1528-7394 print / 1087-2620 online
DOI: 10.1080/15287390902929675

UTEH
Coca (Erythroxylum coca) Control is Affected

by Glyphosate Formulations and Adjuvants

Coca Control is Af1ected by Glyphosates 2 3
E. J. P. Marshall , Keith R. Solomon , and Gabriel Carrasquilla
1Marshall Agroecology Limited, Barton, Winscombe, Somerset, United Kingdom, CentreforToxicology
and Department of Environmental Biology, University of Guelph, Guelph, Ontario, Canada, and
3
Facultad de Salud, Universidad del Valle, Valle, Colombia

particularly aerial application, prior to a recommendation.
The aerial spray program for the eradication of coca in Should the glyphosate prod uct require changing, Roundup
Colombia uses Glyphos, a local formulation of glyphosate tank- Biactive may be considered. Should the adjuvant require chang-
mixed with an adjuvant product, Cosmo-Flux. There are some ing, then on the basis of this research, Silwet L-77 and Mixture B
would be good candidates for further evaluation.
potential risks to amphibians fr om direct overspraying of shal-
low waters. In order to evaluate potential alternative mixtures, a
field experiment was conducted at the Center of National Train-
ing of Police Operations in Tolima province, Colombia. Plants of
coca were established with irrigati on and grown to 75 cm tall. A The control of the drug plant Erythroxylum coca Lam. and
randomized split-plot design experiment was laid out and the closely related E. novogranatense (coca), the sources of

sprayed with a range of glypho sate formulations and different cocaine, is the focus of considerable effort and expenditure in
adjuvants using an experimental ground sprayer. Assessments a number of South American countries. Authorities are tar-
were made of plant vigor, height, and above-ground standing
crop (fresh weight) 3 wk after application. Resprouting of plants geting the removal of the plant and are also engaged in rural
was assessed at 9 wk after trea tment. Unformulated glyphosate development projects to replace the illicit cultivation of this
applied as the product Rodeo gave poorer control of coca than crop. As well as controlling supplies of the drugs, there are

two formulated products, Roundup Biactive (from Europe) and also efforts to reduce global demand for the drug. The illicit
Colombian Glyphos. In general, these products performed well cocaine industry has an estimated production that reached the
without added adjuvants, giving control similar to that of the
Downloadederadication mixture with Cosmo-Flux. There was some evidence streets of 600 tonnes in 2007 with a retail value of approxi-
that addition of the adjuvant Silwet L-77 and to a lesser extent mately $934 million. Retail prices are $2200/kg in Colombia,
Mixture B (from the United Kingdom) gave more rapid herbi- $10,200/kg in the United States, and $85,000/kg in the streets
cide symptoms. There were also indications that glyphosate
of Europe (UNODC, 2007). As part of the supply control
rates of less than 3.69 kg acid equivalents (a.e.)/ha could effort that started in the 1970s, an aerial spraying eradication
give control in the range of 95%. Depending on the environmen- program in Colombia was starte d in 1997 using the herbicide
tal risk requirements, the experiment indicates that, should
other spray mixtures be required, there are potential alterna- glyphosate. The area of coca sprayed with glyphosate has
tives. These would require extensive field testing to cover differ- shown a steady increase over re cent years, reaching 153,000
ent environmental conditions, different coca varieties, and
ha in 2007 (personal communication, National Police of
Colombia, Bogotá, December 2007).
The aerial spray program is conducted with two types of
©General Secretariat of the Organization of American States,
2009. This paper was prepared as part of a Study entitled “Productionommercial crop-spraying aircraft fitted with Accu-Flo air-
of Illicit Drugs, the Environment and Human Health,” financed with induction nozzles. The spray planes, Air Tractor 802 (Air
contributions from the Governments of Colombia and the United
States of America. The conclusions and opinions expressed herein areTractor Inc., Olney, TX) and Rockwell OV-10, are equipped
with high-resolution tracking equipment and positional data
those of the authors and not necessarily those of the Organization orecorders that display position, provide directional guid-
American States and its General Secretariat, which as of the date of
this copyright, have not formulated any opinion with respect to them.nce, and store data for late r analysis (Solomon et al.,
The authors thank Samuel Rodriquez and Oscar Mendez for their 2007).
work in the field and for maintaining the plots.
Address correspondence to Dr. E. J. P. Marshall, Marshall Glyphosate used for the aerial eradication program in
Agroecology Ltd, 2 Nut Tree Cottages, Barton, Winscombe, Somerset, Colombia is the Glyphos product containing 354 g acid equiva-
lents (a.e.)/L sold for agricultural use. Glyphosate was found to
BS25 1DU, UK. E-mail: [email protected] with copy to
[email protected] be effective at controlling both species of coca, E. coca and E.

930

334 Annex 131-C

COCA CONTROL IS AFFECTED BY GLYPHOSATES 931

novogranatense (Ferreira et al., 1997). The aerial application ranges of the Andes, located at N 04º 15.971, W 75º 01.373, and
rate of glyphosate is 3.69 kg a.e./ha in Colombia. To maximize at 517 m above sea level (a.s.l.). The soils are valley alluvia.

penetration and effectiveness of the spray formulation, glypho- The selected area was initially covered with grasses, sedges,
sate is tank-mixed with an adjuvant product (CosmoFlux 411F; and low shrubs. This was mown with a tractor-mounted bush
Cosmoagro, Bogotá). The inclusion of the adjuvant follows the cutter, then sprayed with glyphosate and ploughed. The site was

studies of Collins and Helling (2002), who demonstrated a cultivated and the plants were hand planted in rows 1 m apart
fourfold increase in glyphosate efficacy by including suitable on 1-m spacings in plots of 60 plants, with each plot arranged

adjuvant mixtures in the spray. with 6 rows of 10 plants. The planting and agronomy of the
Surfactants, such as the polyethoxylates in Cosmo-Flux, plants match much of the pattern of illicit growing, where pesti-
enhance efficacy through (1) increasing target surface adher- cides (permethrin and mancozeb) and fertilizer are used to pro-

ence, (2) promoting better droplet spread, (3) better disper- mote crop production. Irrigation was applied approximately
sion, (4) prevention of aggregation, and (5) enhanced twice per week, to enhance survival and growth.
penetration of herbicides into target plant tissues through the

reduction of surface tension on plants. Surfactants also dis-
rupt the water-insoluble wax cuticle, thus increasing the pen- Plant Material
etration of herbicide active ingredient. Base oils, such as the Cuttings of E. coca variety Pajarito were obtained from the

isoparaffins in Cosmo-Flux, are another class of adjuvants Department of Cauca, in the southwest of Colombia. Cuttings
used in pesticide formulations to primarily aid foliar absorp- were initially maintained in a shade house in December 2006,
treated as necessary with fertilizer, fungicide, and insecticide
tion of the pesticide by disrupting the waxy cuticle on the
outer surface of foliage, wh ich increases permeability and then planted out on 15 January and 20 January 2007.
(Solomon et al., 2007). Plants that did not survive were replaced. Plants were allowed
to grow to approximately 75 cm tall. Two weeks prior to spray
Environmental risk assessments of the aerial eradication
program indicated that the relative impact of the glyphosate treatment, taller plants were trimmed to 75 cm.
mixture is small in comparison with the impacts of forest clear-

ance for cropping (Solomon et al., 2007). Nevertheless, there
could be some impact on amphibian species in shallow waters Design Layout and Treatments
There were 49 plots of 60 plants, of which 36 were selected
(<30 cm deep) that are directly oversprayed. Such effects are
unlikely to be due to glyphosate, but to the adjuvants present infor experimental use on the basis of good shrub growth. These
the spray mixture. Further ecotoxicological tests were con- plots, each representing a main plot, were grouped into three

ducted with the spray mix as used in Colombia (Bernal et al., blocks across the experimental area. Main plots (60 plants)
2009a, 2009b). Developments in adjuvant research indicate were allocated to 1 of 12 glyphosate/adjuvant treatments at
that it may be possible to identify an adjuvant mixture or mix- random. Three glyphosate formulations were used: Colombian

Downloaded tures as effective as the current mixture, but with less risk to Glyphos (354 g a.e./L, Monsanto), Roundup Biactive (360 g
native amphibians. a.e./L, Monsanto Europe), and Rodeo ® (479 g a.e./L, Dow
The field experiment described here was conducted as an AgroSciences). Glyphos is formulated with adjuvants based on

initial step to evaluate the efficacy of a range of glyphosate for-lyethoxylated tallowamines (POEA) for agricultural use.
mulations mixed with different adjuvants. It was postulated Roundup Biactive, a European product, uses a patented blend
of surfactants less dependent on tallowamines for use in or near
that alternative combinations might give as good or better con-
trol of coca than the formulation currently used in the aerial water. Rodeo is an unformulated glyphosate product, lacking
eradication program. As noted earlier, reduced toxicity of the adjuvants. Five adjuvants were also used: Cosmo-Flux, Intake,

spray mixture may have environmental benefits, plus there is Mixture B, Silwet L-77, and LI-700. Cosmo-Flux is an agricul-
potential for reducing the application rate of glyphosate when tural adjuvant containing nonionic surfactants (a mixture of
mixed with different adjuvants used in the aerial eradication linear and aryl polyethoxylates: 17% w/v) and isoparaffins

program. With approximately 153,000 ha sprayed in Colombia (83% v/v) (Cosmoagro 2004). Intake (Headland Agrochemi-
in 2007, there would be considerable cost savings if a reduced cals Ltd., Great Chesterford, United Kingdom) is an agricul-
tural penetrant containing 40% (w/w) propionic acid. Mixture
rate gave as good control as current practice.
B (AmegA Sciences, Daventry, United Kingdom) is 50% (w/v)
nonylphenol ethylene oxide condensate and 50% (w/v) primary

MATERIALS AND METHODS alcohol ethylene oxide condensate. Silwet L-77 (GE Silicones,
Wilton, CT) is 99.5% (w/w) polyalkyleneoxide modified
Site heptamethyltrisiloxane, a nonionic organosilicone. LI-700

The field site was located on a finca (ranch) in County San (Nufarm Agriculture, Inc., Calgary, Canada) is a penetrant and
Luis in the Department of Tolima, Colombia. The area is to the acidifier composed of phosphatidylcholine, methylacetic acid,
west of Bogotá between the eastern and central mountain and alkyl polyoxyethylene ether.

335 Annex 131-C

932 E. J. P. MARSHALL ET AL.

A split-plot design was created by dividing each main plot (21 August 2007). At the same time, maximum bush heights
into three subplots, each of two adjacent rows of coca bushes were recorded. All bushes were harvested at ground level 3 wk

(20 plants). Subplots were randomly ascribed to receive one of after treatment (22 August 2007) and total fresh weight for each
three rates of glyphosate (all rates as a.e.): 1 kg/ha, 2 kg/ha,subplot was determined. Six weeks after harvesting (9 wk after
4 kg/ha. Aerial application rates of glyphosate are 3.69 kg/ha ispraying) on 5 October 2007, numbers of sprouts per stem were

Colombia, but 2 lower rates were selected for field testing, in counted to measure plant recovery. These data were used to
order to evaluate efficacy and survival at lower application give a survival rate out of 20 plants per subplot.
rates.

Herbicide and adjuvant treatments were applied with a
three-nozzle plot sprayer, mounted with 8002 Teejet nozzles Statistical Analysis
(Spraying Systems Co., Wheaton, IL) 50 cm above the canopy. Data were analyzedusing analysis of variance (ANOVA) of a

Calibrations showed the sprayer delivered a volume rate of split-plot design. Data were transformed as necessary to comply
200 L/ha at 3 bar (40 psi) boom pressure at the calibrated for- with the need for normality, following examination of residu-
ward speed. These treatments were ground applied, so neces- als; square root or log. transformations (n +0.05) were used.
10
sarily the volume applied was higher than that used in the Plant heights were analyzed using initial bush heights as a
aerial application. This facilitated accurate small plot studies ofariate. Vigor scores were analyzed using bush ellipse area
as a covariate, reflecting potential herbicide coverage and
a manageable size. Windshields were placed between subplots
to prevent any drift between treatments. Applications were therefore dose per bush. The Genstat 9th edition package (VSN
made on 1 and 2 August 2007 with air temperatures increasing International, Hemel Hempstead, UK) was used for analyses

during the day from 25 to 33ºC and relative humidities declin- and data transformations. There were two spraying errors, so
ing from 80% to 40% at midday. the four affected subplots were treated as missing values in the
analyses.

Assessments

Prior to treatment, the maximum height and two widths
(maximum and at 90º; Diameter-A, Diameter-B) of each coca RESULTS
bush were measured, allowing the calculation of bush area
Plant Height and Bush Size
based on an ellipse: Analysis of the initial bush heights prior to treatment

(mean height 80 cm) indicated there were some significant
Ellipse area =Π (Diameter-A /() 2 Diam/e)ter-B 2 (1) differences between treatments. Analysis of the ellipse area
of each bush also indicated some initial and systematic dif-

and bush volume: ferences across the experiment, despite arranging the plots
Downloaded At: 20:07 2 October 2009 into blocks. Plots to receive treatments 4 and 8 (Glyphosate
+ Intake; Roundup Biactive + Silwet L-77) had the
Bush volume = ×ellipse area height (2)
smallest bushes, while plots with treatments 6 and 11
(Glyphosate + Silwet L-77; Rodeo + Silwet L-77) had the
Heights, ellipse area, and volume could be used as covariates largest plants. These results confirmed that it was neces-
in univariate analyses.
Plant symptoms were scored on a 5-point scale (Table 1) for sary to use covariates representing plant size at treatment
in statistical analyses.
individual bushes 1 wk (8 August 2007) and 3 wk after spraying
Three weeks after treatment, on 21 August 2007, plant
heights were measured. Analysis using bush volume as a cova-
riate indicated that treatments and application rate were signifi-
TABLE 1
Symptom Scores for Coca Plant Health cant factors, but there was no interaction. Higher rates of
glyphosate gave shorter plants. Plant heights were not sig-
Score Effect Symptoms nificantly shorter than untreated controls on plots that had

1 None Leaves green, healthy been treated with Glyphos + Silwet L-77 and Rodeo + LI-700
(Figure 1). The shortest plants were found on plots treated
2 Slight A few leaves yellowing, brown with Glyphos + Intake, Roundup Biactive alone, and Roundup
edges; healthy new growth
3 Moderate Half the leaves yellow or brown; Biactive + Silwet L-77.

some recovery
4 Strong Most leaves yellow, brown or
Symptom Scores
fallen; no recovery One week after treatment, symptoms of glyphosate were
5 Killed All leaves brown or fallen
developing on treated plants.There were significant differences

336 Annex 131-C

COCA CONTROL IS AFFECTED BY GLYPHOSATES 933

82 treatment and rate effects and an interaction. All treatments
80 LSD = 2.145 showed glyphosate effects (Table 3) compared with the con-

78 trols. At the lowest rate of glyphosate, the highest symptom
76 scores and therefore the most activity were shown with

74 Glyphos + Intake and Roundup Biactive with Silwet L-77 and
Plant height (cm) Mixture B. At 2 kg /ha, the greatest activity was shown by

70 Glyphos alone, Roundup Biactive alone, Glyphos with Silwet
68 L-77 or Mixture B, and Roundup Biactive with Mixture B. At
nly B eB -77 the highest rate of 4 kg /ha, the poorest treatment was Rodeo +
-lux e L-77itive-77 I-700 L I700
+ L LI-700. When comparing the glyphosate formulations, Rodeo
Glyphos o + Soundup B ictiive + MixRodeo + L gave lower vigor scores than Glyphos or Roundup Biactive
Unsprayed controlGlyphos + Mixtureive + SilwRodeo + Silwet
Glyphos + Cosmolyphos oundup B across the adjuvant treatments, indicating less overall activity.
R Rundup B There was a trend for Roundup Biactive to give higher scores
Roundup B
at the lowest rate, in comparison with the other formulations.
FIG. 1. Mean coca plant height (cm) on polts treated with different glyphosate
formulations and adjuvants 3 wk previously. (SED = 1.094; df = 2041).

Fresh Weights

between treatments and rates of glyphosate (Table 2). All treat- The fresh weights of coca were not significantly different to
ments, apart from the low rate of Roundup Biactive + LI-700 controls on only 6 treatments (coefficient of variation = 21%),

were significantly different from the controls and higher scores all at the 1-kg/ha rate of glyphosate. These treatments were
were found at the higher rates of the herbicide. At the lowest Glyphos alone, Glyphos with Cosmo-Flux, Mixture B, or

rate of glyphosate (1 kg/ha), the treatments with the products Silwet L-77, and the two mixtures of LI-700 with Roundup
Glyphos and Roundup Biactive with Silwet L-77 had higher Biactive and Rodeo. At 2 kg/ha, the lowest standing crop was
scores than other adjuvants. The treatment with Roundup
found on plots treated with Glyphos and Silwet L-77 or Mix-
Biactive + Mixture B also showed good activity. At 2 kg/ha, ture B (1.62 kg). Interestingly, the only 2-kg/ha treatments that
Glyphos with Silwet L-77 and Glyphos alone had highest were statistically greater than this were the two other Silwet

activity, with Roundup Biactive alone or with Mixture B also L-77 treatments and the control. At 4 kg /ha, lowest standing
showing good activity. crop was found with the Glyphos + Cosmo-Flux treatment, but

Three weeks after treatment, on 21 August 2007, glyphosate none of the plots were statistically different from each other.
symptoms were well developed, with plants clearly dying and Only Rodeo + LI-700 plots had fresh weights statistically dif-

leaves dropping. Statistical analyses again showed significant ferent from the Glyphos + Cosmo-Flux at this rate.

Downloaded At: 20:07 2 October 2009
TABLE 2
Symptom Scores (0–5) for Coca Plants 1 wk after Treatment with Different Glyphosate Formulations

and Adjuvants and Three Rates of Glyphosate (SED = 0.11; df = 2041)

Adjuvant Glyphosate rate

Concentration

Glyphosate product Name (%v/v) 1 kg/ha 2 kg/ha 4 kg/ha

None Water control – 2.2 2.2 2.0

Glyphos – – 2.6 3.2 3.3
Glyphos Cosmo-Flux 2.3 2.7 3.0 3.7

Glyphos Intake 0.5 2.8 2.7 3.2
Glyphos Mixture B 2 2.5 3.0 3.2
Glyphos Silwet L-77 1 3.2 3.3 3.5

Roundup Biactive – – 2.7 3.1 2.8
Roundup Biactive Silwet L-77 1 3.0 2.9 3.3

Roundup Biactive LI-700 0.5 2.3 2.9 3.2
Roundup Biactive Mixture B 2 3.1 3.1 3.3

Rodeo Silwet L-77 1 2.9 3.0 3.1
Rodeo LI-700 1 2.5 2.6 2.6

Overall LSD (p = .05) = 0.21.

337 Annex 131-C

934 E. J. P. MARSHALL ET AL.

TABLE 3

Symptom Scores (0–5) for Coca Plants 3 wk after Treatment with Different Glyphosate Formulations and
Adjuvants and Three Rates of Glyphosate (SED = 0.12; df = 2041)

Adjuvant Glyphosate rate

Concentration
Glyphosate product Name (%v/v) 1 kg/ha 2 kg/ha 4 kg/ha

None Water control – 2.2 2.4 2.2
Glyphos – – 2.5 4.1 4.2
Glyphos Cosmo-Flux 2.3 2.5 3.5 4.4

Glyphos Intake 0.5 3.2 3.3 4.3
Glyphos Mixture B 2 2.7 3.7 4.5

Glyphos Silwet L-77 1 3.1 3.8 4.5
Roundup Biactive – – 3.1 3.8 4.1

Roundup Biactive Silwet L-77 1 3.3 3.4 4.4
Roundup Biactive LI-700 0.5 2.7 3.3 4.0

Roundup Biactive Mixture B 2 3.2 3.7 4.3
Rodeo Silwet L-77 1 2.8 3.2 3.9

Rodeo LI-700 1 2.7 3.1 3.4

Overall LSD (p = .05) = 0.23.

Recovery 80
70 1.0 kg/ha
After all coca bushes had been cut, the plots were left to 60 LSD = 22.8 2.0 kg/ha
recover, with occasional watering. On 5 October 2007, the 50 4.0 kg/ha
40
numbers of sprouts per bush showed that most glyphosate 30
treatments gave good control, but these data (not shown) were % survival
rather variable (coefficient of variation >50%). Only the low 20
10
rate of Glyphos + Mixture B was similar to untreated control 0
plots. The Rodeo formulation was less active than other formu- nly -77 -700 eB -7
-lux iactive t I700
lations tested. There were indications that at the 2- and 4-kg/ha Slwet L Silwet L-+7ixtur
Downloadedrates of glyphosate, the Glyphos formulation was as efficacious GlyphosGlyphos + Intake ive Bictive + LI Rodeo + L
Unsprayed controlGlyphos + Mixture B BictRodeo + Silwe
alone, compared with mixtures with adjuvants. At 2 kg/ha, the Glyphos + Cosmolyphos + Rundup
best mixtures were Glyphos and Roundup Biactive with the oundup B Roundup
R
adjuvants Silwet L-77 and Mixture B.
In terms of the numbers of surviving plants 9 wk after FIG. 2. Percentage survival of coca bushes after treatment with different
glyphosate formulations and adjuvants and three rates of glyphosate. Data
treatment, only half of the control plants were actively growing collected 9 wk after application and 6 wk after plants were cut to the ground.
(Figure 2). In terms of survival, glyphosate activity was poor in

the Rodeo formulation, relative to other treatments. The
Glyphos formulation worked well at the 2- and 4-kg/ha rates.
Roundup Biactive gave slightly higher survival overall, but at data, is one measure. However, speed of effect is another factor
to consider. This may be evaluated from vigor at 1 and 3 wk
the 4-kg/ha rate, control was equivalent to the Glyphos formu-
lation, across adjuvants. The standard eradication program after treatment and standing crop at 3 wk. The survival data
(Figure 2) show that the unformulated Rodeo glyphosate did
treatment is Glyphos + Cosmo-Flux, so comparison with this is
appropriate. Glyphos + Silwet L-77 performed the same as the not work as well as the other two products. The standard eradi-
cation program treatment of Glyphos + Cosmo-Flux and
Cosmo-Flux across doses. Roundup Biactive gave similar con-
trol at the two higher doses, when mixed with Silwet L-77 or Glyphos + Silwet L-77 yielded reliable control at 2- and 4-kg/ha
rates. Equivalent control was also given by Roundup Biactive
Mixture B.
when mixed with Silwet L-77 orMixture B at these rates. There
were no great advantages in adding extra adjuvants to the stan-
DISCUSSION dard Glyphos formulation. The adjuvant LI-700 was not particu-

In terms of treatment efficacy, there are a number of factors larly effective in enhancing glyphosate activity. These patterns
to consider. Outright plant kill, interpreted from the survival were repeated in the numbers of sprouts per plant.

338 Annex 131-C

COCA CONTROL IS AFFECTED BY GLYPHOSATES 935

Assessments of plant vigor 1 wk after treatment indicated was only recorded for Glyphos alone or with Cosmo-Flux or
that the addition of Silwet L-77 to Glyphos and of Silwet L-77Silwet L-77. Nevertheless, 95% control might also be achieved

and Mixture B to Roundup Biactive resulted in greater symp- by Roundup Biactive alone or mixed with Silwet L-77 or
toms of herbicide damage to coca plants (Table 2) compared Mixture B. In terms of herbicide efficacy, the results indicate
with other treatments. At spraying, the behavior of Silwet L-7that the advantages of adding alternative adjuvants to the two

was noticeably different to other treatments. The spray mixturformulated glyphosate products are not significant. Indeed,
formed an even film over sprayed leaves, rather than a patternthere appeared to be little advantage in adding Cosmo-Flux to

of spreading droplets. the Glyphos product in this study. This may reflect the conser-
After 3 wk, scores were quantitatively higher (plant vigor vative results of this trial with higher volume spray rates,
was lower) on Glyphos + Silwet L-77 plots compared with compared with the work of Collins and Helling (2002). How-

Glyphos alone or the standard with Cosmo-Flux at the 4-kg/ha ever, in terms of reducing environmental risk from the aerial
rate. The advantages of adding adjuvants to the formulated glyeradication program, the results provide useful data.
phosate products were marginal, in terms of the observed gly- There needs to be careful evaluation of the relative environ-

phosate activity at this stage. The weights of coca bushes 3 wmental risks posed by the different componentsof the glypho-
after application also indicated that the activity of glyphosasate formulations and the added adjuvants in the aerial
was modified only quantitatively by the addition of adjuvants
eradication program. If the greatest risk is presented by Cosmo-
to the formulated products (Table 4). Flux, then alternatives for mixing with Glyphos that may go for
Extensive studies on the interactions between glyphosate further testing are Silwet L-77 and Mixture B.If there are risks

and adjuvants were reported elsewhere (Collins & Helling, associated with formulated Glyphos, then Roundup Biactive
2002). Addition of adjuvants might increase the toxicity of gly- gives similar levels of control, either alone or in mixture with
phosate to coca by a factor of4, compared with unformulated Silwet L-77 or Mixture B. This product is sold in Europe by

glyphosate. Collins and Helling (2002) indicated best results Monsanto Europe S.A., where it is also cleared for use on float-
with two glyphosate-surfactant systems, one being a mixture of ing and emerged aquatic weeds (Monsanto Europe, 2007).
crop-oil concentrate and the organosilicone Silwet L-77, and the Although the results of our research are informative, they

other a mixture of cationic surfactant and anionic surfactants. represent only a single trial, in one year, at one site. Further,
Collins and Helling (2002) noted that the eradication program in the treatments were applied using relatively precise experimen-

Colombia was modified successfully in the light of their studies. tal ground-spraying equipment. The results provide indications
The results here indicate that the unformulated glyphosate of where further development work can be directed, but extrap-
(Rodeo), even with added adjuvants, did not yield adequate olation to aerial application conditions would be premature.

control of coca. In aiming for total control of coca, the rateHerbicide behavior can change with volume rate; studies com-
4 kg/ha of glyphosate gave more consistent effects; total control paring glyphosate applications at 23, 47, 94, or 190 L/ha

Downloaded At: 20:07 2 October 2009 TABLE 4

Fresh Weight (kg) of 20 Coca Bushes Treated with Different Formulations of Glyphosate and Adjuvants and Three
Rates of Glyphosate, for Plants Harvested on 22 August 2007, 3 wk after Treatment (SED = 0.44; df = 64)

Adjuvant Glyphosate rate

Concentration
Glyphosate product Name (%v/v) 1 kg/ha 2 kg/ha 4 kg/ha

None Water control – 3.84 3.63 3.99

Glyphos – – 3.64 1.84 1.68
Glyphos Cosmo-Flux 2.3 3.12 1.83 1.20
Glyphos Intake 0.5 2.38 2.15 1.48

Glyphos Mixture B 2 3.02 1.62 1.38
Glyphos Silwet L-77 1 3.02 1.62 1.48
Roundup Biactive – – 2.59 1.74 1.66

Roundup Biactive Silwet L-77 1 2.59 2.48 1.59
Roundup Biactive LI-700 0.5 3.27 1.96 1.21
Roundup Biactive Mixture B 2 2.24 1.89 1.55

Rodeo Silwet L-77 1 2.81 2.76 1.65
Rodeo LI-700 1 3.47 2.25 2.23

Overall LSD (p = .05) = 0.874.

339 Annex 131-C

936 E. J. P. MARSHALL ET AL.

showed best control of grasses at low volumes (Ramsdale et The current aerial eradication treatments are working well
al., 2003). Low volumes apparently maximize glyphosate effi-
in the field. Before any recommendations to change the spray
cacy, mainly via high herbicide concentration in the spray mixture are made, there needs to be at least a two-stage process
deposit. Higher coca control might therefore be expected at that (a) evaluates the components that drive the key risks

volume rates typical of aerial application and the differences within the current formulation, followed by (b) setting up suit-
between adjuvants may be enhanced. Only one variety of coca able field trials of alternatives applied from the air.

was grown and tested; other varieties might be more or less
susceptible to the tested formulations.

Based on this single field trial, there appears to be some scope REFERENCES
for reducing the rate of glyphosate in the aerial eradication pro- Bernal, M. H., Solomon, K. R., and Carrasquilla, G2.009a. Toxicity of gly-

gram from the current 3.69 kg/ha. Further testing would be phosate and Cosmo-Flux to larval Colombian frogs. 1. Laboratory acute
required to evaluate this point and to evaluate any other changes toxicity. J. Toxicol. Environ. Health A 72:961–965.
Bernal, M. H., Solomon, K. R., and Carrasquilla, G. 2009b. Toxicity of glyphosate
to the current eradication treatment. Such tests would comprise and Cosmo-Flux to larval and juvenile Colombian frogs. 2. Field and labora-
field evaluations in different locations and with different coca tory microcosm acute toxicity.J. Toxicol. Environ. Health A 72:966–973.
varieties, followed by aerial application experiments. The behav- Collins, R. T., and Helling, C. S. 2002. Surfactant-enhanced control of two

ior of the adjuvant mixtures when applied through raindrop noz- Erythroxylum species by ®lyphosate. Weed Technol. 16:851–859.
zles may be different from that from ground application Cosmoagro. 2004. Cosmo-Flux 411F Label, Cosmoagro S.A. Accessed 20
October 2004, www.cosmoagro.com
machinery, so further testing would be essential. Ferreira, J. F. S., Smeda, R. J., and Duke, S. O. 1997. Control of coca plants
(Erythroxylum coca and E. novogranatense) with glyphosate. Weed Sci.
45:551–556.
Conclusions and Recommendations
Monsanto Europe. 2007. Roundup Biactive, Monsanto Europe. Accessed 29
January 2008, http://www.monsanto-ag.co.uk/layout/crop_pro/portfolio/
• Addition of the adjuvants Silwet L-77 or LI-700 to biactive.asp
unformulated glyphosate (Rodeo) was not sufficient Ramsdale, B. K., Messersmith, C. G., and Nalewaja, J. D. 2003. Spray volume,
formulation, ammonium sulfate, and nozzle effects on glyphosate efficacy.
to give acceptable control of coca. Weed Technol. 17:589–598.
• Roundup Biactive would be a suitable formulation of Solomon, K. R., Anadon, A., Carrasquilla, G., Cerdeira, A., Marshall, E. J. P.,

glyphosate to use, if the Colombian Glyphos were to and Sanin, L. H. 2007. Coca and poppy eradication in Colombia: Environ-
be replaced. mental and human health assessment of aerially applied glyphosate. Rev.
• Should Cosmo-Flux need to be replaced, then the Environ. Contam. Toxicol. 190:43–125.
United Nations Office on Drugs and Crime. 2007. World drug report 2007.
adjuvants Silwet L-77 and Mixture B might provide United Nations Office on Drugs and Crime. Accessed 29 January 2008,
suitable replacements. http://www.unodc.org

Downloaded At: 20:07 2 October 2009

340 Annex 131-D

Annex 131-D

R.A.BRAIN ET ., “OMPARISON OF THH AZARDSPOSED TOAMPHIBIANS BY THE
GLYPHOSATESPRAY CONTROL PROGRAM VERSUS THECHEMICAL ANDPHYSICAL
A CTIVITIES COCA PRODUCTION IC OLOMBIA”

(Journal of Toxicology and Environmental Health, Part A, 72:937-948, 2009)

341 Annex 131-D

Journal of Toxicology and Environmental Health, Part A, 72: 937–948, 2009
ISSN: 1528-7394 print / 1087-2620 online
DOI: 10.1080/15287390902929683

UTEH

Comparison of the Hazards Posed to Amphibians by

the Glyphosate Spray Control Program Versus the Chemical

and Physical Activities of Coca Production in Colombia

GLYPHOSATE SPRAY VERSUS COCA PRODUCTION ACTIVITIES
Richard A. Brain and Keith R. Solomon 2
1 2
Department of Environmental Studies, Baylor University, Waco, Texas, USA, and Department
of Environmental Biology, University of Guelph, Guelph, Ontario, Canada

Therefore, the cumulative impacts of coca production, through
This study evaluates the cumula tive multifactorial physical habitat destruction, application of agrochemicals, and poten-
and chemical impacts resultin g from coca production on tial transmission of disease, are judged to pose greater risks to
amphibian populations in comparison with the potential amphibian populations in coca-g rowing regions than the gly-
phosate spray control program.
impacts produced by the herbicide glyphosate (Glyphos),
which, mixed with the surfactant Cosmo-Flux, is used in the
spray control program for illicit crops in Colombia. Using sim-
ilar worst-case assumptions for exposure, several other pesti-
cides used for coca production, including mancozeb, lambda Coca production is a national security issue in Colombia
cyhalothrin, endosulfan, diazinon, malathion, and chlorpyri-
fos, were up to 10- to 100-fold more toxic to frogs than the that has motivated extensive enforcement measures.
Currently the herbicide glyphosate is used to control coca
Glyphos–Cosmo-Flux mixture. Comparing hazard quotients (Erythroxyum coca ) production through a spray eradication
based on application rates, several of these compounds demon-
strated hazards 3–383 times that of formulated glyphosate. program facilitated by the Antinarcoticos Directorate of the
Secondary effects, particularly of insecticides, are also a con- Colombian National Police (DIRAN-CNP). The effort is fur-
cern, as these agents selectively target the primary food source
of amphibians, which may indire ctly impact growth and devel- ther supported through data gathering by other nations in both
opment. Although the potential chemical impacts by other pes- North America and Europe (Solomon et al., 2007). Several
concerns have been raised re garding the spray control pro-
ticides are considerable, physical activities associated with coca
Downloaded production, particularly deforestation of primary forests for gram, ranging from peripheral crop damage to adverse envi-
new coca plots, portend the grea test hazard to amphibian pop- ronmental and human health effects. The government of
ulations. The entire production cycle of cocaine has been linked Colombia has responded by appointing an independent envi-
to ecosystem degradation. The cl earing of pristine forests for
coca propagation in Colombia is well documented, and some of ronmental auditor to review the program (Solomon et al.,
these regions coincide with th ose that contain exceptional 2007). In conjunction with DIRAN-CNP the spray and no-

amphibian biodiversity. This is particularly problematic as spray areas are reviewed, and spray results through field
coca production encroaches more deeply into more remote checks and data analysis are regularly monitored. Addition-
areas of tropical rain forest. Transportation ofdisease, includ- ally, three detailed reviews on the substances used for cocaine
ing the chitrid fungus, to these remote regions via human
intrusion may also adversely affect amphibian populations. production have been conducted for the Inter-American Drug
Abuse Control Commission (CICAD) section of the Organi-
zation of American States (OAS) (Solomon et al., 2007). Of
©General Secretariat of the Organization of American States,
2009. This paper was prepared as part of a Study entitled “Productionrticular concern, and identified through this review, is the
potential toxicity of several formulations of glyphosate to
of Illicit Drugs, the Environment and Human Health,” financed with amphibians.
contributions from the Governments of Colombia and the United
States of America. The conclusions and opinions expressed herein In order to appropriately assess the potential adverse
are those of the authors and not necessarily those of the Organizateffects of glyphosate on frogs and other amphibians, haz-
of American States and its General Secretariat, which as of the date
of this copyright, have not formulated any opinion with respect ards need be evaluated in the context of the effects of other
to them. pesticides and activities asso ciated with the production of
coca on these organisms. Several other pesticides and sub-
The authors thank Kathryh Chin for helping to locate toxicity data.
Address correspondence to Dr. R. A. Brain, Department of stances are used in the production of coca, and many of
Environmental Studies, Baylor University, Waco, TX 76798-7388, these are hazardous to aquatic organisms if the products are
USA. E-mail: [email protected] applied over water at the recommended field application

937

342 Annex 131-D

938 R. A. BRAIN AND K. R. SOLOMON

rates (CICAD/OAS, 2005). Th is assessment specifically measured exposure data, PECs were calculated assuming
considers hazards presented by these other pesticides to the worst-case circumstance; direct overspray of surface water

aquatic stages of amphibians and on their indirect effects on (15 cm deep) with rapid mixing, no absorption to sediments,
the food organisms used by adult amphibians. In addition, and no flow. A depth of 15 cm was used based on assump-
the destructive physical activi ties associated with coca pro- tions of forest pools in Canada, and similar assumptions made

duction are considered. The eff ects of habitat alteration on by the U.S. EPA regarding wetlands (Urban & Cook, 1986).
amphibian populations are well documented (Becker et al., These data were compared to the toxicity of formulated gly-

2007), and the clear-cutting and burning of tropical forests, phosate (mostly Roundup and Vision) and the Glyphos–
such as occur when new coca fields are established, exert Cosmo-Flux mixture as used in Colombia (CICAD/OAS,
profound adverse effects on these organisms, both directly 2006). The toxicity data are summarized in Figure 1 and

and indirectly (Hedges, 1993; Viña et al., 2004). Since habi- Table 1, comparatively illustrating the toxicity data with
tat alteration is considered tbe the greatest factor responsi- associated margins of safety (MOS) and outlining HQs,
ble for global amphibian declines (Hedges, 1993), the respectively. A number of add itional endpoints were reported

extensive loss of tropical rain forest in Colombia due to in the literature. However, because many of these endpoints
coca production is of paramount concern, since these areas are nonstandardized, they were not included in the hazard
of deforestation typically coinci de with or are close to areas
assessment.
containing exceptional amphibian biodiversity (Myers et al., Several of the pesticides used in the production of coca
2000; Etter et al., 2006). Other human activities may also are inherently as toxic as or more toxic to amphibians than
®
contribute to adverse effects on frogs. Diseases have been the Glyphos–Cosmo-Flux mixture used in Colombia (Table 1.).
associated with several frog extinctions, and the potential Both mancozeb and lambda cyhalothrin were inherently
adverse effects of a newly iden tified fungal disease of frogs, more toxic than formulated glyphosate. Endosulfan, diazinon,

chytridiomycosis, are also considered. This virulent disease malathion, and chlorpyrifos were 10- to 100-fold more toxic
is easily spread by human activity and may be carried into to frogs than the Glyphos–Cosmo-Flux mixture. The toxicity
new areas of coca production. Since this disease has caused of endosulfan is particularly relevant because of the detec-

the extinction of several species of frogs (Berger et al., 1998;tion of endosulfan in surfa ce waters from coca-growing
Speare, 2001), its presence and spread within Colombia may regions in Colombia where it is being used illegally

have serious implications for amphibians that will be (Solomon et al., 2007). Endosulfan is not registered for use
exacerbated by expansion of coca production into new in Colombia.
undeveloped areas. Comparison of the worst-case exposure that would result

from overspray of surface waters 15 cm deep at a typical
rate of application (CICAD/OAS, 2006) (Figure 1) shows
that concentrations in surface waters in, or adjoining, coca

Downloaded At: 19:44 2 October 2009ESTICIDES USED fields are predicted to be greater than the LC50 concentra-
IN COCA PRODUCTION ON AMPHIBIANS tions for mancozeb, diazinon, endosulfan, chlorpyrifos, and
Several pesticides are used in the production of coca in malathion. Comparing HQ values based on application

Colombia (CICAD/OAS, 2005) . To assess hazards to rates, these compounds presented 3.2-, 4.3-, 251-, 271-, and
amphibians, the ECOTOX database (U.S. EPA, 2001) and 383-fold higher maximum hazard than formulated glypho-

primary and secondary literature were extensively searched sate. Lambda cyhalothrin and paraquat also demonstrated
to obtain comprehensive and comparable data on the toxicity hazard under these worst-case circumstances. This suggests
of these pesticides to aquatic stages of amphibians. Mortality that these particular pesticides may exert adverse direct

values were the primary data compiled for the purposes of effects on amphibians through their use in the protection of
comparison, where common acute mortality data was avail- coca from pest infestations. Re ports from the literature pro-
able for the vast majority of pesticides considered (Table 1). vide support for the possible adverse effects of these insecti-

Data for a single species were selected for inclusion based on cides on frogs. For example, analysis of historical pesticide
exposure time. For example, toxicity data from 96-h expo- application data has linked organophosphorus (OP) and car-

sures were selected over 48 h, etc. When multiple values bamate insecticides with the declines of four Californian
were reported, the smallest value (most toxic) was selected. amphibians (Davidson, 2004). Toxicity values for amphibi-
In cases where multiple values were reported for a given ans were not available in the literature for several of the

exposure time, a geometric mean was calculated. Hazard quo- other pesticides used in coca production, such as monocro-
tients (HQs) were calculated for multiple amphibian species tophos. These may also present direct hazards to amphibians
for each pesticide according to the following equation: HQ = in or close to coca fields but can not be assessed in the

PEC/LC50, where PEC is the predicted environmental con- absence of data. Several othe r pesticides used in coca
centration and LC50 is the median lethal concentration. production were less toxic to amphibians and present
Values >1 indicate potential hazard. Due to an absence of a smaller hazard (Figure 1 and Table 1). These included

343 Annex 131-D

Continued)
PECμ/L) H2473 1.5 2472472342473 0.5.7 1541541 0.02.00800505 0.08.06 40402 0.0020 0 6.70(6 0.006
(

Reference

ytype
Exposure

g/L)
(

Concentration

TABLE 1 24 2072 Static815 R(Clements et al., 1997)ich et al., 2003) 2473 3.0
time (h)
Exposure

Downloaded At: 19:44 2 October 2009

Larvaeoleoleoleo96a4848v48s41700600300Renewalalwalwal(Edginton et al., 2004)99)99)99)99)222473 0.70.41.01.06)76)6) al., 1998)98)giaarng, 1993)

Scientific name Life stage
Toxicity Data and Hazard Auotients (HQs) for Amphibians Exposed to Various Pesticides
Bufo americanusiferaeies dorsalisaticassusedacrisstssriataperonianusbufo japonicusisocharisatayla

American toadg froglettree frogrroogged frogoongIndian toadan toadogogg frogog frogog
Common namesate formulated 2,4-D Atrazine Carbendazim Carbofuran

939

344 Annex 131-D

PECμ/L) H134103400.80.5 1340 0.1 10015000.1005 1.05 0.4— —— —— —— — 402 16402 0.09 1675 19 675 14
(

Reference

(Cole (Cole and Casida, 1983)sndaC,a183,)1983)

ytype
Exposure injectinjection injectiionnjection

1 NR (Cowman and Mazanti, 2000) 1005 1005
μ/g) Subg/g)neoSubcutaneous Renewal 1.812 (NRrris et al., 1(U.S. EPA, 2001)1)
g/L) μ
(

Concentration

TABL(ontinued) 96
time (h)
Exposure

Downloaded At: 19:44 2 October 2009

Tadpoles
Tadpolesaoeosleoe96l96 96le4apa173047602StaticStitcatic (Zaga et al., 1998)l,)1983)0./gb)(24u-b.c2un.e6o5u(sμg/g)Subcutaneous) NR Static05 0.3

Scientific name Life stage

Xenopus laeviscriorlatiihansudaamysoicasnlayutsccueRana pRipieasipieRnasnRipieasipieRnasna clamitansharistiognriicnuasnaorsitsictus

b
b

-Cypermethrin
African clawed frogrooggogAmMerarcrerasotaeeafdlfopLeopaLrdefrpoagrd frLordefrpoagrdorgeen frogogIndian bullfrogn Indian toad
CommoCarbaryl Chlorpyrifos cis trans Diazinon Endosulfan

940

345 Annex 131-D

13.4 3.438838838 4.2 838 0.40.04 670670 0.02 670670 0.0670 1.370 0.08 670 0.020 0.007003
1675 0.90.2 2412 12 1.7

4 Renewal (Aydin et al., 2005)

24 23,000 NR 24 (Nishiuchi, 1980b)ic (Nishiuchi, 1980a)

Downloaded At: 19:44 2 October 2009

TadpoleoleolTeadp24T96a48ole 11946431906NR NReStatic0.5(U.S. EPA, 2001)001)anKNaisgiaotie,t1.,11)91)675 0.2trls1e.9l2.0),)0200)00) 838 2.0hehrci,k9,19))86) 670 060)

triseria triseria
Bufo vulgaris formosusacashexadactylaseriataataeBfomoaiinusivunlugsaris formosustiisaeriatasnselaevisusei fowleriteetraiataoni
μ/g.

Data represent LC50 (median lethal concentration) values. Only the smallest values (greatest toxicity) are reported. Hazard quotients were calculated as the ratio of the pre-

Toadreen frogAfricanFrogwed frogNorthern chorus frogedriogaoadoadg StriTpiegderrtgh,ndcanruslflggorooaddseronrtcrucshforrougs frogrations of formulated glyphosate are expressed as acid equivalent (AE).
Note.a b
lambda-Malathionin MancozebMethomyl Methyl parathionaquat dicted environmental concentration (PEC) and the LC50, where values >1 indicate potential hazard.
941

346 Annex 131-D

942 R. A. BRAIN AND K. R. SOLOMON

Rangeof worst
caseexposure
Coca mix Glyphosate+onsfor
Glyphosate formulated Cosmo-Flux®
sprayed on coca
Paraquat

Lambda Cyhalothrin
Mancozeb
Diazinon

Endosulfan
Chlorpyrifos
Malathion

Carbaryl
Parathion
2,4-D
Carbendazim

Atrazine
Carbofuran Worst case concentration
Methomyl forotherpesticides

–1 0 1 2 3 4 5 6
10 10 10 10 10 10 10 10
LC50 in µg/L

FIG. 1. Toxicity to amphibians of Glyphos and Cosmo-Flux spray mixture, formulated glyphosate (mostly Roundup and Vision), and other pesticides used in
the production of coca. The data for the mixture of Glyphos and Cosmo-Flux (coca mix) are from tests on Xenopus laevis. Each point on the graph represents a
different species of amphibian. Glyphosate data are all normalized to acid equivalents (a.e.); other data are given as active ingredient (AI). Horizontal dashed

arrows pointing to the left indicate margins of safety between LC50 concentrations for X. laevis (Glyphos plus Cosmo-Flux) or the most sensitive species (othe
products) and estimated exposure; solid arrows pointing to the right indicate a hazard.

2,4-D, atrazine, carbaryl, ca rbendazim, carbofuran, meth-
ratios in Atelopus chiriquiensis and Hyla calypsa frog popula-
omyl, and parathion. tions in 1996, where, incidentally, of the three common chemi-

Davidson (2004) found a strong association between cals sprayed on the apple orchards at Las Tablas, two
upwind pesticide use and amphibian declines in montane (mancozeb and benomyl) are suspected of having reproductive

areas of California, which was consistent across a number of and endocrine-disrupting effects. Different species likely
different species representi ng at least three independent exhibit different symptoms and susceptibility upon exposure to

ranges. For four ranid frogs, pesticides were the single stron- a variety of pesticides, since these parameters vary markedly in
Downloaded gest explanatory variable in model simulations and the rela- amphibians (Berrill et al., 1994).

tionship between declines and upwind pesticide use was
consistent (Davidson, 2004). In the analysis of pesticide
INDIRECT EFFECTS OF OTHER PESTICIDES
classes, acetylcholinesterase (AChE)-inhibiting pesticides
emerged as most strongly associated with declines (Davidson, Many of the pesticides used in the production of coca are

2004). Reduced AChE levels were found in the non-declining insecticides (CICAD/OAS, 2004, 2005). Use of these insecti-
Pacific tree frog ( Hyla regilla) in the Sierra Nevada (down- cides to protect coca from insect damage may inadvertently kill

wind from California’s heavily agricultural Central Valley) insects that are also food items for adult frogs that utilize the
compared with the Coast Range (low pesticide exposure), coca fields and their margins as habitat. Thus, while some of

suggesting that lower AChE leve ls in Sierra Nevada treefrogs the insecticides exert little direct toxicity to the frogs (Figure 1),
may be due to exposure AChE-inhibiting pesticides (Sparling they may have adverse effects through reducing the availability

et al., 2001). of food. However, the indirect effects of food supply on
Lips (1998) conducted surveys of amphibian fauna at Las amphibian population abundance, produced by pesticides, are

Tablas, Puntarenas Province, Costa Rica from 1991 to 1996 poorly studied. Westerman et al. (2003) classify reduced or
and found declining trends or “atypical” fluctuations in frog altered food supply as a biological stressor, defined as a reduc-

and salamander populations, where species with both aquatic tion or change of food supply, such as reduction of insects due
eggs and larvae were most affected. Environmental contamina- to pesticide application or reduction of algae due to aquatic

tion was suggested as a primary factor associated with these herbicide application. Elimination of food base is considered to
declines, particularly agrochemicals, some of which have been be a primary affect of herbicides and insecticides by Henry

banned in the United States and Europe but are still in wide- (2000), though no examples are provided. A small number of
spread use throughout developing countries of the tropics studies document impacts of pesticides on larval stages (Boone

(Lips, 1998). Lips (1998) noted unusual female-biased sex & Semlitsch, 2001) through indirect modification of food

347 Annex 131-D

GLYPHOSATE SPRAY VERSUS COCA PRODUCTION ACTIVITIES 943

sources; however, there is virtually no information available habitat (suffering significant habitat loss), and 207 to enig-
with respect to the effects of pesticides on food sources for matic decline (declining, even where suitable habitat remains)
adult frogs, i.e., insects. Considering the biological target of for reasons that are not fully understood, although disease and

insecticides comprises the primary diet of amphibians, par- climate change are commonly cited causes (Stuart et al., 2004).
ticularly frogs, the lack of food-web cascade data with Surveys conducted by the Research and Analysis Network for
respect to insecticides is of concern. Indirect effects were Neotropical Amphibians (RANA) suggest that intact amphibian

demonstrated for the insecticide carbaryl and the herbicide communities no longer exist throughout most upland (>500 m)
atrazine on body mass, developm ent, and survival of two areas of the Neotropics (Lips et al., 2005). In Latin America,
anuran species (southern leopard frog, Rana sphenocephala ; 107 species of amphibian are identified as in decline, largely

American toad, Bufo americanus ) and two caudate species since the 1980s, though trends continue (Lips et al., 2005).
(spotted salamander, Ambystoma maculatum ; small-mouthed Many extinctions and declines have taken place in seemingly

salamander, A. texanum) reared in outdoor cattle tank meso- pristine and often montane areas (Pounds & Crump, 1994;
cosms (Boone & Semlitsch, 2001). After treatment with car- Pounds et al., 1997; Young et al., 2001).
baryl, zooplankton were eliminated, which likely resulted in

the negative impacts found on growth and development for EFFECTS OF DEFORESTATION FOR COCA
the spotted salamanders. If zooplankton populations are
reduced or eliminated, exposure to insecticides might lead to PRODUCTION ON AMPHIBIANS
Land use change often occurs in temporal waves and in
reproductive failure and subsequent population declines for
carnivorous amphibian species (Boone & Semlitsch, 2001). localized fronts termed “deforestation hotspots” by Myers
(1993) that respond to the pulses of change in land use drivers
(Etter et al., 2006). Of high concern for conservation planning

TRENDS IN AMPHIBIAN DECLINES is the potential for these “deforestation hotspots” to overlap
“biodiversity hotspots” (Myers et al., 2000; Etter et al., 2006).
During the late 1980s, ecologist Norman Myers established The driving forces of land cover change, especially deforesta-
the term “biodiversity hotspots” to distinguish a global set of
high-priority terrestrial ecoregions for conservation (Myers, tion, are reported to result from the complex interaction of
socio-political and economic processes (Etter et al., 2006).
1988). This approach identifies an ecoregion as a “hotspot”
based on the existence of exceptional concentrations of From the late 1980s, three main forces have driven the coloni-
zation process in Colombia: landlessness, illicit crops (largly
endemic species and experiencing exceptional loss of habitat coca), and the presence of rebel armies (Etter et al., 2006).
(Myers, 1988). The lower montane cloud forests (at elevations
of about 1300 to 2000 m) of the eastern Andes are considered a

biodiversity hot spot and amongst the most threatened habitats Deforestation and Amphibians
on earth (Myers, 1988). In terms of species richness, the tropi- Habitat destruction is consideredto be the single major factor
Downloaded cal Andes are considered to have the greatest biodiversity of
responsible for the decline of the earth’s amphibians and other
total species, particularly amphibians, with 604 species of organisms (Hedges, 1993). Deforestation (1) exposes terres-
known endemic amphibians, nearly 13% of the global total trial amphibians to severely altered microclimatic regimes, soil

(Myers, 1988). Colombia is situated in one of the world’s compaction and desiccation, (2) reduces habitat complexity,
biodiversity hotspots, containing 10% of the world’s biodiver- and (3) increases the amount of habitat edge, all of which
sity (CICAD/OAS, 2004) and about 750 species of amphibi- decrease available moisture and elevate extreme temperatures,

ans, half of which are endemic (J. Lynch, personal solar radiation, and wind disturbance compared to forest interiors
communication). Therefore, the contribution of this region to (Alford & Richards, 1999; Boone et al., 2003). Because many
the earth’s total biodiversity is substantial, but at the same tiamphibians spend all or the majority of their life in terrestrial

is under considerable stress due to significant anthropogenic habitats, the outcome of these changes may be the elimination
influences, including coca production. of some species, alterations in abundance, or reduced individ-
According to Stuart et al. (2004), 43% of global amphibian
ual quality of habitat (Hedges, 1993). In addition, aquatic
species are experiencing some form of population decline, stages of amphibians are exposed to stream environments with
32.5% are threatened, 122 species are possibly extinct, and increased siltation and reduced ody debris (Alford & Richards,

most losses have occurred since the 1980s. Declines were 1999). Although populations may recover as regenerating for-
suggested as nonrandom in terms of ecological preferences, ests mature, recovery to predisturbance levels can take many
geographic ranges, and taxonomic associations, which are years and may not occur at all if mixed forests are replaced

most prevalent among Neotropical montane, stream-associated with monocultures (Alford & Richards, 1999). Hedges (1993)
species (Stuart et al., 2004). Of the 435 species listed by the indicated that approximately 65% of tropical forest was
World Conservation Union (IUCN) categorized as being under
destroyed, with a likely proportional decline in the number of
higher threat (rapidly declining) than in 1980, declines of individuals of forest-associated amphibians. Thus far, native
50 species are attributed to overexploitation, 183 to reduced species survival has largely been unaffected, which, unlike the

348 Annex 131-D

944 R. A. BRAIN AND K. R. SOLOMON

decline in individuals, is not expected to be a linear function oflimited ranges, largely due to deforestation and human encroach-
the decline in forest cover (Hedges, 1993). Therefore, the num- ment in those areas, combined with the specialized habitat

ber of extant species may not show significant decline until requirements of the species (Hedges, 1993).
forest cover levels become small, at which time extinctions In Guatemala, there are an estimated 74 threatened species
may occur (Hedges, 1993). Some extinctions will occur prior of amphibians, which are considered to be primarily impacted

to that point due to unequal rates of deforestation in different by the effects of deforestation (NatureServe, 2004). In Brazil,
areas and stochastic effects, while those species that survive habitat loss is the most visible, and probably the main threat to
without forest cover continue to exist (Hedges, 1993). The sen- amphibians (Silvano & Segalla, 2005; Becker et al., 2007).

sitivity of forest-dwelling amphibians to changes in their envi- Deforestation, the advance of the agricultural frontier, mining,
ronment makes them valuable indicators of forest degradation wildfires, and development projects are the main causes of
(Bishop et al., 2003). habitat loss. Although varying in extent, all Brazilian biomes

When considering coca production as the driver, depletion are now severely affected, especially the Atlantic Forest, where
of forest cover to the point of extinction in largely inaccessiblefragmented forest remnants constitute the 8% that currently
montane environments may be argued as unlikely. This conten- remains (Silvano & Segalla 2005).

tion maintains that suitable habitat for refugia may be expected Development of agriculture and other activities can be con-
to remain as forest destruction does not likely occur across a trolled and restricted to areas that are not key amphibian habitat.

broad spatial area greater than the home or migratory range In contrast, it is clear that the uncontrolled deforestation for the
potential of the species in question. However, on average, production of illicit crops such as coca will have a major effect
thousands of square kilometers of forest are destroyed in the on amphibians in Colombia through habitat alteration.

montane forest regions of Colombia each year (UNODC,
2006), which are dominated by species with small ranges: only
tens to hundreds of square kilometers (NatureServe, 2004). Deforestation and Armed Conflict

The Amazonian foothills in the Caquetá and Putumayo The annual net deforestation rate in Colombia peaked from
Departments of Columbia are reported to contain high species 1996 to 1999 at approximately 40,400 ha, which increased
richness and levels of endemism; however, the average annual from 18,600 ha during 1989–1996 (Etter et al., 2006). How-

rate of clearing for Caquetá during 1989–2002 was 25,000 ever, the rate declined more recently to 23,830 ha from 1999 to
hectare (ha), with a peak of 41,000 ha during 1996–1999 (Etter 2002 (Etter et al., 2006). Temporally, the decline in deforesta-
et al., 2006). Furthermore, although tropical rain forests are tion from 1999 to 2002 was largely attributed to peace talks

resilient, where certain aspects can be reestablished within 65 with guerrillas that took place in the Caquetá Department
yr, the time required to reach endemism levels is between 1000 where part of this area was demilitarized (Etter et al., 2006).
and 4000 yr (Liebsch et al., 2008). Thus, given the regional The period of peak deforestation coincides with the period

specificity of coca production, certain areas of intense cultiva- when the illegal economy of narcotics was booming in the
Downloadedtion such as in the Colombian Departments of Nariño and region (UNODC, 2004). Although deforestation rates have

Putumayo (UNODC, 2006) may experience deforestation lev- slowed on average throughout Caquetá, the municipalities of
els commensurate with influencing species eradication. This Macareña and San Vicente del Caguán accounted for 80% of
prospect is enhanced when disease transmission and chemical the regional clearing in 1999–2002 (Etter et al., 2006). During

stress associated with coca production are considered in con- this time, government claims suggest that these municipalities
junction with physical stress of altered habitat. were being used by rebel groups for illegal economic activities
In Hispaniola, several species of frogs associated with during the peace process (Etteret al., 2006). Although it has

streams appear to be reduced innumbers from previous years as been debated as to what effects the Colombian armed conflict
these riparian habitats were highly altered by deforestation has on the deforestation processes (Dávalos, 2001), recent analy-
(Hedges, 1993). The removal of forest results in frequent flood- ses showed that increased deforestation during the period from

ing with intervening dry periods, and clogging of stream beds 1996–1999 was correlated with high guerrilla activity and low
with mud and debris (Hedges, 1993). It is likely that deforesta- government presence (Etter et al., 2006). The magnitude of the
tion affected these stream-associated anurans more than other forest resources threatened by the conflictbetween local authori-

species (Hedges, 1993). This may be a key aspect of potential ties and paramilitary groups in Columbia is significant. About
deleterious effects in montane forests of Colombia, as the local- 33% of the remaining forests are in municipalities with medium

ized effect (deforestation) extends to a potentially greater area to high activity by armed groups, and 20% of them are in munic-
critical habitat for stream-dependent amphibians, particularly if ipalities where both guerrillas and paramilitaries are present
key requirements are for controlled low-level flow and relatively (Álvarez, 2001, 2003). The environmental effects of these con-

sediment-free water (e.g., to prevent washout or siltation of egg tests for land have been identified as a major factor in forest deg-
masses). In Jamaica, four species (Eleutherodactylus caverni- radation (Cavelier & Etter, 1995; Henkel, 1995; Young, 1996;
cola, E. fuscus, E. junori, and E. sisyphodemus) are restricted inÁlvarez, 2006). Etter et al. (2006) suggested that the presence of

distribution and have not been found commonly within their - guerilla armies poses a major obstacle to managing deforestation

349 Annex 131-D

GLYPHOSATE SPRAY VERSUS COCA PRODUCTION ACTIVITIES 945

in a planned manner, and prohibited any form of conservation pressure exerted by armed groups on farmers, the legal econ-
planning and management. omy, and temporary crisis situations all lead to an increase in the

cultivated area (UNODC, 2007). Conversely, factors such as
forced eradication, aerial spraying, improved security condi-
Coca Production and Deforestation tions, and plant diseases contribute to reducing the cultivated

The degradation of ecosystems associated with the produc- area (UNODC, 2007).
tion of coca and its processing into cocaine paste and then into

cocaine hydrochloride constitutes one of Latin America’s most EFFECT OF DISEASE ON AMPHIBIANS
important current environmental issues (Armstead, 1992; Viña Other human activities have been suggested as being partly
et al., 2004). The entire production cycle of cocaine has been
responsible for the extinction of frogs. These range from
linked to ecosystem degradation, and particularly to tropical increased exposure to ultraviolet radiation resulting from the
deforestation (Balslev, 1993; Viña et al., 2004). In Colombia
the most obvious environmental effect of coca cultivation is release of substances that deplete stratospheric ozone, through
the spread of diseases and the interaction of these with climate
the clearing of forests (UNODC, 2006). The tropical rain forests change. Of these, one fungal disease, chytridiomycosis, was
constitute the largest biome in Colombia, though over 11 of the
original 44 million hectares have been lost. The Sub-Andean and identified as being responsible for the extinction of several frog
species. This disease was first described from dead and dying
Andean forests have lost 69 to 76% of their original cover, and frogs at sites of mass deaths inAustralia and Panama from 1993
though these areas are the most densely populated parts of the
country, they are also favored areas for the production of coca to 1998 (Berger et al., 1998). The chytrid that infected the
Australian and Central American amphibians was identified as
(UNODC, 2006). Batrachochytrium dendrobatidis, which has low host specificity
Although coca cultivation is only one factor in deforesta-
tion, the land area affected is significant. Estimates vary con- and is likely to infect any species of amphibian (Longcore et al.,
1999). Infections were detected in 15 amphibian families that
siderably regarding the total area of primary forest loss due to include 94 species (Speare, 2001). Amphibian chytridiomycosis
this activity, however. The most reliable data are provided
is an emerging infectious disease of amphibians that has been
from satellite imagery (UNODC, 2006). From 2000 to 2004, recognized as such on a global scale (Daszak et al., 1999;, 2003).
in total 413,000 ha of coca were planted in Colombia, a quar- This disease was identified as a key threatening process under
ter (97,622 ha) of which was established on land cleared from
the Environment Protection and Biodiversity Conservation Act
primary forest. Although the annual conversion rate has 1999 of New Zealand (Speare 2001; Mendelson et al., 2006).
decreased steadily by 60% during this time, 13,202 ha of pri-
mary forest were still converted in 2004 (UNODC, 2006). It While chytridiomycosis has not yet been described from
frogs in coca-growing areas ofColombia, humans are potential
is likely that several hundred thousand hectares of forest were vectors of the disease though the carrying of spores on clothing
cleared due to the direct and indirect effects of coca cultiva-
Downloaded tion prior to 2000, before remotely sensed data were avail- and equipment (Krajick, 2006). Several studies indicated that the
virulence of the fungal disease chytridiomycosis, one of the most
able, though exact numbers are not known (UNODC, 2006). commonly cited causes of enigmatic declines, is greater at
However, the cumulative area of primary forest lost from
conversion to coca production canbe calculated for the period of higher elevations and among streamside species (Stuart et al.,
2004). Thus, human activities in the growing of coca in remote
1990–2004. Assuming a 13% rate of annual deforestation areas may increase the spread of this disease to new areas of
directly attributable to coca cultivation (UNODC, 2006) and
Colombia. The effects of this onrare or endangered species of
applying this proportion to an annual forest cover change esti- amphibians in Colombia and elsewhere are potentially serious.
mated at 190,470 hectares/year (UNODC, 2006), coca cultiva-
tion accounted for approximately 345,233 deforested hectares
CONCLUSIONS
over this period. This is a conservative estimate, however, since
the actual area of primary forestcleared due to coca cultivation In summary, there are a number of human activities
is greater than the area being directly cultivated for this purpoassociated with the production of coca that present greater

Land used by the coca producers for subsistence farming, aban- risks to amphibians than the glyphosate + Cosmo-Flux mix-
doned after soil becomes infertile, deforested by the farmers who ture used in the aerial eradication spraying. Under worst-
leave areas dominated by drug traffickers and terrorists, defor- case circumstances, several of the pesticides used to protect

ested by the coca producers who are dispersed as a result of coca from pests (mancozeb, lambda cyhalothrin, endosul-
political violence, and cleared for landing strips (of which more fan, diazinon, malathion, and chlorpyrifos) are as toxic, or
than 100 exist at any one time), lab sites, and campsites also con- more toxic, to amphibians than the Glyphos–Cosmo-Flux

tributes to the total deforestedarea (UNODC, 2006). The actual mixture. Furthermore, physical activities such as deforesta-
area deforested is therefore likely to be greater than half a tion pose considerably greater hazards to amphibians in

million hectares for this 14-yr period. Coca cultivation in Colombia. Habitat destruction through the clearing and
Columbia is dynamic, and factors including favorable prices, conversion of primary forests is of paramount concern,

350 Annex 131-D

946 R. A. BRAIN AND K. R. SOLOMON

given the tendency of deforestation hotspots to overlap Cavelier, J., and Etter, A. 1995. Deforestation of montane forest in Colombia
as a result of illegal plantations of opium (Papaver somniferum). In Biodi-
biodiversity hotspots in a country containing the second versity and conservation of Neotropical montane forests, eds. P. Churchill,
largest number of amphibians on earth and concomitantly
H. Baslev, E. Forero, and J. L. Luteyn, pp. 541–549. Bronx: New York
the greatest production of cocaine. The potential for disease Botanical Garden.
transmission (chytridiomycosis ) is also enhanced as coca CICAD/OAS. 2004. The toxicology of chemicals used in the production and
refining of cocaine and heroin: A tier-one assessment. Technical report
production further infiltrates remote areas of rain forest.
Therefore, when considering the cumulative impacts and OAS/CICAD 2004-01, CICAD, Organization of American States.
Washington, DC, USA.
risks of coca production collectively in a multifactorial con- CICAD/OAS. 2005. The toxicology of selected chemicals used in the produc-
tion and refining of cocaine and heroin: A tier-two assessment. Technical
text to amphibian populations in coca growing regions, they report OAS/CICAD 2005-01, CICAD, Organization of American States.
are judged to be greater than those posed by the use of gly-
Washington, DC, USA.
phosate and Cosmo-Flux employed for the spray control CICAD/OAS. 2006. The toxicity of pesticides used in the production of
program. cocaine and heroin to amphibians: A review. Technical report OAS/
CICAD 2005-01, CICAD, Organization of American States. Washington,

DC, USA.
Clements, C., Ralph, S., and Petras, M. 1997. Genotoxicity of select herbicides
REFERENCES in Rana catesbeiana tadpoles using the alkaline single-cell gel DNA elec-
Alam, M. N., and Shafi, M. 1991. Toxicity of two agricultural chemicals trophoresis (Comet) assay. Environ. Mol. Mutagen. 29:277–288.
Metacid 50 and Ekalux EC 25 to tadpoles of Rana tigrina. Environ. Ecol. Cole, L. M., and Casida, J. E. 1983. Pyrethroid toxicology in the frog. Pestic.

9:870–872. Biochem. Physiol. 20:217–224.
Alford, R. S., and Richards, S. J. 1999. Global amphibian declines: A problem Cowman, D. F., and Mazanti, L. E. 2000. Ecotoxicology of “new generation”
in applied ecology. Annu. Rev. Ecol. Syst. 30:133–165. pesticides to amphibians, in Ecotoxicology of amphibians and reptiles, eds.
Álvarez, M. D. 2001. Environmental damages from illicit crops and processing D. W. Sparling, G. Linder, and C. A. Bishop, pp. 233–268. Pensacola, FL:
of prohibited drugs in Colombia, Columbia University. Accessed 2006.
SETAC Press.
http://www.columbia.edu/∼mda2001/osaka.html Daszak, P., Berger, L., Cunningham, A. A., Hyatt, A. D., Green, D. E., and
Álvarez, M. D. 2003. Forests in the time of violence: Conservation implica- Speare, R. 1999. Emerging infectious diseases and amphibian population
tions of the Colombian war. J. Sustain. For. 16:49–70. declines. Emerg. Infect. Dis., 5:745–748.
Álvarez, M. D. 2006. Environmental damages from illicit crops and processing
Daszak, P., Cunningham, A. A., and Hyatt, A. D. 2003. Infectious disease and
of prohibited drugs in Colombia, Columbia University. Accessed 2006. amphibian population declines. Div. Distrib. 9:141–150.
http://www.columbia.edu/∼mda2001/osaka.html Dávalos, L. M. 2001. The San Lucas mountain range in Colombia: How much
Armstead, L. 1992. Illicit narcotics cultivation and processing: The ignored conservation is owed to the violence? Biodivers. Conserv. 10:69–70.
environmental drama. Bull. Narcot. 44:9–20. Davidson, C. 2004. Declining downwind: Amphibian population declines in
Aydin, H., Gungordu, A., and Ozmen, M. 2005. OZM-1117-784489. Toxicity
California and historical pesticide use. Ecol. Appl. 14:1892–1902.
of two different pyrethroid insecticides on Xenopus laevis tadpoles. Edginton, A. N., Sheridan, P. M., Stephenson, G. R., Thompson, D. G., and
SETAC North America 26th Annual Meeting, Baltimore, MD. Boermans, H. J. 2004. Comparative effects of pH and Vision herbicide on
Balslev, H., ed. 1993.Neotropical montane forests boidiversity and conservation. two life stages of four anuran amphibian species. Environ. Toxicol. Chem.
Abstracts from a symposium at the New York Botanical Garden, June
23:815–822.
21–26, 1993. AAU Reports 31. p. 3. Aarhus University Press. Etter, A., Stuart Phinn, C. M., Pullar, D., and Possingham, H. 2006. Unplanned
Becker, C. G., Fonseca, C. R., Haddad, C. F. B., Batista, R. F., and Prado, P. I.land clearing of Colombian rainforests: Spreading like disease? Landscape
Downloaded At: 12007. Habitat split and globladecline of amphibians.Science318: 1775–1777. Urban Plan. 77:240–254.
Berger, L., Speare, R., Daszak, P., Green, D. E., Cunningham, A. A., Goggin, Harris, M. L., Bishop, C. A., Struger, J., Ripley, B., and Bogart, J. P. 1998. The

C. L., Slocombe, R., Ragan, M. A., Hyatt, A. D., McDonald, K. R., Hines, functional integrity of Northern leopard frog (Rana pipiens) and green frog
H. B., Lips, K. R., Marantelli, G., and Parkes, H. 1998. Chytridiomycosis (Rana clamitans) populations in orchard wetlands. II. Effects of pesticides
causes amphibian mortality associated with population declines in the rain and eutrophic conditions on early stage development. Environ. Toxicol.
forests of Australia and Central America. Proc. Nat. Acad. Sci. USA Chem. 17:1351–1363.
95:9031–9036.
Harris, M. L., Chora, L., Bishop, C. A., and Bogart, J. P. 2000. Species- and-
Berrill, M., Bertram, S., McGillivray, L., Kolohon, M., and Pauli, B. 1994. age-related differences in susceptibility to pesticide exposure for two
Effects of low concentrations of forest-use pesticides on frog embryos and amphibians, Rana pipians, and Bufo americanus. Bull. Environ. Contam.
tadpoles. Environ. Toxicol. Chem. 13:657–664. Toxicol. 64:263–270.
Bishop, C. A., Cunington, D. C., Fellers, G. M., Gibbs, J. P., Pauli, B. D., and
Hashimoto, Y., and Nishiuchi, Y. 1981. Establishment of bioassay methods for
Rothermel, B. B. 2003. Physical habitat and its alteration: A common the evaluation of acute toxicity ofpesticides to aquatic organisms.J. Pestic.
ground for exposure of amphibians to environmental stressors. In Amphib- Sci. 6:257–264.
ian decline: An integrated analysis of multiple stressor effects, eds. G. Hedges, B. S. 1993. Global amphibian declines: A perspective from the
Linder, S. K. Krest, and D. W. Sparling, pp. 209–241. Pensacola, FL: Society Carribbean. Biodivers. Conserv. 2:290–303.
of Environmental Toxicology and Chemistry
Henkel, R. 1995. Coca (Erythroxylum coca) cultivation, cocaine production,
Boone, M. D., and Bridges, C. M. 1999. The effect of temperature on the and biodiversity loss in the Chapare region of Bolivia. In Biodiversity and
potency of carbaryl for survival of tadpoles of the green frog (Rana clami- conservation of neotropical montane forests, eds. P. Churchill, H. Baslev,
tans). Environ. Toxicol. Chem. 18:1482–1481. E. Forero, and J. L. Luteyn, pp. 541–549. Bronx: New York Botanical
Boone, M. D., Corn, P. S., Donnelly, M. A., Little, E. E., and Niewiarowski, P.
Garden.
H. 2003. Physical stressors. In Amphibian declines: An integrated analysisHenry, P. F. P. 2000. Aspects of amphibian anatomy and physiology. In
of multiple stressor effects, eds. G. Linder, S. K. Krest, and D. W. SparlingEcotoxicology of amphibians and reptiles, eds. D. W. Sparling, G. Linder,
pp. 129–151. Pensacola, FL: Society of Environmental Toxicology and and C. A. Bishop, pp. 71–110. Pensacola, FL: Society of Environmental
Chemistry (SETAC). Toxicology and Chemistry (SETAC).

Boone, M. D., and Semlitsch, R. D. 2001.Interactions of insecticide with larvaHowe, C. M., Berrill, M., Pauli, B. D., Helbing, C. C., Werry, K., and
density and predation in experimental amphibian communities. Conserv. Veldhoen, N. 2004. Toxicity of glyphosate-based pesticides to four North
Biol. 15:228–238. American frog species. Environ. Toxicol. Chem., 23:1928–1938.

351 Annex 131-D

GLYPHOSATE SPRAY VERSUS COCA PRODUCTION ACTIVITIES 947

Howe, G. E., Gillis, R., and Mowbray, R. C. 1998. Effects of chemical synergy Nishiuchi, Y. 1980b. Toxicity of formualted pesticides to freshwater organisms
and larval stage on the toxicity of atrazine and alachlor to amphibian LXXII. Aquiculture /Suisan Zoshoku 27:238–244.

larvae. Environ. Toxicol. Chem., 17:519–525. Nishiuchi, Y., and Yoshida, K. 1974. Effects of pesticides on tadpoles. Part 3.
Johnson, C. R. 1976. Herbicide toxicities in some Australian anurans and the Noyaku Kensasho Hokoku 14:66–68.
effect of subacute dosages on temperature tolerance. Zool. J. Linn. Soc. Pan, D. Y., and Liang, X. M. 1993. Safety study of pesticides on bog frog, a
59:79–83. predatory natural enemy of pest in paddy field. J. Hunan Agric. Coll.

Khangarot, B. S., Sehgal, A., and Bhasin, M. K. 1985. Man and biosphere— 19:47–54.
Studies on the Sikkim Himalayas. Part 6: Toxicity of selected pesticides to Pawar, K. R., and Matdare, M. 1984. Toxic and teratogenic effects of feni-
frog tadpole Rana hexadactyla (Lesson). Acta Hydochim. Hydrobiol. trothion, BHC and carbofuran on embryonic development of the frog
13:391–394. Microhyla ornata. Toxicol. Lett. 22:7–13.

Krajick, K. 2006. The lost world of the Kihansi toad. Science., 311:1230–1232. Pounds, J. A., and Crump, M. L. 1994. Amphibian declines and climate distur-
Lajmanovich, R. C., Izaguirre, M. F., and Casco, V. H. 1998. Paraquat toler- bance: The case of the golden toad and the harlequin frog. Conserv. Biol.
ance and alteration of internal gill structure of Scinax nasica tadpoles 8:72–85.
(Anura:Hylidae). Arch. Environ. Contam. Toxicol. 34:364–369. Pounds, J. A., Fogden, M. P. L., Savage, J. M., and Gorman, G. C. 1997. Tests

Lajmanovich, R. C., Sandoval, M. T., and Peltzer, P. M. 2003. Induction of of null models for amphibian declines on a tropical mountain. Conserv.
mortality and malformation inScinax nasicus tadpoles exposed to glyphosate Biol. 11:1307–1322.
formulations. Bull. Environ. Contam. Toxicol. 70:612–618. Sanders, H. O. 1970. Pesticide toxicities to tadpoles of the Western chorus frog
Liebsch, D., Marques, M. C. M., and Goldenberg, R. 2008. How long does the Pseudacris triseriata and Fowler’s toad Bufo woodhousii fowleri. Copeia

Atlantic Rain Forest take to recover after a disturbance? Changes in species 2:246–251.
composition and ecological features during secondary succession. Biol. Sheffield, S. R. 2000. (PWA062) Lethal and sublethal effects of the OP insec-
Conserv. 141:1717–1725. ticide chlopyrifos in larval amphibians. SETAC 21st Annual Meeting,
Linder, G., Barbitta, J., and Kwaiser, T. 1990. Short-term amphibian toxicity November, Nashville, TN.

tests and paraquat toxicity assessment. In Aquatic toxicology and risk Silvano, D. L., and Segalla, M. V. 2005. Conservation of Brazilian amphibi-
assessment: Thirteenth volume,eds.W.G.LandisandW.H.vander ans. Conserv. Biol. 19:653–658.
Schalie, p. 371. West Conshohocken, PA: ASTM International. Solomon, K. R., Anadón, A., Brain, R. A., Cerdeira, A. L., Crossan, A. N.,
Lips, K. R. 1998. Decline of a tropical montane amphibian fauna. Conserv. Marshall, A. J., Sanin, L. H., and Smith, L. 2007. Comparative hazard

Biol. 12:106–117. assessment of the substances used for production and control of coca and
Lips, K. R., Burrowes, P. A., Mendelson, J. R. I., and Parra-Oleo, G. 2005. poppy in Colombia. In Rational environmental management of agrochemi-
Amphibian population declines in Latin America: A synthesis. Biotropica cals: Risk assessment, monitoring, and remedial action, ACS Symposium
37:222–226. Series no. 966 (vol. 966), eds. I. R. Kennedy, K. R. Solomon, S. Gee, A. N.

Longcore, J. E., Pessier, A. P., and Nichols, D. K. 1999. Batrachochytrium Crossan, S. Wang, and F. Sanchez-Bayo, pp. 87–99. Washington, DC:
dendrobatidisgen. et sp. nov., a chytrid pathogenic to amphibia. ycologia American Chemical Society.
91:219–227. Sparling, D. W., Fellers, G. M., and McConnell, L. L. 2001. Pesticides and
Mann, R. M., and Bidwell, J. R. 1999. The toxicity of glyphosate and several amphibian population declines in California, USA. Environ. Toxicol.

glyphosate formulations to four species of Southwestern Australian frogs. Chem. 20:1591–1595.
Arch. Environ. Contam. Toxicol. 36:193–199. Speare, R. 2001. Recommendations from Workshop in getting the jump on
Marian, M. P., Arul, V., and Pandian, T. J. 1983. Acute and chronic effects of amphibian disease. In Getting the jump on amphibian disease. Developing
carbaryl on survival, growth, and metamorphosis in the bullfrog (Rana management strategies to control amphibian diseases: Decreasing the risks

tigrina). Arch. Environ. Contam. Toxicol. 12:271–275. due to communicable diseases, ed. R. Speare, pp. 131–147. Townsville,
Mayer, F. L., Jr., and Ellersieck, M. E. 1986. Manual of acute toxicity: New Zealand: School of Public Healthand Tropical Medicine, James Cook
Interpretation and data base for 410 chemicals and 66 species of University.
Downloaded At: 19freshwater animals . Technical report 160. U.S. Fish and Wildlife Stuart, S. N., Chanson, J. S., Cox, N. A., Young, B. E., Rodrigues, A. S. L.,

Service. Washington, DC, USA. Fischman, D. L., and Waller, R. W. 2004. Status and trends of amphibian
Mendelson, J. R., III, Lips, K. R., Gagl iardo, R. W., Rabb, G. B., Collins, J. declines and extinctions worldwide. Science 306:1783–1786.
P., Diffendorfer, J. E., Daszak, P., Ibáñez D, R., Zippel, K. C., Lawson, UNODC. 2004. Colombia: Coca cultivation survey. United Nations Office for
D. P., Wright, K. M., Stuart, S. N., Gascon, C., da Silva, H. R., Burrowes, Drug Control and Government of Colombia. Bogotá Columbia.

P. A., Joglar, R. L., La Marca, E., Lötters, S., du Preez, L. H., Weldon, UNODC. 2006. The environmental effects of illicit drug cultivation and pro-
C., Hyatt, A., Rodriguez-Mahecha, J. V., Hunt, S., Robe rtson, H., Lock, cessing. United Nations Office on Drugs and Crime. Vienna, Austria.
B., Raxworthy, C. J., Frost, D. R., Lacy, R. C., Alford, R. A., Campbell, UNODC. 2007. Columbia coca cultivation survey, Technical. United Nations
J. A., Parra-Olea, G., Bolaños, F., Domingo, J. J. C., Halliday, T., Office on Drugs and Crime. Vienna, Austria.

Murphy, J. B., Wake, M. H., Coloma, L. A., Kuzmin, S. L., Price, M. S., Urban, D. J., and Cook, N. J. 1986. Standard evaluation procedure for ecolog-
Howell, K. M., Lau, M., Pethiyagoda, R., Boone, M., Lannoo, M. J., ical risk assessment. Technical report EPA/540/09-86/167, Hazard Evalua-
Blaustein, A. R., Dobson, A., Griffiths, R. A., Crump, M. L., Wake, D. B., tion Division, Office of Pesticide Programs, U.S. Environmental Protection
and Brodie, E. D., Jr. 2006. Confront ing amphibian declines and extinc- Agency. Washington, DC, USA.

tions. Science 313:48. U.S. Environmental Protection Agency. 2001. Environmental Effects Database
Myers, N. 1988. Threatened biotas: hotspots in tropical forests.Environmentalist (EEDB). ECOTOX Database System, U.S. Environmental Protection
8:178–208. Agency, Office of Pesticide Programs, Environmental Fate and Effects
Myers, N. 1993. Tropical forests: the main deforestation fronts. Environ. Division, Washington, DC. Accessed June 10, 2006. http://www.epa.gov/

Conserv. 20:9–16. ecotox
Myers, N., Mittermeier, R. A., Mittermeier, C., da Fonseca, G. A. B., and Vardia, H. K., Rao, P. S., and Durve, V. S. 1984. Sensitivity of toad larvae to
Kent, J. 2000. Biodiversity hotspots for conservation priorities. Nature 2,4-D and endosulfan pesticides. Arch. Hydrobiol. 100:395–400.
403:853–858. Venturino, A., Gauna, L. E., Bergoc, R. M., and D’Angelo, A. M. P. 1992.

NatureServe. 2004. Global Amphibian Assessment, Center for Applied Biodi- Effect of exogenously applied polyamines on malathion toxicity in the toad
versity Science, Washington, DC. Accessed 2006. www.conservation.org/ Bufo arenarum Hensel. Arch. Environ. Contam. Toxicol. 22:135–139.
xp/news/press_releases/2004/amp_kit/amphibian_factsheet.pdf Viña, A., Echavarria, F. R., and R undquist, D. C. 2004. Satellite change
Nishiuchi, Y. 1980a. Toxicity of formulated pesticides to fresh water organisms. detection analysis of deforestation rates and patterns along the Colombia–

LXXIV, Aquiculture/Suisan Zoshoku 28:107–112. Ecuador border, Ambio 33:118–125.

352 Annex 131-D

948 R. A. BRAIN AND K. R. SOLOMON

Westerman, A. G., van der Schalie, W., Levine, S. L., Palmer, B., Shank, D., Bolaños, F., Chaves, G., and Romo, D. 2001. Population declines and pri-

and Stahl, R. G. 2003. Linking stressors with potential effects on amphiborities for amphibian conservation in Latin America. Conserv. Biol.
ian populations. In Amphibian decline: An integrated analysis of multi- 15:1213–1223.
ple stressor effects , eds. G. Linder, S. K. Kres t, and D. W. SparliYoung, K. R. 1996. Threats to biological diversity caused by coca/cocaine
73–109. Pensacola, FL: Society of Environmental Toxicology and Chem- deforestation in Peru, Environ. Conserv. 23:7–15.
istry (SETAC). Zaga, A., Little, E. E., Rabeni, C. F., and Ellersieck, M. R. 1998. Photoen-

Young, B. E., Lips, K. R., Reaser, J. K., Ibáñez, R., Salas, A. W., Cedeño, J.anced toxicity of a carbamate insecticide to early life stage anuran
R., Coloma, L. A., Ron, S., La Marca, E., Meyer, J. R., Muñoz, A., amphibians. Environ. Toxicol. Chem. 17:2543–3553.

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353354 Annex 131-E

Annex 131-E

L.H.SANIN ET ., “EGIONALDIFFERENCES ITIME TOPREGNANCY AMONG
FERTILE OMAN FROM FIVECOLOMBIAN REGIONS WITHDIFFERENT USE OF
G LYPHOSAT”

(Journal of Toxicology and Environmental Health, Part A, 72:949-960, 2009)

355 Annex 131-E

Journal of Toxicology and Environmental Health, Part A, 72: 949–960, 2009
ISSN: 1528-7394 print / 1087-2620 online
DOI: 10.1080/15287390902929691

UTEH
Regional Differences in Time to Pregnancy Among Fertile

Women from Five Colombian Regions with Different use

of Glyphosate

Regional Differences 1,2,3yphosate Use and Fecundity 4 5
Luz-Helena Sanin , Gabriel Carrasquilla , Keith R. Solomon ,
Donald C. Cole , and E. J. P. Marshall 7
1 2
Universidad Autonoma de Chihuahua, FEN, Mexico, Department of Public Health Sciences,
University of Toronto, Toronto, Ontario, Canada, 3National Institute of Public Health, Mexico City,
4
5exico, Department of Microbiology, School of Health Sciences, University of Valle, Cali, Colombia,
Centre for Toxicology and Department of Environmental Biology, University of Guelph, Ontario,
Canada, 6Institute for Work and Health, Toronto, Ontario, Canada, and Marshall Agroecology

Limited, Winscombe, Somerset, United Kingdom

eradication), displayed minimal risk and was the reference
The objective of this study was to test whether there was an region. Other regions, including Sierra Nevada (control area,
association between the use of glyphosate when applied by aerial organic agriculture), Putumayo and Nariño (illicit crops and
spray for the eradication of illicit crops (cocaine and poppy) and intensive eradication spray program), and Valle del Cauca, dem-
onstrated greater risk of longer TTP, with the highest risk for
time to pregnancy (TTP) among fertile women. A retrospective Valle del Cauca (fOR 0.15, 95% CI 0.12, 0.18), a sugar-cane
cohort study (with an ecological exposure index) of first preg-
nancies was undertaken in 2592 fertile Colombian women from region with a history of use of glyphosate and others chemicals
5 regions with different uses of glyphosate. Women were inter- for more than 30 yr. The reduced fecundability in some regions
viewed regarding potential reproduc tive, lifestyle, and work his- was not associated with the us e of glyphosate for eradication
tory predictors of TTP, wh ich was measured in months. spraying. The observed ecological differences remain unex-
plained and may be produced by varying exposures to environ-
Fecundability odds ratios (fOR) were estimated using a discrete mental factors, history of cont raceptive programs in the region,
time analogue of Cox’s proportional hazard model. There were
differences in TTP between regio ns. In the final multivariate or psychological distress. Fu ture studies examining these or
Downloaded model, the main predictor was the region adjusted by irregular other possible causes are needed.
relationship with partner, maternal age at first pregnancy, and,
marginally, coffee consumption and self-perception of water

pollution. Boyacá, a region with traditional crops and. recently,
illicit crops without glyphosate eradication spraying (manual Glyphosate is one of the most widely used herbicides glo-
bally and has been registered for use in Colombia since 1972

©General Secretariat of the Organization of American States, for weed control in a wide range of crops and in the process of
2009. This paper was prepared as part of a Study entitled “Productiongar cane maturation. Beginning in the early 1980s, it was

of Illicit Drugs, the Environment and Human Health,” financed with used for eradicating the illegal crops of coca (Erythroxylum
contributions from the Governments of Colombia and the United coca) and poppy (Papaver sominferum). Since 2000, it has
States of America. The conclusions and opinions expressed herein arbeen more widely used for the eradication of illicit crops. The
those of the authors and not necessarily those of the Organization of
American States and its General Secretariat, which as of the date ofrea of coca sprayed with glyphosate has shown a steady
this copyright, have not formulated any opinion with respect to them.crease over recent years, reachnig 153,000 ha in 2007 (personal

The authors gratefully acknowled ge the contributions of the communication, National Police of Colombia, Bogotá, December
people of Colombia, Alejandro Ri co, and the epidemiologists who 2007). According to Colombian use data, 10–13% of the total
participated as field coordinators in the five regions, as well as amount of glyphosate purchased in the country is used for
interviewers, local authorities, and communities where the study
was carried out. We thank Sandra Reza, who conducted much aerial spraying of illicit crops; the remainder is used in both
descriptive data analysis, and the Executive Secret ariat of CICAD legal and illegal crop production (Solomon et al., 2007).
for their help in conducting the work.
Colombia is organized into 32 administrative departments
Address correspondence to Dr. Keith R Solomon, Centre for (departmentos). In 12 of them, illicit crops have been sprayed
Toxicology and Department of Environmental Biology, University of with glyphosate by aerial application since 2000. The location
Guelph, Guelph, ON, N1G 2W1, Canada. E-mail: ksolomon@
uoguelph.ca and amounts of glyphosate applied for this purpose are accurately

949

356 Annex 131-E

950 L.-H. SANIN ET AL.

known. Glyphosate is used for other purposes in all depart- Colombia (Figure 1). All participants were informed about the
ments, but actual use statistics are not known as sales data are objectives of the study, and invited to participate if their first

not collected. pregnancy occurred during the last 5 yr (since November 1999)
In developed countries, investigators have increasingly used and they did not use contraceptives during the year prior to
time to pregnancy (TTP) as a sensitive clinical marker of mul- becoming pregnant. The latter was to reduce reporting bias

tiple early adverse reproductive effects (Baird et al., 1986; because there is no accurate method to adjust for the effect of
Joffe 1997, 2000; Joffe & Barnes 2000; Tingen et al., 2004; the use of contraception on fecundity (Tingen et al., 2004).

Joffe et al., 2005). Epidemiological studies examined the role Only data on first pregnancies were used, to reduce recall bias
of agriculture and pesticide exposure in reducing the probabil- and other potential biases that are associated with subsequent
ity of achieving conception in a menstrual cycle (also known as pregnancies. Only one pregnancy was used to maintain out-

fecundability) with mixed results (De Cock et al., 1994; Larsen come independence and minimize the effect of previous repro-
et al., 1998; Curtis et al., 1999; Thonneau et al., 1999; Abell ductive history (Olsen & Skov, 1993).
et al., 2000; Petrelli & Figà-Talamanca, 2001; Sallmén et al., Two days of training were carried out for interviewers and

2003; Idrovo et al., 2005; Bretveld et al., 2006; Lauria et al., supervisors to explain the objectives of the project and the
2006; Bretveld et al., 2008; Joffe et al., 2008). questionnaire to be applied. All interviewers lived in the
There have been some reports in the literature of adverse study area and were supervised by local epidemiologists who

reproductive outcomes associated with pesticide use, most of knew the study area and who were well known to the popula-
which are described in more detail in a recent review (Wigle et tion. In each area, studies started at the closest household

al., 2008). Arbuckle et al. (2001) observed a rise in the risk of where water and sediment samples were taken as part of the
early abortion when preconception self-reported exposures to assessment of aerially applied glyphosate (Solomon et al.,
phenoxyacetic acid herbicides were present (odds ratio [OR] = 2007). From the first household, the interview team moved

1.5, CI95% 1.1–2.1; positive effect if greater than 1) and for lataway (centrifugally), visiting house by house to identify
abortions, self-reported preconception exposure to glyphosate women who met the inclusion criteria until the sample size
(OR = 1.7, CI 1–2.9) was associated with higher risks. In (600 women in each zone) was achieved. Because field work-
95%
another study, Curtis et al. (1999) reported a positive associa- ers were well known by the popu lation, there were no refusals
tion (decrease in fecundability of 20% or more) measured to enter the study, except in Valle del Cauca, where 3% of

through the outcome, TTP, when both spouses reported expo- identified women declined to enter the study, mainly because
sure to pesticide activities, with 5 of 13 pesticides categories their husbands did not allow them to participate. There were
(dicamba, glyphosate, phenoxy herbicides, organophosphorus some differences among the five study sites that required us

insecticides, and thiocarbamates). Garry et al. (2002), studying to visit more households in some areas than in others. For
pesticide applicators in Minnesota through a cross-sectional example, in Boyacá and Nariño, women start families at an
study of 695 workers and 1532 children (offspring), observed early age; thus, when asked about first pregnancy in the last 5

Downloadedthat self-reported use of the herbicide glyphosate yielded an yr there were many who were in the appropriate age group
OR of 3.6 (CI 95% 1.3–9.6) in relation to attention deficit disor-but had their first pregnancy more than 5 yr previously and
der/attention deficit-hyperactivity disorder (ADD/ADHD), and therefore did not meet the inclusion criteria. In Valle del

pointed out that herbicides applied in the spring might be a fac- Cauca, most women had taken oral contraceptives in the last
tor in the birth defects. year, an exclusion criterion for the study. The population of

Our objective in the current study was to test for differences Valle is different because it is a more developed department,
in TTP for first pregnancy among fertile women selected from was one of the first departments (if not the first) where
five regions of Colombia with different use patterns of glypho- extended family planning was initiated in the 1960s, and

sate. This study also took into account other known factors many villages (veredas) needed to be visited in order to
affecting fecundability. A priori, it was postulated that the use obtain the sample size.
of glyphosate in aerial spraying programs for eradication of All women responding to the oral invitation were inter-

illicit crops might be associated with reduced fecundability, viewed in their homes. Those who were confirmed as meeting
and, considering that there are no biomarkers for exposure to the inclusion criteria were informed about the objectives of the

glyphosate, an ecological exposure index was chosen. study. Care was taken to ensure participants that there would
be no reprisal for participation or nonparticipation, and that the
investigators guaranteed the privacy of the information col-

MATERIALS AND METHODS lected. Each participant provided written informed consent, in
keeping with ethical approval by the Ethics Review Board of
Design and Population the Fundación Santa Fé de Bogotá, Colombia. Of a total of

Between August 2004 and Februray 2005, a cross-sectional 3005 women interviewed, 233 women were excluded without
study of first pregnancies was carried out among women based onTTP data and 21 with TTP values greater than 60 mo. Hence,
residence in one of five different regions (departments) from 2751 (91.6%) were included in the analyses. However, for the

357 Annex 131-E

REGIONAL DIFFERENCES IN GLYPHOSATE USE AND FECUNDITY 951

SIERRANEVADA
DESANTAMARTA

MAGDALENA

BOYACÁ

VALLEDELCAUCA

NARIÑO

PUTUMAYO

FIG. 1. Location of the study areas in Colombia (departments).

Downloaded At: 20:07 2 October 2009
multiple regression and the alternative models, a restricted you having sexual intercourse before you became pregnant for
analysis was conducted without the 159 women who reported the first time?” The questionnaire was field tested in the five

consultation with a physician because of fertility problems. different regions to ensure the question was clearly understood
This removed potential bias that may have been introduced by in all areas since the departments are far from each other and

those who suspected themselves to be subfertile (Tingen et al.there are subtle differences in understanding some terms. TTP
2004; Idrovo et al., 2005; Joffe et al., 2005) was defined as duration in months, not divided by menstrual
cycle duration in days, because women are more able to recall

Exposure Assessment time in months than in cycles (Joffe, 1997). In this case,
months and cycles were treated as equivalents.
As exposure could not be measured directly, an ecological
design was used in which five different regions in the country,

with different levels of exposure, were selected according to Potential Confounders
agricultural practices and presence or not of the aerial spray
program for eradication of illicit crops with glyphosate. Table 1During the interview, participants also provided information
on potential confounders, including age at which the woman
shows the characteristics of the study areas.
started trying to become pregnant, age at first pregnancy, and
current age; relationship with partner; work history and gyne-
Outcome Measurement cologic and medical history prior to first pregnancy; x-ray

Valid data on TTP can be derived retrospectively, with a exposure in the year prior to conception; body image percep-
recalltimeof14yrormore (Joffe et al., 1995). A modified tion prior to conception as a proxy for body mass index (Singh,

version of the key question from the questionnaire of Baird 1994; Madrigal-Fritsch et al., 1999; Romieu et al., 2004); and
et al. (1986) was used to elicit TTP: “How many months were lifestyle practices in the year prior to conception, such as

358 Annex 131-E

program

eracdrcaptsi)on of illicit glycpohnoaufetutosancoN%rcoieogh.nftheo–s2p4Pyutngiofycsoccinea.eitn,einnthsievsetudy glymphaosearfofonhrwnekesting.
GlyphosateNone (manualy Aerial spraying of In 2003 and 2004, None Aerial spraying of

use

inserctdiidtiecuaeGddetsuhhdoanduisnesreritbdetfeuoeeleclmpuoseunds.duisnsdretitieestuatadbifaylc.fiaafnolhntrctnotps.boeo.loegsuseininclyginhrgocsaantee.
UsuallevelHerbicides, fungicides, Herbicides, fungicides, Herbicides, fungicides, No pesticide use for Substantial herbicide

Most common crops

canaer,ectrhhst,hlvanraen,iagecoMenoacaicccaiicaiitsoiolnngdecoasu)lu1tan,rmanidaacerreroeyction ofpessnreee,fheahiroer,noan.taistudplalnertnir,xtebnrat.nae.areoaffe,grown to a
Vegetables, potato, maize, baAgriculture m Foarkeessliiitacrotpefsrevpeobrteaesouwrce ionf the areafiene, apnladntations are thMain crop in Valle del Cauca and in the
TABLE 1

Downloaded At: 20:07 2 October 2009

2) located at 2 m a.s.l., has

” (not large enough to be a

Geographic, Population, Agriculture, and Pesticide Use Characteristics in the Study Regions

Description of the study area corregimiento

wes2t4z°Cnuaecsiati8itf5bfealsnl.an aloncntarmeastuAroeoc.h0tetstiije.ner3o6nthoefereonftlltinnd,bfoafeett.ectilev(an–ely9ofraaTshliteeeseelives iAfrican origin.uy,,taaAitnceo0piraaraetc.eaál..ys.1
Boyacá has 1,404,309 inhabitants, 36,136 of which reside in the The population of Putumayo and iTnhteheetTada dshuiedcaurdoeihaspThe population in the area includes mestizos as well as people of

Nev daeMaaanrttaa Cauca
Departmentcá Boyacá is located 130 km northwest of Bogotá (Figure 1), and the)The project was carried out in the munV icipllaelidtyelof Puerto Asis located

952

359 Annex 131-E

REGIONAL DIFFERENCES IN GLYPHOSATE USE AND FECUNDITY 953

smoking, drug, coffee, and alcohol consumption. Data on life RESULTS
style practices and work status for the father were also col- TTP showed large differences in different regions (Table 2).

lected. A variable for self-perception of pollution of water waThe Department of Valle del Cauca displayed a low percentage
included, as well as one related to the source of water con- for the first month and Boyacá and Nariño were exceptionally
sumption in the current domicile.
high for the twelfth month (Figure 2).
Participating women were generally young (mean and
median age 21 yr, range 15–48 yr, but there was one of 54 yr of
Statistical Analysis
age) and had completed at least some secondary education
For analysis purposes, if TTP was reported as zero months (Table 3). The vast majority had regular menstrual cycles
(or “unexpected”), the answer was interpreted as 1 mo. Cut (96.7%); a substantial proportion had irregular partner relation-

points for categorization of continuous variables were set as fships. Most became pregnant first at young ages (73.6% at 20
lows: age at time of interview at≤25 yr; age when attempting toyr of age or less). During the year before first pregnancy
get pregnant and age when first becoming pregnant was set at
(YBF), most were free of illness (84.3%), had not had x-rays
≤20 yr. For each exposure and potential confounder variable, (95.4%), and did not smoke tobacco (95.1%). Alcohol and cof-
analysis of variance (ANOVA) of mean TTP was conducted. fee consumption were 51.8% and 80.3%, respectively.
Among the 2592 women, 2477 pregnancies and 12,393
In the crude analyses (Table 3), longer TTP was associated
months (11,033 for final model) were included in multivariate with region, older maternal age, ethnic group, irregular men-
models. Each month was classified according to the relevant strual cycles, and irregular partner relationship. Previous visits
exposure and confounder variables and an indicator variable
to physician for problems related with fertility, x-rays taken in
was generated for every month, giving information on whether the year before pregnancy (YBP), and coffee consumption in
the cycle under this exposure resulted in a pregnancy or not. the YBP were associated with longer TTP. A significant trend

Fecundability odd ratios (fOR) were calculated with 95% con- between coffee consumption and longer TTP was observed.
fidence intervals (95% CI) using a discrete time analogue of Maternal overweight showed a borderline significant associa-
Cox’s proportional hazard model (Baird et al., 1986; Curtis
tion with a longer TTP.
et al., 1999; Zhou & Weinberg, 1999). Because TTP was The majority of women were housekeepers at the time they
assessed for a period of 12 mo, a separate censor variable was become pregnant. A tendency to longer TTP was observed among
introduced if a woman took >12 mo to conceive. A value of 0
those engaged in some waged work and with higher education.
(noncensored) was used if TTP was ≤12 mo and 1 if TTP was Paternal unemployment or self wrok, were associated with longer
>12 mo. fOR below unity indicate subfertility. All analyses TTP. No other paternal data were related to the outcome.

were performed using Stata 7.0 (Stata Corporation, College Self-perception about bad quality of water was associated
Station, TX) with macros developed by Dinno (2002). with longer TTP, and all sources of water presented risk when
The initial saturated multivariate model included all vari-
they were compared with pure water (“nacimiento”), except
Downloaded At: 20:07 2 October 2009ariate analysis (p < .10) and variablessome few cases that used carried water (“carro-tanque”).
prime biological importance (age at time of trying to become
pregnant). Several goodness-of-fit statistics for logistic regres-

sion were checked: Pearson chi-square, deviance, and Hosmer– TABLE 2
Lemeshow statistics (Hosmer & Lemeshow, 1989). The final
model consisted of only those variables that contributed to the Time to Pregnancy and Percentage of Pregnancy
by Month in the Study Regions
explanatory value of the model at a .05 level of significance
(coefficient of determination). Collinearity was tested with VIF Regions

(variance inflation factor). The assumption that the fecundabil- Sierra
ity odds ratio was constant across time (Weinberg & Wilcox,
1998) was tested graphically and by including an interaction Nevada Valle
de Santa del
term between months to pregnancy and exposure or confounder Months Boyacá Nariño Marta Putumayo Cauca Total
variables in the final model. The latter were not significant,
implying that the proportional assumption was not violated.
1 69.2 21.2 25.5 49.4 17.0 36.8
Finally, to evaluate a possible selection bias based on wanted-3 82.5 62.9 52.9 56.1 28.7 57
ness, the analyses were repeated excluding the pregnancies 6 88 94.8 72.1 74.9 45.2 75.2

occurring in the first month (Weinberg et al., 1994). No signif12 96.9 99.3 87.3 89 73.5 89.4
cant changes in the final model were observed. MTTP a 3 3.3 8.6 6.4 14 7
An alternative model without perfect fitting is presented for b
MTTP 3 3.3 7.1 6 12.6 6.3
the sake of research interest, even though it had some marginal a
variables (p values >.05). bTTP, mean time to pregnancy in months.
Censored to 60 mo (see text).

360 Annex 131-E

954 L.-H. SANIN ET AL.

100

Boyacá

Cu20lativepercent of pregnancies Nariño
Sierra Nevada
Putumayo
Valle del Cauca

0
2 4 6 8 10 12

Timeto first pre gnancy (months)

FIG. 2. Unadjusted cumulative percentage of pregnancies over time for the five study regions in Colombia.

In the final multivariate model (Table 4), the main predictor problem in many epidemiological studies is the lack of appro-
was region adjusted by irregular relationship with partner and priate exposure data based on actual measurements (Arbuckle

maternal age at first pregnancy. Boyacá displayed minimal risk et al., 2002; Harris et al., 2002; Coble et al., 2005; Ritter et al.,
and was used as the reference. Nariño, Sierra Nevada de Santa 2006; Firth et al., 2007). In most cases, exposures are approxi-

Marta, and Putumayo showed higher risk, with the highest risk mated through questionnaires, geographical regions, type of
in Valle del Cauca. Goodness-of-fit statistics for the final crop, season of application, ch emical group, or classifica-

model were optimal when adjustment for maternal age when tion according to mode of acti on (herbicides, insecticides,
the first pregnancy had occurred was carried out. Table 4 fungicides, etc). This is done because most pesticides lack a
shows the analysis without including 159 women who reported persistent biomarker, which prevents a measurement-based char-

Downloaded At: 20:07 2 October 2009use of fertility problems. In the crude acterization of exposure for the majority of the pesticide prod-
analysis, irregular cycles and medication for this purpose were ucts, including glyphosate (Acquavella et al., 2004).

associated with longer TTP, but when potentially subfertile For this reason, the acute effects of this herbicide are the
couples were excluded, these two variables were no longer most extensively documented (Acquavella et al., 1999) with

included in the final model. Age at first pregnancy and irregu- predominant manifestations being eye irritation and other tem-
lar relationship remained in the model after excluding those porary dermal effects. Whether pneumonitis occurs is contro-

with fertility problems. Table 5 shows that coffee consumption versial (Pushnoy et al., 1998), and fatal cases have been
and perception of contamination of water, although no longer recorded only with accidents or when glyphosate was ingested
significant, were borderline. When categorized in number of with the purpose of committing suicide (Williams et al., 2000).

cups, coffee consumption still showed a positive trend; the Some cases of Parkinson’s disease have been associated with
greater the number of cups, the longer was the TTP. acute intoxication with glyphosate (Barbosa et al., 2001), but

An alternative model is presented in Table 5 because that the small number of cases and lack of laboratory animal analo-
model includes variables such as coffee consumption and gies do not allow assignment of causality.

water pollution with marginal statistical significance but with Some authors have made efforts to identify the compounds
strong biological and environmental significance. used by study subjects. Several studies on different populations

that specifically addressed the use of glyphosate were found
and published since the last major reviews (Williams et al.,
DISCUSSION 2000; Solomon et al., 2007). Studies related to cancer and to

This was the first study performed in Colombia with the adverse reproductive and developmental effects reported
objective of assessing whether an association existed between equivocal and unclear relationships between glyphosate use

use of aerially applied glyphosate for eradication of illicit croand some reproductive outcomes (Curtis et al., 1999; Arbuckle
and subchronic effects on reproduction, such as TTP. A major et al., 2001; Garry et al., 2002; De Roos et al., 2005).

361 Annex 131-E

REGIONAL DIFFERENCES IN GLYPHOSATE USE AND FECUNDITY 955

TABLE 3

Mean Time to Pregnancy (Without Censoring) and Crude Fecundability Odds Ratio (fORc)
Analyzed by Different Sociodemographic Characteristics

Time to pregnancy (mo), b
Variable n X (SD) a fORc (CI 95%) p

Region

Boyacá 582 3 (4.7) 1 –
Nariño 552 3.3 (3.3) 0.72 (0.62, 0.83) <.01
Sierra Nevada de Santa Marta 551 7.1 (10.3) 0.41 (0.35, 0.48) <.01

Putumayo 535 6 (8.3) 0.44 (0.38, 0.51) <.01
Valle del Cauca 531 12.6 (13.5) 0.2 (0.17, 0.24) <.01

Maternal age (yr)
≤25 2356 5.7 (8.2) 1 –
>25 395 10 (14 ) 0.64 (0.56, 0.73) <.01

Age to first pregnancy (yr)
≤20 2026 5.5 (8 ) 1 –

>20 725 8.6 (12.3 ) 0.69 (0.62, 0.76) <.01
Age at start TTP study period (yr)
≤20 2094 6.2 (9) 1 –

>20 657 6.8 (10.5) 0.98 (0.88, 1.1) .69
Ethnic group
Mestizo 2121 6.5 (9.6) 1 –

Negro 508 6.3 (9.6) 1. (0.9, 1.14) .83
Indígena 49 3.7 (4.1) 1.37 (0.9, 1.94) .08

Zambo 41 3.6 (2.6) 1.38 (0.95, 2.01) .09
Mulato 32 3.3 (2.6) 1.6 (1.05, 2.51) .03
Grouped ethnic group

Mestizo and Negro 2629 6.45 (9.6) 0.7 (0.56, 0.87) <.01
Indígena, Zambo, and Mulato 122 3.54 (3.2) 1 –
Education

None 42 4.9 (9.1) 1 –
Downloaded AtIncomplete elementary school 582 4.6 (7.1) 0.93 (0.63, 1.39) .74

Complete elementary school 526 5.6 (7.7) 0.76 (0.51, 1.13) .17
Incomplete high school 459 7 (10.3) 0.66 (0.45, 0.98) .04
Complete high school 130 9.7 (12.5) 0.47 (0.31, 0.73) <.01

Zone
Urban 5 15.2 (18.6) – –

Rural c 2743 6.3 (9.4) – –
Marital status
Common law 1010 5.1 (8.3) 1 –

Not common law 1741 7 (9.9) 0.71 (0.64, 0.78) <.01
Socioeconomic statusc,d
0 38 4.8 (5.8) 1.1 (0.75, 1.63) .62

1 2013 6.4 (9.4) 1 –
≥2 493 6.6 (10.1) 1.03 (0.91, 1.17) .60
e
Nutritional status
Low weight 111 7.1 (10.9) 0.91 (0.72, 1.15) .42
Normal weight 2453 6.2 (9.3) 1 –

Overweight 184 7.2 (10.0) 0.83 (0.69, 1.00) .04
Maternal work c
Administrative, teacher, or student 678 6 (8.6) 0.99 (0.89, 1.11) .92

(Continued)

362 Annex 131-E

956 L.-H. SANIN ET AL.

TABLE 3

(Continued)

Time to pregnancy(mo),
Variable n X (SD) a fORc (CI )b p
95%

Home, no work, housekeeper 1631 6 (9.2) 1 –
Community mother, mining, various, 229 8.7 (11.9) 0.7 (0.59, 0.83) <.01
other, occasional

Health worker, independent 126 8.7 (12.3) 0.74 (0.59, 0.93) .01
worker, seller
Agriculture and floriculture 86 4.2 (4.6) 1.14 (0.87, 1.48) .34
c
Maternal work in cocaine
No 2743 6.3 (9.4) 1 –

Yes 8 7.6 (7.4) 0.67 (0.29, 1.54) .35
Menarche age (yr)
<12 1031 6.6 (10) 1 –

13 802 5.8 (8.6) 1.06 (0.94, 1.19) .32
14 523 6.4 (9.3) 1.02 (0.89, 1.16) .81

15 392 6.6 (9.4) 0.94 (0.82, 1.09) .43
Menstrual cycle
Regular 2612 6.2 (9.3) 1 –

Irregular 88 9.5 (12.3) 0.64 (0.49, 0.84) <.01
Previous consultation for pregnancy problems

No 2592 5.8 (8.7) 1 –
Yes 159 15.2 (14.4) 0.33 (0.27, 0.41) <.01
Smoking c

No 2616 6.3 (9.5) 1 –
Yes 135 6.2 (7.9) 0.95 (0.77, 1.17) .63
c
Alcohol consumption
No 1325 6.2 (9.4) 1 –
Yes 1425 6.3 (9.4) 0.97 (0.88, 1.06) .52
c
Coffee consumption
Downloaded ANo 20:07 2 October 2009 543 5.3 (8.1) 1 –

Yes c 2208 6.6 (9.7) 0.81 (0.72, 0.91) <.01
Number of coffee cups/day **
0 543 5.3 (8.1) 1 –

1 to 3 1916 6.4 (9.5) 0.83 (0.73, 0.93) <.01
≥4 292 7.4 (10.7) 0.73 (0.61, 0.87) <.01
c
X-rays
No 2616 6.2 (9.2) 1 –
Yes 125 9.4 (12.5) 0.67 (0.54, 0.84) <.01
c
Any kind of illness
No 2316 6.3 (9.3) 1
Yes 432 6.4 (9.7) 0.97 (0.86, 1.10) .68
c,e
STD
No 2717 6.3 (9.3) 1 –

Yes 27 7.3 (10.4) 0.84 (0.52, 1.38) .50
Medication for regularizing menses
No 2721 6.3 (9.4) 1 –

Yes 30 11.2 (10) 0.45 (0.28, 0.71) <.01
Medication for “sugar in blood”

No 2735 6.3 (9.4) 1 –

(Continued)

363 Annex 131-E

REGIONAL DIFFERENCES IN GLYPHOSATE USE AND FECUNDITY 957

TABLE 3

(Continued)

Time to pregnancy(mo),
Variable n X (SD) a fORc (CI )b p
95%

Yes c 16 7.6 (6.5) 0.7 (0.39, 1.24) .22
Other medications
No 2027 6.8 (9.7) 1 –

Yes 703 5 (8.3) 1.36 (1.22, 1.51) <.01
Paternal workc
Administrative or student 160 6.3 (8.2) 0.86 (0.7, 1.05) .14

No work, occasional 212 8.4 (12.5) 0.74 (0.61, 0.88) <.01
Carpenter, driver, construction, mining, 507 5.7 (9) 1.02 (0.89, 1.16) .80

mechanic, industrial timbering
Other, health worker, independent 713 7.7 (10.8) 0.75 (0.67, 0.84) <.01
worker, vendor

Agriculture, floriculture, livestock 1157 5.4 (7.9) 1 –
Paternal work in cocainec

No 2457 6.4 (9.6) 1 –
Yes 292 5.5 (7.8) 1.07 (0.92, 1.24) .40
Any disease of the father

No 2398 6.4 (9.5) 1 –
Yes 248 6.8 (9.4) 0.93 (0.80, 1.1) .41
c,f
STD of the father
No 2608 6.4 (9.5) 1 –
Yes 37 6.1 (8.2) 0.99 (0.66, 1.47) .94
c
Paternal alcohol consumption
No 1325 6.2 (9.4) 1 –

Yes c 1425 6.4 (9.4) 0.97 (0.88, 1.06) .52
Paternal smoking
No 2143 6.2 (9.3) 1 –

Yes 538 6.9 (10) 0.9 (0.80, 1.01) .08
Downloaded Paternal use of psychotropic drugs

Yes 2619 6.4 (9.5) 1 –
No 54 4.5 (5.7) 1.23 (0.88, 1.72) .22
Perceived contamination of water

No 1218 6 (9.2) 1 –
Yes 1533 6.6 (9.5) 0.9 (0.82, 0.98) .02

Source of drinking water
Municipal tap water 598 7.2 (11) 1.13 (1, 1.28) .05
Rain water 65 5.3 (8) 1.38 (1.02, 1.88) .04

Stream, ravine, or creek 257 6.1 (10.2) 1.36 (1.14, 1.61) <.01
“Carried water” 10 14.8 (16.5) 0.44 (0.19, 1.05) .07
“Pure water” 311 3.7 (5.7) 2.03 (1.73, 2.39) <.01

Deep well 1040 7.4 (10) 1 –
River 470 4.6 (6.5) 1.52 (1.33, 1.74) <.01

**There is a significant trend when p < .05.
a
Mean and standard deviation.
bCrude fecundability odds ratio; 95% confidence interval.
cDuring the year prior to pregnancy.
d
eThe population is classified in 6 socioeconomic strata, from 1 being the lowest to 6the highest. A zero indicates extreme poverty.
Based on self-reporting images scaled from 1 to 9. Low weight 1 to 4, normal 5 to 7, overweight 8 and 9 (BMI≥25) (Madrigal-Fritsch et al.,
1999).
fSTD, sexually transmitted disease.

364 Annex 131-E

958 L.-H. SANIN ET AL.

TABLE 4 TABLE 5
Causes of Fecundability Adjusted a for the Relationship a
b Causes of Fecundability Adjusted for the Relationship
Between Time to Pregnancy (TTP) and Region Between Time to Pregnancy (TTP) and Region b
Based on an Alternative Model
Variable fRMa c EE d IC e p
95% c d e
f Variable fRMa EE IC 95% p
Region
Nariño 0.53 0.044 0.45, 0.63 <.01 Region f
Sierra Nevada 0.36 0.030 0.30, 0.42 <.01
Nariño 0.56 0.048 0.47, 0.66 <.01
Putumayo 0.34 0.029 0.29, 0.41 <.01 Sierra Nevada 0.36 0.031 0.31, 0.43 <.01
Valle del Cauca 0.15 0.013 0.12, 0.18 <.01 Putumayo 0.35 0.029 0.29, 0.41 <.01

Age at first pregnancy 0.81 0.048 0.72, 0.91 <.01 Valle del Cauca 0.15 0.014 0.13, 0.18 <.01
>20 yr g Age at first pregnancy 0.81 0.048 0.73, 0.91 <.01
g
Irregular relationship 0.76 0.041 0.68, 0.84 <.01 >20 yr
with fatherh Irregular relationshiph 0.76 0.041 0.68, 0.84 <.01

Note. n = 2592 mothers 11,270 cycles. Consumption of coffee i
a
Proportional risk model of Cox, modified after Dinno, (2002). Medium (1–3 cups 0.91 0.059 0.81, 1.04 .15
bRestricted to those mothers who did not consult a physician per day)
regarding problems in conceiving. High (4 and more 0.84 0.083 0.69, 1.02 .08
c
fRMa Adjusted cause of fecundability. cups per day)
dStandard error. Perception of 0.91 0.51 0.81, 1.01 .08
e95% Confidence interval. j
f contamination of water
Compared to Boyacá as reference.
gCompared to ≤20 years as reference. Note. n = 2592 mothers, 11,270 cycles.
hCompared to regular relationship as reference. aProportional risk model of Cox, modified after Dinno (2002).
b
Restricted to those mothers who did not consult a physician
regarding problems in conceiving.
cfRMa Adjusted cause of fecundability.
Other risk factors present in the rural and agricultural envi- d
ronment of the women studied and individual characteristics Standard error.
(genetic, for example) may be associated with TTP. Longer e95% Confidence interval.
fCompared to Boyacá as reference.
TTP were observed in some populations with higher physical g
activity (Florack et al., 1994) or psychological distress Compared to ≤20 years as reference.
hCompared to regular relationship as reference.
(Hjollund et al., 1999). Further, TTP may be influenced by iCompared to no consumption as reference.
Downloaded knowledge and behavior, such as patterns of intercourse as j
Compared to no contamination as reference and based on self-
well as biologic factors (Joffe et al., 2005), and these need to perception and source of water normally consumed.
be considered as potential confounders (Tingen et al., 2004;

Stanford & Dunson, 2007) The potential effect of these factors
on TTP could not be isolated in this study, even though the Pesticides in general are likely not the cause either, as large
differences in TTP were observed between two regions of high
fOR was adjusted for most known confounders and indepen-
dent predictors. to moderate pesticide use, Valle del Cauca and Boyacá. The
As shown in Figure 2, there was no difference in cumulative observed ecological differences remain unexplained, but may

TTP between Putumayo, where illicit crops were sprayed, and be produced by varying exposures to environmental factors,
Sierra Nevada, where there was no herbicide use. In turn, the history of contraceptive programs in the region, or psychologi-

latter region showed lower cumulative percent pregnancies cal distress. Future studies examining these causes are needed.
than Nariño, an eradication spray area, and Boyacá, where Table 3 shows crude associa tion between coffee consump-

there is agricultural herbicide use but manual eradication of tion and longer TTP with a signifi cant trend. This association
illicit crops. Although classification of exposure may be a is not significant in the adjusted model but the level of signif-
icance was borderline. Published results regarding coffee or
source of bias in this type of study, no relationship between
reduced fecundability in the studied regions and use of glypho- caffeine consumption and TT P are not conclusive. Some
sate specifically for spray eradication or use of pesticides in studies showed no association (J oesoef et al., 1990; Alderete

general can be established from our data. Prospective studies et al., 1995), but other investigators found that coffee drink-
that prevent or reduce classification bias of exposures are rec- ers have a lower risk of pregnancy (Wilcox et al., 1988;

ommended to further elucidate relationships between aerial Christianson et al., 1989; Williams et al., 1989; Hatch &
spraying of glyphosate for eradication, agricultural pesticide Bracken, 1993; Curtis et al., 1997). This relationship needs to

use, and human health indicators. be further investigated.

365 Annex 131-E

REGIONAL DIFFERENCES IN GLYPHOSATE USE AND FECUNDITY 959

Distribution of pregnancies in relation with months in dif- REFERENCES

ferent regions showed great differences (Table 2). In a previ- Abell, A., Juul, S., and Bonde, J. P. E. 2000. Time to pregnancy among female
ous study in Colombia (Idrovo et al., 2005), the percentage for greenhouse workers. Scand. J. Work Environ. Health 26:131–136.
Acquavella, J. F., Alexander, B. H., Mandel, J. S., Gustin, C., Baker, B.,
first month was close to 30%, which is lower than more than Chapman, P., and Bleeke, M. 2004. Glyphosate biomonitoring for farmers
40% reported from a Danish study (Joffe et al., 2005). In our and their families: Results from the farm family exposure study. Environ.
Health Perspect., 112:321–326.
study, the region of Valle del Cauca showed a low percentage Acquavella, J. F., Weber, J. A., Cullen, M. R., Cruz, O. A., Martens, M. A.,
and Boyacá exceptionally high for first and twelfth months
(Figure 2). The mean for 12 mo in developed countries is Holden, L. R., Riordan, S., Thompson, M., and Far®er, D. 1999. Human
ocular effects from self-reported exposures to Roundup herbicides. Hum.
between 85 and 90%. These results are consistent with the Exp. Toxicol., 18:479–486.
National Survey of Demography and Health (Ojeda et al., Alderete, E., Eskenazi, B., and Sholtz, R. 1995. Effect of cigarette smoking
and coffee drinking on time to conception. Epidemiology 6:403–408.
2005) that showed Boyacá as th e Department with the lowest Arbuckle, T. E., Burnett, R., Cole, D., Teschke, K., Dosemeci, M., Bancej, C.,
proportion of women who reported fertility problems (4.2%).
and Zhang, J. 2002. Predictors of herbicide exposure in farm applicators.
Valle del Cauca (11.2%) and Magdalena (16.1%), where Int. Arch. Occup. Environ. Health 75:406–414.
Sierra Nevada is located, were above the national average Arbuckle, T. E., Cole, D. C., Ritter, L., and Ripley, B. D. 2004. Farm chil-
dren’s exposure to herbicides: Comparison of biomonitoring and question-
(10.6%). naire data. Epidemiology 15:187–194.
A retrospective assessment of TTP as an outcome variable Arbuckle, T. E., Lin, Z., and Mery, L. S. 2001. An exploratory analysis of the

was conducted to evaluate ecological exposure to glyphosate. effect of pesticide exposure on the risk of spontaneous abortion in an
Although it is widely recognized that retrospective studies for Ontario farm population. Environ. Health Perspect. 109:851–857.
Axmon, A., Rylander, L., Albin, M., and Hagmar, L. 2006. Factors affecting
TTP can be carried out, they are prone to some biases that need time to pregnancy. Hum. Reprod. 21:1279–1284.
to be taken into account in the interpretation of our results. Dif- Baird, D. D., Wilcox, A. J., and Weinberg, C. R. 1986. Use of time to preg-
nancy to study environmental exposures. Am. J. Epidemiol. 124:470–480.
ference in sexual behavior between exposed and nonexposed
subjects, particularly in frequency of intercourse, has been Barbosa, E. R., Leiros da Costa, M. D., Bacheschi, L. A., Scaff, M., and Leite,
C. C. 2001. Parkinsonism after glycine-derivate exposure. Movement
pointed out as source of bias (Tingen et al., 2004; Stanford & Disorders 16:565–567.
Dunson, 2007). Women of reproductive age differed in report- Bonde, J. P., Joffe, M., Sallmén, M., Kristensen, P., Olsen, J., Roeleveld, N.,
and Wilcox, A. 2006. Validity issues relating to time-to-pregnancy studies
ing intercourse in the last 4 wk, from 48.8% in Boyacá and of fertility. Epidemioogy 17:347–349.
neighboring departments to 53.8% in the Pacific region where
Tumaco (Nariño) is located (Ojeda et al., 2005). Couples who Bretveld, R., Kik, S., Hooiveld, M., van Rooij, I., Zielhuis, G., and Roeleveld,
N. 2008. Time-to-pregnancy among male greenhouse workers. Occup.
had not used contraception in the last year were included and, Environ. Med. 65:185–190.
in the multivariate analysis, those who had had consultation Bretveld, R., Zielhuis, G. A., and Roeleveld, N. 2006. Time to pregnancy
among female greenhouse workers. Scand. J. Work Environ. Health
because of fertility problems were excluded. These two criteria 32:359–367.
excluded those who may have become pregnant while using
Christianson, R. E., Oechsli, F. W., and van den Berg, B. J. 1989. Caffeine
contraception (highly fecund couples) and subfertile couples beverages and decreased fertility. Lancet 335:378–378.
Downloaded A(Bonde et al., 2006). Studies also evaluated whether there were Coble, J., Arbuckle, T., Lee, W., Alavanja, M., and Dosemeci, M. 2005. The
validation of a pesticide exposure algorithm using biological monitoring
other sources of bias such as pregnancy recognition (Joffe et results. J. Occup. Environ. Hyg. 2:194–201.
al., 2005) by asking whether a miscarriage occurred, and thus it Curtis, K. M., Savitz, D. A., and Arbuckle, T. E. 1997. Effects on cigarette

was possible to control for this variable. However, biological smoking, caffeine consumption, and alcohol intake on fecundability. Am. J.
factors such as age at first pregnancy and use of contraception Epidemiol., 146:32–41.
Curtis, K. M., Savitz, D. A., Weinberg, C. R., and Arbuckle, T. E. 1999.
in the past were taken into account as these appear to be more The effect of pesticide exposure on time to pregnancyEpidemiology
important than lifestyle factors in assessing TTP (Axmon et al., 10:112–117.
De Cock, J., Westveer, K., Heederik, D., te Velde, E., and van Kooij, R. 1994.
2006).
Classification of exposure was by location of residence. Time to pregnancy and occupational exposure to pesticides in fruit growers
in the Netherlands. Occup. Environ. Med. 51:693–699.
Nonexposed participants were those who lived in the region De Roos, A. J., Blair, A., Rusiecki, J. A., Hoppin, J. A., Svec, M., Dosemeci,
where organic crops were produced and who, in addition, did M., Sandler, D. P., and Alavanja, M. C. 2005. Cancer incidence among
not report any use of pesticides in the interview. In the other glyphosate-exposed pesticide applicators in the Agricultural Health Study.
Environ. Health Perspect. 113:49–54.
four departments, there was exposure not only to glyphosate,
but also to other herbicides and pesticides. Although place of Dinno, A. 2002. DTHAZ: Stata module to compute discrete-time hazard and
survival probability. Accessed March 15, 2005. http://ideas.repec.org/c/
residence is not an accurate surrogate for exposure to pesti- boc/bocode/s420701.html
cides and may generate misclassification (Arbuckle et al., Firth, H. M., Rothstein, D. S., Herbison, G. P., and McBride, D. I. 2007. Chem-
ical exposure among NZ farmers.Int. J. Environ. Health Res.17:33–44.
2004), this ecological assessment is useful to explore, at the Florack, E. I., Zielhuis, G. A., and Rolland, R. 1994. The influence of occupational
population level, whether an association exists between the
physical activity on the menstrual cycle and fecundabilityE.pidemiology
putative exposure (aerial spraying of glyphosate) and outcome 5:14–18.
(Ritter et al., 2006). However, in this study, aerial spraying of Garry, V. F., Harkins, M. E., Erickson, L. L., Long-Simpson, L. K., Holland,
S. E., and Burroughs, B. L. 2002. Birth defects, season of conception, and
glyphosate was not consistently associated with delayed time sex of children born to pesticide applicators living in the Red River Valley
to pregnancy. of Minnesota, USA. Environ. Health Perspect. 110(Suppl. 3):441–449.

366 Annex 131-E

960 L.-H. SANIN ET AL.

Harris, S. A., Sass-Kortsak, A. M., Corey, P. N., and Purdham, J. 2002. Devel- Petrelli, G., and Figà-Talamanca, I. 2001. Reduction in fertility of male green-
opment of models to predict dose of pesticides in professional turf applicators.use workers exposed to pesticides. Eur. J. Epidemiol. 17:675–677.

J. Expos. Anal. Exp. Epidemiol. 12:130–144. Pushnoy, L. A., Avnon, L. S., and Carel, R. S. 1998. Herbicide (Roundup)
Hatch, E. E., and Bracken, M. B. 1993. Association of delayed conception with pneumonitis. Chest 114:1769–1771.
caffeine consumption. Am. J. Epidemiol. 138:1082–1092. Ritter, L., Goushleff, N. C. I., Arbuckle, T., Cole, D., and Raizenne, M. 2006.
Hjollund, T. B., Jensen, T. K., Bonde, J. P., Henriksen, T. B., Andersson, A. Addressing the linkage between exposure to pesticides and human health

M., Kolstad, H. A., Ernst, E., Giwercman, A., Skakkebaek, N. E., and effects— Research trends and priorities for research 1. J. Toxicol. Environ.
Olsen, J. 1999. Distress and reduced fertility: A follow up study of first- Health B 9:441–456.
pregnancy planners. Fertil. Steril. 72:47–53. Romieu, I., Lazcano-Ponce, E., Sanchez-Zamorano, L. M., Willett, W., and
Hosmer, D. W., and Lemeshow, S. 1989.Applied logistic regression. New York: Hernandez-Avila, M. 2004. Carbohydratesand the risk of breast cancer

Wiley. among Mexican women.Cancer Epidemiol. Biomarkers Prev1.3:1283–1289.
Idrovo, J., Sanin, L. H., Cole, D., Chavarro, J., Caceres, H., Narváez, J., Sallmén, M., Liesivuori, J., Taskinen, H., Lindbohm, M. L., Anttila, A., Aalto, L.,
and Restrepo, M. 2005. Time to first pregnancy among women work- and Hemminki, K. 2003. Time to pregnancy among the wives of Finnish
ing in agricultural production. Int. Arch. Occup. Environ. Health 78: greenhouse workers. Scand. J. Work Environ. Health 29:85–93.

493–500. Singh, D. 1994. Waist-to-hip ratio and judgment of attractiveness and healthi-
Joesoef, M. R., Beral, V., Rolfs, R., Aral, S. O., and Cramer, D. W. 1990. Are ness of female figures by male and female physicians. Int. J. Obesity Rel.
caffeinated beverages risk factors for delayed conception? Lancet Metab. Disorders 18:731–737.
335:136–137. Solomon, K. R., Anadón, A., Carrasquilla, G., Cerdeira, A., Marshall, J., and

Joffe, M. 1997. Time to pregnancy: A measure of reproductive function in Sanin, L.-H. 2007. Coca and poppy eradication in Colombia: Environmental
either sex. Occup. Environ. Med. 54:289–295. and human health assessment ofaerially applied glyphosate.Rev. Environ.
Joffe, M. 2000. Time trends in biological fertility in Britain. Lancet Contam. Toxicol. 190:43–125.
355:1961–1965. Stanford, J. B., and Dunson, D. B. 2007. Effects of sexual intercourse patterns

Joffe, M., and Barnes, I. 2000. Do parenteral factors affect male and female. in time to pregnancy studies. Am. J. Epidemiol. 165:1088–1095.
Epidemiology 11:700–705. Thonneau, P., Abell, A., Larsen, S. B., Bonde, J. P., Joffe, M., Clavert, A.,
Joffe, M., Key, J., Best, N., Keiding, N., Scheike, T., and Jensen, T. K. 2005. Ducot, B., Multigner, L., and Danscher, G., for the ASCLEPIOS Study
Studying time to pregnancy by uesof a retrospective design.Am. J. Epidemiol. Group. 1999. Effects of pesticide exposure on time to pregnancy: Results of

162:115–124. a multicenter study in France and Denmark.Am. J. Epidemiol.150:157–163.
Joffe, M., Paranjothy, S., Fielder, H., Lyons, R., and Palmer, S. 2008. Use of Tingen, C., Stanford, J. B., and Dunson, D. B. 2004. Methodologic and statisti-
time to pregnancy in environmental epidemiology and surveillance.J. Public cal approaches to studying human fertility and envir onmental exposure.
Health (Oxford)30:178–185. Environ. Health Perspect. , 112:87–93.

Joffe, M., Villard, L., Li, Z., Plowman, R., and Vessey, M. 1995. A time to Weinberg, C. R., Baird, D. D., and Wilcox,A. J. 1994. Sources of bias in studies
pregnancy questionnaire designed for long term recall: Validity in Oxford, of time to pregnancy.Stat. Med. 13:671–681.
England. J. Epidemiol. Commun. Health 49:314–319. Weinberg, C. R., and Wilcox, A. J. 1998. Reproductive epidemiology. in
Larsen, S. B., Joffe, M., Bonde, J. P., and the Asclepios Study Group. 1998. Modern epidemiology, 2nd ed., eds. K. J. Rothman and S. Greenland,

Time to pregnancy and exposure to pesticides in Danish farmers. Occup. pp. 585–608. Philadelphia, PA: Lippincott-Raven.
Environ. Med. 55:278–283. Wigle, D. T., Arbuckle, T. E., Turner, M. C., Berube, A., Yang, Q., Lui, S., and
Lauria, L., Settimi, L., Spinelli, A., and Figà-Talamanca, I. 2006. Exposure to Krewski, D. 2008. Epidemiologic evidence of relationships between repro-
pesticides and time to pregnancy among female greenhouse workers. ductive and child health outcomes and environmental chemical contami-

Reprod. Toxicol. 22:425–430. nants. J. Toxicol. Environ. Health B 11:373–517.
Madrigal-Fritsch, H., de Irala-Estevez, J., Martinez-Gonzalez, M. A., Kearney, Wilcox, A., Weinberg, C., and Baird, D. 1988. Caffeinated beverages and
J., Gibney, M., and Martinez-Hernandez, J. A. 1999. [The perception of decreased fertility. Lancet 312:1453–1456.
Downloaded At: 2body image as a qualitative approach to nutritional status]. Salud Pública Williams, G. M., Kroes, R., and Munro, I. C. 2000. Safety evaluation and risk
®
Mex. 41:479–486. assessment of the herbicide Roundup and its active ingredient, glypho-
Ojeda, G., Ordoñez, M., and Ochoa, L. H. 2005. Salud Sexual y Reproductiva sate, for humans. Regul. Toxicol. Pharmacol. 31:117–165.
em Colombia, Encuesta Nacional de Demografia y Salud. Accessed June Williams, M. A., Monson, R. R., Goldman, M. B., and Mittendorf, R. 1989.
2, 2008. http://www.profamilia.org.co/encuestas/index_ends.htm Coffee and delayed conception. Lancet 335:1603–1603.

Olsen, J., and Skov, T. 1993. Design options and methodological fallacies in thZhou, H., and Weinberg, C. R. 1999. Potential for bias in estimating human
studies of reproductive failures.Environ. Health Perspect. 101(Suppl. 2): fecundability parameters: A comparison of statistical models. Stat. Med.
145–152. 18:411–422.

367368 Annex 131-F

Annex 131-F

M.H. BERNAL ET A., “OXICITY OFFORMULATED G LYPHOSATE(G LYPHOS)AND
COSMO -FLUX TOLARVAL COLOMBIAN F ROGS1.LABORATORY ACUTE TOXICIT”

(Journal of Toxicology and Environmental Health, Part A, 72:961-965, 2009)

369 Annex 131-F

Journal of Toxicology and Environmental Health, Part A, 72: 961–965, 2009
ISSN: 1528-7394 print / 1087-2620 online
DOI: 10.1080/15287390902929709

UTEH
Toxicity of Formulated Glyphosate (Glyphos)

and Cosmo-Flux to Larval Colombian

Frogs 1. Laboratory Acute Toxicity

M. H. Bernal , K. R. Solomon , and G. Carrasquillaratory 3
1
Laboratory of Herpetology, Eco-Physiology & Ethology, Universidad del Tolima, Barrio Santa Elena,
Ibagué, Tolima, Colombia, 2Centre for Toxicology and Department of Environmental Biology, University
3
of Guelph, Guelph, Ontario, Canada, and Facultad de Salud, Universidad del Valle, Cali, Colombia

in the glyphosate formulation, as the addition of Cosmo-Flux did
®
The spraying of coca (Erythroxylum coca) with glyphosate in not enhance toxicity above those reported for Vision = Roundup.
Colombia has raised concerns about possible impacts on amphibi-
ans. There are few toxicity data for species other than those from
temperate regions, and these have not been generated with the

combination of formulated glyphosate (Glyphos) and the adju- Extensive reviews of the effects of glyphosate and its for-
vant, Cosmo-Flux (coca mix) as used in coca control in Colombia. mulated products on aquatic organisms concluded that glyphosate
In order to characterize toxicity of the spray mixture to frogs
from Colombia, Gosner stage-25 tadpoles of Scinax ruber, presents a negligible risk to aquatic organisms (World Health
Dendropophus microcephalus, Hypsiboas crepitans, Rhinella gran- Organization International Program on Chemical Safety, 1994;
ulosa, Rhinella marina, Rhinella typhonius, Centrolene prosoble-
pon, and Engystomops pustulosus were exposed to the coca mix at Giesy et al., 2000; Solomon & Thompson, 2003). Although
amphibians are physiologically unique and ecologically impor-
concentrations of glyphosate ranging from 1 to 4.2 mg a.e./L tant, no regulatory agencies currently require amphibian toxic-
diluted in dechlorinated tap water in glass containers. Cosmo-Flux
was added to Glyphos in the proportion of 2.3% v/v, as used in ity data as part of their registration requirements. Determining
aerial application for coca control. Exposures were for 96 h at 23±direct and indirect effects of agrochemicals on amphibian spe-
1.5°C with 12:12-h light/dark cycle. Test solutions were renewed
every 24 h. Concentrations, measured within the first hour and at cies continues to be identified as a general research need
Downloaded At: 20:08 2 October 2009 (Linder et al., 2003).
24 and 96 h using enzyme-linked immunosorbent assay (ELISA) Several recent publications reported that glyphosate (active
(Abraxis, LLC), ranged from 70 to 130% of nominal values. LC50
values ranged from 1200 to 2780 mg glyphosate acid equivalents ingredient) exerts low toxicity to larval amphibians. The 48-h
(a.e.)/L for the 8 species tested. Data suggest that sensitivity toLC50 values of technical-grade glyphosate isopropylamine
Roundup-type formulations of glyphosate in these species is simi- (IPA) salt to larval Australian frogs (Litoria moorei, Crinia
lar to that observed in other tropical and temperate species. In
addition, sensitivity of larvalamphibians to Roundup-type formu- insignifera, Lymnodynastes dorsalis, and Heleioporus eyrei)
were reported to range from >343,000 to >466,000μg glyphosate
lations spans a relatively narrow range. Finally, toxicity of the
mixture as used to spray coca was likely driven by the surfactant acid equivalents (a.e.)/L (Mann & Bidwell, 1999). The 96-h
LC50 of glyphosate IPA in Rana clamitans was reported to be
>38,900 μg a.e/L from a static exposure study (Howe et al.,

2001). From this limited data set, it appears that glyphosate
©General Secretariat of the Organization of American States, IPA is essentially nontoxic to amphibians.
2009. This paper was prepared as part of a Study entitled The toxicity of some formulated glyphosate products to

“Production of Illicit Drugs, the Environment and Human Health,” amphibians is greater than that of the active ingredient. A study
financed with contributions from the Governments of Colombia and by Mann and Bidwell (1999) examined the acute toxicity of
the United States of America. The conclusions and opinions
expressed herein are those of the authors and not necessarily thoseRoundup herbicide (MON 2139) for C. insignifera, H. eyrei,
of the Organization of American States and its General Secretariat,L. dorsalis, and L. moorei tadpoles and reported 48-h LC50
which as of the date of this copyright, have not formulated any values ranging between 2900 and 11,600 μg a.e./L glyphosate.
opinion with respect to them.
Using a formulation of glyphosate (Vision containing glypho-
Address correspondence to Dr. M. H. Bernal, Laboratory of sate and ethoxylated tallowamine surfactant [POEA] and
Herpetology, Eco-Physiology & Ethology, Universidad del Tolima,
Barrio Santa Elena, Ibagué, Tolima, Colombia. E-mail: mhbernal@ equivalent to Roundup), 96-h LC50 values as low as 880 μg
ut.edu.co a.e./L were reported for tadpoles of Xenopus laevis, Bufo

961

370 Annex 131-F

962 M. H. BERNAL ET AL.

americanus, Rana clamitans, and Rana pipiens (Edginton Cosmo-Flux did not alter the toxicity of the mixture. However,
et al., 2004). Embryo stages were less sensitive than Gosner there are few data in the literature on the susceptibility of tropi-

stage 25 larvae, and toxicity was affected by the pH of the cal frog species to formulations of glyphosate and there are no
exposure medium, although not in a consistent manner. A data for species native to Colombia. Because of this, acute labo-
study on R. clamitans, R. pipiens, Rana sylvatica, and B. amer-ratory tests on larvae of native Colombian species of frogs were

icanus (Howe et al., 2004) reported 96-h LC50 values for conducted. This study describes the acute toxicity of the Gly-
Roundup Original of 2200, 2900, and 5100 μg a.e./L, respec- phos and Cosmo-Flux mixture to tadpoles (Gosner stage 25) of

tively. A study on Rana cascadae reported a 48-h LC50 for the frog species, Hypsiboas crepitans (Wied-Neuwied, 1824),
Roundup of 2336 μg a.e./L using static exposures in glass Rhinella granulosa (Spix, 1824), Engystomops pustulosus
tanks (Cauble & Wagner, 2005). In a study carried out with (Cope, 1864), Rhinella marina (Linnaeus, 1758), Scinax ruber

R. catesbeiana, R. clamitans, Hyla versicolor, R. pipiens, (Laurenti, 1768), Dendropsophus microcephalus(Cope, 1886),
B. americanus, and R. sylvatica, 384-h LC50 for Roundup Rhinella typhonius (Linnaeus, 1758), and Centrolene prosoble-
were reported to range from 977 to 1865 μg a.e /L (based on pon (Boettger, 1892), during a 96-h exposure period under

the assumption that the reported concentration of the AI was asstatic-renewal test conditions in the laboratory.
the IPA; Relyea 2005). It is not clear why only the 16-d LC50
values were calculated when it appeared that mortality

occurred early in the exposure period; however, the reported
LC50s were not greatly different from those reported by other MATERIALS AND METHODS

authors (discussed earlier). Test Organisms
The toxicity of some other formulations of glyphosate is Species that occur in locations where coca is grown (≤ 1000
less than that of Roundup. Roundup Biactive (MON 77920)
m a.s.l.) were the focus of the study. Embryos of the test spe-
was practically nontoxic to tadpoles, producing 48-h LC50 val- cies were collected in the locations shown in Table 1 and trans-
ues of 328,000 μg a.e./L for L. moorei and >360,000 μg a.e./L
for C. insignifera, H. eyrei, and L. dorsalis (Mann & Bidwell, ported to the University of Tolima where they were raised to
Gosner stage 25 at a temperature of 23–25ºC in tanks contain-
1999). It is clear that components of the formulation other thaing city water that was dechlorinated by continuous aeration
the active ingredient are drivers of acute toxicity.
for at least 48 h prior to use. Embryos were not fed while they
Aerial applications of glyphosate to control illicit coca developed to stage 25. Only the tadpoles of R. typhonius and
(Erythroxylum coca) and poppy (Papaver sominferum) crops have S. ruber were caught directly in field in stage 25. Tadpoles
been made in Colombia since 1997. Since 2006, poppy has not
were not fed for 24 h before or during the test.
been grown to a significant extent in Colombia and is no longer
sprayed. It has been pointed out that the glyphosate–Cosmo-
Flux mixture as used in the spray program in Colombia could Testing Procedures

Downloadedpresent a risk to native frog species (Solomon et al., 2007). The Formulated glyphosate (Glyphos, a product sold in Colombia
96-h LC50 for the spray mixture as used in Colombia to larvae but similar to Roundup in terms of active ingredient and POEA
of X. laevis was 1300 and 1100 μg a.e./L for the poppy and surfactant) and Cosmo-Flux as used in the spray program were

coca mixtures (Solomon et al., 2007), respectively (Wildlife obtained and stored separately at room temperature in the dark.
International, 2006a, 2006b). This was not greatly different Glyphos contains 354 g glyphosate a.e./L (as the IPA) and
between 10 and 15% ethoxylated tallowamine (POEA) surfactant.
from the values reported in the literature for Roundup in the
same species, suggesting that the addition of the adjuvant Cosmo-Flux contains a mixture of linear and aryl polyethoxylates

TABLE 1

Species of Larval Frogs Used in the Acute Toxicity Studies and Their Location of Collection

Stage Altitude
Species collected Location (m a.s.l.)

Hypsiboas crepitans Gosner 10–11 Potrerillo (4º14’N; 74º58’W) 430

Rhinella granulosa Gosner 10–11 Payandé (4º19’N; 75º06’W) 630
Engystomops pustulosus Gosner 10–11 Ibagué (4°21’N; 75°06’W) 827

Rhinella marina Gosner 10–11 Payandé (4º19’N; 75º06’W) 630
Scinax ruber Gosner 25 Potrerillo (4º14’N; 74º58’W) 430
Dendrosophus microcephalus Gosner 10–11 Potrerillo (4º14’N; 74º58’W) 430

Rhinella typhonius Gosner 25 Ibagué (4º25’N; 75º12 W) 1200
Centrolene prosoblepon Gosner 10–11 Falan (5°07’N; 74°58’W) 1100

371 Annex 131-F

FORMULATED GLYPHOSATE TOXICITY TO FROGS 1. LABORATORY 963

(17% w/v) and isoparaffins (83% v/v) (Cosmoagro, 2004). The Analytical Methods
water used for testing was the same as that used for raising the The ELISA test kit manufactured by Abraxis, LLC

tadpoles (described earlier). Specific conductance, hardness, (Warminster, PA), was used to measure the concentration of
alkalinity, and pH of the water were measured. glyphosate in the test solutions. Calibration standards of gly-
Test chambers were 2-L glass jras containing 1 L of test solu-
phosate solutions, ranging in concentration from 0.15 to 5 μg/
tion. The chambers were indisrciminately positioned by treatment L, were prepared from the sta ndard supplied with the test kit
group in an air-conditioned area of the laboratory designed to and used to construct the standard curve. A standard curve

maintain the test temperature and day length throughout the test was prepared with each set of samples analyzed. The standard
period. A primary stock solution was prepared by dissolving the curve was constructed by plotting the %B/Bo (absorbance
Glyphos and Cosmo-Flux in dilution water to obtain a nominal value for each standard/absorbance value for the zero stan-

concentration of 100,000μg glyphosate a.e./L. All solutions were dard) against the corresponding glyphosate concentration.
prepared using a positive displcaement pipette. Cosmo-Flux was Concentrations of glyphosate were determined by interpola-
added in the proportion of 0.023μl per 1μl of Glyphos (2.3% v/v)
tion from the standard curve. Final concentration of glypho-
to obtain the proportion of spray mix as used in the field. Nominsate in the exposure solution was calculated by correcting for
test concentrations were selected based upon the results of explothe mean quality control (QC) percent recovery based on anal-
atory range-finding toxicity tests. Two replicates of each test solu-
yses of two replicates of one concentration (0.5 μg/L) of the
tion were prepared by serial dilution of the stock solution with standard. The Abraxis glyphosate assay has an estimated min-
dilution water to yield a range of nominal concentrations in 1 L imum detectable concentration based on a 90% B/Bo of 0.μ1g/L.

solution of 4200 to 1000μg glyphosate a.e./L. Each solution was The method limit of quantitation (LOQ) for these analysis was
mixed by inversion. Ten tadpoles were impartially placed in each defined as the lowest calibration standard, 0.15 μg/L. Two
of the two test chambers for a total of 20 tadpoles per concentramatrix blank samples were analyzed to determine possible

tion. The rate of biomass loading (defined as total wet weight ofinterferences. No interferences were detected above the LOQ
10 tadpoles/L test water) was below 0.6 g/L as recommended in during the samples analysis.
ASTM guidelines (ASTM, 1998), with the exception oSf. ruber,
It was not logistically possible to measure concentrations of
which were about 0.7 g/L. Test solutions were renewed daily by Cosmo-Flux; however, the proportions of Glyphos and Cosmo-
transferring the test organisms to freshly prepared solutions. MeFlux were kept constant and were the same as those used in the

measured test concentrationswere determined from samples of aerial spraying of coca. The results are thus representative of
test water collected from the pooled replicates for each treatmenrealistic field exposures.
and control group at the beginning of the test (0 h) and from the

test solutions for each treatment and control group at 24 h and at
test termination (96 h). All samples were collected at mid-depthStatistical Analyses
and were analyzed immediately without storage.
For consistency with other studies, the mortality data
Downloaded At:Fluorescent lights that emit wavelengths similar to natu- were analyzed using the U.S. EPA Probit Program Version
ral sunlight (Phill ips TLT 20W/54RS) were used for illumi- 1.5 (U.S. EPA, 1994). The LC1 was estimated as a regres-
nation of the test chambers. A photoperiod of 12:12-h light/
sion-derived approximation of the no-observed-effect con-
dark cycle was controlled with an automatic timer. The tem- centration, and the LC50 was calculated for comparison to
perature during the study was 23 ± 1.5°C. Temperature was other literature values. The LC1, which is derived from the

measured in each test chambe r at the beginning and end of response data, is preferred as an indicator of the low effect
the test and at approximately 24-h intervals during the test, concentration as it is independent of the experimental design
including before and after rene wals, using a liquid-in-glass (Crane & Newman, 2000). Where insufficient data were

thermometer. Temperature also was measured continuously available for the Probit program (no or one concentration
during the test in one negative control test chamber using a with a response between 0 and 100%), LC50 values were
maximum and minimum digital thermometer. Dissolved
estimated by interpolation from a graph of percent concen-
oxygen and pH were measured in each test chamber at the tration versus response.
beginning and end of the test and at approximately 24-h

intervals during the test, including before and after renew-
als. Dissolved oxygen was measured using a portable dis- RESULTS AND DISCUSSION
solved oxygen meter Hanna HI 8043, and measurements of
Measurement of Test Concentrations and Water Quality
pH were made using a Hanna HI 8314 membrane pH meter.
Observations of mortality and other signs of toxicity, such Samples collected at test initiation had measured concentra-
as unusual swimming activity, were observed approxi- tions of glyphosate that ranged from 75 to 125% of the nominal
concentrations. Samples collected prior to the renewal of the
mately 4, 24, 48, 72, and 96 h after test initiation. The
cumulative percent mortality observed in the treatment test solutions at 24 h contained measured concentrations that
groups was used to estimate LC50 values at 96 h. ranged from 74 to 112% of the nominal concentrations. Samples

372 Annex 131-F

964 M. H. BERNAL ET AL.

collected at test termination contained measured concentra- Toxicity values for the eight species of frogs (Table 3) are

tions that ranged from 71 to 130% of the nominal concentra- presented in terms of glyphosate concentration (a.e.) to allow
tions. When the measured concentrations of samples collected for comparison to data from the literature. The most sensitive
at 0, 24, and 96 h were averaged, the mean measured concen- species was D. microcephalus and the least sensitive was

trations ranged from 85 to 105% of nominal concentrations. E. pustulosus. The slopes of the concentration-response rela-
Because measured values were close to nominal, the nominal tionships ranged from 7.18 to 13.47. These are large slopes and
concentrations were used to determine the LC1 and LC50 val- are consistent with literature data where such slopes have been

ues in order to compare responses of larval anurans reported reported for formulated glyphosate (Perkins et al., 2000) and
under field conditions (Bernal et al., 2009) and those in othesurfactants (Dorn et al., 1993; Wong et al., 1997).

studies. Water temperatures were within the 23 ± 1.5°C range The LC50 values for these eight species of frogs were com-
established for the test. Mean dissolved oxygen concentrationsbined with those from the literature (Brain & Solomon, 2009) in
were about 6.85 mg/L. The 95%confidence interval of measure- a species sensitivity distribution (SSD). Plotting positions were

ments of oxygen concentration, hardness, alkalinity, specific calculated using standard procedures (Solomon & Takacs,
conductance, and pH in the dilution water at test initiation a2002). The SSD data (Figure 1) illustrate that the larvae of frogs
summarized in Table 2. from Colombia are not more nor are they less sensitive than

Signs of toxicity, such as slow swimming and remaining on other frogs tested in other locations are to glyphosate formula-
bottom with no movement, were generally noted at lower expo-tions such as Roundup and Vision. This is consistent with the

sure concentrations, and uncontrolled fast swimming and remain-observed toxicity of the mixture to X. laevis (Wildlife Interna-
ing in a vertical position were evident at concentrations close tional, 2006a, 2006b) where values were similar but slightly
to and exceeding the LC50 concentartion. In general, most of the greater (less toxic) than thosereported with the Vision formula-

toxic responses were expressed within 24 to 48 h of test initiation.tion of glyphosate (Edginton et al., 2004). The 5th centile of the
toxicity distribution was 692μg a.e./L, suggesting that overall,

95% of larval frogs would have LC50s greater than this value.

TABLE 2
Hardness, Alkalinity, Specific Conductance, and pH in the CONCLUSIONS
The acute toxicity values determined in Colombian species
Dilution Water at Test Initiation
of frogs suggest that sensitivity to Roundup-type formulations
95% of glyphosate in these species is similar to that observed in
Confidence
other species tested in other locations (Brain & Solomon,
Parameter Mean interval 2009). There is no underlying assumption that would suggest
that tropical species, such as those tested in Colombia, would
Oxygen concentration(mg/L) 6.85 (n = 96) 6.60–7.10
DownloadedHardness (mg/L as CaCO009) 112 (n = 6) 97.1–126.8 have different sensitivity to pesticides such as those containing
3 glyphosate and our observations are consistent with other observa-
Alkalinity (mg/L as CaCO 3) 89.3 (n = 6) 70.2–108.4
Specific conductance (mS/cm) 263.2 (n = 6) 197.4–328.9 tions on tropical and temperate species (Maltby et al., 2005).
pH 8.23 (n = 96) 8.20–8.25 These data add to those currentlyin the literature and suggest that
sensitivity of larval amphibians to Roundup-type formulations

TABLE 3
Toxicity Values for Colombian Frog Species Exposed to Formulated Glyphosate and Cosmo-Flux

95% Confidence 95% Confidence
Species Slope b Interceptb LC1 (μg a.e./L) interval (μg a.e./L) LC50 (μg a.e./L) interval (μg a.e./L)

D. microcephalus a – – – – 1200 –
a
R. typhonius – – – – 1500 –
S. ruber 13.47 2.09 1103 716–1294 1642 1470–1783
H. crepitans 7.23 2.72 984 645–1225 2064 1835–2285

R. granulosa 9.09 1.62 1300 737–1632 2348 2036–2588
C. prosoblepon 7.18 2.24 1145 2414

R. marina 9.75 0.74 1578 1122–1874 2733 2473–2982
E. pustulosus 8.78 1.09 1514 1040–1827 2787 2510–3057

aLC50 values estimated from a graph of concentration vs. percent response. Slope could not be calculated.
bSlope and intercept in log probit units.

373 Annex 131-F

FORMULATED GLYPHOSATE TOXICITY TO FROGS 1. LABORATORY 965

Cosmoagro. 2004. Cosmo-Flux 411F Label, Cosmoagro S.A. Accessed October
99
98 20, 2004, www.cosmoagro.com
Crane, M., and Newman, M. C. 2000. What level of effect is a no observed
95 H.eyrei effect. Environ. Toxicol. Chem. 19:516–519.

90 R. sylvatica Dorn, P. B., Salanitro, J. P., Evans, S. H., and Kravetz, L. 1993. Assessing the
C. insignifera aquatic hazard of some branched and linear surfactants by biodegradation
80 E. pustulosus and toxicity. Environ. Toxicol. Chem. 12:1751–1762.
70 L. dorsalis
L. moorei Edginton, A. N., Sheridan, P. M., Sptheenson, G. R., Thompson, D. G., and
R. granulosaon Boermans, H. J. 2004. Comparative effecotfspH and Vision herbicide on two life
50 H. crepitansana stages of four anuran amphibian specieEsn. viron. Toxicol. Chem2.3:815–822.
B. americanus
30 R. typhonius Giesy, J. P., Dobson, S., and Solomon, K. R. 2000. Eoctoxicological risk assess-
Percentrank R . clamitans ment for Roundup herbicide.Rev. Environ. Contam. Toxicol. 67:35–120.
20 D. microcephalus Colombian species Howe, C., Berrill, M., and Pauli, B. D. 2001. The acute and chronic toxicity of
R. pipiens Non-Colombian
10 S. nasicus glyphosate-based pesticides in northern leopard frogs. SETAC Annual
Meeting, Baltimore, MDAbstract PH058.
5 X. laevis Howe, C. M., Berrill, M., Pauli, B. D., Helbing, C. C., Werry, K., and

2 Veldhoen, N. 2004. Toxicity of glyphosate-based pesticides to four North
1 American frog species. Environ. Toxicol. Chem., 23:1928–1938.
10 3 10 4 Linder, G., Krest, S. K., and Sparling, D. W., eds. 2003. Amphibian decline:

LC50 (µgglyphosatea.e./L ) An integrated analysis of multiple stressor effects. Pensacola, FL: Society
of Environmental Toxicology and Chemistry.
Maltby, L., Blake, N. N., Brock, T. . ., and van den Brink, P. J. 2005. Insecti-
FIG. 1. Species sensitivity distribution of LC50 values for glyphosate plus
Cosmo-Flux as used in the spraying of coca in Colombia in larval amphibians cide species sensitivity dsitributions: The importance oftest species selection
from Colombia and LC50 values for frogs from other locations to Roundup and relevance to aquatic ecosystems.Environ. Toxicol. Chem.24:379–388.
Mann, R. M., and Bidwell, J. R. 1999. The toxicity of glyphosate and several
and Vision. Data for Roundup and Vision are from (Brain & Solomon, 2009).
glyphosate formulations to four species of Southwestern Australian frogs.
Arch. Environ. Contam. Toxicol. 36:193–199.
spans a relatively narrow range. Toxicity of the mixture as used Perkins, P. J., Boermans, H. J., and Stephenson, G. R. 2000. Toxicity of gly-

to spray coca was likely driven by the surfactant in the phosate and triclopyr using the frog embryo teratogenesis assay—Xenopus.
Environ. Toxicol. Chem. 19:940–945.
glyphosate formulation as the addition of Cosmo-Flux did not Relyea, R. A. 2005. The lethal impacts of Roundup and predatory stress on six
increase toxicity above those reported for Vision = Roundup
species of North American tadpoles. Arch. Environ. Contam. Toxicol.
(discussed earlier). The relatively low toxicity of Cosmo-Flux is 48:351–357.
Rondon-Barragan, I. S., Ramirez-Duarte, W. F., and Eslava-Mocha, P. R.
consistent with the LC50 values reported in the fish,Piaractus
brachypomus, which was 4,417 mg formulation/L in juveniles of 2007. Evaluación de los efectos tóxicos y concentración letal 50 del surfac-
tante Cosmoflux ® 411F sobre juveniles de cachama blanca (Piaractus
40 g mass (Rondon-Barragan et al., 2007). brachypomus). Rev. Col. Cienc. Pec. 20:431–446.

Extrapolation of these toxicity values directly to the envi- Solomon, K. R., Anadón, A., Carrasquilla, G., Cerdeira, A., Marshall, J., and
ronment is inappropriate except for simple hazard ranking. In Sanin, L.-Hw. 2007. Coca and poppy eradication in Colombia: Environ-
mental and human health assessment of aerially applied glyphosate. Rev.
realistic environmental conditions where sediments and
Downloaded At: 20:08 2 October 2009 Environ. Contam. Toxicol. 190:43–125.
organic detritus are present in pools inhabited by amphibian Solomon, K. R., and Takacs, P. 2002. Probabilistic risk assessment using spe-
larvae, concentrations of glyphosate and the POEA surfactant cies sensitivity distributions. In Species sensitivity distributions in ecotoxi-

will decrease rapidly due to binding to sediments (Solomon cology, eds. Posthuma, L., Suter, G. W., and Traas, T. pp. 285–313. Boca
Raton, FL: CRC Press.
et al., 2007) and this will likely reduce exposures and risks to Solomon, K. R., and Thompson, D. G. 2003. Ecological risk assessment for
amphibians. These hypotheses were tested and results are
aquatic organisms from over-water uses of glyphosate. J. Toxicol. Environ.
reported in a companion article (Bernal et al., 2009). Health B 6:211–246.
U.S. Environmental Protection Agency. 1994. Probit Program, Ecological Moni-

toring Research Division, Environmental Monitoring Systems Laboratory,
U. S. Environmental Protection Agency. http://www.epa.gov/eerd/stat2.htmS
REFERENCES Wildlife International. 2006a. Glyphosate–Cosmo-Flux–coca mix: A 96-hour

ASTM. 1998. Standard guide for conducting the frog embryo teratogenesis static-renewal acute toxicity test with the African clawed-frog tadpoleX(eno-
assay—Xenopus (vol. E 1439-91), pp. 825–836. West Conshohocken, PA: pus laevis). Final report. Technical report 628A-101, Wildlife International.
ASTM International. Easton, MD, USA.
®
Bernal, M. H., Solomon, K. R. and Carrasquila, G. 2009. Toxicity of formu- Wildlife International. 2006b. Glyphosate–Cosmo-Flux –poppy mix: A
lated glyphosate (Glyphos ) and Cosmo-Flux ® to larval and juvenile 96-hour static-renewal acute toxicity test with the African clawed-frog
Colombian frogs 2. Field and laboratory microcosm acute toxicity. J. Toxi- tadpole (Xenopus laevis ). Final report. Technical report 628A-102,

col. Environ. Health A 72:966–973. Wildlife International. Easton, MD, USA.
Brain, R. A., and Solomon, K. R. 2008. Comparative hazards of glyphosate, Wong, D. C. L., Dorn, P. B., and Chai, E. Y. 1997. Acute toxicity and
other pesticides, and other human activities to amphibians in the produc- structure–activity relationships of nine alcohol ethoxylate surfactants to

tion of coca. J. Toxicol. Environ. Health A 72:937–948. fathead minnow and Daphnia magna . Environ. Toxicol. Chem.
Cauble, K., and Wagner, R. S. 2005. Sublethal effects of the herbicide glyphosate16:1970–1976.
on amphibian metamorphosis and development.Bull. Environ. Contam. World Health Organization International Program on Chemical Safety. 1994.

Toxicol. 75:429–435. Glyphosate (vol. 159). Geneva: WHO IPCS.

374 Annex 131-G

Annex 131-G

M.H. BERNAL ET A., OXICITY OFFORMULATED GLYPHOSATE (GLYPHOS)AND
COSMO-FLUX TOLARVAL AND JUVENILECOLOMBIAN FROGS 2.IELD AND
L ABORATORY MICROCOSM A CUTETOXICITY”

(Journal of Toxicology and Environmental Health, Part A, 72:966-973, 2009)

375 Annex 131-G

Journal of Toxicology and Environmental Health, Part A, 72: 966–973, 2009
ISSN: 1528-7394 print / 1087-2620 online
DOI: 10.1080/15287390902929717

UTEH
Toxicity of Formulated Glyphosate (Glyphos)

and Cosmo-Flux to Larval and Juvenile Colombian

Frogs 2. Field and Laboratory Microcosm Acute Toxicity

M. H. Bernal , K. R. Solomon , and G. Carrasquillad 3
1
Laboratory of Herpetology, Eco-Physiology & Ethology, Universidad del Tolima, Barrio Santa Elena,
Ibagué, Tolima, Colombia, 2Centre for Toxicology and Department of Environmental Biology,
3
University of Guelph, Guelph, Ontario, Canada, and Facultad de Salud, Universidad del Valle, Cali,
Colombia

The bottom of the container was filled with moistened soil and
The spraying of coca (Erythroxylum coca) with glyphosate leaf litter to a depth of 1 cm a nd 0.5 cm, respectively. Mortality
(coca mixture, a combination of formulated glyphosate, Gly- in the controls was low, from 0 to 10%, and from 0 to 35% at the
normal application rate. LC50 values ranged between 4.5 kg
phos, and an adjuvant, Cosmo- Flux) in Colombia has raised a.e./ha and 22.8 kg a.e./ha, 1.5- to 6-fold greater than the normal
concerns about possible impacts on amphibians. Although acute
LC50 for 8 species of Colombian frogs ranged from 1.2 to 2.78 application rate. Data indicate that, under realistic worst-case
mg acid equivalents (a.e.)/L, these exposures were conducted in exposure conditions, the mixture of Glyphos and Cosmo-Flux as
the laboratory in the absence of sediments and organic matter used for control of coca in Colombia exerts a low toxicity to
such as would occur under realistic field conditions. In order to aquatic and terrestrial stages of anurans and that risks to these
organisms under field conditions are small.
assess the effects of overspray of frog habitat under field condi-
tions, Gosner stage 25 tadpoles of Rhinella granulosa, R. marina,
Hypsiboas crepitans, and Scinax ruber were placed in outdoor
microcosms made from polyethylene plastic fish ponds (2.07 m
in diameter, 37 cm high) in an experimental area in Tolima,
Colombia. The bottoms of the microcosms were covered with a
3-cm layer of local soil and they were filled to a depth of 15 cm Although the toxicity of glyphosate formulations to lar-
val amphibians has been relativ ely well characterized in the
Downloaded At: 20:08 2 October 2009 local spring water. After up to 100 laboratory (Bernal et al., 2009), few studies have actually
tadpoles of each frog species were placed in the microcosms,
they were sprayed with the coca mixture at concentrations examined the effects of gly phosate-based herbicide formu-
greater and less than the normal application rate (3.69 kg gly- lations under realistic conditions or on terrestrial stages of
phosate a.e./ha). Mortality at 96 h in the control microcosms was
between 0 and 16% and LC50 valu es were between 8.9 and 10.9 amphibians. Field studies (Wojtaszek et al., 2004) con-
kg glyphosate a.e./ha (equivalent to initial concentrations of ducted on larvae of Rana clamitans and Rana pipiens with
the Vision formulation of glyphosate (equivalent to
5963 to 7303 mg glyphosate a.e./L). Mortality >LC50 was only
observed in the tested species wh en the application rate was >2- Roundup) showed that acute toxicity was strongly influ-
fold the normal application rate. In other experiments, juvenile enced by natural fa ctors such as presence of sediment,
and adult terrestrial stages of frogs were exposed by direct aquatic macrophytes, and pH, generally resulting in lower
spraying to a range of concentrat ions of coca mixture. Juveniles
and adults were exposed in pl astic food containers (19 ´ 19 cm). toxicity relative to laboratory studies. Wojtaszek et al.
(2004) reported 96-h LC50 values ranging from 2700 to

11,470 μg acid equivalents (a.e.)/L under conditions more
©General Secretariat of the Or ganization of American States, relevant to the field. In addition, no significant differences
2009. This paper was prepared as part of a Study entitled in mean growth rates or maximum size at metamorphosis
“Production of Illicit Drugs, the Environment and Human Health,”
financed with contributions from the Governments of Colombia and were observed for these two larval species even when
the United States of America. The conclusions and opinions exposed to concentratio ns as great as 14,300 μg a.e./L.
expressed herein are those of the authors and not necessarily thoseThese observations were consistent with an operational

of the Organization of American States and its General Secretariat,study (Thompson et al., 2004) where biomonitoring with
which as of the date of this copyright, have not formulated any caged amphibian larvae showed no significant responses of
opinion with respect to them.
Address correspondence to Dr. M. H. Bernal, Laboratory of mortality (48 h) of either R. pipiens (p = .194) or R. clamitans
Herpetology, Eco-Physiloogy & Ethology, Universidad del Tolima, Barriovae (p = .129). Mortality was al so not significantly corre-
Santa Elena, Ibagué, Tolima, Colombia. E-mail: [email protected] lated with exposure concentrations for either amphibian

966

376 Annex 131-G

FORMULATED GLYPHOSATE TOXICITY TO FROGS 2. FIELD 967

species tested. Results sugges ted that exposures typically a.e./m2), resulting from application of 6.5 ml of a formulation

occurring in forest wetlands we re insufficient to induce sig- containing 1.9% glyphosate (IPA assumed) per tub (91 mg
nificant acute mortality in amphibian larvae. a.e./tub). Although the area of the tubs was not reported, to
In contrast, Relyea (2004) reported mortality and growth achieve the stated application rate with the volume of formula-
2
effects of Roundup Weed and Grass Killer on several amphibian tion used, a surface area somewhat greater than 75 m /tub
species following exposures to four commercial formulations of would have been required. It was recently stated (Relyea, per-
pesticides (diazinon, carbaryl, malathion, and glyphosate) either sonal communication, September 2008) that there was a typo-

alone or in combination. The glyphosate formulation used con- graphical error in the description of the experiment in the
tained the ethoxylated tallowamine (POEA) surfactant and was paper. This sentence has the typo: The “1.6 mg AI/m 2” should
2
tested at concentrations equivalent to either 740 or 148μ0g a.e./L,read “1.6 mL AI/m ” and the rates of application were as rec-
with Gosner stage 25 larvae exposed in 10-L plastic tubs housed ommended on the product label. However, active ingredient
outdoors for a period of 16 d. No significant effects on growth (AI) is normally expressed in terms of mass, not volume, and

were observed for any species at ≥740 μg a.e./L. Significant lev- details of the methods are still incomplete. In addition, frogs
els of mortality were observed for R. clamitans, R. catesbiana, were exposed on paper towels, a scenario not highly represen-
and B. americanus at 1480 μg a.e./L, whileHyla versicolor and tative of field conditions.

R. pipiens showed no significant mortality at this exposure. In a A review of environmental effects of aerial applications of
subsequent paper (Relyea, 2005a), tadpoles of several amphib- glyphosate to control illicit coca (Erythroxylum coca) pointed

ian species were exposed to the same glyphosate formulation out that the Glyphos–Cosmo-Flux mixture as used in the spray
already described (Relyea, 2004) in 1000-L cattle tanks serving program in Colombia could present a risk to native frog species
as field microcosms. As Thompson et al. (2006) noted, this study exposed in shallow waters typically associated with wetland

utilized a single test concentration of 3800μg a.e./L, equivalent frog habitat (15 cm deep) (Solomon et al., 2007). However,
to an application rate of 16 kga.e./ha over water of 15 cm depth, this did not consider absorption and/or degradation of the

both of which are in excess of typical environmental exposure glyphosate (AI) and the POEA surfactant in the presence of
concentrations or application rates. In a similar study, Relyea sediments, which was found to reduce exposures and, there-
(2005b) examined the effects of the same formulation of glypho- fore, risk (Tsui & Chu, 2004; Wang et al., 2005). The study

sate as reported in Relyea (2005c) at the same nominal concen- reported here was conducted to compare the toxicity of the
tration on tadpoles of three species when exposed in 1200-L mixture of Glyphos and Cosmo-Flux to larval amphibians as
cattle tanks with no soil, 19 L of sand, or 19 L of loam soil. The reported under laboratory conditions (Bernal et al., 2009) to

author reported no effect on the toxicity of glyphosate formula- responses under similar field conditions where sediments and
tion by soil interaction. At this high test concentration, over a 20-dpended particles are present in shallow water systems. In

exposure period, the glyphosateformulation againresulted in a addition, toxicity of the coca mixture was assessed in terrestrial
significant reduction in survival (<4%) in all 3 amphibian test stages of representative species under more realistic conditions
Downloaded species regardless of the soil treatment. Substantial mortality in the presence of soil and leaf litter.

occurred within the first 24 h of exposure. This result contrasts
sharply with other observations in the literature (Tsui & Chu,
2004; Wang et al., 2005) that demonstrated substantial reduction MATERIALS AND METHODS

in acute toxicity of Roundup and the surfactant ethoxylated tal- Test Organisms
lowamine (POEA) to test organisms in the presence of sediment.
Four of the species previously tested in the laboratory
The discrepancy between these observations may have been the (Bernal et al., 2009) were used for the field microcosm studies.
result of differences in the water–sediment ratios and the depth They were collected from Potrerillo, Tolima, Colombia
of the water above the sediment, which was greater in the study
(4°14’N; 74°58’W) at 430 m altitude.
by Relyea (2005c).
Few studies reported toxicity of glyphosate formulations to
Testing Procedures—Microcosms
terrestrial stages of amphibians. In a study in Australia, the
48-h LC50 values for Roundup herbicide tested against adult Six outdoor in-ground microcosms, constructed from food-
and newly metamorphosed C. insignifera ranged from 49,400 grade high-density polyethylene plastic cattle tanks/fish ponds

to 51,800 μg a.e./L and were greater than those reported for (2.07 m in diameter, 37 cm high), were placed in dug holes in
larvae (Gosner stage 25 for this species) (Mann & Bidwell, the experimental area in Tolima. Microcosms were placed in a
1999). In a laboratory study (Relyea 2005b) in which juvenile tree-shaded area where they were protected uniformly from

terrestrial stages of 3 different species (R. sylvatica, B. wood- full sunlight. Soil was backfilled around the pools (Figure 1)
housii fowleri, and H. versicolor) were exposed to direct appli- but the upper 22 cm of the wall was left uncovered to prevent
2
cations (1.2 mg a.e./m ) in 10-L plastic tubs, 79% mortality water running into the pools. The bottoms of the microcosms
was observed after only 24 h. The author stated that the formu- were covered with a 3-cm layer of local soil and then filled to a
lated glyphosate was applied at a rate of 1.6 mg AI/m 2 (1.2 mg depth of 15 cm (above the sediment) with local spring water

377 Annex 131-G

968 M. H. BERNAL ET AL.

from microcosms, and the tadpoles were transferred to a

smaller container, identified, and counted. Missing animals
207 cm were assumed to be lost in the recovery process or by decom-
position after dying. Mortality was based on the number of

Screen dead and missing animals compared to the total animals added
to the microcosm at the initiation of the exposure.

Overflow Water For the second experiment, the water and sediment were
Sediment 20 cm removed from the microcosms and replaced with fresh mate-
rial. The preceding procedure was repeated but with larvae of

15 cm H. crepitans (100 animals per each treatment) and S. ruber (65
animals per each treatment) with a rate of biomass loading
3 cm
Soil approximately of 0.0153 g/L. Because of particulates present
in the water, enzme-linked immunosorbent assay (ELISA)
FIG. 1. Diagram of aquatic microcosm design. analysis was not possible and nominal initial concentrations

were calculated from the application rates for comparison to
the laboratory observations (Bernal et al., 2009).
with hardness equivalent to 41 mg CaCO and an alkalinity
3/L
equivalent to 51 mg CaCO 3/L. A screened overflow opening at
15 cm above the sediment allowed overflow in the case of rain- Testing Procedures—Terrestrial Stages of Frogs
fall. After filling with water, microcosms were covered with
Juvenile terrestrial stages of frogs were obtained by raising
black plastic shade cloth (3 mm mesh) stapled to a wooden frogs collected in the field under laboratory conditions as pre-
frame to exclude predators such as dragonflies and birds. The viously described (Bernal et al., 2009). In addition, small

microcosms were allowed to stabilize for 4–5 d. Prior to add- adults of two species of frogs were collected from the field for
ing the tadpoles, a fine screen nylon cloth (0.5 mm mesh) was these experiments. Species used and sources are listed in Table 1.
placed in the microcosms and pressed into the sediment. This
Polyethylene plastic food containers, 19 × 19 cm (internal
was done to facilitate collection of larvae after exposure. surface area = 361 cm 2) and about 2.3 L capacity and with a
In the first experiment, 100 Gosner stage 25 larvae of tight-fitting lid in which a 7× 7 cm mesh-screened ventilation

Rhinella granulosa and R. marina each were placed in each opening was made, were used as terrestrial exposure micro-
microcosm with a rate of biomass loading approximately of cosms. Soil and leaf litter, obtained from the botanical garden of
0.0122 g/L, below the 0.6 g/L as recommended in ASTM
the University of Tolima, a nonagricultural area, was added to a
guidelines (ASTM, 1998). Application rates of Glyphos– depth of 1 cm and 0.5 cm, respectively. The soil and leaf litter
Cosmo-Flux mixture (coca mixture as is fully described in the were moistened with distilled water to about 90% of holding

Downloaded At: 20:08 2 October 2009 et al., 2009) were selected on the capacity, giving a relative humidity from 80 to 95% in the con-
basis of preliminary experiments. Immediately after adding the tainers. The chambers were indiscriminately positioned by treat-
frog larvae, the microcosms were sprayed with the coca mix-
ment group in an air-conditioned laboratory where they were
ture at a range of concentrations equivalent to 0 (control) up to maintained at a temperature of 25± 2°C and 12:12 h light/dark
29.52 kg glyphosate a.e./ha. The Glyphos and Cosmo-Flux for- cycle throughout the test period.

mulations were those used previously (Bernal et al., 2009), and Based on preliminary tests, frogs were exposed to a geomet-
amounts were measured with a positive displacement pipette ric series of Glyphos–Cosmo-Flux concentrations higher and
and then mixed with 500 ml water. The entire amount was
lower than the field application rate (3.69 kg a.e. glyphosate/
sprayed over the pools, at a height of approximately 50 cm, ha). A primary stock solution was freshly prepared by mixing
with a small hand-held pump-up garden sprayer. Spray drift 30 ml Glyphos and 0.69 ml Cosmo-Flux (delivered by positive

was minimized by (1) setting the sprayer to apply large drop- displacement pipette) with 500 ml water by inversion to give a
lets, (2) spraying in the absence of wind, and (3) spraying in concentration that would provide an application rate of 29.52
circle from the center of the pool to the rim, but never over the kg a.e. glyphosate/ha when 5 ml was sprayed over the area of
2
edge. In addition, 20 cm of the wall of the pool was above the the container (361 cm ). The stock solution was serially diluted
water level, which would have reduced loss of pesticide though to provide application rates equivalent to 14.76, 7.38, 3.69, and
spray drift. Based on spray drifttables for hand-held (pedestrian)
1.85 kg a.e. glyphosate/ha and one water control.
sprayers using small droplet sizes for insecticides and fungi- Two duplicate test chambers were used for each treatment
cides (European Commission, 2000), loss to drift is estimated rate and the control group. Groups of 10 frogs (R. typhonius,

to be less than 5%. Water in the microcosms was sampled at 2, R. granulosa), 9 frogs (R. marina and S. ruber), and 5 frogs
24, 48, and 96 h after application and the concentration of dis- (C. prosoblepon, E. pustulosus, P. taeniatus, and D. truncatus)
solved oxygen, pH, and temperature were measured. After 96 h
were impartially assigned to each test chamber for a total of
of exposure, the 0.5-mm nylon mesh was carefully removed 20, 18, and 10 frogs per concentration, respectively. Because

378 Annex 131-G

FORMULATED GLYPHOSATE TOXICITY TO FROGS 2. FIELD 969

TABLE 1

Juvenile and Adult Frogs Used in the Terrestrial Microcosm Exposures

Mean mass in mg
Species Location of collection (95% CI)

Rhinella typhonius (Linnaeus, 1758) Ibagué (4°25’N; 75°12 W) 0.047 (0.042–0.051)

R. granulosa (Spix, 1824) Potrerillo (4°14’N; 74°58’W) 0.063 (0.060–0.066)
R. marina (Linnaeus, 1758) Potrerillo (4°14’N; 74°58’W) 0.114 (0.110–0.117)
Engystomops pustulosus (Cope, 1864) Ibagué (4°21’N; 75°06’W) 0.175 (0.165–0.185)

Scinax ruber (Laurenti, 1768) Potrerillo (4°14’N; 74°58’W) 0.182 (0.172–0.191)
Centrolene prosoblepon (Boettger, 1892) Falan (5°07’N; 74°58’W) 0.251 (0.241–0.261)
Pristimantis taeniatus (Boulenger, 1912) Ibagué (4°25’N; 75°12 W) 0.853 (0.821–0.884)
a
Dendrobates truncatus (Cope, 1861) Potrerillo (4°14’N; 74°58’W) 1.583 (1.514–1.651)

aAdult frogs.

of lack of availability, not all species could be tested in larvaglyphosate a.e./ha. The reason for this is uncertain but may be

as well as terrestrial stages. For quality control purposes, frogdue to increased oxygen demand as a result of the death of lar-
were weighed as a group before being placed in the test cham- vae at these treatment rates or because the large amounts of
bers. At the end of the study, they were weighed individually to glyphosate stimulated microbiological activity and produced

calculate the average body mass. Body mass was approxi- an oxygen demand. This was only observed at rates of applica-
mately similar between treatments. There are no guidelines for tion that were greater than those used in the field (3.69 kg gly-
biomass loadings when testing terrestrial stages of frogs, but phosate a.e./ha). The pH in the microcosms was generally

mortality was <10% in the controls in these studies, suggesting more consistent over the length of the exposures (Figure 2).
that the loadings were not excessive. Test solutions were Temperatures in the microcosms were consistent between

freshly prepared and sprayed on the frogs using a modified microcosms (Figure 2). As expected, temperature showed no
domestic hand sprayer fitted to a test-tube reservoir to allow response to treatment concentration.
complete spraying of a total volume of 5 ml. This volume of

spray solution was sprayed on the frogs, starting with the water
control and then from the lowest to highest concentration. The Responses in the Aquatic Microcosms
sprayer was rinsed with clean water between tests. Observa- Mortality or loss of frog larvae in the control microcosm in all
experiments was between 0 and 16%. InR. granulosa,there was
tions of mortality and other signs of toxicity were made at 4,
Downloaded 24, 48, 72, and 96 h after test initiation. All frogs were fed ada greater mortality (or loss), probably due to its smaller body size,
libitum with small insects once per day during the experiments which may have reduced efficiency of recovery of larvae. LC50

values in the treated microcosms were approximately similar
between species (Table 2).Rhinella marinawas the most sensi-
Statistical Analyses tive species, but large mortalities were not observed until the
As in the laboratory studies (Bernal et al., 2009), LC1 and
application rate exceeded fourfold the normal rate of 3.69 kg gly-
LC50 values were calculated using the U.S. EPA Probit phosate a.e./ha, when more than 80% mortality was observed. An
Program Version 1.5 (U.S. EPA, 1994). Where insufficient LC50 could not be calculated forS. ruberbut was estimated from
data were available for the Probit program (no or only one con-
graphical interpolation to be about 10.3 kg glyphosate a.e./ha (ini-
centration with a response between 0 and 100%), the LC50 tial concentration of 6900μg glyphosate a.e./L), almost threefold
was estimated by interpolation from a graph of percent concen-
greater than the normal application rate. Larvae ofR. granulosa
tration versus response. and H. crepitans were less sensitive (Table 2,) and significantly
higher mortalities were not observed until the application rate

RESULTS AND DISCUSSION was about 14.8 kg glyphosate a.e./ha (initial concentration of
9916 μg glyphosate a.e./L).
Water Quality In all cases, sensitivity of frog larvae in the field microcosms

The concentration of dissolved oxygen in the microcosms was less than in laboratory toxicity tests (Bernal et al., 2009). This
(Figure 2) was different in the two experiments. Concentration is consistent with observations in the literature where both
at 0 h was generally lower in experiment 1 than in experiment 2. Roundup (Tsui & Chu, 2004) and surfactant POEA (Wang et al.,

However, in both experiments, oxygen concentration declined 2005) were less toxic in the presence of sediment, likely as a result
over the exposure period in the microcosms treated at the of rapid binding to sediment and/or breakdown by microbes, thus

greater rates of glyphosate, particularly at 14.7 and 29.5 kg reducing exposures (and apparenttoxicity) in the water column.

379 Annex 131-G

970 M. H. BERNAL ET AL.

Experiment 1 Experiment 2

and and

10 10
9 9

8 8

7 7
6 6

5 5

4 4
3 3
oxygen (mg/L)
2 2

1 1
Conce0tration of dissolved 0

0 20 40 60 80 100 0 20 40 60 80 100
8 8

pH 7 7

6 6

0 20 40 60 80 100 0 20 40 60 80 100
28 28

26 26

Downloaded At: 20:08 2 October 2009 24

kg a.e./ha
22 22 0.0
1.85
3.69
20 20
7.38
Temperature (C) 14.76
18 18 29.52

16 16

0 20 40 60 80 100 0 20 40 60 80 100
Time after application (h) Time after application (h)

FIG. 2. Concentration of dissolved oxygen, pH, and temperature in the aquatic microcosms after treatment with various rates of the coca mixture of Glyphos
and Cosmo-Flux. Application rates are of glyphosate (a.e.) in the mixture.

Responses in the Terrestrial Microcosms the terrestrial stages of the eight species of frogs (Table 3) are
Observations of frogs revealed signs of toxicity such as lackresented in terms of nominal application rate (kg a.e. glypho-

of normal movement or slow movement. In some frogs, the sate/ ha) to allow for comparison to field application rates.
hind limbs were extended and thus were unable to walk. In S. Mortality in the controls was low, from 0 to 10%.

ruber and P. taeniatus, a milky secretion from the skin was The range of LC50 values for adults (5-fold) was greater
observed. In general, most of the adverse responses were than that observed (Bernal et al., 2009) in larvae (2.3-fold), and

expressed within 24 to 48 h of test initiation. Mortality data formulative distribution of LC50 values (Figure 3) suggested

380 Annex 131-G

FORMULATED GLYPHOSATE TOXICITY TO FROGS 2. FIELD 971

TABLE 2

Mortality for Colombian Tadpole Species Exposed to Mixtures of Glyphos and Cosmo-Flux in the Aquatic Field Microcosms

LC values in kg a.e./ha
Percent mortality at glyphosate application rates (kg a.e./ha) (μg a.e./L)

Species 0 1.85 3.69 7.38 14.76 29.52 LC1 LC50

R. marina 10 13 25 41 82 100 2.4 8.9 (5963)

S. ruber 4.6 4.6 15.4 12.3 100 100 — 10.3 (6900)
R. granulosa 16 8 21 19 94 100 6.4 10.7 (7169)

H. crepitans 0 0 3 3 86 100 4.8 10.9 (7303)
a
Nominal initial concentration estimated from application rate and depth (15 cm) of water with complete mixing but no adsorption to sedi-
ment or particulates.

TABLE 3

Mortality of Juvenile and Adult Stages of Colombian Frog Species Exposed to Mixtures
of Glyphos and Cosmo-Flux in Terrestrial Microcosms

LC values

Percent mortality at glyphosate application rates (kg a.e./ha) (kg a.e./ha)

Species 0 1.85 3.69 7.38 14.76 29.52 LC1 LC50

C. prosoblepon 0 0 30 90 100 100 1.97 4.5
P. taeniatus 0 0 20 70 — — 1.93 5.6

R. granulosa 10 35 35 55 90 100 — 6.5
S. ruber 0 17 33 50 56 95 0.32 7.3
R. typhonius 10 10 15 35 50 80 1.56 14.8

E. pustulosus 0 0 0 0 30 80 7.02 19.6
R. marina 0 0 0 6 22 67 5.08 22.8
a
D. truncatus 0 0 0 0 — — >7.38 >7.38
a
Downloaded At: 20:08 2 October 2009

2 groupings of species in terms of sensitivity. Among the that of R. granulosa. In this case, it is also possible that the
juveniles, the most sensitive was C. prosoblepon and the leincreased sensitivity is related to its thin skin. The lower

sensitive wasR. marina. Of the adults,P. taeniatuswas sensisensitivity of R. marina may be attributable to the greater
and D. truncatus was very tolerant to the mix of Glyphos– body mass of juveniles (Table 2) and to its thicker skin, and
Cosmo-Flux (Table 3). for D. truncatus to the greater body mass. There was also no

There are a number of possible reasons for these differ-obvious relationship between sensitivity and preferred habitat,
ences in sensitivity. It is possible that the higher sensitwhich is related to ability to tolerate loss of water (Figure 1), but

C. prosoblepon and P. taeniatus is due to their thinner skiD. truncatus may be generally less sensitive to pesticides as it
which is smooth, translucent, and probably more permeable. is routinely collected from agri cultural areas where pesticides
In particular, this may also explain the greater sensitivitare used.

C. prosoblepon, the internal organs of which are visible Overall, in 5 of 8 experimental species, mortality at the rate
through the transparent underside, a feature that gives theof 3.69 kg a.e/ha used in eradication spraying was between
common name of glass frogs to this species and others of th15% and 35% and LC50 values were between 1.2- and 6-fold

same family. However, D. truncatus also has thin skin and greater than the application rate for eradication spraying.
was insensitive. The higher sensitivity for R. granulosa maExtrapolation of the species sensitivity distribution (Figure 3)

be attributable to small body mass (Table 1) and greater sugave a 5th centile intercept of 2.2 kg glyphosate a.e./ha (for the
face area to volume ratio . Scinax ruber had a larger body coca mixture), suggesting that >95% of the LC50 values for
mass (Table 1) and smaller su rface area to body mass ratioterrestrial frogs would not be exceeded at application rates less

but its sensitivity to glyphosate-Cosmo-Flux was similar tothan this value.

381 Annex 131-G

972 M. H. BERNAL ET AL.

99 The differences in sensitivity between species of terrestrial
98 Open area habitat stages of frogs were possibly due to a combination of differ-
95 Forest onlyorests
Tolerant of habitat change ences in body mass (surface area to volume ratio) and skin per-
90 Thick skin meability. However, these factors and others, such as habitat
80 Thin skin preference, did not fully explain the apparent bimodal distribu-

70 tion of sensitivity (Figure 3). A greater understanding of the
toxicokinetics of uptake of the formulation may explain rea-
50
sons for differences in susceptibility between species and
Percent rank would be an interesting subject for future work that may
20
10 explain these differences.
For all species tested, the responses to the mixture of
5
2 Glyphos–Cosmo-Flux were not large at the rate of 3.67 kg
1 a.e./ha as used in Colombia for eradication spraying. The
terrestrial microcosm experiments showed that juveniles of
1 10 100
LC50 for terrestrial stages (kg glyphosate a.e./ha) S. ruber and R. granulosa were relatively less tolerant to
glyphosate–Cosmo-Flux mixture than their tadpoles. How-
FIG. 3. Cumulative frequency distribution of acute LC50 values for
terrestrial stages of frogs sprayed with the mixture of Glyphos and Cosmo-he results of R. marina (Tables 2 and 3) showed the
opposite. In Australian frogs, Mann and Bidwell (1999)
Flux used in eradication spraying. The relative mass of the frogs (Table 1) is
indicated by the size of symbols. The value for D. truncatus, which was veryt adults and new metamorphs of C. insignifera
insensitive (Table 3), is plotted for information but was not used in theess sensitive to Roundup than tadpoles, but this study
regression. was carried out under laboratory conditions; thus, these

results are not directly comparable with ours. Data showed
that mortalities ≥50% for tadpoles, juveniles, and adults

were only observed at rates of application greater than the
CONCLUSIONS 3.69 kg glyphosate a.e./ha used in eradication spraying in
The results of the aquatic and terrestrial microcosm stud-
Colombia.
ies show that responses of frogs under realistic field exposure Risk in the field will likely be lower because of less expo-
conditions are less than would be predicted from laboratory sure. Sites >5 m outside the spray swath and with no vegeta-

toxicity studies and less than reported by some authors for tive cover would receive deposition rates (Hewitt et al., 2009)
other species. The reason for this is likely that glyphosate and less than the LC1 observed in microcosms. For terrestrial

the surfactant POEA adsorb rapidly to sediments and organic species, assuming no vegetativ e cover, none of the observed
matter that is present in na tural systems or are rapidly LC50 values would be exceeded directly under the spray

Downloaded At: 20:08 2 October 2009s on the influence of the depth of swath. A 10-m margin downwind of the spray swath would
the surficial water on the fate of glyphosate and POEA have be protective of all measured LC1 values. Under actual con-
not been conducted, this process is probably more rapid in ditions of use, interception by trees and other vegetation, such

shallow waters where diffusion and mixing would be as observed in forestry spraying (Thompson et al., 2004) and
expected to be more rapid. This may explain differences also occurs under Colombian conditions (Hewitt et al., 2009),

between the results observed in this study and observations would further reduce exposures and subsequent risks. Multi-
of mortality in deeper wate r reported elsewhere (Relyea, ple applications of eradication sprays are unlikely in the time-

2005a, 2005c). Mixing in deeper water would be expected to frame for complete dissipation of glyphosate and surfactants
be less rapid and adsorption to sediments and organic matter from water and/or soil. Sprays are very accurately applied
would be delayed for long enough that exposures in the and only one pass is made over the field (Solomon et al.,

surfice water might exceed thresholds of toxicity. Direct 2007), minimizing the likelihood of a double application.
comparison of the two studies is not possible as the depths of While there may be small risks to frogs in unvegetated areas

water were different. The importance of the interactions in coca fields that are directly oversprayed, the risks from
between water depth, type of sediment, content of organic exposures to other more toxic pesticides used by coca grow-

matter, and microbiological activ ity needs to be addressed in ers are much greater (Brain & Solomon, 2009). When consid-
future environmental fate studies with pesticides and surfac- ering the small area of Colombia that is actually sprayed each
tants that adsorb strongly to sediments. Interactions between
year (< 0.1% Solomon et al., 2007) and that frogs are not
water-quality parameters such as concentrations of divalent exclusively associated with coca growing (Lynch & Arroyo,
cations, oxygen, and hydrogen ion (pH) may be an important 2009), it can be concluded that the Glyphos–Cosmo-Flux

area of general research as suggested in laboratory investiga- mixture as used in Colombia fo r eradication of coca presents
tions on pH interactions with Vision in frog larvae (Edginton a slight but not ecologically significant risk to larval and ter-

et al., 2004). restrial stages of anurans.

382 Annex 131-G

FORMULATED GLYPHOSATE TOXICITY TO FROGS 2. FIELD 973

REFERENCES Relyea, R. A. 2005b. The lethal impact of Roundup on aquatic and terrestrial
amphibians. Ecol. Appl. 15:1118–1124.
ASTM. 1998. Standard guide for conducting the frog embryo teratogenesis
assay—Xenopus, (vol. E 1439-91), pp. 825–836. West Conshohocken, PA : Relyea, R. A. 2005c. The lethal impacts of Roundup and predatory stress on
ASTM International. six species of North American tadpoles. Arch. Environ. Contam. Toxicol.
Bernal, M. H., Solomon, K. R. and Ca rrasquila, G. 2009. Toxicity of for- 48:351–357.
Solomon, K. R., Anadón, A., Carrasquilla, G., Cerdeira, A., Marshall, J., and
mulated glyphosate (Glyphos) and Cosmo-Flux to larval Colombian
frogs 1. Laboratory acute toxicity. J. Toxicol. Environ. Health A 72: Sanin, L.-H. 2007. Coca and poppy eradication in Colombia: Environmental
961–965. and human health assessment of aerially applied glyphosate. Rev. Environ.
Brain, R. A., and Solomon, K. R. 2009. Comparative hazards of glyphosate, Contam. Toxicol. 190:43–125.
Thompson, D. G., Solomon, K. R., Wojtaszek, B. F., Edginton, A. N., and
other pesticides, and other human activities to amphibians in the produc- Stephenson, G. R. 2006. The impact of insecticides and herbicides on the
tion of coca. J. Toxicol. Environ. Health A 72:937–948.
Edginton, A. N., Sheridan, P. M., Stephenson, G. R., Thompson, D. G., and biodiversity and productivity of aquatic communities. Ecol. Appl.
Boermans, H. J. 2004. Comparative effects of pH and Vision herbicide on 16:2022–2027.
two life stages of four anuran amphibian species. Environ. Toxicol. Chem.Thompson, D. G., Wojtaszek, B. F., Staznik, B., Chartrand, D. T., and
Stephenson, G. R. 2004. Chemical and biomonitoring to assess potential
23:815–822.
European Commission. 2000. Guidance document on aquatic ecotoxicology. acute effects of Vision herbicide on native amphibian larvae in forest
Technical report 8075/VI/97 rev 7, Directorate General for Agriculture. wetlands. Environ. Toxicol. Chem. 23:843–849.
Brussels, Belgium. Tsui, M. T. K., and Chu, L. M. 2004. Comparative toxicity of glyphosate-based
herbicides: Aqueous and sediment porewater exposures. Arch. Environ.
Hewitt, A. J., Solomon, K. R., and Marshall, E. J. P. 2009. Spray droplet
size, drift potential, and risks to non-target organisms from aerially- Contam. Toxicol.46:316–323.
applied glyphosate for coca control in Columbia. J. Toxicol. Environ. U.S. Environmental Protection Agency. 1994. Probit Program, Ecological
Health A 72:921–929. Monitoring Research Division, Environmental Monitoring Systems Labo-
ratory, U. S. Environmental Protection Agency. http://www.epa.gov/eerd/
Lynch, J. D., and Arroyo, S. 2009. Risks to Colombian amphibian fauna from
cultivation of coca (Erythroxylum coca): A geographical analysis.J. Toxicol. stat2.htmS
Environ. Health A72:974–985. Wang, N., Besser, J. M., Buckler, D. R., Honegger, J. L., Ingersoll, C. G.,
Mann, R. M., and Bidwell, J. R. 1999. The toxicity of glyphosate and several Johnson, B. T., Kurtzweil, M. L., MacGregor, J., and McKee, M. J. 2005.
Influence of sediment on the fate and toxicity of a polyethoxylated tallowa-
glyphosate formulations to four species of Southwestern Australian frogs. mine surfactant system (MON 0818) in aquatic microcosms. Chemosphere
Arch. Environ. Contam. Toxicol. 36:193–199.
Relyea, R. A. 2004. Growth and survival of five amphibian species exposed to 59:545–551.
combinations of pesticides. Environ. Toxicol. Chem. 23:1737–1742. Wojtaszek, B. F., Staznik, B., Chartrand, D. T., Stephenson, G. R., and
Thompson, D. G. 2004. Effects of Vision herbicide on mortality, avoidance
Relyea, R. A. 2005a. The impact of insecticides and herbicides on the response, and growth of amphibian larvae in two forest wetlands. Environ.
biodiversity and productivity of aquatic communities. Ecol. Appl.
15:618–627. Toxicol. Chem. 23:832–842.

Downloaded At: 20:08 2 October 2009

383384 Annex 131-H

Annex 131-H

J. D.LYNCH ET AL ., “ISKS TOC OLOMBIAN AMPHIBIAN FAUNA FROM CULTIVATION
OF COCA (E RYTHROXYLUM COCA ): AGEOGRAPHICAL ANALYSIS ”

(Journal of Toxicology and Environmental Health, Part A, 72:974-985, 2009)

385 Annex 131-H

Journal of Toxicology and Environmental Health, Part A, 72: 974–985, 2009
ISSN: 1528-7394 print / 1087-2620 online
DOI: 10.1080/15287390902929733

UTEH

Risks to Colombian Amphibian Fauna from Cultivation

of Coca (Erythroxylum coca): A Geographical Analysis

Risks to Colombian Amphibian Fauna from Coca Cultivation
J. D. Lynch and S. B. Arroyo
Laboratorio de Anfibios, Instituto de Ciencias Naturales, Universidad Nacional de Colombi,

Bogotá, Colombia

species of frogs at risk. These include Ameerega bilingua, Den-
The Colombian amphibian fauna is among the richest known dropsophus bifurcus, Pristimantis colomai, P. degener, P. diade-
matus, P. quaquaversus, P. variabilis, and Trachycephalus
in the world, with about 20 species of salamanders (order Cau- jordani. Other species may also be at risk but exact numbers are
data), 35 of the limbless caecilians (order Gymnophiona), and unknown since few investigations were undertaken in these
more than 700 species of frogs and toads (order Anura) recorded
from localities within the country. The potential effects of expo- areas during the past 30 yr. Th e main ranges for these species
sure to glyphosate on amphibians arising from production of were assumed to be in Ecuador.
illegal crops (coca) were examined. The analysis was based on
(1) behavior and ecology of species and (2) proximities of actual
museum records to localities in which illegal crops are being

grown and the subset of those that have been sprayed with gly- Currently the herbicide glyphosate (Glyphos) is used to
phosate. Based on data on the lo cation of amphibians collected
in Colombia, records were obtained for 193 species (28% of the control coca (Erythroxylum coca) production through an aerial
national diversity) of frogs and toads found in localities within spray eradication program (Solomon et al., 2007). Several
10 km of areas where coca is gr own. Further analyses with ARC concerns have been raised regarding the spray control program,
MAP software allowed for measurement of the direct distance
separating collection locations for frogs, known coca fields, and ranging from peripheral crop damage to adverse environmental
effects. As larval stages of amphibians are sensitive to formu-
areas where aerial spraying was being conducted. Records in or lated glyphosate (Bernal et al., 2009a), these organisms may be
near coca fields included data for 11 of 13 families of frogs and
toads known to be present in Colombia. Only Ceratophryidae particularly at risk. In addition, the pesticides used by growers in
and Pipidae were not reported fr om these locations and appear the production of coca also present a risk to larval amphibians,
Downloaded not to be at risk. For eight species ( Dendrobates truncatus , Craushould they be found in or close to the edge of coca fields
gastor raniformis, Pristimantis gaigeae, Smilisca phaeota, Ela-
chistocleis ovale, Hypsiboas crepitans, Trachycephalus venulosus , (Brain & Solomon, 2009).
All amphibians have permeable skins and hence are espe-
and Pseudis paradoxa) selected to represent several habitat pref-
erences and life-cycle strategies, large areas of their distribu- cially susceptible to application of extraneous chemicals, such
tions lie outside coca production regions and their populations as pesticides, to their environment (Stebbins & Cohen, 1995).
as a whole are at low risk. For a limited number of species that Pathways of exposure in terrestrial amphibians include direct
barely enter Colombian territory, the consequences of coca
production may be more serious and may have placed several contact of spray droplets on the exposed skin or through contact
with sprayed surfaces, such as the animals moving over leaves
that have been sprayed. In addition, contamination of water
©General Secretariat of the Organization of American States,
through direct overspray or spray drift may result in exposure
2009. This paper was prepared as part of a Study entitled “Productito aquatic larval stages.
of Illicit Drugs, the Environment and Human Health,” financed with
contributions from the Governments of Colombia and the United The behavior of most amphibians reduces the risk of direct
States of America. The conclusions and opinions expressed herein exposure to spray. With few exceptions, Colombian amphibians
are those of the authors and not necessarily those of the Organizatare nocturnal organisms, emerging when the air temperatures
of American States and its General Secretariat, which as of the date
of this copyright, have not formulated any opinion with respect have fallen and the relative humidity of the air exceeds the dew
point. Aside from Dendrobatids, a frog family composed of
to them. nearly exclusively diurnal species, the nocturnal activity cycles
The authors gratefully acknowledge the contributions of Policia
Antinarcótica de Colombia, which provided mapping data on the of amphibians render them leastlikely to experience direct appli-
location of coca production and aerial eradication spraying. cation of the sprays. Exceptions are some species that “sleep” in
Address correspondence to J. D. Lynch, Laboratorio de Anfibios, exposed positions—the Rana platanera, known to Colombians
Instituto de Ciencias Naturales, Universidad Nacional de Colombi,
Bogotá, Colombia. E-mail: [email protected] visiting the lowlands adjacent to the eastern cordillera, is a

974

386 Annex 131-H

RISKS TO COLOMBIAN AMPHIBIAN FAUNA FROM COCA CULTIVATION 975

species (actually, a pair of species) with many casual observa- installations. In military installations more removed from natural
tions of frogs “sleeping” on leaves in full sunlight—as well as forests (for example, those immediately adjacent to Leticia),

juveniles of many species and tadpoles of some species. Since only 12 species of frogs and toads were found. In the parks of the
eradication spraying is only conducted in daylight and early in city (and in undeveloped lots), only 6 species of frogs were
the day (Solomon et al., 2007), the most likely pathway of detected.

exposure is via contact with sprayed surfaces. Glyphosate and These data provide a necessary background against which
the polyethoxylated talowamine (POEA) surfactant with which one can assess the effect of coca production (as an illegal crop)

it is formulated are strongly adsorbed to soils and organic mat- on the Colombian amphibian fauna. Cultivations of coca dis-
ter (Tsui & Chu, 2004; Wang et al., 2005), reducing likelihood play alterations that are even more severe than chagras of the
of exposure from these sources; however, contact with wet indigenous communities in the area of Leticia (Lynch, 2005).

leaves may allow exposure. Other more toxic pesticides, such Using this as a basis for comparison, cultivation of coca is
as are used by coca growers (Brain & Solomon, 2009), may likely to reduce the resident frog fauna by approximately 90%
not be strongly absorbed and may be more bioavailable by (see earlier discussion), prior to consideration of any effects of

this route. glyphosate spraying upon the amphibian fauna.
Habitat alterations sometimes favor some amphibians.
* (Lynch, 1998) in the Cordillera
COCA PRODUCTION Based on a study ofPristimantis
Occidental (an area that was originally covered with evergreen
In Colombia, coca plants grow in areas that were once wet montane forests) and on the fauna in the immediate environs of
lowland tropical forest. The plant is a shrub and grows to
between 1.5 and 2 m. In most cases the coca plants are grown Leticia (Lynch, 2005), an area that originally was wet lowland
forests, there are some species in each fauna that appear to
in clear-cut areas, but this illicit crop may be found in the have benefitted from human intervention (or habitat modifica-
middle of other crops such as banana plantations with the
tion). These species become common locally in areas that have
objective of avoiding detection. The most obvious character- experienced complete or partial removal of the natural vegeta-
istic of coca cultivation is the degradation of ecosystem. The tion, but that are rarely encountered during intensive searches
farmers deforest from the canopy to the leaf litter in order to
within natural (forested) habitats. These species (including
prepare the land for a coca plantation (see Anonymous, 2006, Pristimantis brevifrons, P. erythropleura, P. palmeri, and
p. 49), leaving the habitat completely destroyed by physical
activities. Craugastor raniformis) perhaps become more abundant
because they have been relieved of the effect of competition
Coca production results in the alteration of the natural habitatwith other species (most species in each community react
(Brain & Solomon, 2009), which may exert significant adverse
effects on frogs. Illustrations in the book Colombia/Monitoreo adversely, in terms of numbers or visibility, to habitat alter-
ation). However, additional clues are available from other spe-
de Cultivos de Coca (Anonymous, 2006, pp. 21, 23, 25, 27, 29, cies, such as Trachycephalus venulosus, a canopy-dwelling
31, 33–35) graphically show the degree of environmental alter-
Downloaded At: 20:08 2 October 2009 frog in lowland forests in Colombia. Although the adults live
ations and/or destruction that accompanies coca plantations. within the confines of the forest, the species appears to need
Alteration in the habitat may exert significant adverse effects “open” habitats in which to breed. Presumably, in times before
on frogs, and these are illustrated in a study conducted in the
human intervention (or habitat modification) became common,
immediate vicinity of Leticia (the southeastern most city of these frogs used clearings generated by the occasional tree-
Colombia (Lynch, 2005). A number of species of frogs and
toads were detected from undisturbed habitats within a small falls in natural forest ecosystems; now, of course, they have
many more opportunities granted by the creation of pastures
area (approximately 5 km× 5 km) following an intensive inven- and croplands.
tory conducted during 3 mo early in 2003. The inventory
revealed the presence of 96 species and additional efforts during The objective of this study was to evaluate the likely risks to
the Colombian amphibian fauna from cultivation of coca.
the next 3 yr raised the number of captured species to 98. Extensive occurrence records of the Colombian amphibian
Beyond localities representing “natural” habitats, occasional
fauna are held at the Instituto de Ciencias Naturales, Bogotá.
samples were gathered along a transect defined by degree of
intervention (or habitat modification): (1) Least intervened were
the chagras (small holdings) of the indigenous communities of *The classification of amphibians is subject to much debate and
instability at this time and the authors acknowledge that nomenclature
Huitotos and Ticunas, with more intervention (or habitat modifi- will change in the near future. The nomenclature used in this paper is
cation) evident at (2) abandoned farms of colonos (areas current at this time except that Hedges et al. (2008a) is not followed in
recovering to the native habitat), (3) active farms folonos, (4) their partition of the family Brachycephalidae into four families
because that study was a phenetic one masquerading as a phylogenetic
military installations, and (5) the green spaces within the urban study (furthermore, all new names proposed therein are not available
center of Leticia. Thirty species were found in human com-
because the paper did not meet Article 13.1.1; ICZN, 1999), and
munities as well as in areas undergoing recovery. About 20 spe- Wiens et al. (2005, 2007) rather than Frost et al. (2006) as concerns
cies were found in active farms and in some areas of military the Hemiphractidae were followed.

387 Annex 131-H

976 J. D. LYNCH AND S. B. ARROYO

In order to evaluate the likely risks of coca production to this but, aside from the Pacific lowlands of Colombia, the species
faunal group, the spatial records of occurrence for all species are rarely encountered (e.g., are not captured and/or seen

were compared with known areas of coca production and loca- every night). At least 90% of the Brachycephalids of Colom-
tions of eradication spraying. In addition, the known ecological bia are restricted in distribution to the Andean cordilleras (and
and behavioral traits of species were reviewed, in relation to Sierra Nevada de Santa Marta) and have small distributional

habitat loss and potential exposure to chemicals used for coca areas, arranged in altitudinal bands on the flanks of the moun-
production. tains (Lynch, 1998, 1999b). Brachycephalids are also atypical

of the North Temperate vision of proper frogs and toads in
that these frogs are direct deve lopers (no tadpole stages). The
remaining species of Colombian frogs and toads are divided
MATERIALS AND METHODS
among 12 families, most of which include substantial frac-
Identification of Groups of Amphibians at Risk tions of the local diversity with “typical” life histories (e.g.,
Based on habitat use and behavior, some amphibians are aquatic tadpoles) and may be exposed to pesticides in both the

unlikely to be affected by coca production or eradication sprayingterrestrial and aquatic stages. The following sections review
Caecilians are unlikelyto be affected by application of glyphosatethe distribution of Colombian frogs and toads in relation to
because most are subterranean species, surfacing especially on
general distribution.
rainy nights in search of earthworms. Three species of caecilians About 60% of the Colombian frog fauna is confined to the
are aquatic and are distributed in larger rivers as well as Andes and Sierra Nevada de Santa Marta (Lynch et al., 1997)
streams and ciénagas, areas not used for coca production and
and nearly all of the species endemic to the country occur in
hence unlikely to be exposed to pesticides. the Andean zone or in the Chocó biome. Those high levels of
The salamander fauna of Colombia is small and these spe- endemicity result mostly from the fact that Andean species

cies are distributed in the lowlands up to the páramos, about tend to have small distributional areas, whereas lowland
3200 m above sea level (a.s.l.). Aside from a few species species tend to be more widely distributed. Coca cultivation (as
restricted (or nearly restricted) to the treeless páramo habitats,an illegal crop) is not randomly distributed in the country

all of these animals require dense forest environments, rendering (Figures 1–4 in this article; and see Anonymous, 2006, figures
them relatively protected from pesticides used in coca production on pp. 9 and 12 and see p. 80 for the geographical pattern of
or eradication. Similarly, all salamanders are species with
distribution for aerial spraying) and is more prevalent in the
direct development, and embryos (or “eggs”) are found in leaf lowlands. Those cases in which a high proportion of known
litter or in epiphytes (again, with a minimal environmental localities for some frogs species (in Colombia) are at, near, or

exposure to extraneous chemicals). surrounded by coca production are restricted to species having
The largest proportion of amphibians in Colombia is made only a small part of their distributions within Colombia. For
up of frogs and toads, distributed across the full spectrum of these species, the high density of coca production in the low-

Downloaded At: 20:08 2 October 2009nd from sea level to at least 4350 m lands of western Caquetá and Putumayo plus eastern Cauca
a.s.l. Of the families of frogs and toads currently recognized poses a high risk of exposure to pesticides used for production
(Wiens et al., 2005; Frost et al., 2006; Grant et al., 2006) for and eradication of coca. The same is true for the lowlands of

Colombia, the largest is the Brachycephalidae (38%), followed western Nariño.
by the Hylidae (19%), and the Bufonidae (9%), Centrolenidae For the other 12 families of frogs and toads, Lynch (1999a)
(10%), and Dendrobatidae (10%). The other 8 families are
noted that most groups of frogs and toads were sorted into
small (2 to 30 species) with a sharply reduced proportionality— Andean (including the lowlands of the Pacific) or lowland
each less than 1% to as much as3% of the Colombian frog fauna groups and that this sorting extended virtually to the level of

(Aromobatidae, Ceratophryidae, Hemiphractidae, Leiuperidae, genus. Three other families (Aromobatidae, Bufonidae, and
Leptodactylidae, Microhylidae, Pipidae, and Ranidae). Dendrobatidae) have significant representation in both elevated
More than one-third of Colombian frog species belong to zones and in lowlands, whereas the remaining families may be

the family Brachycephalidae. Brachycephalid frogs are sensi- considered as either Andean or of lowlands. The dominant mode
tive to available moisture and are absent (or nearly so) from the of reproduction for these 12 families (excepting Hemiphractidae)
dry areas of Colombia (eastern Llanos, Caribbean lowlands,
includes an aquatic tadpole stage.
inter-Andean valleys, and the rain-shadows of the dry enclaves The predominant species family (Hylidae) for the lowlands
distributed along the Andean cordilleras). Aside from species has a minor Andean component; this family also has a signifi-

restricted in distribution to above tree line (thepáramo species),cant representation (19% of species recorded for Colombia) in
all native species of Brachycephalids are denizens of forests. the country. All species of Hylidae also are “typical” frogs for
With these exceptions, Brachycephalids in Colombia are usually having aquatic tadpoles. Within this family, many species

found in forested habitats, substantial modification of which exhibit a low fidelity to forests, especially during their reproduc-
results in the species becoming rare (or disappearing). In tive seasons. This lack of fidelity to conditions that might reduce
Colombia, there are species of Brachycephalids in the lowlands, exposure to pesticides used in the production and eradication

388 Annex 131-H

RISKS TO COLOMBIAN AMPHIBIAN FAUNA FROM COCA CULTIVATION 977

FIG. 1. Distribution of Dendrobates truncatus (a Colombian endemic) and Craugastor raniformis based on records documented in the Collection of
Amphibians of the Instituto de Ciencias Naturales.

Downloaded At: 20:08 2 October 2009
of coca raises their likelihood of experiencing negative The Glass frogs (family Centrolenidae) are typical of
effects from coca production. In addition, there are many spe- streams in the Andes and the Sierra Nevada de Santa Marta. In

cies “typical” of open (naturally or produced by intervention addition, these are organisms that require some measure of the
(or habitat modification)) situations, that is, not covered by original vegetation (but are tolerant of gallery forests within
forests. pasturelands). The majority of species deposit their embryos on

The true toads (family Bufonidae) are better represented in vegetation (above a stream) where the initial week or so of
the Andes (and Sierra Nevada de Santa Marta) than in low- development occurs. Subsequently, the tadpoles fall into a

lands. The Andean species either have tadpoles adapted to cur- stream and resist the current by hiding in vegetation within the
rents (Atelopus) or exhibit direct development (Osornophryne, stream or by burrowing into the substrate. These tadpoles are
Rhamphophryne). Atelopus deposit their egg masses in the active at night. Centrolenid frogs are also are present in most

water, usually attached to the undersurfaces of rocks. The hab- wet lowland forests of Colombia but are rarely collected there,
itat (above the altitude where coca is grown) and reproductive presumably because the adults occupy the canopy. Once again,
biology reduce exposure to pesticides used in production and the requirement of forest cover reduces the likelihood of expo-

eradication of coca. In the lowlands (and the lower parts of the sures to pesticides used in production and eradication of coca.
mountain slopes), one finds the more “typical” toads (Chau- However, the fact that they survive (apparently well) in mere
nus, Dendrophryniscus, Incilius, Rhaebo, and Rhinella) with remnants (gallery forests) might result in exposure of egg

tadpoles either in ponds or in slow-moving streams. Except for masses.
the Chaunus, these are organisms of heavy native forests and, The Dendrobatidae family is widely distributed in Colombia

by virtue of living beneath an undisturbed canopy, are unlikely (sea level to just above 4000 m.a.s.l.) and nearly always associated
to be exposed to pesticides used in production and eradication with areas with abundant moisture. Virtually all of the species
of coca. of this family are exclusively diurnal (in contrast to nearly all

389 Annex 131-H

978 J. D. LYNCH AND S. B. ARROYO

FIG. 2. Distribution of Elachistocleis ovale, Pristimantis gaigeae, Pseudis paradoxa, and Smilisca phaeota based on records documented in the Collection of
Amphibians of the Instituto de Ciencias Naturales. Additional records of P. gaigeae are available in Lynch (1999b) and for S. phaeota in (Duellman & Trueb,
1986).

Downloaded At: 20:08 2 October 2009

other frog species in Colombia), which indicates that they may The smaller families of frogs and toads (Aromobatidae,

be exposed to pesticides used in production and eradication of Ceratophryidae, Hemiphractidae,Leiuperidae, Leptodactylidae,
coca that are applied during daylight hours. The exceptional Microhylidae, Pipidae, and Ranidae) include one (Hemiphractidae)

species (exceptional for living in relatively dry environments) confined to forested regions with little or no intervention (or
Dendrobates truncatusis widely distributed across the Caribbean habitat modification). Hemiphractid frogs are mostly (or
lowlands and up the Madgalena valley in areas that were once exclusively) species without larval stage. The family Aromo-

dry tropical forest. The reproductive biology of frogs of this batidae was separated recently (Grant et al., 2006) from the
family is unusual in that the terrestrial eggs are large and few inDendrobatidae and shares the sa me ecological and reproduc-
number (deposited in leaf litter and guarded by a parent). Large tive characteristics.

eggs are usually associated with direct development, but in the The Ceratophryidae, Leiuperidae, Leptodactylidae, Micro-
case of Dendrobatidae they develop into tadpoles. The attend- hylidae, Pipidae, and Ranidae are families that in Colombia are
ing parent permits the hatched tadpoles to climb onto its back exclusive to the lowlands (or nearly so), and some species are

and the tadpoles are transported to bodies of water and present in open habitats, indicating that some of these frogs
released. The bodies of water vary from arboreal and terrestrial cope well with a degree of human intervention (or habitat

bromeliads to cavities in fallen logs or small ponds and slug- modification) of the habitat. Pipids are aquatic and rarely venture
gish streams. Given that species of the family have terrestrial onto the land. Eggs of Ceratophryids, Microhylids, and Ranids
eggs, they are obliged to live in wetter areas and their distribu- are deposited in water (or on water in the case of Microhylids),

tions are markedly associated with evergreen forests, offering whereas the eggs of Leiuperids and Leptodactylids are depos-
some protection from pesticides used in production and eradi- ited in foam nests floating on the surface of ponds or concealed
cation of coca. within burrows constructed by the frogs themselves. All of

390 Annex 131-H

RISKS TO COLOMBIAN AMPHIBIAN FAUNA FROM COCA CULTIVATION 979

FIG. 3. Distribution of Hypsiboas crepitans and Trachycephalus venulosus based on records documented in the Collection of Amphibians of the Instituto de
Ciencias Naturales. Additional records for H. crepitans are available in (Lynch & Suarez-Mayorga, 2001).

Downloaded At: 20:08 2 October 2009
these have aquatic tadpoles. The foam nests presumably repre- these are aquatic frogs, they appear to exhibit relatively little
sent an adaptation to seasonally dry conditions and provide a fidelity to forested habitats where they are less likely to be

moist environment in which the embryo develops. In some exposed to pesticides used in production and eradication of coca.
species the foam nests are found within the cover of forests,

but in most of these species (Leiuperids and Leptodactylids),
foam nests may be observed in areas of intense intervention (or Specific Locations of Frogs in Relation to the Production
habitat modification) (e.g., agricultural lands and pasture- of Coca

lands). There are no data in the literature on the penetration of The largest and most complete repository of amphibians
pesticides into the foam nest. One of the two species of Cer- from Colombia is the Instituto de Ciencias Naturales (ICN),
atophryids is confined to natural forests in Amazonia; the other with more than 55,000 records including specimens of at least

occurs in nonforested habitats along the Caribbean coast. 98% of the species known for Colombia. This study permitted
Some species of Leiuperids and Leptodactylids are denizens us to systematize this large collection (i.e., generate an elec-
of deep, undisturbed forests, but in each group one finds spe- tronic database) and, for most locality records, to determine the

cies commonly seen in areas subject to human intervention (or geographic coordinates of the collection locality. This activity
habitat modification). At least three species of Microhylids are allowed us to characterize the expected (and documented)

animals associated with nonforest environments, and although geographic distribution for 750 species with a high level of pre-
the adults are burrowers, the fact that all produce a floating surcision. Further, working with the Antinarcotics Police, it was
face film of eggs means that this sensitive stage of the life possible to localize all known cultivations of coca (also geo-

cycle may be at risk from pesticides used in production and referenced), so as to measure with precision the geographic
eradication of coca. The four aquatic Pipids include two with separations of populations of amphibians from areas where
direct development and two with tadpoles. Perhaps because pesticides are used in production and eradication of coca.

391 Annex 131-H

980 J. D. LYNCH AND S. B. ARROYO

To illustrate the issue of co-location of frogs and coca pro-

duction and aerial eradication spraying, eight species of frogs
well known in Colombia were selected to represent several
habitat preferences and lifecycle strategies. Dendrobates trun-

catus (Figure 1) was selected because it is diurnal and not
closely tied to deep forests. Craugastor raniformis (Figure 1),
Pristimantis gaigeae (Figure 2), and Smilisca phaeota, Figure

2) were selected because each tolerates habitat modification
well. Elachistocleis ovale (Figure 2), Hypsiboas crepitans

(Figure 3), and Trachycephalus venulosus (Figure 3) were
selected because (1) these species either prefer open habitats or
(2) also reproduce there, and (3) each deposits the eggs as films

on water surfaces, and Pseudis paradoxa (Figure 2) because
this species inhabits open areas and is aquatic. The effect of
local extinctions upon the integrity of the entire species is

beyond the scope of this study; however, one might view with
alarm how much (approximately 25%, Figure 1) of the known

distribution of an endemic Dendrobatid (D. truncatus) is over-
lapped by coca production and/or aerial eradication spraying,
given that this is a diurnal frog and may be more exposed to

pesticides used in production of coca, though it is relatively
insensitive to the glyphosate formulation used in the eradica-

tion of coca (Bernal et al., 2009a). Among the other species,
there is some co-occurrence with coca production, but most
species are also recorded from coca-free areas. Location of

other frogs in relation to coca production and eradication
spraying are summarized in the following subsections.

FIG. 4. Locations in Colombia where amphibians are at risk from thFamily Aromobatidae
production of coca. Heavy lines (marked with arrows) enclose two areasRheobates palmatus has 6 records within 10 km of coca
harboring all known Colombian records for a variety of Brachycephalid,
Dendrobatid, and Hylid frogs adversely affected by habitat destruction and0.2 to 9 km from zones of aerial aerial eradication spray-
Downloaded At: 20:08 2 October 2009ticides used in the production and eradicaing). Four other species (Allobates “brunneus,” probably = A.
coca (however, intensive inventory work has not been done in these areas).
trilineatus], A. femoralis, A. picachos, and A. talamancae) have
one or two records within 9 km of coca production 1.7 to 6 km
away. Aromobatids may be relatively tolerant of human inter-
Frogs and toads were recorded from localities that are within
10 km of those areas where cocais grown. For many of these vention (or habitat modification) but their sensitivity to pesti-
records, the original collectionswere made before significant cides is unknown. The eggs are terrestrial and are hidden in
leaf litter. Some species require heavy forest to sustain popula-
plantations of coca (as an illegal crop) appeared. For reasons of
personal security, none of these sites was revisited by herpetolo- tions, but others seem able to cope with modest intervention
(or habitat modification).
gists from the ICN. Analyses were conducted with Arc Map soft-
ware version 9.2 (ESRI, 2006) so that direct distance separating
collection locations for frogs, known coca fields, and areas where
Family Brachycephalidae
aerial spraying was being conducted could be measured.
Forty-seven species (all formerly of the genus Eleuthero-
dactylus; see earlier footnote) have between 1 (23 species) and
RESULTS 8 records within 10 km of coca production [or in three cases

Based on data from the ICN amphibian database, records (E. boulengeri, E. scopaeus, and E. w-nigrum) close to poppy
exist for 193 species (28% of the national diversity) of anurans production]. Data are given in Table 1. The proximity records

that are within 10 km of those areas where coca is cultivated. for E. mantipus and E. permixtus involve altitudinal differ-
Records in or near crops of illegal plants include those for 11 ences as well (coca fields lie at elevations ≤1000 m a.s.l.
of the 13 families of frogs and toads known to be present in whereas these frog species occur above 1500 m a.s.l.); hence, it

Colombia. Only Ceratophryidae and Pipidae were not reported is inappropriate to consider these two records as pertinent to
from these locations and would not be at risk. the investigation.

392 Annex 131-H

RISKS TO COLOMBIAN AMPHIBIAN FAUNA FROM COCA CULTIVATION 981

TABLE 1 TABLE 1

Records of the Family Brachycephalidae Found Within 10 km (Continued)
of Coca and Their Proximity to Production Areas or
Eradication Spraying Number Distances to
of coca production
Number Distances to Species records or spraying

of coca production
Species records or spraying Strabomantis sulcatus 3 1, 2 (spray) and 3 km
Pristimantis taeniatus 8 1 to 7.5 km
Pristimantis achatinus 4 1 to 8 km Pristimantis tamsitti 2 3.5 and 6.5 km

Strabomantis anatipes 2 at 9 km and in a coca Diasporus tinker 2 1 and 2 km
field Pristimantis thectopternus 1 1 km
Pristimantis anomalus 4 1 to 9 km Pristimantis viejas 3 1–10 km

Diasporus anthrax 1 7.5 km Pristimantis vilarsi 3 1 (spray), 1.7 and
Hypsodactylus babax 2 2.6 to 7 km 3.0 km

Pristimantis boulengeri 3 0.25 to 7.6 km Pristimantis w-nigrum 6 0 to 9 km (spray at
Pristimantis brevifrons 1 7.6 km 9 km)
Strabomantis bufoniformis 5 1 to 9 km Strabomantis zygodactylus 1 9 km

Pristimantis caprifer 1 9 km Note. Formerly in the genus Eleutherodactylus but now partitioned
Strabomantis cerastes 1 9 km
Pristimantis chalceus 2 2.6 and 3 km into four families and seven genera by Hedges et al. (2008a, 2008b).

Pristimantis colomai 1 4.3 km
Pristimantis conspicillatus 2 1 km and eradication

spray at 2 km Family Bufonidae
Pristimantis cruentus 1 1 km Fourteen species (Andinophryne atelopoides, A. olallai,
Pristimantis degener 1 2.2 km Atelopus famelicus, A. sanjosei, Chaunus granulosus, C. mari-

Pristimantis diogenes 1 9 km (spray) nus, Dendrophryniscus minutus, Rhaebo blombergi, R. gutta-
Hypsodactylus dolops 2 3.5 and 6.5 km tus, R. haematiticus, R. hypomelas, and Rhinella dapsilis, and
Pristimantis epacrus 2 3.5 and 6.5 km two species termed to date Rhinella sp.) have records (1 to 14)

Pristimantis erythropleura 2 2.6 and 9 km (spray) within 10 km of coca crops. Coca production and aerial eradi-
Pristimantis fallax 1 4.5 km cation sprays 2 km away are known for Rhinella dapsilis, 9 km
Craugastor fitzingeri 2 1.6 and 7.5 km
for Rhaebo blombergi and R. haematiticus, 0.6 and 9 km for
Pristimantis gaigeae 7 0.5 to 7.5 km Atelopus sanjosei and A. famelicus, 2 and 6 km for Den-
Downloaded Diasporus gulariser 2009 5 0.5 to 9 km (9 km drophryniscus minutus, and 2 to 11 km for Chaunus granulo-

spray) sus and C. marinus. Most of the species discussed earlier
Pristimantis hectus 1 7.6 km require dense forest and do not react well to intervention or
Pristimantis jaimei 2 2.6 and 9 km (spray) habitat modification. However, the two Chaunus appear to pre-

Pristimantis labiosus 1 4.3 km fer open (= nonforested) habitats. In the case of C. marinus, the
Pristimantis lanthanites 2 1 km, 2 km (spray) “preference” is so marked that a collector is well advised to
Pristimantis latidiscus 3 1.6 to 9 km
search for specimens in villages or on farms, where they are
Craugastor longirostris 6 0.5 to 8.7 km very common. Given that coca production involves a marked
Hypsodactylus mantipus 1 4.5 km alteration of the environment, these toads are likely to be

Pristimantis ockendeni 3 0.6, 1 and 2 km (spray) among the few native Colombian frogs to be found (and to
Strabomantis opimus 1 9 km breed) within such enclaves and are likely to be exposed to
Pristimantis palmeri 1 9 km (spray) pesticides used for coca production and eradication spraying.

Pristimantis parvillus 3 2.2 to 4.3 km
Pristimantis penelopus 1 2.2 km
Pristimantis permixtus 1 4.5 km Family Centrolenidae
Twenty-one species of this family have 1–4 records within
Pristimantis pugnax 2 3.5 and 6.5 km
Craugastor raniformis 8 0.2 to 7.5 km (spray at 10 km of coca cultivation and 3 of these records are within 0.1
6 km) km of areas where eradication spraying occurred. The species
involved areCentrolene grandisonae (record within a cultivation
Pristimantis ridens 2 1 and 9 km
Pristimantis scopaeus 1 0.25 km plot), C. hybrida, C. ilex, C. litorale, C. medemi, C. peristictum, C.
prosoblepon(four records), Cochranella chami, C. griffithsi, C.
(Continued) orejuela, C. oreonympha, C. punctulata, C. ramirezi, C.

393 Annex 131-H

982 J. D. LYNCH AND S. B. ARROYO

rosada, C. savagei, C. susatamai, C. xanthocherida, Hyalino- Six species of Gastrotheca(G. angustifrons, G. argenteovi-
batrachium aureoguttatum, H. colymbiphyllum, H. fleischmanni, rens, G. bufona, G. dendronastes, G. espeletiaa,nd G. guentheri)

and H. valerioi. Centrolenid frogs are probably protected from each have 1 known locality within 10 km of a coca field with
adverse effects of glyphosate by the fact that they are coca production and aerial eradication sprays between 3 and
extremely rare (or absent) when no trace of the forest cover 9 km away. Gastrotheca either are common animals or are

remains. However, apparently healthy populations with obvi- extraordinarily rare in collections. The genus Flectonotus
ous evidence of reproduction—egg masses applied to the sur- barely enters Colombia along the mountain border with

faces of leaves—have been encountered in the narrow gallery Venezuela, where it is locally abundant but rare in collections.
forests that border streams passing through pasturelands on the The Colombian species either occur in treeless areas (páramos)
Andean slopes. above the altitudes at which coca are grown or occupy dense

forest areas where the forest offers protection from pesticides
used in coca production and eradication spraying. The physical
Family Dendrobatidae alteration of the habitat by cocaleros likely results in local
Nineteen species of these small diurnal frogs have 1–6
extinction of populations of these frogs near coca crops.
records within 10 km of coca production: Ameerega hahneli A single species (Hemiphractus fasciatus) has two records
(cultivation within 1 km, aerial eradication spraying within 2 km),
A. ingeri (cultivation within = 0.6 km, eradication spraying at (1 to 9 km distant) near coca production. Hemiphractus appear
to be among the most sensitive of frog species to habitat alter-
6 km), A. trivittata (2 records at 1–3 km from coca crops and ation because all accumulated records of the five species
coca production and aerial eradication spraying at 6 km),
Colostethus fraterdanieli (2 records, cultivations at 1.6 and known for Colombia come from pristine forest habitats—and
subsequent visits to those sites (by experienced collectors)
5.6 km), C. inguinalis (2 records at 0.2 and 2 km, eradication after habitat alterations failed to detect the animals.
spraying at 6 km), C. pratti (cultivation at 1 km), C. ruthveni

(cultivation at 2 km), D. truncatus (6 records at 0.2 to 4 km
from cultivation, eradication spraying at 6 km), Epipedobates Family Hylidae
boulengeri (cultivation at 3.5 km), Hyloxalus fascianigrus
At least 49 species of this predominantly lowland family
(cultivation and eradication spraying at 9 km), H. fuliginosus have 1 to 8 records within 10 km of coca production, as listed
(2 records at 3.5 and 6.5 km), H. lehmanni (2 records at 2.2 and
9 km, eradication spraying at 9 km), H. saltuarius (cultivation in Table 2. Aerial eradication spraying occurred between 0.1
and 11 km from the locality records, potentially influencing
at 6.5 km), Hyloxalus sp. (cultivation at 6.5 km), Oophaga his- Dendropsophus ebraccatus, D. leucophyllatus, D. mathiassoni,
trionica (5 records at 0.5 to 9 km from crops, eradication
spraying at 0.1 to 3 km), Phyllobates bicolor (2 records at D. microcephalus, D minutus, D. parviceps, D. triangulum,
Hyloscirtus alytolylax, H. palm eri, H. simmonsi, Hypsiboas
3.5 km from cultivation with eradication spraying at 4.5 km), boans, H. cinerascens, H. crepitans, H. geographicus,
Ranitomeya opisthomelas(1 km from cultivation),R. virolinensis
Downloaded At: 20:08 2 October 2009 H. lanciformis, H. pugnax, H. punctatus, Osteocephalus taurinus,
(2.5 km from cultivation), andSilverstoneia nubicola(at 1 km). P. paradoxa, Scarthyla vigilans,Scinax blairi, S. cruentommus,
Dendrobatids may be relatively tolerant of human intervention S. garbei, S. ruber, Smilisca phaeota, S. sila, Sphaenorhynchus
(or habitat modification) and D. truncatus adults were found to
carneus, and T. venulosus. Although the common name given
be the most tolerant of 8 species to glyphosate (Bernal et al., to the family (treefrogs) suggests that these animals absolutely
2009b). However, if habitat de struction is extensive, the
peculiarities of the reproducti ve mode (terrestrial eggs in require forests, the reality is somewhat different. Many species
are obligate denizens of intact forests and a small portion of
leaf litter—that are moisture-sensitive) may result in local these appear to not come to the ground surface even for repro-
extinctions. Some species require heavy forest for their per-
sistence but others seem able to cope with modest interven- duction (Lynch, 2005), while others require that some part of
the forest persist for the continued existence of the local
tion (or habitat modification). The fact that these are diurnal population (e.g., Dendropsophus ebraccatus, D. triangulum,
frogs increases the risk of direct exposure to eradication
Hypsiboas boans, Osteocephalus taurinus, T. venulosus). Yet
spraying. other species appear to do even better (in terms of population

size) in open habitats than in dense forests (e.g.,Dendropsophus
Family Hemiphractidae minutus, D. triangulum, Hypsiboas lanciformis, H. punctatus,
Two species (Cryptobatrachus boulengeriand C. fuhrmanni) Scinax ruber). Lastly, there are lowland species that appear to

have 1 or 3 records within 10 km of coca crops. Frogs of this not use the forests at all (e.g., Dendropsophus mathiassoni,
genus are strict denizens of forested streams. As such, they D. microcephalus, H. crepitans, H. pugnax, Phyllomedusa
hypocondrialis, P. paradoxa, Scarthyla vigilans, Scinax blairi,
probably behave in much the same way as do Centrolenids (a
relatively high tolerance of intervention (or habitat modifica- S. rostratus, and S. wandae). In a study of the frogs found in
tion)) as long as the intervention (or habitat modification) does the vicinity of Villavicencio (Lynch, 2006), these species were
characterized best able to tolerate human intervention (or habitat
not remove all the trees.

394 Annex 131-H

RISKS TO COLOMBIAN AMPHIBIAN FAUNA FROM COCA CULTIVATION 983

TABLE 2 TABLE 2
Records of the Family Hylidae Found Within 10 km (Continued)

of Coca and Their Proximity to Production Areas
or Eradication Spraying Number Distances to coca
of production or
Number Distances to coca Species records spraying

of production or
Species records spraying Scinax ruber 3 2.7 and 3 km,
spraying at 11 km
Agalychnis spurrelli 1 9 km Smilisca phaeota 7 0.2 to 9 km

Cruziohyla calcarifer 1 9 km Smilisca sila 2 0.2 and 1 km
Dendropsophus columbianus 1 2.6 km Sphaenorhynchus carneus 2 1 km and 2.5 km
Dendropsophus ebraccatus 1 0.2 km (spray)

Dendropsophus leucophyllatus 1 1 km Sphaenorhynchus lacteus 1 5 km
Dendropsophus marmoratus 1 3 km Trachycephalus venulosus 3 1, 2, and 2.5 km

Dendropsophus mathiassoni 5 1 to 10 km (spray)
Dendropsophus microcephalus 5 1 to 6.4 km
Dendropsophus minutus 1 1 km
modification). However, their tolerance also renders them most
Dendropsophus parviceps 1 1 km likely to be adversely affected by coca production and aerial
Dendropsophus triangulum 1 1 km
Hylomantis buckleyi 1 6.5 km eradication spraying. Among these tolerant species one finds
species that deposit their eggs as films on the surface of water
Hyloscirtus alytolylax 1 9 km (e.g., H. crepitans, H. pugnax, T. venulosus) and others depos-
Hyloscirtus lindae 2 3.5 to 6.5 km
Hyloscirtus palmeri 9 0.5 to 9 km iting their eggs on the surfaces of leaves (e.g., Dendropsophus
mathiassoni, D. microcephalus, Phyllomedusa hypocondria-
Hyloscirtus platydactylus 1 1 km
Hyloscirtus simmonsi 1 9 km lis)—habits that may expose the spawn to adverse effects of
coca production and aerial eradication spraying.
Hyloscirtus torrenticolus 2 3.5 to 6.5 km
Hypsiboas boans 4 0.2 to 7.5 k
Hypsiboas calcaratus 1 5 km Family Leiuperidae

Hypsiboas cinerascens 3 0.6 to 2.7 km Five species of this family have records within 10 km of
Hypsiboas crepitans 4 1 to 7.5 km coca production: Engystomops petersi (at 1 km of production,
Hypsiboas geographicus 1 1 km
within 2 km of aerial eradication spraying), E. pustulosus
Hypsiboas lanciformis 4 1 to 10 km (7 records at 0.2–5 km from coca production and aerial eradi-
DownloadedHypsiboas picturatus 2009 1 9 km cation spraying at 6 km), Pleurodema brachyops (at 2 km of

Hypsiboas pugnax 3 within 1 km production), Pseudopaludicola pusilla (3 records 1–2 km from
Hypsiboas punctatus 8 1 to 10 km production with coca production and aerial eradication spray-
Hypsiboas rosenbergi 1 1 km ing at 6 km), and Physalaemus fischeri (at 5 km of production).

Osteocephalus carri 1 6.5 km Some Leiuperids are restricted to undisturbed forests (e.g.,
Osteocephalus oophagus 1 4 km Edalorhina, Pseudopaludicola ceratophyes), but most are
Osteocephalus taurinus 4 1 to 6.5 km and either tolerant of some intervention (or habitat modification)

spraying at 2.5 k (Engystomops petersi) or virtually restricted to open habitats.
Osteocephalus verruciger 2 3.5 and 6.5 km Most species of this family employ foam nests in which the
Phyllomedusa hypocondrialis 2 5 to 10 km
embryos are protected from desiccation and it remains to be
Phyllomedus perinesos 2 3.5 and 6.5 km determined whether glyphosate penetrates the foam surface.
Phyllomedusa venustus 1 2 km

Pseudis paradoxa 1 10 km Family Leptodactylidae
Scarthyla vigilans 4 1 to 10 km
Scinax blairi 4 1 to 9.3 km These abundant lowland animals include 14 species with
records within 10 km of coca production: Leptodactylus
Scinax cruentommus 2 1 and 3 km
Scinax garbei 2 1 and 2.5 km (spray) andreae (at 1 km from coca production and 2 km from aerial
Scinax ictericus 2 3.5 and 6.5 km eradication spraying), L. colombiensis (at 5 km), L. fragilis
(2 records at 0.5 and 2 km from cultivati, . fuscus(6 records
Scinax rostratus 6 1 to 10 km
Scinax x-signatus 1 10 km at 0.8 to 9.3 km from production with aerial eradication spraying
at 6 to 8 km), L. hylaedactylus (at 0.6 km from production with
(Continued ) aerial eradication spraying at 6 km), L. insularum (5 records at

395 Annex 131-H

984 J. D. LYNCH AND S. B. ARROYO

2 to 10 km from production and aerial eradication spraying at (a) amphibians are especially diverse in Colombia and (b)
6 to 11 km), L. knudseni(2 records at 0.8 to 3 km),L. mystaceus amphibians do not necessarily have the integument protection

(at 1.7 km with aerial eradication spraying at 1 km), available to other terrestrial vertebrates.
L. poecilochilus(at 1 km), L. rhodomerus (at 8.7 km), L. riveroi However, those concerns need to be related to the magni-
(3 records at 0.8 to 4 km from production), L. savagei tude of the problem. Coca production and aerial eradication

(3 records at 1 km from production and aerial eradication spray sprays are restricted geographically to areas where this activity
at 9 km), L. ventrimaculatus (at 2.2 km), and L. wagneri can be accomplished without government intervention and do

(2 records at 3.5 and 6.5 km). The habitat preferences and the not cover the country’s latitudinal extent or the altitudinal
reproductive biology of this family parallel closely those of the dimension—coca is usually “commercially” grown at ≤1000 m
preceding family with only some notable differences. Several a.s.l. The 153,134 ha of land thatwas subject to aerial eradica-

species of Leptodactylus deposit their eggs in foam nests tion spraying in 2007 (personal communication, National Police
within burrows, possibly a physical protection from pesticides of Colombia, Bogotá, December, 2007) represents only about
used for coca production and eradication spraying. 0.15% of the total land area of Colombia (Solomon et al.,

2007). Reviewing the anuran faunas of lowland Colombia, one
notes three areas that might be identified as diversity
Family Microhylidae
Eight species of this small family (small for Colombia, but a “hotspots”—one in the extreme southeast, a second in the
northern half of the Pacific lowlands, and the third a lens lying
large family for the Old World) have records within 10 km of along the eastern base of the Andes in southwestern Colombia.
coca production: Chiasmocleis bassleri (at 1.7 km with aerial
Coca production is modest within the first two of these
eradication spraying at 1 km), C. panamensis (at 7.5 km), “hotspots” and apparent in the third (Figure 4).
Elachistocleis ovale (1 record within 6 km of coca production Vast areas of the distributions of most species lie outside of
and aerial eradication spray), Otophryne pyburni (at 3 km),
the efforts of Colombian authorities to control coca production
Nelsonophryne aterrima (at 7.5 km), Relictivomer pearsei (Figures 1–3). Further, the consequences of coca production
(3 records at 2 to 9.3 km, aerial eradication spraying at 8 km), may be more serious for only a limited number of species that
Synapturanus mirandaribeiroi (at 3 km), and S. rabus (at 1 km
barely enter Colombian territory. Especially in Nariño, in western
with aerial eradication spraying at 2 km). Most of the species Putumayo and adjacent areas of Cauca and Caquetá, the effects
(15) of this family in Colombia are denizens of forests and
appear to be confined to undisturbed forests. Nonetheless, four of coca production and aerial eradication sprays have placed
several species of frogs at risk (Figure 4), at least in terms of
species occupy open and/or heavily intervened habitats (Chias- their distribution in Colombia. These includeAmeerega bilingua,
mocleis panamensis, E. ovale, Nelsonophryne aterrima, and
Dendropsophus bifurcus, Pristimantis colomai, P. degener,
Relictivomer pearsei) and thus may be at risk from pesticides P. diadematus, P. quaquaversus, P. variablis,and Trachyceph-
used for coca production and eradication spraying because all alus jordani. Other species may also be at risk, but exact num-
deposit their embryos as films on tehsurface of ponds. In contrast,
Downloaded At: 20:08 2 October 2009 bers are unknown because few investigations were undertaken
the adults are burrowers, are active on the ground surface only in these areas during the past 30 yr. As these species are mostly
at night, and do not climb vegetation (except for one species of found in Ecuador, it is assumed that healthy populations persist
deep forests in Amazonia—Syncope antenori).
there.
Overall, the risks from pesticides used for coca production

Family Ranidae and eradication spraying must be placed in the context of the
Two species (Lithobates palmipes and L. vaillanti) have 4 greater toxicity of products used by growers (Brain &
Solomon, 2009) and the sensitivity of frogs from Colombia to
and 8 records within 10 km of coca production. Two of these
records are within 2.5 km of aerial eradication spraying. the mixture of glyphosate and Cosmo-Flux as used in the aerial
eradication spraying. Laboratory-based toxicity studies showed
Although these two species occupy forested areas, they also are that aquatic larval stages of Colombian species are not differ-
common in open areas, including those with significant human
intervention (or habitat modification). Reproduction is confined entially sensitive as compared with frogs from other locations
(Bernal et al., 2009a). When tested under realistic conditions—
largely to water courses with a current and eggs are submerged
in the water. in shallow water (15 cm deep) in the presence of sediment and
particulates that absorb glyphosate and the more toxic
surfactant—toxicity was reduced (Bernal et al., 2009a).

CONCLUSIONS Terrestrial stages were less susceptible than aquatic stages
Anyone concerned about the persistence of natural habitats (Bernal et al., 2009b). In contrast, some of the products used
must be concerned by the effects of (1) ecological damage by growers may be more bioavailable in the environment

done in order to facilitate the illegal production of coca and and risks to these may not be mitigated. Of greater signifi-
(2) the potentially negative effects on biodiversity of efforts atcance is the effect on amphibians and other fauna and flora
eradication. These concerns are greater for amphibians because of habitat change associated with forest clearance for coca

396 Annex 131-H

RISKS TO COLOMBIAN AMPHIBIAN FAUNA FROM COCA CULTIVATION 985

production. Some insight into this was provided by Lynch ICZN (International Commision on Zoological Nomenclature). 1999. Interna-
tional Code of Zoological Nomenclature adopted bf the International
(2005) in the region of Le ticia, where species richness Union of Biological Sciences. Fourth Edition. International Trust for
declined from 98 species in rural areas to 6 species in the
Zoological Nomenclature, London.
city parks, reflecti ng the effects of increasing disturbance Lynch, J. D. 1998. New species of Eleutherodactylus from the Cordillera
and habitat fragmentation. Occidental of western Colombia with a synopsis of the distributions of spe-
cies in western Colombia. Rev. Acad. Colomb. Cienc. Exact. Fís. Nat.

22:117–148.
Lynch, J. D. 1999a. La riqueza de la fauna anfibia de los Andes colombianos.
REFERENCES Innov. Cienc., 7:46–51.
Anonymous. 2006. Colombia/Monitoreo y Cultivos de Coca, Technical. Lynch, J. D. 1999b. Ranas pequeñas, lageometría de evolución, y la especiación
United Nations, Office on Drugs and Crime & Gobierno de Colombia. en los andes Colombianos. Rev. Acad. Colomb. Cienc. Exact. Fís. Nat.

http://www.biesimci.org/Documento s/archivos/Colombia_coca_survey_ 23:143–159.
es05.PDF Lynch, J. D. 2005. Discovery of the richest frog fauna in the world—An
Bernal, M. H., Solomon, K. R., and Carrasquilla, G. 2009a. Toxicity of formu- exploration of the forests to the north of Leticia,. Rev. Acad. Colomb.
lated glyphosate (Glyphos) and Cosmo-Flux to larval Colombian frogs. 1. Cienc. Exact. Fís. Nat., 29:581–588.

Laboratory acute toxicity, J. Toxicol. Environ. Health A 72:961–965. Lynch, J. D. 2006. The amphibian fauna in the Villavicencio region of eastern
Bernal, M. H., Solomon, K. R., and Carrasquilla, G. 2009b. Toxicity of Colombia. Caldasia 28:135–155.
formulated glyphosate (Glyphos) and Cosmo-Flux to larval and juvenile Lynch, J. D., Ruiz-Carranza, P. M., and Ardila-Robayo, M. C. 1997. Biogeo-
Colombian frogs. 2. Field and lratory microcosm acute toxicity.J. Toxicol. graphic patterns of Colombian frogs and toads. Rev. Acad. Colomb. Cienc.
Environ. Health A72:966–973. Exact. Fís. Nat. 21:237–248.

Brain, R. A., and Solomon, K. R. 2009. Comparative hazards of glyphosate, Lynch, J. D., and Suarez-Mayorga, A. M. 2001. The distributions of the gladiator
other pesticides, and other human activities to amphibians in the production frogs (Hyla boansgroup) in Colombia, withcomments on size variation and
of coca. J. Toxicol. Environ. Health A 72:937–948. sympatry. Caldasia 23:491–507.
Duellman, W. E., and Trueb, L. 1986. Biology of amphibians. Baltimore, MD: Solomon, K. R., Anadón, A., Carrasquilla, G., Cerdeira, A., Marshall, J., and

John Hopkins University Press. Sanin, L.-H. 2007. Coca and poppy eradication in Colombia: Environmental
ESRI. 2006. ArcMap, ESRI. Inc. Procalculo Prosis S.A., Bogota. http:// and human health assessment ofaerially applied glyphosate.Rev. Environ.
www.esri.com Contam. Toxicol. 190:43–125.
Frost, D. R., Grant, T., Faivovich, J., Bain, R., Haas, A., Haddad, C. F. B., debbins, R. C., and Cohen, N. W. 1995. A natural history of amphibians.

Sá, R. O., Donnellan, S. C., Raxworthy, C. J., Wilkinson, M., Channing, Princeton, NJ: Princeton University Press.
A., Campbell, J. A., Blotto, B. L., Moler, P., Drewes, R. C., Nussbaum, Tsui, M. T. K., and Chu, L. M. 2004. Comparative toxicity of glyphosate-based
R. A., Lynch, J. D., Green, D., and Wheeler, W. C. 2006. The amphibian herbicides: Aqueous and sediment porewater exposures. Arch. Environ.
tree of life. Bull. Am. Mus. Nat. Hist. 297:1–370. Contam. Toxicol.46:316–323.
Grant, T., Frost, D. R., Caldwell, J. P., Gagliardo, R., Haddad, C. F. B., Kok,ng, N., Besser, J. M., Buckler, D. R., Honegger, J. L., Ingersoll, C. G., Johnson,

P. J. R., Means, B. D., Noonan, B. P., Schargel, W., and Wheeler, W. C. 2006.B. T., Kurtzweil, M. L., MacGregor, J., and McKee, M. J. 2005. Influence of
Phylogenetic systematics of dart-poison frogs a nd their relatives (Anura: sediment on the fate and toxicity ofa polyethoxylated tallowamine surfactant
Athesphatanura: Dendrobatidae). Bull. Am. Mus. Nat. Hist. 299:1–26. system (MON 0818) in aquatic microcosmsC . hemosphere59:545–551.
Hedges, S. B., Duellman, W. E., and Heinicke, M. P. 2008a. New world direct- Wiens, J. J., Fetzner, J. W., Parkinson, C. L., and Reeder, T. W. 2005. Hylid

developing frog (Anura: Terrarana): Molecular phylogeny, classification, frog phylogeny and sampling strategies for speciose clades. Syst. Biol.
biogeography, and conservation, Zootaxa 1737:1–182. 54:719–748.
Hedges, S. B., Duellman, W. E., and Heinicke, M. P. 2008b. A replacement Wiens, J. J., Kuczynski, C., Duellman, W. E., and Reeder, T. W. 2007. Loss
Downloaded At: 20name for Isodactylus Hedges, Duellman, and Heinicke, 2008. Zootaxa and re-evolution of complex life cycles in marsupial frogs: Can ancestral
1795:67–68. trait reconstruction mislead? Evolution 61:1886–1899.

397398 Annex 131-I

Annex 131-I

C.BOLOGNESI ET A., IOMONITORING OFGENOTOXICRISK IA GRICULTURAL
WORKERS FROM FIVECOLOMBIAN REGIONS: SSOCIATION TOCCUPATIONAL
EXPOSURE TOGLYPHOSATE”

(Journal of Toxicology and Environmental Health, Part A, 72:986-997, 2009)

399 Annex 131-I

Journal of Toxicology and Environmental Health, Part A, 72: 986–997, 2009
ISSN: 1528-7394 print / 1087-2620 online
DOI: 10.1080/15287390902929741

UTEH
Biomonitoring of Genotoxic Risk in Agricultural Workers

from Five Colombian Regions: Association to Occupational

Exposure to Glyphosate

C. Bolognesi , G. Carrasquilla , S. Volpi , K. R. Solomon , and E. J. P. Marshall 4
1
Environmental Carcinogenesis Unit. Department of Epidemiology and Prevention, National Cancer
Research Institute, Genoa, Italy, 2Facultad de Salud, Universidad del Valle, Cali, Colombia, Centre

for Toxicology and Department of Environmental Biology, University of Guelph, Guelph, Ontario,
Canada, and 4Marshall Agroecology Limited, Barton, Winscombe, Somerset, United Kingdom

direct contact with the erad ication spray showed a higher
In order to assess possible human effects associated with quantitative frequency of BNMN compared to those without
glyphosate formulations used in the Colombian aerial spray glyphosate exposure. The increase in frequency of BNMN
observed immediately after the glyphosate spraying was not
program for control of illicit cr ops, a cytogenetic biomonitor- consistent with the rates of application used in the regions and
ing study was carried out in subjects from five Colombian
regions, characterized by different exposure to glyphosate and there was no association between self-reported direct contact
other pesticides. Women of reproductive age (137 persons 15– with eradication sprays and fr equency of BNMN. Four months
49 yr old) and their spouses (137 persons) were interviewed to after spraying, a statistically si gnificant decrease in the mean
obtain data on current health st atus, history, li festyle, includ-frequency of BNMN compared wi th the second sampling was
observed in Nariño, but not in Putumayo and Valle del Cauca.
ing past and current occupational exposure to pesticides, and Overall, data suggest that ge notoxic damage associated with
factors including those known to be associated with increased glyphosate spraying for control of illicit crops as evidenced by
frequency of micronuclei (MN). In regions where glyphosate
was being sprayed, blood sample s were taken prior to spraying MN test is small and appears to be transient. Evidence indi-
(indicative of baseline exposure ), 5 d after spraying, and 4 mo cates that the genotoxic risk po tentially associated with expo-
after spraying. Lymphocytes were cultured and a cytokinesis- sure to glyphosate in the areas where the herbicide is applied
block micronucleus cytome assay was applied to evaluate chro- for coca and poppy eradication is low.

Downloaded At: 20:08 2 October 2009xicity. Compared with Santa
Marta, where organic coffee is grown without pesticides, the
baseline frequency of binucleated cells with micronuclei
(BNMN) was significantly greater in subjects from the other
four regions. The highest frequency of BNMN was in Boyacá, Glyphosate (N-phosphonomethyl glycine), a nonselective
where no aerial eradication sp raying of glyphosate was con- herbicide, is the active ingredient of a number of herbicide
ducted, and in Valle del Cauca, where glyphosate was used for
formulations and one of the most widely used pesticides on a
maturation of sugar cane. Region, gend er, and older age ( ³35 global basis (Baylis, 2000; Woodburn, 2000; Duke & Powles,
yr) were the only variables as sociated with the frequency of
BNMN measured before spraying. A significant increase in fre- 2008). It is a postemergence herbicide, effective for the con-
quency of BNMN between first and second sampling was trol of annual, biennial, and perennial species of grasses,
observed in Nariño, Putumayo, and Valle immediately (<5 d) sedges, and broadleaf weeds. The relatively high water solu-
after spraying. In the post-spray sample, those who reported
bility and the ionic nature of glyphosate retard penetration
through plant hydrophobic cuticular waxes. For this reason,
glyphosate is commonly formulated with surfactants that
©General Secretariat of the Or ganization of American States,
2009. This paper was prepared as part of a Study entitled decrease the surface tension of the solution and increase pen-
“Production of Illicit Drugs, the Environment and Human Health,” etration into the tissues of plants (World Health Organization
financed with contributions from the Governments of Colombia and
the United States of America. The conclusions and opinions International Program on Chemical Safety, 1994; Giesy et al.,
expressed herein are those of the authors and not necessarily those2000).
A large number of glyphosate-based formulations are reg-
of the Organization of American States and its General Secretariat,
which as of the date of this copyright, have not formulated any istered in more than 100 countries and are available under dif-
opinion with respect to them. ferent brand names. One of the most commonly applied
Address correspondence to K. R. Solomon, Centre for Toxicology
and Department of Environmental Biology, University of Guelph, glyphosate-based products is Roundup, containing glyphosate
Guelph, ON, N1G 2W1, Canada. E-mail: [email protected] as the active ingredient (AI) and polyethoxylated tallowamine

986

400 Annex 131-I

BIOMONITORING GENOTOXIC RISK IN AGRICULTURAL WORKERS 987

(POEA) as a surfactant. Glyphosate and its formulations have chromosome mutation, DNA damage and repair) using bac-
been extensively investigated for potential adverse effects in teria and mammalian somatic cells (Williams et al., 2000).

humans (Williams et al., 2000). This pesticide was reported to The active ingredient did not induce any relevant genotoxic
exert a low acute toxicity to different animal species. Experi- effects such as gene mutations in a variety of in vitro bacte-
mental evidence showed that glyphosate did not bioaccumulate rial assays including the Salmonella typhimurium reversion

in any animal tissues (Williams et al., 2000). Chronic feeding assay, with and without metabolic activation (Wildeman &
studies in rodents did not find evidence of carcinogenic activityNazar 1982; Moriya et al., 1983; Li & Long, 1988) and

or any other relevant chronic effects (U.S. EPA, 1993; World Escherichia coli WP-2 (Moriya et al., 1983; Li & Long,
Health Organization International Program on Chemical 1988). The active ingredient wa s also negative in the Chi-
Safety, 1994). nese hamster ovary cell HGPRT gene mutation assay and in

With in vitro studies with tissue cultures or aquatic organ- primary hepatocyte DNA repair assay (Li & Long, 1988).
isms, several of the formulated products are more toxic than The genotoxic potential of the formulation Roundup was
glyphosate AI (Giesy et al., 2000; Williams et al., 2000). Dif- investigated in a number of studies evaluating various

ferences in the response of test organisms to the AI and the genetic endpoints in different biological systems and was
commercial formulation, e.g., Roundup, are likely due to the (1) negative in the S. typhimurium reversion assay (Kier
toxicity of different formulants and surfactants contained in
et al., 1997), (2) negative in the sex-linked recessive lethal
commercial products. There is a general agreement that adju- assay with Drosophila melanogaster (Gopalan & Njagi,
vants may be more toxic for animals than glyphosate itself 1981), and (3) negative for in vivo micronucleus (MN)

(Giesy et al., 2000; Williams et al., 2000; Richard et al., induction in mouse bone marrow (Rank et al., 1993; Kier
2005). Cytotoxicity of the commercial formulation Roundup et al., 1997; Dimitrov et al., 2006). The Roundup formula-
to human peripheral mononuclear cells was 30-fold higher tion was reported in a number of studies to exert weak geno-

(LC 50 = 56 mg/L) than for the AI (LC 50= 1640 mg/L) (Mar- toxic effects in short-term assays.
tinez et al., 2007). Several in vitro and in vivo studies with Differences in the response of test organisms to the
parallel testing of glyphosate AI and Roundup showed that active ingredient glyphosate and the commercial formula-

only the commercial formulation was genotoxic (Rank et al., tion Roundup might be due to the toxicity of different
1993; Bolognesi et al., 1997b; Gebel et al., 1997; Grisolia co-formulants and surfactants contained in commercial

2002). Cytotoxic and genotoxic effects were observed with products. Several studies with parallel testing of glyphosate
Roundup and other formulations of glyphosate, but not with and Roundup showed that only the commercial formulation
glyphosate AI alone in comparative studies involving differ- was genotoxic (Rank et al., 1993; Bolognesi et al., 1997b;

ent experimental systems (Peluso et al., 1998; Richard et al., Gebel et al., 1997; Grisolia 2002). A recent study on the
2005; Dimitrov et al., 2006). The observed differences were genotoxic potential of glyphosate formulations found that in
attributed to some ingredients of Roundup, mainly surfac- some cases the genotoxic eff ects were obtained under expo-

Downloaded tants, and/or to a synergic effect of glyphosate and compo- sure conditions that are not relevant for humans (Heydens
nents of the formulation (Sirisattha et al., 2004; Peixoto et al., 2008).
2005). An in vitro study described a concentration-dependent

Epidemiological studies generally showed no consistent or increase of DNA single-strand breaks (SSB), evaluated by comet
strong relationships between human exposure to glyphosate assay, in two different human cell lines treated with glyphosate

or glyphosate-containing products and health outcomes in at sublethal concentrations (Monroy et al., 2005). Roundup for-
human populations. No statistically significant association in mulations were shown to affectthe cell cycle by inhibiting the
humans was found with spontane ous abortion, fetal deaths, G2/M transition and DNA synthesis leading to a genomic insta-

preterm birth, neural tube defe cts (Rull et al., 2006), and can-bility (Marc et al., 2004a, 2004b). Evidence of DNA damage in
cer incidence overall, alt hough a suggested association peripheral lymphocytes from a small group of subjects
between cumulative exposure to glyphosate and the risk of potentially exposed to glyphosate was reported in a recent paper

multiple myeloma was reported (De Roos et al., 2005).The (Paz-y-Miño et al., 2007). The number of subjects (21 control
epidemiologic evidence is insufficient to verify a cause– and 24 exposed) was small and there were 23 females and only

effect relationship for childhood cancer (Wigle et al., 2008). 1 male in the exposed group, making interpretation of the results
Four case-control studies suggested an association between difficult.
reported glyphosate use and the risk of non-Hodgkin’s lym- Frequency of MN in human lymphocytes has been widely

phoma (NHL) in age groups from 20 to 70 yr (Hardell & used for biomonitoring exposure to pesticides (Bolognesi,
Eriksson, 1999; McDuffie et al., 2001; Hardell et al., 2002; 2003; Costa et al., 2006; Montero et al., 2006). The MN test,
De Roos et al., 2003; Eriksson et al., 2008). an index of chromosomal damage, is one of the most appro-

Glyphosate AI and Roundup we re extensively tested for priate biomarkers for monitoring a cumulative exposure to
genotoxicity in a wide range of in vitro and in vivo systems genotoxic agents. Chromosomal damage, as a result of ineffi-
evaluating different genetic endpoints (gene mutation, cient or incorrect DNA repair, is expressed during the cell

401 Annex 131-I

988 C. BOLOGNESI ET AL.

division and represents an index of accumulated genotoxic reproductive age in each area and, where possible, the same
effects. The cytokinesis-block micronucleus (CBMN) meth- couples in the study conducted by Sanin et al. (2009) were

odology (Fenech & Morley, 1985) allows a distinction to be sampled. In Putumayo, Nariño, and Valle del Cauca, the pop-
made between a mononucleated cell that did not divide and a ulation was selected based on the scheduled aerial spraying of
binucleated cell that has divided once, expressing any glyphosate. This schedule was confidential and provided

genomic damage associated to recent exposure. The test in its exclusively for the purpose of the study by the Antinarcotics
comprehensive application, as was proposed by Fenech Police (Putumayo and Nariño) or ASOCAÑA (Valle del

(2007) including a set of markers of gene amplification, cel- Cauca). In Valle del Cauca, a sa mple size of 30 couples could
lular necrosis, and apoptosis, allows evaluation of genotoxic not be achieved because spraying was not carried out in pop-
and cytotoxic effects induced by exposure to a genotoxic ulated areas of the study region. Most spraying during the

agent. study period was carried out on sugar cane crops where no
Colombia’s anti-drugs strategy includes a number of mea- inhabitants were found. All reported areas to be sprayed in
sures ranging from aerial spraying of a mixture of a commer- Valle del Cauca were visited to search for couples; however,

cial formulation of glyphosate (Glyphos) and an adjuvant, only 14 could be included.
Cosmo-Flux (Solomon et al., 2007b), to manual eradication, In Sierra Nevada de Sant a Marta and Boyacá, the same
including alternative development and crop substitution pro-
areas investigated in a previous study (Sanin et al., 2009)
grams (UNODC, 2007). In order to assess the potential geno- were identified, although, due to the instability of the popula-
toxic risk associated with the aerial spraying program with tion and high migration, most couples from the previous

the glyphosate mixture, a cytogenetic biomonitoring study study were not located. In all regions, the same strategy as
was carried out in subjects from five Colombian regions, described before (Sanin et al., 2009) was followed, visiting
characterized by different exposure to glyphosate formula- household by household until completing 30 couples who ful-

tions and other pesticides. filled the inclusion criteria, women of reproductive age (15–49 yr
of age) and their spouses, who voluntarily accepted to partici-
pate in the study.

MATERIALS AND METHODS
The study was carried out in five regions of Colombia, with
Field Data Collection
different potential exposure to glyphosate as reported by Sanin
et al. (2009). Briefly, the characteristics of the study areas are Field data collection was carried out between October
2006 and December 2007. Epidemiologists and interviewers
described here:
in the five regions who participated in the Sanin et al. (2009)
Sierra Nevada de Santa Marta—where organic coffee is grown study were informed about the objectives of the study and
without use of pesticides. trained for data collection. The Ethical Committee of Funda-

Downloaded At: 20:08 2 October 2009 crops, where manual eradication is cion Santa Fe de Bogotá approved the study protocol and the
performed and the use of pesticides and other chemical informed consent form s used for the study. All the subjects
agents is common. were informed about the aims of the study. All of them gave

Putumayo and Nariño—where aerial spraying of glyphosate their informed consent and volunteered to donate blood for
is performed for coca and poppy eradication. The aerial sampling. They did not self-report illness at the time of
application rate for eradi cation of coca is 3.69 kg blood sampling and interviews. Every volunteer was inter-

glyphosate a.e. (acid equivalents)/ha (Solomon et al., viewed with a standardized que stionnaire, designed to obtain
2007b). In order to maximize penetration and effective- relevant details about the current health status, history, and
ness of the spray formulation, Glyphos is tank-mixed
lifestyle. This included information about possible con-
with an adjuvant (Cosmo-Flux® 411F; Cosmoagro, founding factors for chromosomal damage: smoking, use of
Bogotá). medicinal products, severe infections or viral diseases during

Valle del Cauca—where glyphosate is applied through aerial the last 6 mo, recent vaccinatio ns, presence of known indoor/
spraying for sugar cane maturation. Roundup 747 is the outdoor pollutants, exposure to diagnostic x-rays, and previ-
most commonly used product and is applied at a rate of 1 ous radio- or chemotherapy. A simplified food frequency

kg a.e./ha, and has no additional adjuvant (personal com- questionnaire that had already been used in other regions of
munication, ASOCAÑA, the Colombian Association for Colombia was also applied, in order to evaluate dietary folic
Sugar Growers, December 2008).
acid intake. Folic acid intake was characterized because of
the role of folic acid deficiency in baseline genetic damage
in human lymphocytes (Fenech & Rinaldi, 1994). Specific

Study Population information about exposure at the time of aerial spraying in
Two hundred and seventy-four individuals were included Putumayo, Nariño, and Valle de l Cauca was addressed in the
in the study. The objective was to sample 30 couples of questionnaire.

402 Annex 131-I

BIOMONITORING GENOTOXIC RISK IN AGRICULTURAL WORKERS 989

Blood Sampling and Cell Culture as proportions. Dependent variables, micronuclei per binucle-
Blood samples were collected twice in Boyacá, at the begin- ated cell (BNMN), and differences in MN between sampling

ning of the study and 1 mo after the first survey, and at 3 differ-re square-root transformed where required to comply with
ent times in Nariño, Putumayo, and Valle del Cauca: the required assumptions of normal distribution and equal vari-
immediately before spraying, within 5 d after spraying, and 4 ances. Comparison of MN between areas was made by one-way

mo later. A sample of 10 ml whole blood was collected from analysis of variance (ANOVA). A significance level at 5% was
each subject, by venipuncture, using heparinized Vacutainer used to assess differences among areas. For multiple compari-
tubes kept at room temperature and sent within 24 h for the sons, the Bonferroni test was applied (a = .05). Significance of

establishment of the lymphocyte cultures. The samples were differences in frequency of BNMN between first and second,
coded before culturing. The modified cytokinesis-blocked and second and third sampling were tested by the unpaired
method of Fenech and Morley (1985) was used to determine t-test with equal variances. Difference and 95% confidence

frequency of MN in lymphocytes. Whole blood cultures were interval were used to compare between samplings.
set up for cytogenetic analysis in Bogotá (Colombia) by per- Bivariate analysis between dependent variables and putative
risk factors was performed by one-way ANOVA, comparing
sonnel specifically trained by cytogeneticists from Environ-
mental Carcinogenesis Unit of the National Cancer Research exposed and nonexposed subjects. In cases where risk factor
Institute (Genoa, Italy). was continuous, such as age, folic acid intake, alcohol con-

Three sterile cultures of lymphocytes were prepared. A 0.4-ml sumption, and coffee consumption, the correlation coefficient
aliquot of whole blood was incubated at 37ºC in duplicate in was used.
4.6 ml RPMI 1640 (Life Technologies, Milano, Italy) supple- A multiple linear regression was conducted to assess associ-

mented with 10% fetal bovine serum (Gibco BRL, Life Tech- ation with BNMN at the first sampling with different variables:
nologies SrL, Milano, Italy), 1.5% phytohemoagglutinin region, age (as continuous variable as well as categorical age),
(Murex Biotech, Dartford, UK), 100 units/ml penicillin, and ethnicity as a dichotomous variable, exposure to genotoxic

100 μg/ml streptomycin. After 44 h, cytochalasin B (Sigma, products as defined earlier, gender (female vs. male), and
Milano, Italy) was added at a concentration of 6 μg/ml. At the intake of folic acid (categorized in quartiles). Regression anal-
end of incubation at 37°C for 72 h, cells were centrifuged (800 ysis was conducted with transformed variables, with square

× g, 10 min), then treated with 5 ml of 0.075 mM KCl for 3 min root transformation of BNMN and natural logarithm of age, to
at room temperature to lyse erythrocytes. The samples were obtain a normal distribution.

then treated with pre-fixative (methanol:acetic acid 3:1) and
centrifuged . The cellular pellets were resuspended in 1 ml RESULTS
methanol. At this step the samples were sent to the Environmental
Demographic characteristics and habits of the study groups
Carcinogenesis Unit (National Cancer Research Institute, are described in Table 1. The study population comprised 274
Genoa, Italy). All the samples were centrifuged in methanol.
Downloaded Treatment with fixative (methanol:acetic acid, 5:1) followed subjects (137 female and 137 male; average age 30.4 ± 7.8 yr).
The mean age of the subjects was similar in the different
by centrifugation was repeated twice for 20 min. Lymphocytes regions. A large part of the studied population was mestizo,
in fresh fixative were dropped onto clean iced slides, air-dried,
and stained in 2% Giemsa (Sigma, Milano, Italy). MN analysis with the exception of the Nariño area consisting of individuals
of African origin. In the total population, 38% of interviewees
was performed blind only on lymphocytes with preserved cyto- had not completed primary education. Putumayo had the larg-
plasm. On average, 2000 cells were analyzed for each subject.
Cells were scored cytologically using the cytome approach to est proportion with education and Valle del Cauca the lowest
as shown in Table 1. Only 10% of all subjects were smokers,
evaluate viability status (necrosis, apoptosis), mitotic status (20% in Putumayo); a large majority of subjects were drinkers
(mononucleated, binucleated, multinucleated) and chromo-
somal damage or instability status (presence of micronuclei, of beer or liquor with a consistent consumption of guarapo (tra-
ditional alcoholic beverage prepared by fermentation of maize)
nucleoplasmic bridges, nucleoplasmic buds) (Fenech 2007).
The proliferation index (PI) was calculated as follows: in Santa Marta and Boyacá. No statistically significant differ-
ences of folic acid intake were observed between different
regions (the mean values ranged from 750 and 1189 mg/wk).
PI = (number of mononucleated cells + 2
One hundred and nine (39.8%) of 274 participants reported
× number of binucleeated cells + 3 current use of pesticides in their occupation or other activities.
Nariño (76.6%) and Putumayo (61.7%) were the two regions
× number of polynucleated cells)/ total umber of cells.
where prevalence of use of genotoxic pesticides was higher;
Boyacá (24.2%) and Valle del Cauca (28.6%) reported lower
Statistical Analysis use. None of the study subjects in Santa Marta reported use of

Continuous variables were characterized using mean and pesticides. No data regarding quantity of pesticide used were
standard deviation, while categorical variables were expressed available. Fifty (18.3%) out of 273 who gave information

403 Annex 131-I

990 C. BOLOGNESI ET AL.

TABLE 1
Demographic Characteristics and Possible Confounding Exposures in the Study Populations

Santa Valle del
Area Marta Boyacá Putumayo Nariño Cauca

Number of subjects 60 62 60 64 28

Age (mean (SD)) 27.0 (5.6) 29.1 (8.8) 31.4 (7.2) 32.5 (7.4) 33.4 (8.7)
Ethnicity (%)

Mestizo 100 100 88.3 3.1 60.7
African 6.7 96.9 39.3
Indian 5.0

Education (%)

None 4.8 1.7
Primary incomplete 26.7 38.7 53.3 42.2 21.4
Primary complete 21.7 29.0 20.0 23.4 32.1

High school incomplete 25.0 8.1 20.0 25.0 28.6
High school complete 26.7 19.4 3.3 9.4 17.9

Technical 1.7
Occupation (%)

Agriculture 10.0 41.9 60.0 62.5 7.1
Housewife 40.0 50.0 38.3 34.4 50.0
Other 50.0 8.1 1.7 3.1 42.9

Health insurance (%)

Uninsured 50.0 9.7 36.7 71.9 7.1
Subsidized 38.3 83.9 60.0 18.7 50.0
Insured 11.7 6.4 3.3 9.4 42.9

Coffee consumption (cups/day)

Mean (SD) 1.8 (2.3) 1.7 (0.8) 2.3 (4.1) 1.3 (0.4) 1.7 (1.2)
Percent of population 80.0 67.7 88.3 76.6 82.1

Smoking (%)
Downloaded ANonsmokers October 2009 91.7 95.2 80.0 87.5 92.9

Alcohol (%)
Liquor 28.3 25.8 53.3 78.1 78.6

Beer 51.6 67.7 63.1 82.8 64.3
Guarapo 6.7 59.7 1.7 3.2 10.7
Users of illicit drugs (%) 6.7 0 5.0 7.8 0

Diet

Folic acid intake (μg/wk) 1189 873 750 1160 812

about x-ray examination reported to having been exposed at and presented graphically in Figure 1. Compared with Santa
some time; however, only 21 out of 46 who gave information Marta, where people grow organic coffee without the use of
on dates of x-ray reported exposure in the last 6 mo before thpesticides and which is considered as a reference area, the

interview and first blood sample. Sixty-one percent of popula-baseline frequency of BNMN was significantly greater in sub-
tion reported viral infections, the highest prevalence in Narijects from the other four regions. The highest frequency of
(89.5%) and the lowest in Putumayo (49.2%). However, 89.3% BNMN was in Boyacá, where no aerial eradication spraying of

of viral infections were the common cold and 6.1% dengue glyphosate was carried out, and Valle del Cauca, where aerial
fever. Hepatitis was reported by six interviewees without any spraying was for maturation of sugar cane. There was no

specification of the type of the infection. significant difference between mean frequency of BNMN in
The means and standard deviations of frequency of MN and Boyacá and Valle del Cauca. There was no significant differ-
related parameters according to regions are shown in Table 2 ence in frequency of BNMN between Putumayo and Nariño,

404 Annex 131-I

BIOMONITORING GENOTOXIC RISK IN AGRICULTURAL WORKERS 991

TABLE 2

Mean (SD) Frequency of Binucleated Cells with Micronuclei (BNMN), Total Micronuclei (MNL) per 1000 Binucleated
Peripheral Lymphocytes, Frequency of Mononucleated Cells per 1000 Lymphocytes (MNMO), and Proliferation Index (PI)
by Region before the Exposure (Phase 1), 5 d after Spraying (Phase 2) and 4 mo Later (Phase 3)

Region Santa Marta Boyacá Putumayo Nariño Valle del Cauca

Phase 1
Number of subjects 60 62 58 63 28
BNMN 1.83 (0.97) 5.64 (1.72) 3.61 (1.51) 4.12 (1.65) 5.75 (2.48)

MNL 1.97 (1.05) 6.16 (1.91) 3.90 (1.66) 4.36 (1.85) 6.02 (2.50)
MNMO 0.41 (0.44) 0.99 (0.64) 0.47 (0.51) 0.51 (0.39) 1.12 (0.88)
PI 1.54 (0.14) 1.45 (0.14) 1.68 (0.15) 1.47 (0.12) 1.51 (0.15)

Phase 2
Number of subjects ND 55 53 55 27

BNMN 4.96 (2.00) 4.64 (2.45) 5.98 (2.03) 8.64 (2.81)
MNL 5.41 (2.25) 5.02 (2.95) 6.35 (2.18) 8.98 (2.93)
MNMO 0.87 (0.65) 0.44 (0.46) 0.70 (0.45) 1.65 (0.62)

PI 1.72 (0.14) 1.66 (0.20) 1.40 (0.18) 1.51 (0.14)
Phase 3
Number of subjects ND ND 50 56 26

BNMN 5.61(3.08) 3.91 (1.99) 7.38 (2.41)
MNL 5.96 (3.23) 4.13 (2.20) 8.17 (2.72)

MNMO 0.82 (0.54) 0.55 (0.42) 0.98 (0.60)
PI 1.43 (0.17) 1.41 (0.14) 1.45 (0.20)

Downloaded At: 20:08 2 October 2009

FIG. 1. Box plot of frequency of BNMN in the five study regions with samples taken prespray, 4–5 d post-spray, and 4 mo post-spray. Box plots: The center
horizontal line marks the median of the sample. The length of each box shows the range within which the central 50% of the values fall, with the top and bottom
of the box at the first and third quartiles. The vertical T-lines represent intervals in which 90% of the values fall. The ❍ symbols show outliers. See text for

description of statistically significant differences.

405 Annex 131-I

992 C. BOLOGNESI ET AL.

although Boyacá and Valle del Cauca showed a significantly low (∼10% in the total population). A higher baseline fre-
higher frequency than Nariño and Putumayo. A higher fre- quency of MN was observed in subjects of African origin, sug-

quency of BNMN in Boyacá was also observed in a second gesting greater susceptibility. Other lifestyle factors such as
sampling 1 mo later. alcohol, coffee consumption, or illicit drug intake were not
There were differences in frequency of BNMN between associated with initial measures of BNMN and MOMN.

sampling periods. A statistically significant difference in fre- One hundred and thirty-four of the 152 subjects in Nariño,
quency of BNMN between first and second sampling was Putumayo, and Valle reported information on contact with

observed in Valle, Putumayo, and Nariño immediately (<5 d) Glyphos and Cosmo-Flux after eradication spraying. The other
after spraying. Four months after spraying in Nariño, there was 18 did not provide information in the second survey or blood
a statistically significant decrease in the mean frequency of samples were inadequate for testing micronuclei. Sixty-six

BNMN compared with the second sampling, but in Valle del (49.2.0%) reported no contact with the spray and 68 (50.8%)
Cauca the decrease was not significant nor was the increase reported coming into contact with the spray because they
observed in Putumayo significant (Figure 1 and Table 2). entered sprayed fields or reported contact with the spray drop-

The frequency of mononucleated cells with micronuclei lets. The mean BNMN in Nariño and Putumayo was greater in
(MOMN) was used as an index of background level of chro- respondents who self-reported exposure, but differences were
mosomal damage accumulated in vivo (Table 2). The lowest
not statistically significant (Table 4). In Valle, only one
frequency of MOMN for the first sampling was observed in respondent reported contact with glyphosate.
Santa Marta; however, there was no marked difference in fre- Region, gender, and older age (≥35 yr) were the only vari-

quency of MOMN in Santa Marta, Putumayo, and Nariño and ables associated with the frequency of BNMN before spraying
no statistically significant difference between Valle and (Table 5). In fact, using Santa Martha, where no use of pesti-
Boyacá. However, Valle and Boyacá had a significantly higher cides was reported, as reference, Boyacá, Valle del Cauca,

frequency of MOMN than Putumayo, Nariño, and Santa Marta Putumayo, and Nariño showed a statistically significant higher
at first sampling. Immediately after spraying, Valle showed a mean frequency of BNMN. There were also significant differ-
significantly higher frequency of MOMN compared to Putumayo ences between Boyacá and Valle and Putumayo and Nariño.

and Nariño, and Nariño was also higher than Putumayo. Females had a statistically higher mean frequency of BNMN
Between first and second sampling, the increase in frequency than males after adjusting for all other variables. Greater age

of MOMN in Nariño and Valle was statistically significant, but was also associated with greater frequency of BNMN. Neither
there was no difference in Putumayo nor in Boyacá 4 mo after exposure to genotoxic products, nor ethnicity, nor intake of
the first sampling. Data suggest greater exposure to genotoxic folic acid was associated with frequency of BMMN at the first

agents in these populations is independent of the exposure to sampling. The multiple linear regression analysis of difference
glyphosate products. between second and first sampling only demonstrated statisti-
The proliferation index (PI) in all the studied groups was in cally significant association with region after adjusting for all

Downloadedthe range of normal values described in the literature. No sig- other variables, indicating that Putumayo, Nariño, and Valle
nificant reduction of PI was observed in association with envi- had significantly greater differences between second and first
ronmental exposures in groups of subjects from the different sampling than Boyacá.

regions. A statistically significant correlation coefficient
(0.288) between PI values from the first and the second sam-
DISCUSSION
plings was observed, confirming the association with individ-
ual characteristics and not with any toxicity related to the The main objective of this study was to test whether there
exposure or to the culture techniques. Due to the low frequency was an association between aerial spraying of glyphosate and

observed, data with respect to other nuclear alterations, includ-cytogenetic alterations, evaluated as frequency of MN in
ing in cytome analysis (Fenech, 2007), are not described in peripheral leukocytes. Biomonitoring was carried out in three
Table 2: the mean frequency of nucleoplasmic bridges (NPB) regions of Colombia in populations exposed to aerial spraying

for all subjects was 0.010 per 1000 cells, that of nuclear buds of glyphosate: Putumayo and Nariño, where the application
was 0.022 per 1000 cells, and only rare necrotic and apoptotic was performed for eradication of coca and poppy, and Valle del

cells were found in some samples. Cauca where the herbicide was used for maturation of sugar
Gender was the most important demographic variable cane. Two control populations not exposed to aerial spraying of
affecting the BNMN index. Frequencies of BNMN in females glyphosate were also selected: the first one from Sierra Nevada

were greater than those in males (mean 4.43 ± 2.36 vs. 3.61 ± de Santa Marta, where organic coffee is grown without the use
1.82, respectively, in total population) (Table 3). The groups ofof any pesticides, and the other from Boyacá, with a region of
subjects were evenly matched for gender by including only illicit crops, where manual eradication is performed and sub-

couples in the study. No association was found between fre- jects were potentially exposed to several pesticides but not
quency of MN and age as a categorical variable, nor was there glyphosate for aerial eradication. The ex vivo analysis of leu-
an association with smoking, but prevalence of smoking was kocytes in the presence of cytochalasin B, added 44 h after the

406 Annex 131-I

BIOMONITORING GENOTOXIC RISK IN AGRICULTURAL WORKERS 993

TABLE 3

Association of Mean (SD) Frequency of Binucleated Cells (First Sampling) with Micronuclei
(BNMN/1000 Binucleated Lymphocytes) and Demographic Variables

Variable Santa Marta Boyacá Putumayo Nariño Valle del Cauca Total

Sex

Females 1.98 (1.03) 6.22 (1.79) 3.91 (1.71) 4.57(1.77) 6.45 (2.82) 4.43 (2.36)
Males 1.68 (0.90) 5.06 (1.46) 3.31 (1.25) 3.66 (1.39) 5.05 (1.94) 3.61 (1.82)
p .236 .007 .131 .028 .138 .002

Age

18–24 yr 2.00 (1.14) 5.50 (1.96) 3.32 (1.25) 3.64 (1.72) 6.19 (2.15) 3.67 (2.16)
25–34 yr 1.66 (0.87) 5.70 (1.66) 3.53 (1.17) 4.20 (1.77) 4.20 (0.76) 3.97 (2.08)
35 yr and older 1.93 (0.67) 5.62 (1.73) 3.84 (1.86) 4.25 (1.52) 6.04 (2.84) 4.41 (2.19)

p .438 .929 .574 .564 .313 .093
Ethnicity

Mestizo 1.83 (0.97) 5.64 (1.72) 3.72 (1.52) 4.75 (1.06) 5.82 (2.44) 3.94(2.24)
Africa and 0 0 2.86 (1.31) 4.10 (1.66) 5.64 (2.65) 4.20(1.90)

Indian
p .162 .588 .850 .368

Smoking
Yes 2.00 (1.06) 5.33 (0.76) 3.31 (1.00) 4.77 (1.51) 4.50 (1.41) 3.83 (1.60)

No 1.82 (0.97) 5.65 (1.76) 3.80 (1.56) 4.03 (1.66) 5.90 (2.57) 4.07 (2.20)
p .693 .756 .395 .233 .459 .592

Folic acid intake (quartiles)
1 1.92 (0.99) 6.11 (1.95) 3.23 (1.12) 4.50 (1.75) 5.86 (2.34) 3.89 (2.23)
2 1.64 (0.66) 5.70 (1.75) 3.47 (1.49) 3.80 (1.47) 5.86 (2.74) 3.97 (2.21)

3 1.69 (0.92) 5.69 (1.82) 4.00 (1.37) 3.85 (2.04) 6.58 (2.84) 4.47 (2.22)
4 1.94 (1.20) 4.94 (1.13) 3.69 (2.429) 4.28 (1.51) 4.63 (2.05) 3.75 (1.89)

p .779 .399 .515 .645 .612 .220

Downloaded At: 20:08 2 October 2009

TABLE 4
Mean Frequency of Binucleated Cells with Micronuclei (BNMN) at the Second Sampling per 1000 Binucleated Lymphocytes

and Self-Reported Exposures to the Glyphosate Spray in Three Areas Where Aerial Application Had Occurred

Nariño (n = 55) Putumayo (n = 53) Valle del Cauca (n = 26)

Route of exposure n Mean BNMN (SD) n Mean BNMN (SD) n Mean BNMN (SD)

No exposure 28 5.81 (1.85) 13 3.84 (1.30) 25 8.56 (2.90)
Spray in air 5 7.30 (0.57) 1 5.50 (0)

Spray on skin 8 5.62 (1.60) 15 4.90 (1.87) 1 9.50 (0)
Entered sprayed field 14 6.06 (2.77) 24 4.87 (3.18)

p Value (ANOVA) 0.472 0.612 0.760
Any exposure 27 6.16 (2.22) 40 4.90 (2.69) 1 9.50 (0)
p Value (no exposure 0.525 0.181 0.760

vs. any exposure)

Note. The data comprise respondents in the second survey from which blood samples were obtained.

407 Annex 131-I

994 C. BOLOGNESI ET AL.

TABLE 5 There was no clear relationship between BNMN and the
reported use of pesticides classified as genotoxic. Participants
Multiple Linear Regression Analysis Adjusted for Region,
Age, Gender, Ethnicity, and Folic Acid Intake in Boyacá and Valle del Cauca showed higher frequency of
BNMN than those in Putumayo and Nariño. However, a
Variable Coefficient p 95% CI greater proportion of participants in the latter regions self-

Region reported the use genotoxic pesticides (76.6% in Nariño and
61.7% in Putumayo). There is no information available on
Boyacá 3.75 ≤.0001 3.19, 4.31
Putumayo 1.58 ≤.0001 1.00, 2.16 other relevant factors such as frequency of use, rate applied,
Nariño 2.06 ≤.0001 1.49, 2.64 time of exposure, and protective measures used, and we could
therefore not characterize exposures to explain the differences.
Valle del Cauca 3.65 ≤.0001 2.92, 4.39
There were further inconsistencies; for example, in Boyacá,
Age (yr) where more frequent use of pesticides was expected, only
25–34 0.28 .250 –0.20, 0.76 24.2% of participants self-reported use, compared with the
35 and older 0.75 .008 0.20, 1.31
greater values in Nariño and Putumayo. However, it is possible
Gender that in areas such as Boyacá, individuals might be potentially

Females 1.00 ≤.0001 0.60, 1.40 exposed to persistent pesticides applied in the past and still
present in the environment.
There was no evidence of an association between BNMN
start of cultivation, made it possible to distinguish between non-
and folic acid deficiency. An assessment of folic acid intake
dividing mononucleated cells—as an index of accumulated from the semiquantitative food frequency questionnaire
chromosomal damage—and binucleated cells, which had com- showed that, according to accepted recommendations (Herbert,
pleted one nuclear division during in vitro culture and expressed
1987), the diet of the study populations was not deficient in
MN associated with recent exposure to genotoxic agents. folic acid and there were only small differences between
The baseline level of chromosomal damage, evaluated as regions. Consistent with these data, no association was found

frequency of BNMN, was associated with the different regions between MN and folic acid intake, either as a continuous vari-
considered in our study. The frequency of BNMN before able or by quartiles.
spraying was also associated with region, gender, and age.
The frequency of BNMN increased after spraying with
Gender difference in the background incidence of MN in glyphosate but not consistently. The results obtained with a
peripheral leukocytes, with the frequency being consistently second sampling, carried out immediately after the glyphosate
higher in females, and a strong correlation between MN fre-
spraying, showed a statistically significant increase in fre-
quency and increasing age are well documented (Bonassi et al., quency of BNMN in the three regions where glyphosate was
1995, 2001; Bolognesi et al., 1997a). sprayed. However, this was not consistent with the rates of

Downloaded At: 20:08 2 October 2009ignificant effect of smoking, con- application use in the regions. The increase in frequency of
firming findings from the literature (Bonassi et al., 2003) BNMN in Valle (application rate = 1 kg a.e. glyphosate/ha)
although prevalence of smoking in our study population was was greater than that in Nariño and Putumayo (3.69 kg a.e.

small (7–20%, Table 1). No association with alcohol consump- glyphosate/ha).
tion was observed. A higher susceptibility of people of African There was no significant association between self-reported
origin compared to the mestizo group was suggested by a
direct contact with eradication sprays and frequency of
greater baseline frequency of BNMN and increased frequency BNMN. The frequency of BNMN in participants who self-
at the second sampling period. reported that they were exposed to glyphosate because they
There was some indication of an association between
entered the field immediately after spraying (to pick the coca
BNMN and exposure to pesticides in general. The lowest fre- leaves), felt spray drops in their skin, or they thought they were
quency of BNMN was observed in Sierra Nevada de Santa exposed because they had contact with the chemical in the air,

Marta, where people self-reported that they did not use pesti- was not significantly greater than in subjects living in the same
cides. The mean frequency of BNMN in this group of subjects areas but who were not present during spraying. Decreases in
(1.83 ± 0.97) was similar to that observed in healthy unexposed
frequency of BNMN in the recovery period after glyphosate
subjects for the same range of age (Bolognesi et al., personal spraying were not consistent. The third sampling, 4 mo after
communication). The higher mean frequency of BNMN spraying, demonstrated a statistically significant decrease in
observed in Boyacá and Valle del Cauca (5.64 ± 1.72 and 5.75
frequency of BNMN only in Nariño.
± 2.48, respectively) and that in Nariño and Putumayo (4.12 ± Overall, these results suggest that genotoxic damage associ-
1.65 and 3.65 ± 1.51, respectively), compared to Santa Marta, ated with glyphosate spraying, as evidenced by the MN test, is

are in agreement with similar biomonitoring studies carried outsmall and appears to be transient. The frequencies of BNMN in
in subjects exposed to pesticides using the MN test or other Nariño and Putumayo during the second and the third sampling
genetic endpoints (Bolognesi, 2003; Bull et al., 2006). fell within the range of values observed in Boyacá, an area

408 Annex 131-I

BIOMONITORING GENOTOXIC RISK IN AGRICULTURAL WORKERS 995

where people were exposed to a complex mixture of different of skin exposed, region of skin exposed, differences in rates of
pesticides (including glyphosate). A greater increase in fre- penetration, or personal hygiene.
quency of BNMN was observed in Valle del Cauca, but it can- Given the situation, the best approach possible, a prospec-

not be attributed only to the glyphosate exposure, because the tive cohort, was used but the need to use better procedures to
application rate of the herbicide in this area was one-third com- estimate the exposure is acknowledged. Based on the applica-

pared with that in Nariño and Putumayo. This conclusion is ble Bradford–Hill guidelines (Hill, 1965), it is not possible to
further supported by the frequency of MN in mononucleated assign causality to the increases in frequency of BNMN

cells (MOMN), which provides an indication of the back- observed in our study. There was a smaller frequency of
ground level of chromosome/genome mutations accumulated BNMN and MOMN in the region of no pesticide use com-
in vivo (Manteuca et al., 2006). A statistically significant pared with the regions where pesticides (including glypho-

increase of MOMN was observed in Boyacá and Valle del sate) were used, which is consistent with other reports in the
Cauca before and after the aerial spraying, suggesting exposure literature. Although temporality was satisfied in the increase

to other genotoxic compounds in these populations was inde- in frequency of BNMN after spraying, this response did not
pendent of the exposure to glyphosate. Evidence indicates that show strength as it was not co nsistently correlated with the
the genotoxic risk potentially associated with exposure to rate of application. Recovery was also inconsistent with

glyphosate in the areas where the herbicide is applied for erad- decreases in frequency of BNMN in the areas of eradication
ication of coca and poppy is of low biological relevance. One spraying but not in the area where lower rates were applied

of the strengths of our study was the detection of a transient on sugar cane.
chromosomal damage, evaluated as MN frequency in periph- Further studies are needed to better characterize the poten-

eral blood of the exposed subjects, since it was possible to tial genotoxic risk associated with the application of glypho-
compare the baseline before spraying with the effects detected sate for sugar cane maturation. The smaller number of subjects
immediately after spraying. Glyphosate persists in the environ- recruited in this study and small amount of information about

ment for only a short time (half-life for biological availability the exposure precluded any conclusions. Many pesticides are
in soil and sediments is hours, and 1-3 d in water; Giesy et al., used in conventional agriculture in Colombia and many pesti-

2000), is rapidly excreted by mammals and other vertebrates cides are used in the production of coca (Solomon et al., 2007a,
(Williams et al., 2000; Acquavella et al., 2004) and chronic 2007b); however, there is not sufficient information to corre-

effects, if any, would not be expected. late the frequency of MN to the pesticide exposure.
One of the major drawbacks of environmental epidemiol-
ogy studies is the characterization of exposures to the agents

being investigated. In this study two approaches were used to REFERENCES
characterize exposures to glyphosate: ecological and self- Acquavella, J. F., Alexander, B. H., Mandel, J. S., Gustin, C., Baker, B.,
Chapman, P., and Bleeke, M. 2004. Glyphosate biomonitoring for farmers
reported. In the ecological study design, frequency of BNMN and their families: Results from the farm family exposure study. Environ.
Downloaded Ain participants was compared from regions with different pat- Health Perpect. 112:321–326.
terns of pesticide use. As previously discussed (Sanin et al.,
Arbuckle, T. E., Cole, D. C., Ritter, L., and Ripley, B. D. 2004. Farm
2009), this ecological design may result in misclassification of children’s exposure to herbicides: Comparison of biomonitoring and
exposures (Arbuckle et al., 2004), but as an exploratory assess- questionnaire data. Epidemiology 15:187–194.
Baylis, A. D. 2000. Why glyphosate is a global herbicide: Strengths,
ment of exposure it is useful (Ritter et al., 2006). weaknesses and prospects. Pestic. Manage. Sci. 56:299–308.
Others have attempted to improve assessment of exposure Bolognesi, C. 2003. Genotoxicity of pesticides: A rveiew of human biomonitoring
studies.Mutat. Res. 543:251–272.
to pesticides in epidemiological studies. One study used a self- Bolognesi, C., Abbondandolo, A., Barale, R., Casalone, R., Dalpraà, L., De
administered questionnaire for the assessment of exposure to
glyphosate, which was defined as (a) ever personally mixed or Ferrari, M., Degrassi, F., Forni, A., Lamberti, L., Lando, C., Migliore, L.,
Padovani, D., Pasquini, P., Puntoni, R., Sbrana, I., Stella, M., and Bonass, S.
applied products containing glyphosate; (b) cumulative life- 1997a. Age-related increase of baseline frequencies of sister chromatid
time days of use, or “cumulative exposure days” (years of use exchanges, chromosome aberrations, and micronuclei in human lympho-
cytes. Cancer Epidemiol. Biomarkers. Prev. 6:249–256.
times days/year); and (c) intensity-weighted cumulative expo- Bolognesi, C., Bonatti, S., Degan, P., Gallerani, E., Peluso, M., Rabboni, R.,
sure days (years of use times days/year times estimated inten- Roggieri, P., and Abbondandolo, A. 1997b. Genotoxic activity of glypho-
sity level) (De Roos et al., 2005). A pesticide exposure score sate and its technical formulation, Roundup. J. Agric. Food. Chem.

based on self-reported work practices was recently developed 45:1957–1962.
to estimate annual exposure level (Firth et al., 2007). Based on Bonassi, S., Bolognesi, C., Abbondandobo, A., Barale, R., Bigatti, P., Camurri,
L., Dalpra, L., De Ferrari, M., Forni, A., Lando, C., Padovani, P., Pasquini,
an algorithm to estimate lifetime exposure to glyphosate from R., Stella, M., and Puntoni, R. 1995. Influence of sex on cytogenetic end-
questionnaire information, a moderate correlation was found points: Evidence from a large human sample and review of the literature.
Cancer Epidemiol. Biomarkers. Prev. 4:671–679.
with concentrations of glyphosate in urine and no significant Bonassi, S., Fenech, M., Lando, C., Lin, Y. P., Ceppi, M., Chang, W. P.,
correlation with self-reported exposure (Acquavella et al., 2004). Holland, N., Kirsch-Volders, M., Zeiger, E., Ban, S., Barale, R., Bigatti, M.,
In our study, questions related to whether there was direct Bolognesi, C., Jia, C., Di Giorgio, M., Ferguson, L. R., Fucic, A., Lima, O.

contact with the spray were used but this did not consider area G., Hrelia, P., Krishnaja, A. P., Lee, T. K., Migliore, L., Mikhalevich, L.,

409 Annex 131-I

996 C. BOLOGNESI ET AL.

Mirkova, E., Mosesso, P., Müller, W. U., Odagiri, Y., Scarffi, M. R., Kier, L. D., Stegeman, S. D., Dudek, S., McAdams, J. G., Flowers, F. J.,

Szabova, E., Vorobtsova, I., Vral, A., and Zijno, A. 2001. HUman Micro- Huffman, M. B., and Heydens, W. F1. 997. Genotoxicity studies of glyphosate,
Nucleus project: International database comparison for results with the alachlor and butacholr formulations.Fundam. Appl. Toxicol. 36:305.
cytokinesis-block micronucleus assay in human lymphocytes: I. Effect of Li, A. P., and Long, T. J. 1988. .An evaluation of genotoxic potential of
laboratory protocol, scoring criteria, and host factors on the frequency of glyphosate. Fundam. Appl. Toxicol. 10:537–546.

micronuclei. Environ. Mol. Mutagen. 37:31–45. Manteuca, R., Lombaert, N., V, A. P., Decordier, I., and Kirsch-Volders, M.
Bonassi, S., Neri, M., Lando, C., Ceppi, M., Lin, Y.-P., Chang, W. P., Holland, 2006. Chromosomal changes: induction,detection methods and applicability
N., Kirsch-Volders, M., Zeiger, E., Fenech, M., and The HUMN collabora- in human biomonitoring.Biochimie 88:1515–1531.
tive group. 2003. Effect of smoking habit on the frequency of micronuclei Marc, J., Bellé, R., Morales, J., Cormier, P., and Mulner-Lorillon, O. 2004a.

in human lymphocytes: Results from the Human MicroNucleus project. Formulated glyphosate activates the DNA-response checkpoint of the cell
Mutat. Res. 543:155–166. cycle leading to the prevention of G2/M transition.Toxicol. Sci. 82:436–442.
Bull, S., Fletcher, K., Boobis, A. R., and Battershill, J. M. 2006. Evidence for Marc, J., Mulner-Lorillon, O., and Bellé, R. 2004b. Glyphosate-based pesti-
genotoxicity of pesticides in pesticide applicators: A review. Mutagenesis cides affect cell cycle regulation. Biol. Cell. 96:245–247.

21:93–103. Martinez, A., Reyes, I., and Reyes, N. 2007. Cytotoxicity of the herbicide glypho-
Costa, C., Teixeira, J. P., Silva, S., Roma-Torres, J., Coelho, P., Gaspar, J., sate in human peripheralblood mononuclear cells.Biomédica27:594–604.
Alves, M., Laffon, B., Rueff, J., and Mayan, O. 2006. Cytogenetic and McDuffie, H. H., Pahwa, P., McLaughlin, J. R., Spinelli, J. J., Fincham, S.,

molecular biomonitoring of a Portuguese population exposed to pesticides. Dosman, J. A., Robson, D., Skinnider, L., and Choi, N. W. 2001. Non-
Mutagenesis 21:343–350. Hodgkin’s lymphoma and specific pesticide exposures in men: Cross-
De Roos, A. J., Blair, A., Rusiecki, J. A., Hoppin, J. A., Svec, M., Dosemeci, Canada study of pesticides and health. Cancer Epidemiol. Biomarkers.
M., Sandler, D. P., and Alavanja, M. C. 2005. Cancer incidence among Prev. 10:1155–1163.

glyphosate-exposed pesticide applicators in the Agricultural Health Study. Monroy, C. M., Cortes, A. C., Sicard, D. M., and Groot, H. 2005. Citotox-
Environ. Health Perspect. 113:49–54. icidad y genotoxicidad de células humanas espuestas in vitro a glifosato.
De Roos, A. J., Zahm, S. H., Cantor, K. P., Weisenburger, D. D., Holmes, F. Biomédica 25:335–345.
F., Burmeister, L. F., and Blair, A. 2003. Integrative assessment of multipleMontero, R., Serrano, L., Araujo, A., Davila, V., Ponce, J., Camacho, R.,

pesticides as risk factors for non-Hodgkin’s lymphoma among men. Morales, E., and Mendez, A. 2006. Increased cytogenetic damage in a zone
Occup. Environ. Med. 60:E11. in transition from agricultural to industrial use: Comprehensive analysis of
Dimitrov, B. D., Gadeva, P. G., Benova, D. K., and Bineva, M. V. 2006. Com- the micronucleus test in peripheral blood lymphocytes. Mutagenesis
parative genotoxicity of the herbicides Roundup, Stomp and Reglone in 21:335–342.

plant and mammalian test systems. Mutagenesis 21:375–382. Moriya, M., Ohta, T., Watanabe, K., Miyasawa, T., Kato, K., and Shirasu, Y.
Duke, S. O., and Powles, S. B. 2008. Glyphosate: A once-in-a-century herbi- 1983. Further mutagenicity studies on pesticides in bacterial reversion
cide. Pestic. Manage. Sci. 64:319–325. assay systems. Mutat. Res., 116:185–216.
Eriksson, M., Hardell, L., Carlberg, M., and Akerman, M. 2008. Pesticide Paz-y-Miño, C., Sánchez, M. E., Arévalo, M., Muñoz, M. J., Witte, T., De-la-

exposure as risk factor for non-Hodgkin lymphoma including histopatho- Carrera, G. O., and Paola, L. E. 2007. Evaluation of DNA damage in an
logical subgroup analysis. Int. J. Cancer 123:1657–1663. Ecuadorian population exposed to glyphosate.Genet. Mol. Biol.30:456–460.
Fenech, M. 2007. Cytokinesis-block micronucleus cytome assay. Nat. Prot. Peixoto, F. 2005. Comparative effects of the Roundup and glyphosate on mito-
2:1084–1104. chondrial oxidative phosphorylation. Chemosphere 61:1115–1122.
32
Fenech, M., and Morley, A. A. 1985. Measurement of micronuclei in lympho- Peluso, M., Munnia, A., Bolognesi, C., and Parodi, S. 1998. P-postlabeling
cytes. Mutat. Res., 147:29–36. detection of DNA adducts in mice treated with the herbicide Roundup.
Fenech, M., and Rinaldi, J. 1994. The relationship between micronuclei in Environ. Mol. Mutagen. 31:55–59.
human lymphocytes and plasma levels of vitamin C, vitamin E, vitamin Rank, J., Jensen, A. G., Skov, B., Pedersen, L. H., and Jensen, K. 1993. Geno-

Downloaded At: 20:08 2 October 2009 Carcinogenesis 15:1405–1411. toxicity testing of Roundup and its active ingredient glyphosate isopropy-
Firth, H. M., Rothstein, D. S., Herbison, G. P., and McBride, D. I. 2007. lamine using the mouse bone marrow micronucleus test,Salmonella
Chemical exposure among NZ farmers. Int. J. Environ. Health. Res. mutagenicity test andAlliumanaphase-telophase test.Mutat. Res.300:29–36.
17:33–44. Richard, S., Moslemi, S., Sipahutar, H., Benachour, N., and Seralini, G.-E.

Gebel, T., Kevekordes, S., Pav, K., Edenharder, R., and Dunkelberg, H. 1997. 2005. Differential effects of glyphosate and Roundup on human placental
In vivo genotoxicity of selected herbicides in the mouse bone marrow cells and aromatase. Environ. Health Perpect. 113:716–720.
micronucleus test. Arch. Toxicol. 71:193–197. Ritter, L., Goushleff, N. C. I., Arbuckle, T., Cole, D., and Raizenne, M. 2006.

Giesy, J. P., Dobson, S., and Solomon, K. R. 2000. Ecotoxicological risk Addressing the linkage between exposure to pesticides and human health
assessment for Roundup® herbicide. Rev. Environ. Contam. Toxicol. effects—Research trends and priorities for research 1. J. Toxicol. Environ.
167:35–120. Health B 9:441–456.
Gopalan, H. N. B., and Njagi, G. D. E.1981. Mutagenicity testing of pesticides: Rull, R. P., Ritz, B., and Shaw, G. M. 2006. Neural tube defects and maternal

III. Drosophila: Recessive sex-linked lethals.Genetics97(Suppl.):S44. residential proximity to agricultural pesticide applications. Am. J. Epide-
Grisolia, C. K. 2002. A comparison between mouse and fish micronucleus test miol. 163:743–753.
using cyclophosphamide, mitomycin C and various pesticides. Mutat. Res. Sanin, L.-H., Carrasquilla, G., Solomon, K. R., Cole, D. C., and Marshall, E. J.
518:145–150. P. 2009. Regional differences in time to pregnancy among fertile women

Hardell, L., and Eriksson, M. 1999. A case-control study of non-Hodgkin lym- from five Colombian regions with different uses of glyphosate. J. Toxicol.
phoma and exposure to pesticides. Cancer 85:1353–1360. Environ. Health A 72:949–960.
Hardell, L., Eriksson, M., and Nordstrom, M. 2002. Exposure to pesticides Sirisattha, S., Momse, Y., Kitagawa, E., and Iwahashi, H. 2004. Genomic pro-
as risk factor for non-Hodgkin’s lymphoma and hairy cell leukemia: file of roundup treatment of yeast using DNA microarray analysis. Envi-

Pooled analysis of two Sw edish case-control studies. Leuk. Lymphoma ron. Sci. 11:313–323.
43:1043–1049. Solomon, K. R., Anadón, A., Brain, R. A., Cerdeira, A. L., Crossan, A. N.,
Herbert, V. 1987. Recommended dietary intakes (RDI) of folate in humans. Marshall, A. J., Sanin, L. H., and Smith, L. 2007a. Comparative hazard
Am. J. Clin. Nutr. 45:661–670. assessment of the substances used for production and control of coca and

Heydens, W. F., Healy, C. E., Hotz, K. J., Kier, L. D., Martens, M. A., Wilson, poppy in Colombia. In Rational environmental management of agro-
A. G. E., and Farmer, D. R. 2008. Genotoxic potential of glyphosate formu- chemicals: Risk assessment, moni toring, and remedial action. ACS
lations: Mode-of-action investigations.J. Agric. Food. Chem.56:1517–1523. Symposium Series no. 966 (vol. 966), eds. Kennedy, I. R., Solomon, K.
Hill, A. B. 1965. The environment and disease: association or causation? Proc. R., Gee, S., Crossan, A. N., Wang, S., and Sanchez-Bayo, F. pp. 87–99.

R. Soc. Med. 58:295–300. Washington, DC: American Chemical Society.

410 Annex 131-I

BIOMONITORING GENOTOXIC RISK IN AGRICULTURAL WORKERS 997

Solomon, K. R., Anadón, A., Carrasquilla, G., Cerdeira, A., Marshall, J., and reproductive and child health outcomes and environmental chemical con-
Sanin, L.-H. 2007b. Coca and poppy eradication in Colombia: Environ- taminants. J. Toxicol. Environ. Health B 11:373–517.
mental and human health assessment of aerially applied glyphosate. Rev. Wildeman, A. G., and Nazar, R. N. 1982. Significance of plant metabolism in
Environ. Contam. Toxicol. 190:43–125. the mutagenicity and toxicity of pesti. an. J. Genet. Cytol.24:437–449.
UNODC. 2007. World drug report 2007. United Nations Office on Drugs and Williams, G. M., Kroes, R., and Munro, I. C. 2000. Safety evaluation and risk

Crime. Accessed January 29, 2008. http://www.unodc.org assessment of the herbicide Roundup® and its active ingredient, glypho-
U.S. Environmental Protection Agency. 1993. R.E.D. Facts Glyphosate. Tech- sate, for humans. Regul. Toxicol. Pharmacol. 31:117–165.
nical report EPA 738-R-93-014. Washington, DC: U.S. Environmental Woodburn, A. T. 2000. Glyphosate: production, pricing and use worldwide.
Protection Agency. Pestic. Manage. Sci. 56:309–312.
Wigle, D. T., Arbuckle, T. E., Turner, M. C., Berube, A., Yang, Q., Lui, S.,orld Health Organization International Program on Chemical Safety. 1994.

and Krewski, D. 2008. Epidemiologic evidence of relationships between Glyphosate (vol. 159). Geneva: WHO IPCS.

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411412 Annex 132

CENTRE FORT OXICOLOGY ANDENVIRONMENTAL H EALTH,L.L.C.,NIVERSITY OF
ARKANSAS FORM EDICALSCIENCES,GLYPHOSATEF REQUENTLY ASKED
QUESTION, 2009

(Available at:
http://www.akrr.com/pdf/PR_2335%20Glyphosate_Toxocologist%20FAQ.pdf (last

visited 7 March 2010))

413Annex 132

Center for Toxicology and Environmental Health, L.L.C.

5120 North Shore Drive North Little Rock, AR 72118 Phone: 501.801.8500 Fax: 501.801.8501 www.cteh.com

GLYPHOSATE
Frequently Asked Questions

What is glyphosate?
Glyphosate is N‐phosphonomethyl glycine. Glyphosate agricultural products are broad‐spectrum,

non‐selective herbicides used on most species of green plants. You may know the product name, or
have purchased the product for your own home use by one of its common names i.e., Roundup,

WeatherMax or Roundup Ultramax. Glyphosate has been used for more than 35 years and is
probably the world’s most widely used herbicide. It is registered in more than 130 countries and is

approved for weed control in more than 100 crops. The glyphosate formulation that will be used by
the Alaska Railroad Corporation (hereafter referred to as “ARRC”) is called Aquamaster Herbicide

and contains 53.8% glyphosate (isopropylamine salt). In addition to glyphosate, the formulation
typically includes water and a surfactant system. The surfactant system enables the product to

adhere to the plant surfaces. A few days after treatment, the plant wilts and yellows. In addition to
being approved for use on land, Aquamaster is approved for weed control in aquatic environments,

including ponds and reservoirs, waterfowl sanctuaries, and recreational waterways. Only a few
herbicides have the favorable toxicological and environmental characteristics that allow them to be

directly applied to a quatic vegetation. Most recently, AquaMaster was selected by the State of
Florida to rid the Everglades of invasive weeds. The University of Florida in their publication on

glyphosate, discuss why land managers should continue to use glyphosate containing products to
protect managed habitats from weeds without concern for unreasonable adverse environmental

impacts.

Glyphosate is water soluble and binds tightly to soil. The product works by disrupting a plant

enzyme essential for plant growth. The enzyme is called EPSP synthetase and is not present in
humans or animals. Therefore, the biochemical pathway affected is specific to plant species (not

humans, mammals, or fish) contributing to the low risk to human and animal health from use of
glyphosate according to the label directions used by ARRC.

In addition to agricultural use, glyphosate is used to control weeds in utility right‐of‐ways, on

roadsides, along railways or in places around the home such as sidewalks and gardens. Glyphosate
is also used by Wildlife organizations to protect and restore wildlife habitats threatened by invasive,

non‐native vegetation. Conservation groups have chosen glyphosate formulations because of their
effectiveness against most weeds and because they have very low toxicity to wildlife.

How o d glyphosate compare in toxicity to other commonly used chemicals?

Glyphosate has been the subject of hundreds of health, safety, and environmental studies. To get a
clear picture of the environmental and toxicological characteristics of glyphosate it is important to

consider the total weight of evidence from scientific studies provided by regulatory agencies,

University of Arkansas for Medical Sciences Bioventures Program Associate

414 Annex 132

industry, universities, governmental agencies, and scientists from around the World. The U.S

Environmental Protection Agency (EPA), Health Canada, European Commission, U.S. Department of
Agriculture Forest Service, World Health Organization and other scientists have reviewed this data.

Those reviews applied internationally accepted methods, principles and procedures in toxicology
and have determined that there are no grounds to suggest concern for human health. Their

overwhelming consensus is that glyphosate when used according to label directions, poses no
unreasonable risk to people, wildlife, or the environment. Researchers from New York Medical

College similarly conclude that glyphosate does not have the potential to produce adverse effects in
humans. Short‐term exposure to glyphosate and its breakdown products is estimated to be 7,350

to 1,730,000 times lower than their corresponding median lethal dose (LD50) values, thus
demonstrating that potential short‐term exposure is not a health concern. Several toxicology text

books have developed a relative ranking system for chemicals by their LD50 values to help people
answer the question “how toxic is this stuff”? The ranking system listed below puts glyphosate in

the category of slightly toxic and is less toxic than table salt.

Table 1.
General Toxicity Ranking Categories

Toxicity ranking Dose (mg/kg) For Average Adu lt

Practically nontoxic >15,000 > 1 quart

Slightly Toxic 5,000‐15,000 1 pint to 1 quart

Moderately toxic 50 – 5,000 1 ounce to 1 pint

Very toxic 50 – 500 1 teaspoon to 1 ounce

Extremely toxic 5 – 50 7 drops to 1 teaspoon

Supertoxic < 5 < 7 drops

415Annex 132

Table 2
Relative Toxicity Ranking of Glyphosate Compared to Other Compounds

Actual Ranking # LD50(mg/kg) Chemical

14 10,000 Alcohol (ethanol)

13 >5,000 Glyphosate

12 4,000 Sodium Chloride (table salt)

11 2,400‐3,480 biphenyl

10 1,500 Ferrous Sulfate (iron supplement)

9 1,375 Malathion (pesticide)

8 900 Morphine (opiate analgesic)

7 150 Phenobarbital (sedative)

6 142 Tylenol (acetaminophen)

5 2 Strychnine (rat poison)

4 1 Nicotine (stimulant found in
cigarettes)

3 0.5 Curare (arrow poison)

2 0.001 2,3,7,8‐TCDD (dioxin)

1 0.00001 Botulinum toxin (food poison)

Can glyphosate cause genetic damage?

Genetic toxicity tests are performed to provide information on the production of heritable changes
(mutations) that could lead to further adverse biological consequences. In other words, will the

chemical cause DNA damage? Glyphosate has been studied extensively in a wide battery of genetic
toxicity tests. Such extensive data sets are sometime difficult to interpret but for glyphosate this is

not the case. The overwhelming evidence indicates that glyphosate does not damage DNA. No
genotoxic activity is observed in standard assays conducted according to international guidelines. A

number of regulatory agencies, scientists, and researchers conclude that glyphosate is neither
mutagenic nor clastogenic (causing chromosome breaks). Thus, glyphosate does not pose a risk of

heritable (passed from parent to child) or somatic (body cell) mutations in humans.

416 Annex 132

Can glyphosate cause cancer?
The long‐term toxicity and cancer potential of glyphosate has been evaluated in studies with mice

and rats. Glyphosate was not carcinogenic to either species. These studies and results have been
evaluated by a number of regulatory agencies and scientific organizations. Each group has

concluded that glyphosate is not carcinogenic. The EPA uses a summary ranking system for human
and animal cancer studies. These rankings place the overall evidence in classification Groups A

through D. Group A rankings are chemicals that are known human carcinogens, whereas Group D
chemicals are not classifiable as to human carcinogenicity. Accordingly, EPA has classified

glyphosate as Group D, “Not classifiable as to human carcinogenicity” because there is inadequate
evidence that glyphosate causes cancer in animals and no evidence that it causes cancer in humans.

Have other studies indicated health issues with Glyphosate?

A basic principle of toxicology was first stated by the 16th century physician Paracelsus, who said
“all substances are poisons; there is none which is not a poison. The right dose differentiates a

poison and a remedy.” In other words, all chemicals are toxic at some dose, and conversely, there is
some dose at which no toxicity i apparent. For all chemicals, the toxicity resulting from exposure is

determined by the dose of chemical absorbed by the organism. As the dose of a chemical
increases, its effects increase in magnitude and severity; conversely, as the dose of a chemical

decreases, so does the magnitude and severity of its effects. This concept is termed “dose‐
response,” and forms one of the basic foundations of toxicology. For all chemicals there is a dose

(termed the “threshold dose”) below which no effect is elicited. This is true for both the beneficial
(pharmacological) and harmful (toxicological) effect of chemicals. Some studies show toxicity of

glyphosate at very high concentrations. Given the fact that there are literally hundreds of
toxicology studies on glyphosate, the question should not be “have other studies indicated health

issues with glyphosate?”, but rather “would health effects be expected from AKRR’s use of
glyphosate along railroad right‐of‐ways?”

The answer to this question is that glyphosate poses no substantial concern for systemic toxic

effects in workers or the general public at the recommended application rate. Adult applicators

and children have been identified as the most sensitive subpopulations because they have the
highest potential exposures to glyphosate. Estimates o f exposure to these two subpopulations are

typically evaluated using a “Margin of Exposure” or MOE analysis where toxicologists compare the
lowest No Observable Adverse Effect Levels (NOAELs) determined from animal and human studies

to worst‐case levels of human exposure. MOEs of greater than 100 are considered by authoritative
bodies to indicate confidence that no adverse health effects would occur. The MOEs for worst‐case

chronic exposure to glyphosate ranged from 3,370 to 5,420. Based on these values, it can be
concluded that glyphosate does not have the potential to produce adverse effects in humans. The

only real risk from glyphosate exposure is possible skin or eye irritation from direct contact with the

417Annex 132

liquid by those individuals preparing the spray for weed control. Irritant effects are considered to

be transient and reversible.

What is the environmental fate of glyphosate?
As with the human health risks described above, there is very little indication that glyphosate will

cause adverse effects in the environment at the anticipated levels of exposure from use of this
product on railroad right‐of‐ways. Glyphosate binds readily with soil particles, which limits its

movement in the environment; therefore it does not have herbicidal activity once it contacts soil.
Glyphosate is also not absorbed from the soil by plant roots. Glyphosate has a low Koc (measure of

soil adsorption) which is an indication that glyphosate will not move readily through soil and is
unlikely to leach into non‐target areas. When applied to foliage, glyphosate is readily absorbed and

translocated to various parts of plants via the phloem. Glyphosate is readily degraded by soil
microbes with an average half‐life of two months in soil and two to 10 weeks in water. The major

degradation product is aminomethylphosphonic acid (AMPA). AMPA is further degraded in the
environment to carbon dioxide and phosphate. In plants, glyphosate is slowly metabolized. The

potential for glyphosate to leach into groundwater has been evaluated in a number of studies which
reveal that glyphosate is unlikely to cause groundwater contamination. Most studies show no

adverse effect on soil microorganisms.

How rapidly does glyphosate break down in the Alaska environment?
The University of Alaska with assistance from the Alaska Railroad and the US Department of

Agriculture is conducting a study on the environmental fate and soil dissipation of glyphosate in the
Alaskan environment. The first phase of the study near Seward included two separate rail bed sites;

one site in the Seward Yard and the other near the Upper Trail Lake area. Monitoring wells were
installed and samples of glyphosate were taken over time from surface soils, root zone soils, below

root zone soils, and groundwater. Results agree with the discussion above in that glyphosate is
degraded relatively rapidly in Alaskan soil and does not migrate to a significant degree in the soil

environment. In addition, the results indicate that the levels of glyphosate detected in soil and
groundwater after treatment would not be toxic to humans or animals including fish. The second

phase of the study is being concluded near Fairbanks.

What effect does Glyphosate have on birds, insects, and mammals?

Glyphosate has been tested on a variety of wildlife birds and mammals in both laboratory and wild
land environments. Furthermore, there are several available field studies that examine the effects

of glyphosate application comparable to those that will be used by AKRR. The toxicity studies on
terrestrial animals are generally consistent with those on experimental mammals. The available

field studies clearly show that at plausible levels of ambient exposure, direct toxic effects are
unlikely. In fact, if any effects are seen in terrestrial mammals after the application of glyphosate,

they are most likely to be associated with changes in habitat rather than direct toxic effects. The
changes would be no different than from mechanical clearing of vegetation. Data for single

418 Annex 132

exposures classify glyphosate as practically non‐toxic to tested insects and birds. Glyphosate is no

more than slightly toxic to mammals. EPA does not expect that most endangered terrestrial
organisms will be affected by the registered uses of glyphosate. The small mammal is a

conservative target species for characterizing risks because small organisms, compared with large
organisms generally receive higher doses at fixed levels of exposure in environmental media (e.g.,

contaminated food, water). Also, available toxicity data does not suggest systematic differences in
sensitivity to glyphosate among species. The primary route of exposure for a terrestrial animal is

from contaminated vegetation. For this source, levels of exposure remain below those of concern
even at the highest allowed application rates of glyphosate. At application rates that ARRC would

use, levels of exposure are substantially below those of concern. This analysis is consistent with
field studies on glyphosate that indicate that it would be unlikely for glyphosate to have direct toxic

effects on wildlife. Based on current data, EPA has determined that the effects of glyphosate on
birds and mammals are minimal. The available data indicate that glyphosate does not

bioaccumulate in terrestrial species including carnivores, herbivores, and omnivores.

What effect does Glyphosate have on aquatic species including fish?
There is not much evidence that aquatic animals or plants will be adversely affected by normal

applications of glyphosate. Although glyphosate is registered for use as an aquatic herbicide, it is
only effective on plants with vegetation growing above the water level. Most species of algae and

macrophytes do not appear to be more sensitive than fish or aquatic invertebrates to glyphosate.
For most aquatic species, glyphosate levels of 1 mg/L are not likely to cause adverse effects. Field

studies indicate that maximum initial concentrations of glyphosate in water after aerial or ground
applications is considerably less than 1 mg/L.

A review of the published toxicity studies on fish indicate that glyphosate is relatively non‐toxic to

fish with 24‐96 hours LC50 values ranging from approximately 10 mg/L to >200 mg/L. EPA and
USDA examined the toxicity of glyphosate to a variety of fish species including rainbow trout,

various salmonid species including chinook, coho, sockeye, as well as fingerlings, fry, and early life
stages. EPA and USDA determined that glyphosate effects on fish would not be expected based on

registered application rates. Glyphosate does not bioaccumulate in fish.

What happens f I iat berries from along the tracks?

Glyphosate application along right‐of‐ways creates the potential for accidental overspray of wild
foods such as berries that could be later collected for consumption. Consideration of actual

glyphosate use patterns, the percentage of forests or roadsides that actually receive treatment, and
the resulting phytotoxic effects on the sprayed plants, suggests that inadvertent exposure will be

extremely unlikely. Residual levels of glyphosate arising from mock overspray of berries have been
measured and the potential dietary exposure quantified. Peak glyphosate residue levels were 19.5

ug/g and it was estimated that maximal berry consumption for an individual might be 150 g for an
adult and 30 g for a 1‐6 year old child. These parameters predict an exposure of 45 ug/kg body wt

419Annex 132

for both subgroups and rely on the assumption that the surface residues were not reduced by

washing before consumption. This dose is considerably below the NOAEL for chronic toxicity and
more than two times below the EPA reference dose for glyphosate, indicating that occasional eating

of berries containing a glyphosate residue would not result in adverse human health effects.

What can you tell me about the toxicity of the “inert ingredients” i.e., the surfactant Agri‐Dex?
Agrid‐Dex is a trade name of a product and is approved for use in aquatic applications. Agri‐Dex is a

surfactant used with glyphosate. The surfactant system enables the products to adhere to the
surface of leaves so the active ingredient (glyphosate) can penetrate. AGRI‐DEX is designed to be

compatible with a wide range of pesticides and form stable emulsions in their tanks mixes. Agri‐Dex
is classified as practically non‐toxic to both fish and vertebrates.

What is the overall Risk of Herbicide Program Proposed by ARRC?

To address the overall risk and potential toxicity impact of glyphosate uses specific to the state of
Alaska ARRC contracted a study with the University of Alaska Fairbanks to address the transport nd a

degradation of glyphosate under real‐world conditions that mimic use by the ARRC. As previously
mentioned the preliminary results of this study indicate that glyphosate has low mobility on the

environment and is being degraded in the soil. Concentrations of glyphosate measured in soil and
water were well below those that would harm humans or animals, including fish. The results of this

study build on the large knowledge base of the environmental impacts of glyphosate which indicate
that the herbicide program proposed by ARRC would not have an adverse impact on the health of

humans or the environment.

420 Annex 132

REFERENCES

Barnes, D. L. (Water and Environment Research Center, Department of Civil and Environmental
Engineering, University of Alaska Fairbanks). Letter To: Matthew Kelzenberg, Alaska Railroad

Corporation. Glyphosate Sampling. 2009 Jun 9.

Benachour, N. and Seralini, G. E. Glyphosate formulations induce apoptosis and necrosis in human
umbilical, embryonic, and placental cells. Chem Res Toxicol. 2009 Jan; 22(1):97‐105.

CCME. Canadian Water Quality Guidelines for the Protection of Aquatic Life: GlyphosateIn: CCME.
Canadian Environmental Quality Guidelines. Winnepeg: Canadian Council of Ministers of

the Environment.; 1999.

Durkin, P. R. Glyphosate‐‐Human and Ecological Risk Assessment Final Report. Arlingotn, VA: USDA,
Forest Service, Forest Health Protection; 2003 Mar 1; SERA TR 02‐43‐09‐04a.

FSH 6709.11 Health and Safety Code Handbook. Washington, DC: U.S. Department of Agriculture,
Forest Service; 1999.

Helena Chemical Company. MSDS: Agri‐Dex. Collierville, TN: Helena Chemical Company; 2005 Apr

29.

HSDB for Glyphosate. In: Hazardous Substance Data Bank [TOXNET]. Bethesda, MD: National Library

of Medicine, National Toxicology Information Program; 2009.
http://toxnet.nlm.nih.gov/

Johnson, W. W. and Finley, M. T. Glyphosate. In: Johnson, W. W. and Finley, M. T. Handbook of

acute toxicity of chemicals to fish and aquatic invertebrates: summaries of toxicity tests
conducted at Columbia National Fisheries Research Laborator, 1965‐1978. Washington, DC:
U.S. Department of the Interior, Fish and Wildlife Service; 1980; p. 43.

Kuppelwieser, H. Vegetation control as part of environmental strategy of Swiss Federal Railways.
Japan Railway & Transport Review. 1998 Sep; 17:8‐11

Mark Twain National Forest 2005 Forest Plan. Milwaukee, WI: U.S. Department of Agriculture,

Forest Service, Eastern Region Office; 2005 Sep.

Monsanto Company. MSDS: AquaMaster Herbicide. St. Louis, MO: Monsanto Company; 2005 Feb
3.

Schmidt, B.; Siegemund, B.; Ehses, H.; Zietz, E., and Miersch, H. Risk of groundwater pollution from
weed control on railway tracks. In: Del Re, A., Editor. Human and environmental exposure to
xenobiotics : proceedings of the XI Symposium Pesticide Chemistry; Cremona, Italy. Pavia: La

goliardica pavese; 1999: 591‐597.

Schuette, J. Environmental fate of glyphosate. Sacramento, CA: California Department of Pesticide

Regulation; 1998 Nov; www.cdpr.ca.gov/docs/emon/pubs/fatememo/glyphos.pdf .

421Annex 132

Specimen Label: Agri‐Dex: a non‐ionic spray adjuvant. Collierville, TN: Helena Holding Company;

2005.

Specimen Label: Aquamaster Herbicide. St. Louis, MO: Monsanto Company; 2006.

Syracuse Environmental Research Associates. Selected commercial formulations of glyphosate‐‐

Accord, Rodeo, Roundup and Roundup Pro Risk Assessment. Final Report. Riverdale, MD:
USDA, Forest Service; 1996 Jun 30; SERA TR 96‐22‐02‐01c.

Title 2100‐‐Environmental Management, Amendment No. 2100‐94‐7. Forest Service Manual 2150.

Washington, DC: U.S. Forest Service; 1994 Dec 6.

Tu, M.; Hurd, C., and Randall, J. M. Glyphosate. Weed Control Methods Handbook: Tools &

Techniques for Use in Natural Areas. Davis, CA: The Nature Conservancy; 2001 Apr; p. 7E.1 ‐
7e.10.

U.S. Forest Service. Glyphosate Herbicide Information Profile. Portland, OR: U.S. Forest Service,

Pacific Northwest Region; 1997 Feb.

USEPA. R.E.D. Facts: Glyphosate. Washington, D.C.: U.S. Environmental Protection Agency, Office of
Prevention, Pesticides and Toxic Substances; 1993 Sep; EPA/738/F‐93/011.

USEPA. Reregistration Eligibility Decision (RED): Glyphosate. Washington, D.C.: U.S. Environmental
Protection Agency, Office of Prevention, Pesticides and Toxic Substances; 1993 Sep;
EPA/738/R‐93/014.

WHO. Glyphosate and AMPA in drinking water.  Background document for development of WHO
Guidelines for Drinking‐water Quality.  Geneva: World Health Organization; 2005;
WHO/SDE/WSH/03.04/97.

WHO. Glyphosate. Geneva: World Health Or ganization; 1994. (Environmental Health Criteria; 159).

WHO. WHO/FAO Data Sheets on Pesticides: No. 91, Glyphosate. Geneva: World Health

Organization; 1996 Jul.

Williams, G. M.; Kroes, R., and Munro, I. C. Safety evaluation and risk assessment of the herbicide
Roundup and its active ingredient, glyphosate, for humans. Regul Toxicol Pharmacol. 2000

Apr; 31(2 Pt 1):117‐65.

422 Annex 133

TECHNICAL DATA SHEET FOR ROUNDUP SL

(Agriculture and Livestock National Exchange Market (Bolsa Nacional Agropecuaria S.A.)

Agriculture and Livestock FORMAT PRODUCT CODE
National Exchange Market TECHNICAL DATA VALIDITY SINCE
SHEET VERSION

PRODUCT TECHNICAL DATA SHEET
PRODUCT NAME ROUNDUP SL
1. to have valid ICA registration number as producer and/or
seeds distributor
2. Comply with Resolution 148 issued by ICA

3. to have adequate places for seeds storage
SUPPLIER
REQUIREMENTS Valid ICA registration, for offered species that have it. Seeds
must be treated to prevent insects and pathogens.
To have more than 5 years’ operation in the market
To have made transactions for at least 950 minimum monthly

wage in force the previous year

TYPE Agricultural Herbicide, Soluble Liquid (certificate).

Glyphosate, Isopropyl

Active ingredient amine Salt
State Liquid
Appearance and smell Amber viscous liquid with
a slight amine smell
Solubility Very soluble in water
TECHNICAL
pH 4,4 to 4,9 (Solution at 1%)
CHARACTERISTICS Crystallization point < 0 °C
OF THE PRODUCT ICA Registration Number 756-R
Roundup SL is not flammable, nor explosive. It is considered
harmful if ingested; it might be irritant in contact with eyes

Additional information Acute toxicity: It is very
Risks Identification unlikely that exposure to
Roundup SL poses risk of

423Annex 133

acute toxicity. It may cause
slight irritation on skin and
moderate to severe in eyes.

Chronic Toxicity: Chronic
effects on humans have not
been documented attributable
to chronic exposure; studies
on mutagenicity and
carcinogenicity have resulted

negative.
Ecotoxicity: It is not harmful
to aquatic organisms (middle
toxicity to rainbow trout, not
toxic to birds). It must not be
poured near channels, drains,

nor running water or water
reservoirs. It is not toxic to
bees under recommended use
directions.
PRESENTATION Plastic container of 1, 4 or 10 liters; metal container of 10 and
20 liters

QUANTITY 50 liters
LIFE SPAN 1 year (validity)
ADDITIONAL Refusal conditions: Not compliance with any of the conditions
INFORMATION of this technical data sheet.
Submit in person the product in 22 Mayors’ Office or

UMATAS [Municipal Agricultural Technical Assistance
Units] in the municipalities (except Puerto Colombia
Municipality) in which productive projects of medicine plants,
or in the rural producers associations determined by the
Economic Development Office, according to the list of
beneficiaries, term, and Schedule set by the mentioned audit

and/supervising office.
The supplier pays on the cost of transport, downloading and
delivery of the product.

Prepared by: Ivette Salas Rodriguez
Reviewed by: Martin Atencio Garcia

424 acute toxicity. It may cause
slight irritation on skin and
moderate to severe in eyes.
Chronic Toxicity: Chronic

effects on humans have not
been documented attributable
to chronic exposure; studies
on mutagenicity and
carcinogenicity have resulted
negative.
Ecotoxicity: It is not harmful

to aquatic organisms (middle
toxicity to rainbow trout, not
toxic to birds). It must not be
poured near channels, drains,
nor running water or water
reservoirs. It is not toxic to
bees under recommended use

directions.
PRESENTATION Plastic container of 1, 4 or 10 liters; metal container of 10 and
20 liters
QUANTITY 50 liters
LIFE SPAN 1 year (validity)

ADDITIONAL Refusal conditions: Not compliance with any of the conditions
INFORMATION of this technical data sheet.
Submit in person the product in 22 Mayors’ Office or
UMATAS [Municipal Agricultural Technical Assistance
Units] in the municipalities (except Puerto Colombia
Municipality) in which productive projects of medicine plants,
or in the rural producers associations determined by the

Economic Development Office, according to the list of
beneficiaries, term, and Schedule set by the mentioned audit
and/supervising office.
The supplier pays on the cost of transport, downloading and
delivery of the product.

Prepared by: Ivette Salas Rodriguez

Reviewed by: Martin Atencio Garcia426 Annex 134

LABEL AND SAFETY D ATAS HEET FORGLY-41

(Colombian Agriculture & Livestock Institute (ICA), Compañía Agrícola
Colombiana, Ltda. y Cia. S.C.A.)

SECTION 1

READ THE LABEL BEFORE USING THIS PRODUCT
KEEP OUT OF CHILDRENS REACH

CAUTION AND USE WARNINGS

Spray solutions of this product should be mixed, stored, and applied using only stainless
steel, alumina, fiberglass or plastic-lined steel containers.

DO NOT MIX, STORE OR APPLY THIS PRODUCT OR SPRAY SOLUTIONS IN
GALVANIZED STEEL OR UNLINED (EXCEPT STAINLESS STEEL) STEEL
CONTAINERS OR SPRAY TANKS. This products or spray solutions of this product
react with such containers and tanks to produce hydrogen gas which may form a highly
combustible gas mixture. This gas mixture could flash or explode causing serious

personal, if ignited by open flame, spark or a welder’s torch, a lighted cigarette or other
ignition source.

AVOID CONTACT OF HERBICIDE WITH FOLIAGE, GREEN STEMS, EXPOSED
ROOTS OR FRUIT OF CROPS, DESIRABLE PLANTS AND TREES, BECAUSE
SEVERE INJURY OR DESTRUCTION MAY RESULT

CAUTION:
THIS PRODUCT CAUSES EYE IRRITATION. Avoid contact with eyes and clothes.

FIRST AID

Call a medical center or doctor for treatment advice
IF IN EYES:
- Hold eye open and rinse slowly and gently with water for 15 - 20
minutes.
- Remove contact lenses if present after the first 5 minutes then continue

rinsing eye.
TOXICOLOGICAL EMERGENCIES PHONE NUMBER
Have the product container or label with you when calling a poison control center or

427Annex 134

doctor, or going for treatment
EMERGENCY NUMBERS 24 HOURS
In Bogotá:
Outside Bogotá

CISPROQUIM 288 6012 01 8000 9160
12

diarrhea, colic, etc.). If such symptoms are observed, provide the animal with plenty of
fluids to prevent dehydration. Call a veterinarian if symptoms persist for more than 24
hours

STORAGE AND DISPOSALWASTE
When storing or disposing the product, do not contaminate water, foodstuffs, animal

foodstuff or seeds. Do not store at home. DO not transport or store in vehicles or places
where seeds or foodstuffs for human consumption are transported or stored. Keep
container closed to prevent spills and contamination. DESTROY THIS CONTAINER
AFTER USING THIS PRODUCT. No container that has contained herbicides should
be used to store water or food for human or animal consumption.

Approval date by ICA: 25/11/2008

[Caution symbols]

[ICA’s Approval seal]

SECTION 2

GLY-41 SL

428doctor, or going for treatment
EMERGENCY NUMBERS 24 HOURS
In Bogotá:
Outside Bogotá
CISPROQUIM 288 6012 01 8000 9160
12

DOMESTIC ANIMALS: This product is considered to be relatively nontoxic to dogs
and other domestic animals; however, ingestion of this product or large amounts of
freshly sprayed vegetation may result in temporary gastrointestinal irritation (vomiting,
diarrhea, colic, etc.). If such symptoms are observed, provide the animal with plenty of
fluids to prevent dehydration. Call a veterinarian if symptoms persist for more than 24

hours

STORAGE AND DISPOSALWASTE
When storing or disposing the product, do not contaminate water, foodstuffs, animal
foodstuff or seeds. Do not store at home. DO not transport or store in vehicles or places
where seeds or foodstuffs for human consumption are transported or stored. Keep
container closed to prevent spills and contamination. DESTROY THIS CONTAINER

AFTER USING THIS PRODUCT. No container that has contained herbicides should
be used to store water or food for human or animal consumption.

Approval date by ICA: 25/11/2008

[Caution symbols]

[ICA’s Approval seal]

SECTION 2

GLY-41 SLAnnex 134

MIXTURE:

Clean thoroughly all parts of application equipment after using this product, rinsing
with abundant water.

NOTICE: RESULTS WILL NOT BE OPTIMAL IF DIRTY WATER, OR WATER
FROM PONDS AND DRAINS THAT IS NOT CLEAR IS USED.

Mixing with water: This product mixes readily with water. Mix spray solutions of this
product as follows: Begin filling the mixing tank or spray tank with the required amount
of clean water. Add the recommended amount of this product near the end of the filling
process and mix well. Use caution to avoid siphoning back into the carrier source. Use
approved anti-back siphoning devices. During mixing and application, foaming of the
spray solution may occur. To prevent or minimize foam, avoid the use of mechanical
agitators, terminate by-pass and return lines at the bottom of the tank and, if needed, use

an approved anti-foaming agent.

Surfactants.

Non-ionic surfactants may be used to improve wetting on foliage. Do not reduce rates
of this product when adding surfactant. Read and carefully observe all caution

statements and other information on the surfactant label.

WEED CONTROL

Annual weeds

Apply from 2 to 3 litres of GL Y-41 SL per hectare to control the following species:
Amaranthus spp (pigweed), Bidens pilosa (Spanish needle), Boerhaavia erecta (erect
spiderling), Borreris laevis (purple-leaved button weed), Cenchrus brownii (Brown’s
burgrass), Commoiina diffusa (climbing dayflower), Cyperus diffuses (Dwarf Umbrella
Grass), Digitaria sanguinalis (hairy crabgrass), Echinochloa coIonum (Jungle rice),
Eleusine indica (wire grass), Euphorbia hirta (asthma weed), Fimbrisilys annua (annual

fimbry), Galinsoga ciliata (hairy galingosa), Ipomoea spp (morning glory), Ischaemum
rugosum (ribbed muriana grass),
Jussiaea linifolia, Kallstroemia maxima (big caltrop), Oryza sativa* (Asian rice),
Portulaca oleracea (common purslane), Physalis angulata (cutleaf groundcherry),
Rottboellia exaltata (itch grass), Tradescantia cumanensis* (spiderwort).

430MIXTURE:

Clean thoroughly all parts of application equipment after using this product, rinsing

with abundant water.

NOTICE: RESULTS WILL NOT BE OPTIMAL IF DIRTY WATER, OR WATER
FROM PONDS AND DRAINS THAT IS NOT CLEAR IS USED.

Mixing with water: This product mixes readily with water. Mix spray solutions of this
product as follows: Begin filling the mixing tank or spray tank with the required amount

of clean water. Add the recommended amount of this product near the end of the filling
process and mix well. Use caution to avoid siphoning back into the carrier source. Use
approved anti-back siphoning devices. During mixing and application, foaming of the
spray solution may occur. To prevent or minimize foam, avoid the use of mechanical
agitators, terminate by-pass and return lines at the bottom of the tank and, if needed, use
an approved anti-foaming agent.

Surfactants.

Non-ionic surfactants may be used to improve wetting on foliage. Do not reduce rates
of this product when adding surfactant. Read and carefully observe all caution
statements and other information on the surfactant label.

WEED CONTROL

Annual weeds

Eleusine indica (wire grass), Euphorbia hirta (asthma weed), Fimbrisilys annua (annual
fimbry), Galinsoga ciliata (hairy galingosa), Ipomoea spp (morning glory), Ischaemum
rugosum (ribbed muriana grass),
Jussiaea linifolia, Kallstroemia maxima (big caltrop), Oryza sativa* (Asian rice),
Portulaca oleracea (common purslane), Physalis angulata (cutleaf groundcherry),
Rottboellia exaltata (itch grass), Tradescantia cumanensis* (spiderwort).Annex 134

treatments to control weed that regrow from the roots or runners or when new weeds
germinate by seed below the surface of the soil. You can repeat the treatments up to an
annual maximum download of 27.7 litres of this product per hectare. You can use the
product of 10 to 12 L/has for best results t control perennial weeds, semiligneous shrubs

and trees of difficult control, when the plants grow in poor conditions, or when the
infestation is dense.Do not apply this product through an irrigation systemTHE
HERBICIDE SOLUTION WITH WELL MAINTAINED AND CALIBRATED
EQUIPMENTS, CAPABLE OF SPRAYING THE DOSES IN DESIRED VOLUMES.

This product can be applied using the following equipments:

Implementation with land equipments: Systems with or without boom and other
equipment for the implementation on the ground.

To control annual weeds or perennial with equipment for the implementation on the
ground, use the recommended doses per hectare for this product in volumes of 30 to 350
litres of solution. As it increases the population density of the weeds, the volume of
implementation must be increased within the range recommended to obtain full

coverage with the recommended dose per hectare. Select carefully the nozzle to prevent
that the application is too thin and causes drift. For better results in terrestrial
application use flat spray nozzle.

Hand-held or high-volume spray equipment. Knapsack and backpack sprayers, pump-
up pressure sprayers, handguns, handwands, mistblowers, lances or other hand-held or
motorized spray equipment used to direct the spray onto weed foliage.

Apply the spray solution onto vegetation foliage to be controlled. For applications made
on a spray-to-wet basis, spray coverage should be uniform and complete. Do not spray
to the point of runoff. Use coarse sprays only. Prepare the desired volume of the spray
solution by mixing the amount of this product with the volume water, as shown in the
following table:

Desired Quantity of GLY-41 herbicide
volume ½ % 1% 1½ % 2% 5% 10%
4 L 20 cc 40 cc 60 cc 80 cc 200 cc 400 cc
20 L 100 cc 200 cc 300 cc 400 cc 1 L 2 L
200 L 1 L 2 L 3 L 4 L 10 L 20 L

432treatments to control weed that regrow from the roots or runners or when new weeds
germinate by seed below the surface of the soil. You can repeat the treatments up to an
annual maximum download of 27.7 litres of this product per hectare. You can use the
product of 10 to 12 L/has for best results t control perennial weeds, semiligneous shrubs

and trees of difficult control, when the plants grow in poor conditions, or when the
infestation is dense.Do not apply this product through an irrigation systeTHE
HERBICIDE SOLUTION WITH WELL MAINTAINED AND CALIBRATED
EQUIPMENTS, CAPABLE OF SPRAYING THE DOSES IN DESIRED VOLUMES.

This product can be applied using the following equipments:

Implementation with land equipments: Systems with or without boom and other
equipment for the implementation on the ground.

Desired Quantity of GLY-41 herbicide
volume ½ % 1% 1½ % 2% 5% 10%
4 L 20 cc 40 cc 60 cc 80 cc 200 cc 400 cc
20 L 100 cc 200 cc 300 cc 400 cc 1 L 2 L
200 L 1 L 2 L 3 L 4 L 10 L 20 LAnnex 134

users must notify opportunely this Company any complaint, either based on contract,
negligence, strict obligation, injury or otherwise. The buyer and all users are responsible
for the loss or damage by use or handling, resulting from conditions outside the control

of the Company, including but not limited and incompatibility with products other than
the indicated on the label, implementation or contact with desirable vegetation, unusual
climate conditions, other than the conditions that are considered normal for the site and
during the period of the application, as well as weather conditions other than those
specified in the label and other implementation than the explicitly specified in the label.
This company does not guarantee any product reformulated or re-packaged from this

product, except in accordance with the requirements of this Company and express
permission in writing, granted by this Company.

THE WAY OF COMPENSATE THE USER OR THE BUYER DAMAGE AND THE
LIABILITY LIMIT OF THIS COMPANY OR ANY OTHER SELLER IN REGARD
TO ANY AND ALL THE LOSSES, INJURIES OR DAMAGES RESULTING FROM
THE USE OR HANDLING OF THIS PRODUCT (INCLUDING CLAIMS BASED

ON CONTRACT, NEGLIGENCE, STRICT OBLIGATION, OTHER DAMAGE, OR
OTHERWISE), WILL BE THE PURCHASE PRICE PAID BY THE USER OR THE
BUYER FOR THE RESPECTIVE QUANTITY OF THIS PRODUCT OR AS THIS
COMPANY OPTION OR ANY OTHER SELLER, THE REPLACEMENT OF THE
QUANTITY, IF NOT BOUGHT, THE REPLACEMENT OF THE QUANTITY IN
ANY CASE SHALL THIS COMPANY WILL BE RESPONSIBLE OR ANY OTHER

SELLER FOR ANY INCIDENTAL, CONSEQUENTIAL OR SPECIAL PREJUDICE
Once the product IS open and used, it is understood that the buyer and users have
accepted the terms of this limited warranty and liability, which may not be altered by
any verbal or written agreement. If these terms are unacceptable, immediately return the
product without opening it.

434users must notify opportunely this Company any complaint, either based on contract,
negligence, strict obligation, injury or otherwise. The buyer and all users are responsible
for the loss or damage by use or handling, resulting from conditions outside the control

during the period of the application, as well as weather conditions other than those
specified in the label and other implementation than the explicitly specified in the label.
This company does not guarantee any product reformulated or re-packaged from this
product, except in accordance with the requirements of this Company and express

permission in writing, granted by this Company.

THE WAY OF COMPENSATE THE USER OR THE BUYER DAMAGE AND THE
LIABILITY LIMIT OF THIS COMPANY OR ANY OTHER SELLER IN REGARD

TO ANY AND ALL THE LOSSES, INJURIES OR DAMAGES RESULTING FROM
THE USE OR HANDLING OF THIS PRODUCT (INCLUDING CLAIMS BASED
ON CONTRACT, NEGLIGENCE, STRICT OBLIGATION, OTHER DAMAGE, OR

OTHERWISE), WILL BE THE PURCHASE PRICE PAID BY THE USER OR THE
BUYER FOR THE RESPECTIVE QUANTITY OF THIS PRODUCT OR AS THIS
COMPANY OPTION OR ANY OTHER SELLER, THE REPLACEMENT OF THE
QUANTITY, IF NOT BOUGHT, THE REPLACEMENT OF THE QUANTITY IN

ANY CASE SHALL THIS COMPANY WILL BE RESPONSIBLE OR ANY OTHER
SELLER FOR ANY INCIDENTAL, CONSEQUENTIAL OR SPECIAL PREJUDICE
Once the product IS open and used, it is understood that the buyer and users have
accepted the terms of this limited warranty and liability, which may not be altered by

any verbal or written agreement. If these terms are unacceptable, immediately return the
product without opening it.Annex 134

cholinesterase. Atropine and oximes must not be used as an antidote. s open

given
Section 5: MEASURES TO EXTINGUISH FIRES
Glyphosate herbicide is neither flammable nor explosive. In case of fire,
combustion of the product may produce toxic vapors such as carbon oxides and
nitrogen.
Extinguishing: Containers exposed to heat must be kept cool. It is extinguished
with carbon dioxide, foam, dry chemical powder or water spray in limited

quantities. Water used to extinguish it must not be allow to flow to superficial
water nor must it be allowed to flow to sewage systems. Water must be collected
and kept as special residues. Autonomous breathing protection equipment must be
used.
Section 6: MEASURES IN CASE OF ACCIDENTAL SPILL
Spills are contained with sand or earth dikes. It is pick up by suction or vacuuming
or by absorption using dry sand or earth and the material collected is packed in a

sealed container duly labeled. The contaminated sites must be decontaminated by
washing it with industrial detergent and it is handled as special residues in burning
devices or approved safety landfills.
Recommended personal protection: See section 8
Section 7: HANDLING AND STORAGE:
Handling: Eating, drinking or smoking not allowed during handling or application
of this product. Was hands after being in contact with this product.

Storage: Keep the product in original packaging and containers. No smoking in the
place. Keep out of children and animals’ reach. Stored in a ventilated place, away
from food, drink, hay, or concentrated food for animals.
Compatible materials for storage: Stainless steel, aluminum, fiber glass or plastic.
Section 8: EXPANSION CONTROLS AND PERSONAL PROTECTION:
Chemical safety gloves and eye protection must be worn while handling.
Occupational health and safety general rules must be observed.

Section 9 PHYSICAL-CHEMICAL PROPERTIES:
State: Liquid
Appearance and color: Amber viscous liquid, practically odorless
Density: 1,17 g/mL pH: 4.99 (Solution at 1%)
Section 10: STABILITY AND REACTIVITY:
Glyphosate herbicide is stable under normal storage and manipulation conditions.

There is no probability of dangerous reaction in the original containers. It reacts
with galvanized steel or soft steel without covering layer, producing hydrogen, a
very flammable gas that may cause explosion.
Section 11: TOXICOLOGICAL INFORMATION:
ACUTE TOXICITY:
It is very unlikely that exposure to glyphosate herbicide poses risk of acute toxicity
(LD50 Oral (rats): > 5.000 mg/Kg; LD50 Dermal (rats): > 5.000 mg/Kg. It may

cause slight irritation on skin and from moderate to severe in the eyes.
CHRONICLE TOXICITY:
No chronicle effects on humans have been documented, attributable to chronicle
exposure. Mutagenicity and carcinogenicity studies have resulted negative.
Section 12: ECOTOXICOLOGICAL INFORMATION:
ECOTOXICITY:

436cholinesterase. Atropine and oximes must not be used as an antidote. s open
given

Section 5: MEASURES TO EXTINGUISH FIRES
Glyphosate herbicide is neither flammable nor explosive. In case of fire,
combustion of the product may produce toxic vapors such as carbon oxides and
nitrogen.
Extinguishing: Containers exposed to heat must be kept cool. It is extinguished
with carbon dioxide, foam, dry chemical powder or water spray in limited
quantities. Water used to extinguish it must not be allow to flow to superficial
water nor must it be allowed to flow to sewage systems. Water must be collected

and kept as special residues. Autonomous breathing protection equipment must be
used.
Section 6: MEASURES IN CASE OF ACCIDENTAL SPILL
Spills are contained with sand or earth dikes. It is pick up by suction or vacuuming This information is applicable only to the purposes stated for the product. It
or by absorption using dry sand or earth and the material collected is packed in a
sealed container duly labeled. The contaminated sites must be decontaminated by complements the technical information on the label but does not substitute it in any
washing it with industrial detergent and it is handled as special residues in burning matter. It is based on the best information available at the time of issuing; it does not
devices or approved safety landfills.
imply insurance or warranty and it is provided on good faith basis.
Recommended personal protection: See section 8
Section 7: HANDLING AND STORAGE:
Handling: Eating, drinking or smoking not allowed during handling or application
of this product. Was hands after being in contact with this product.
Storage: Keep the product in original packaging and containers. No smoking in the
place. Keep out of children and animals’ reach. Stored in a ventilated place, away
from food, drink, hay, or concentrated food for animals.
Compatible materials for storage: Stainless steel, aluminum, fiber glass or plastic.

Section 8: EXPANSION CONTROLS AND PERSONAL PROTECTION:
Chemical safety gloves and eye protection must be worn while handling.
Occupational health and safety general rules must be observed.
Section 9 PHYSICAL-CHEMICAL PROPERTIES:
State: Liquid
Appearance and color: Amber viscous liquid, practically odorless
Density: 1,17 g/mL pH: 4.99 (Solution at 1%)

Section 10: STABILITY AND REACTIVITY:
Glyphosate herbicide is stable under normal storage and manipulation conditions.
There is no probability of dangerous reaction in the original containers. It reacts
with galvanized steel or soft steel without covering layer, producing hydrogen, a
very flammable gas that may cause explosion.
Section 11: TOXICOLOGICAL INFORMATION:
ACUTE TOXICITY:

It is very unlikely that exposure to glyphosate herbicide poses risk of acute toxicity
(LD50 Oral (rats): > 5.000 mg/Kg; LD50 Dermal (rats): > 5.000 mg/Kg. It may
cause slight irritation on skin and from moderate to severe in the eyes.
CHRONICLE TOXICITY:
No chronicle effects on humans have been documented, attributable to chronicle
exposure. Mutagenicity and carcinogenicity studies have resulted negative.
Section 12: ECOTOXICOLOGICAL INFORMATION:
ECOTOXICITY:438 Annex 135

ANDEAN R EGULATION FOR THE R EGISTRATION AND CONTROL OF C HEMICAL
PESTICIDES FOR A GRICULTURAL U SE,ANDEAN C OMMUNITY , ECISION 436 OF 2000

(Norma Andina para el Registro y Control de Plaguicidas Químicos de Uso Agrícola, Arts. 57- 59;
Annex 1 – Glossary, “Mean Lethal Dose, LD 50” (Comunidad Andina, Decisión 436 de 2000). Available
at: http://www.comunidadandina.org/normativa/dec/D436.htmted 19 February 2010))

[…]

Article 57.- The information contained in the files of the National Registers of chemical
pesticides for agricultural use shall be public. However, the Competent National
Authority will refrain from circulating the information received, when the individual or
legal entity that has provided such information requested a confidential treatment.

The documents which contain confidential information will be kept separate from the
main file.

Article 58.- In no case will the following information be labeled as confidential:

- The name and content of the active substance or substances and the pesticide´s
name;

- The name of other substances considered as hazardous;

- The physical and chemical data of the active substance, the formulated product and
the additives of toxicological importance;

- The methods used to inactivate the technical-grade active ingredient or the
formulated product.

- The summary of the results of the tests to determine the product´s efficacy and its

toxicity for human beings, animals, plants and the environment;

- The recommended methods and precautions to reduce the risks of handling, storage,
transport, and fire;

- The methods for eliminating the product and its containers;

- The decontamination measures to be adopted in case of accidental spill or leak.

439Annex 135

- The first aid and medical treatment to be given in case of bodily injury;

- The data and information that appears on the label and instructions sheet.

Article 59.- The interested party requiring confidential treatment of specific information
shall state the reasons for such a request and include a non-confidential summary of
such information, or an explanation of any reasons why it is not possible to summarize
it.

In case the requesting party does not comply with the aforementioned paragraph or if
the information does not qualify as confidential, the Competent National Authority shall
give reasoned notification of such circumstance to the requesting party, and grant it a
reasonable time-limit so that it may withdraw the documents containing the information
subject to the denial. Following that time-limit period the documents will be included
in the public record.

[…]

ANNEX 1
GLOSSARY

[…]

Mean Lethal Dose, LD 50, statistical estimate of the minimum dose required to kill
fifty percent of a population of laboratory test animals in controlled conditions. It is
stated in milligrams of toxic [substance] per kilogram of animal weight indicating the
species, gender and age of the animals used for experimenting. It is administrated via

oral, dermal, mucosal or parenteral channels.

440 Annex 136

R EPORT OF THE FIFTH M EETING OF VICE-M INISTERS OFF OREIGN AFFAIRS OF THE
A NDEAN COMMUNITY , CARACAS , 16-17 CTOBER 2000

(Comunidad Andina, Consejo Andino de Ministros de Relaciones Exteriores, Informe de la Quinta
Reunión de Viceministros de Relaciones Exteriores de la Comunidad Andina, Caracas, Venezuela, 16-17
October 2000, 6 November 2000)

Confidential Report

The Vice-Ministers of Foreign Affairs of the five member countries, who met on the 16
and 17 October 2000 in the framework of the Andean Council of Ministers of Foreign
Affairs in Caracas, requested the General Secretariat to prepare a Confidential Report
th
about the debate that arose from the 5 point of the Provisional Agenda: “Early warning
mechanisms for the detection of biological control agents in the eradication of illicit
crops, which affect Andean ecosystems”. This point had been removed from the
approved agenda.

Referring to this topic, the Colombian Vice-Minister, Clemencia Forero, expressed her
gratitude for the welcome given by the government of Venezuela and particularly by
Vice-Minister Valero. With regards to the proposed agenda and in reference to point 5,
she explained that in the previous days a Note had been sent to the Minister of Foreign

Affairs of Venezuela containing preliminary comments about this point. It explained
Colombia’s point of view that the treatment of this topic belonged to the established
forum, namely, that of the Andean Committee of Environmental Authorities.

In that sense, Colombia considered that the Vice-Ministers should not express their
direct views on the draft Decision prepared by the Secretary-General which their
respective environmental authorities were not acquainted with in great detail. She
mentioned that this project could be further studied at the CAAM meeting to be held in
Venezuela during the month of November, taking into account that this was a matter of

instances and one where the Committee of Environmental Authorities existed.

She requested that the more specific opinions that their environmental authorities may
have on this matter be taken into account and, thus, [deemed] it would be difficult to
consider supporting or endorsing the draft Decision.

She offered to provide two documents to the General Secretariat containing the
response given by the Minister of Foreign Affairs Fernández de Soto to a questionnaire
formulated by some Colombian senators. She requested that such document be

441Annex 136

included in the Confidential Record of this meeting and, hence read the response stating
the position of the Colombian government regarding this topic in the following terms:

“The Government of Colombia has stated that it does not approve under any
circumstance experimenting with myco-herbicides exogenous to our ecosystems and
which may affect the environmental balance and the population’s health. In particular,
it has rejected experimentation with fusarium oxysporum. It is important to clarify that
no myco-herbicide is currently being studied.”

She pointed out that the position of the Colombian government is unequivocal since the

use of chemical and now biological agents for crops eradication has had a long history
in the countries that have suffered this scourge.

She stated that Colombia, on several occasions and under intense pressure, has been
subject to questioning, and even to demands, in relation to the use of new chemical
herbicides. She explained that on each of those occasions, the National Narcotics

Council of her country, which gathers all the Ministers involved in the fight against
drugs, has publicly referred to the matter and has never allowed the application of any
herbicide or biological agent that is harmful to the environment or to health. Thus the
answer of the Colombian government to this topic is unequivocal.

She made reference to a public statement by the Minister of Environment, Juan Mayer
[sic], in October 2000. It stated that, in relation to the possible experimentation, use or

application of fusarium oxysporum as a mechanism to eradicate illicit crops in the
country, the Ministry of Environment, as supreme environmental authority, did not
approve the proposal made by United Nations International Drug Programme [sic] to
conduct tests with fusarium oxysporum, as it considered that any agent, external to the
native ecosystems of the country, could cause serious risks to the environment and to
normal health.

She reaffirmed the high sensitivity of this matter for Colombia’s Government and that
her country is not experimenting with this fungus, nor will it do so in the future, for
which reason she reiterated her request for this item not to be considered in the agenda.

On the other hand, the Vice-Minister of Ecuador, Gonzalo Salvador Holguín, expressed

his gratitude to the government of Venezuela for the opportunity of having this meeting
and, in connection to the issue raised by the Vice-Minister of Colombia, he expressed
his full agreement with the removal of this item from the agenda. He further expressed
that in the course of the last two months his country has held intensive conversations
with Colombia with respect to this matter, the results of which were reflected in a

442 Annex 136

Presidential Declaration recently signed by the Presidents of both nations, on the
occasion of the last visit of President Pastrana to Ecuador.

He stated that complete and absolute assurances were received from the authorities of
the Colombian Government to the effect that no use or experimenting with the fungus
fusarium oxysporum or any other type of myco-herbicide will be undertaken for the
illicit crops eradication programs. Thus, the Government of Ecuador deems it
unnecessary to bring this issue to the next meeting of the Council.

In turn, the Vice-Minister of Bolivia, Ana María Solares, thanked the Government of
Venezuela, through its Vice-Minister, for its hospitality, and in relation to the discussed
issue, reiterated that the Government of her country opposes any type of practice or uses
of this nature. But in any case, she requested further details of the background of this
draft Decision to the General Secretariat.

Likewise the Vice-Minister of Peru, José Antonio Arróspide, took the floor and thanked
Ambassador Valero for the invitation and pleasant opportunity of being in Caracas for
such an important meeting. He recalled the topics that had been the subject of work
between the Presidency of Peru and the General Secretariat and, with regards to the
subject, he stated that he fully shared the position of Colombia.

He expressed that in the case of Peru, his country has a legal norm that prohibits the use
of any kind of biological agent of this type and therefore, he considered that the matter
had no direct link with the work of the Ministers of Foreign Affairs. Like Colombia, he
stated his preference for the item not to be included in the Council’s Agenda.

He further recalled that a written communication had been sent to the Ministry of
Foreign Affairs of Venezuela with regards to another item of the agenda, i.e. item 4, on
the Latin-American Monetary Fund.

The meeting’s secretary, Carlos Longa, explained that the draft Decision initially
referred to, came from the Andean Committee of Environmental Authorities, and that
the idea of the Committee in bringing this proposal to the Vice-Ministers’ meeting had

the purpose of assessing the possibility of submitting it to the Ministers of Foreign
Affairs. That Committee considered the advisability of developing an early warning
mechanism for possible uses of biological control agents and, given that such
Committee is an advisory body with no power to adopt binding decisions in the
framework of the Agreement, it chose to submit this proposal to determine the
feasibility of adopting this mechanism by Decision of the Ministers of Foreign Affairs.

443Annex 136

Vice-Minister Solares deemed it important to continue dialogue on this early warning

mechanism once the issue is more evolved and stated her wish to continue receiving
information about this sort of sectional topics, such as those coordinated by other organs
of the System or other bodies of the [Andean] Community, which would allow for their
adequate treatment by the Andean Council of Ministers of Foreign Affairs and the
Presidents.

444 Annex 137

ANDEAN COOPERATION PLAN FOR THC ONTROL OFILLEGALD RUGS ANDRELATED
OFFENSES, NDEANC OMMUNITY, DECISIO505OF 2001

(Andean Community, Decision 505 of 2001)

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Decision 505
Andean Cooperation Plan for the Control of Illegal Drugs and Related
Offenses

THE ANDEAN COUNCIL OF FOREIGN MINISTERS,

HAVING SEEN: Article 16 of the Cartagena Agreement and Decision 458

"Common Foreign Policy Guidelines";

BEARING IN MIND: The agreement approved by the Presidents of the Andean
Countries at the meeting held on April 17, 2001 in Cartagena, Colombia;

WHEREAS: Illegal drug production, traffic, and consumption, asset laundering,
diversion and smuggling of chemical precursors, and arms trafficking seriously

threaten the development and security of the Andean countries;

The efforts being made today by the Andean countries under their respective
national programs for the control of illegal drugs and related offenses can be

significantly boosted and supplemented through an Andean Cooperation Plan to
intensify national, bilateral, and Community measures in this area;

The international community can wage an integrated campaign against the
worldwide illegal drug problem that could cover all of the aspects involved in
illegal drug production, traffic, and consumption and related offenses, based on
the principle of shared responsibility;

The Andean strategy for the control of illegal drugs and related offenses, as
well as international efforts in this area, should be carried out with full respect

for national law and sovereignty.

DECIDES:

Sole article. – To approve the Andean Cooperation Plan for the Control of
Illegal Drugs and Related Offenses set out in the document attached to this
Decision.

Signed in the city of Valencia, Venezuela, on the twenty-second of June of two
thousand and one.

ANDEAN COOPERATION PLAN FOR THE CONTROL OF ILLEGAL DRUGS
AND RELATED OFFENSES

Principles and Objectives

The Andean Cooperation Plan for the Control of Illegal Drugs and Related
Offenses:

1. Is grounded in the conviction that illegal drug production, trafficking, and
consumption is a worldwide problem that seriously threatens the development

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and security of the Andean countries and of the international community.

Recognizes that this is one of the most harmful and dangerous forms of
organized transnational crime, that makes use of the globalizing logic of the
markets, disrupts the social dynamic, distorts the economy, undermines the

state of law, and subverts the public order.

2. Is comprehensive and encompasses all of the aspects of the worldwide drug

problem and related offenses: prevention, interdiction, reduction of illicit crops,
and alternative development, as well as the control of the diversion of chemical
precursors, asset laundering, and the traffic in arms, ammunition, and
explosives.

3. In this connection, it is based on the shared responsibility of each and every
one of the actors involved in creating the problem and, as a result, in the

search for solutions to it at both the Andean Subregional level and the South
American, hemispheric, and world levels.

4. Is respectful of national legislation and territorial sovereignty and integrity,
as well as of the strict observance of International Law and is implemented
through cooperation and solidarity. That cooperation is inherent to the shared
nature of the problem and, consequently, should be mutual, voluntary and not

subject to limitations or requirements that set conditions on the unhampered
involvement of the wills of the parties. The solidarity, for its part, is an
expression of the recognition that national and Subregional drug control efforts

should be accompanied by international cooperation in order to reinforce
democracy and the exercise of human rights and at the same time promote the
sustainable economic and social development of the Andean Countries.

5. Seeks to strengthen and step up national programs in each of the Andean
Countries through coordination, cooperation, and the exchange of experiences

among the Member Countries and by taking joint action in dealing with third
countries and in international forums.

6. Is a key issue of Andean political cooperation that links up the common

foreign policy, Community border integration and development policy,
sustainable development policy, the Andean Social Agenda, and security and
confidence-building efforts in the Subregion, as applicable.

7. It is proposed that the Andean Community be consolidated as the moving
force for a South American and hemispheric strategy for the control of illegal
drugs and related offenses.

Mechanisms

1. The Andean Council of Foreign Ministers is the body responsible for defining,
coordinating, and following up the Andean Cooperation Plan for the Control of
Illegal Drugs and Related Offenses.

2. An Executive Committee will be set up, formed of high-level officials of the
Ministries of Foreign Affairs and the national officers responsible for controlling
illegal drugs and related offenses, including representatives of the security

bodies.

3. The Executive Committee may establish subcommittees and working groups

specializing in the different aspects of the Andean Plan. The Subcommittees
and working groups may be made up of two or more Member Countries and will
be open to the participation of the others.

4. The Andean Community General Secretariat will act as the Executive
Committee’s Technical Secretariat.

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Program of Action

1. The Program of Action will be approved by decision of the Andean Council of

Foreign Ministers and will cover a two-year period.

2. The Executive Committee will draw up the Operating Plans for carrying out
the Program of Action and every two years will propose the updating of that

program to the Andean Council of Foreign Ministers.

3. It will be the Executive Committee’s responsibility to coordinate, oversee,

and evaluate the Program of Action, with the assistance of the General
Secretariat.

PROGRAM OF ACTION

I. REINFORCEMENT OF NATIONAL STRATEGIES

A. Control of the production, smuggling, and diversion of chemical
precursors

1. Implement the mechanism for reporting the export of controlled chemical
substances prior to their shipment and the timely answers from the intended
country of destination, pursuant to article 12 of the 1988 Vienna Convention.

2. Reinforce the monitoring of enterprises that produce, use, and/or sell
controlled chemical substances in order to avoid their diversion for the
production of illegal drugs.

3. Strengthen the mechanisms for detecting controlled chemical substances and
train the pertinent officials to identify those substances.

4. Make the measures for controlling the illegal production and smuggling of
controlled chemical substances stronger.

5. Set up and/or reinforce the mechanisms for controlling the transport and
diversion of chemical substances in the national territory through the following,
among other things:

a) The use of customs documents containing the generic name and
corresponding tariff code for trade in the controlled chemical
substances.

b) The establishment of a system of labeling and safety sealing that
would make it difficult to divert packaged or bottled products by

replacing them.

6. Create and/or update the national register of firms that import and export
controlled chemical substances. That information will be made available to

whoever asks for it.

7. Identify the needs of the legitimate industries dealing with potassium

permanganate, acetic anhydride, and the substances included in the Andean
Community’s control operations.

8. Develop inter-sector coordination for the purpose of identifying new
substances used in drug production for possible inclusion on the list of
controlled substances following a study of their level of use.

B. Technical eradication of illegal crops

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1. Build up national capacities to implement programs for the technical
eradication of illegal crops by hand or by air, in the countries that need it,
keeping in mind the environmental standards established by the competent

authorities.

2. Identify illegal crop growing areas, their size, evolution, features, zones of

influence, and any other relevant information.

3. Contribute to the coordination between the authorities responsible for
execution of the eradication programs and those in charge of the alternative

development programs.

4. Promote the establishment of agreements between local communities and

public authorities as a means of reducing crops grown for illegal purposes and
introducing alternative products.

C. Alternative Development.

1. Create the necessary economic, social, and cultural conditions to make it
feasible to replace the illegal crop production-based economy and to bring

farmers into the legal economy.

2. Identify and establish the characteristics of the populations of the illegal crop
growing areas and their zones of influence.

3. Design and put into use financial, economic, and technological instruments
for supporting national alternative development programs, including private

investment promotion, market opening, and the incorporation into alternative
production of products with a larger value added.

4. Boost the strengthening of grass-roots organizations and give them support
through consciousness-raising, training, and communication processes.

5. Step up the national capacity to offer basic social services and develop the

economic infrastructure in the illegal crop growing areas and their zones of
influence.

6. Design and implement social communication strategies to discourage the
involvement of farm and native families with illegal crops and promote
alternative crops.

D. Dismantling of the production and transport infrastructure and
organizations

1. Reinforce the mechanisms for coordination among the police and military
forces and state security bodies, the Public Ministry and/or the Attorney
General’s Office, and the Judiciary.

2. Build up the capacity for action of the intelligence units specialized in the
control of drug trafficking and ensure the timely exchange of information
among the various competent national authorities.

3. Increase the human, material, financial, and technological resources
allocated to the specialized units responsible for controlling drug trafficking

organizations.

4. Strengthen the mechanisms for detecting illegal drug production laboratories
and hidden airstrips.

5. Buttress the system for controlling illegal drug trafficking by sea, river, air,

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and land.

6. Create and/or reinforce the control mechanisms in order to impede illegal
trafficking in arms, ammunition, explosives, and other similar materials.

E. Asset laundering

1. Establish and/or build up the national intelligence and financial analysis

units.

2. Identify the existing types or methods of asset laundering and create the

corresponding control mechanisms.

3. Train specialized personnel in the agencies responsible for detecting and
controlling asset laundering operations.

4. Investigate the sectors capable of use for activities connected with asset
laundering and link them up with the intelligence and financial analysis units.

5. Design and implement mechanisms for administrative control of international
currency transactions.

6. Criminalize asset laundering as an autonomous offense, so that it will
encompass other criminal behavior (vehicle theft, extortion, kidnapping, white
slavery, trafficking in human organs, and arms trafficking).

7. Strengthen the application of provisions on the seizure of goods procured as
a result of drug trafficking or related offenses.

8. Establish regulations on the prevention of asset laundering in free trade
areas and at free ports.

F. Reduction of the demand

1. Put a stop to the rising trend in illegal drug consumption, especially among

children and young people, with schooling or not, through programs targeting
the family, community, and school.

2. Develop a mass media strategy to inform, sensitize, and educate young

people about the consequences of drug consumption, giving special emphasis to
the synthetic or designer drugs that have recently appeared on the scene.

3. Incorporate more information about prevention in the curriculums at the
different educational levels and educate parents and educational agents in the
new trends in illegal drug consumption.

4. Promote programs for the rehabilitation and social reinsertion of
drug-dependent individuals.

5. Implement mechanisms for overseeing and evaluating programs to cut down
the demand for drugs.

6. Design and launch programs to give human resources preparation and
training in prevention and rehabilitation.

7. Boost and support the participation of civil organizations in prevention and

rehabilitation activities.

II. REINFORCEMENT OF BINATIONAL STRATEGIES

1. Evaluate the existing bilateral drug control agreements, update and perfect

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them, and put them into force.

2. Promote and strengthen bilateral mechanisms, such as the mixed

commissions, border workshops, and neighborhood committees, in order to
draw up border action plans for:

a. Coillg thii drugs and lld cialc

substances.
b. Giving border authorities training in subjects connected with
drug control.

c. Carrying out combined interdiction operations.
d. Stepping-up the exchange of information and coordination of
logistics among border authorities.

e. Controlling the illegal traffic in firearms, ammunition, and other
similar materials.

3. Incorporate alternative development projects in the Border Integration
Zones and include them in the Project Bank to be set up as part of the Andean
Integration and Development Policy.

4. Institute effective mechanisms to control trafficking in illegal drugs,
controlled chemical substances, arms, ammunition, and other related materials,
through the National and Binational Border Service Centers (NBSC and BBSC).

5. Periodically examine and evaluate the execution and efficiency of the
binational cooperation measures that are carried out under this Andean

Cooperation Plan.

III. COMMUNITY STRATEGY

1. Establish an Andean mechanism for exchanging information through the
Andean Community website about the methods of trafficking in and diversion of
controlled chemical substances, use of new substances, successful control

operations, updated national registers of enterprises that import and export
controlled chemical substances, and changes in the importance and use of
border crossings for the illegal trafficking in those substances, and promote the

use of other national, regional and international computerized systems, such as
Unios contra Drogas (UCD) and the Venezuelan, inter-American, and
European drug observatories.

2. Step up the exchange of intelligence among the competent authorities of the
Andean countries, among others, making more use of existing communication

mechanisms, such as the Regional Liaison Offices of the World Customs
Organization (RILO) and the Inter-American Telecommunications Network for
Drug Control (RETCOD), in order to back regional efforts to control drugs,

related offenses, and the arms traffic.

3. Establish closer coordination among the national authorities responsible for

drug control in the Member Countries, among others, by appointing national
liaison officers in the respective institutions and assigning new duties to the
police and military attachés’ offices, as applicable, to support this Andean
Cooperation Plan.

4. Promote the training in common of national drug control officials through,
among others, the Andean Community’s Regional Antidrug Intelligence School

(ERCAIAD), ensuring its appropriate funding and adjusting its curriculum to the
priorities of the Andean Strategy, and supporting the establishment and
activation of the Andean Anti-drug Canine Training School.

5. Contribute to the signing of legal assistance agreements on criminal matters
and step up the execution of existing agreements, including procedures for the

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extradition of defendants accused of drug trafficking or related offenses under

the existing accords.

6. In each Member Country, appoint as liaison officers investigating judges who

are empowered to answer requests for reciprocal legal assistance in drug
trafficking cases or to remit them to the competent authorities for compliance.

7. Contribute to harmonizing national criminal and procedural legislation
through periodic meetings of the Ministers of Justice of the Andean Community,
bearing in mind the work that is being done under the aegis of the mechanism
between the European Union and the Andean Community on coordination and

cooperation in drug matters.

8. Promote the exchange of experiences and undertake joint actions to back

alternative development programs, incorporating for that purpose the Andean
Committee for Alternative Development (CADA) as the Andean Cooperation
Plan’s specialized body on the subject and supporting its efforts.

9. Reinforce cooperation in order to prevent and control asset laundering at the
Andean level through the exchange of experiences and interlinkage of the
Financial Intelligence Analysis Units of the countries in the Subregion and other

competent bodies.

10. Implement the guidelines established by the Inter-American System of

Standardized Drug Consumption Data (SIDUC) and entrust the analysis to the
Hipólito Unanue Convention (CONHU), so that the Andean Community can
have data on which it can draw to prepare prevention strategies that are

attuned to its situation.

11. Develop a joint strategy for preventing drug consumption and production
and controlling synthetic and designer drugs.

12. Identify the international technical cooperation requirements and capacities
and establish a mechanism for horizontal cooperation among the Member

Countries.

13. Design and carry out joint strategies for mobilizing international technical
and financial cooperation to support the measures provided for in the Andean

Strategy for the control of drugs and related offenses, as well as debt-for-
alternative development program support swaps.

14. Apply for the renewal and expansion of the programs of trade preferences
in support of drug control that benefit the Member Countries and obtain
conditions for preferential access to other markets and the removal of

restrictions on their full use.

15. Promote the mobilization of international cooperation for programs to
prevent and alleviate the environmental impact of the illegal drug problem,

including the recovery of ecosystems and conservation of the biodiversity.

16. Further international cooperation, in particular through the organization of

donor groups in order to boost alternative development, create jobs in
production, and alleviate poverty in illegal drug crop growing areas, in areas
from which labor is migrating, and in areas that are highly prone to use for

drug cultivation.

17. Coordinate joint drug control positions in dealing with third countries and in
international forums and organizations as part of the Andean Common Foreign

Policy.

18. Update the "Rodrigo Lara Bonilla" Convention on cooperation for preventing

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drug abuse and for suppressing the illegal traffic in narcotic drugs and
psychotropic substances, in order to adjust it to the needs created by this
Andean Cooperation Plan.

19. Examine and evaluate, as a Community, the implementation and
effectiveness of the measures that are carried out under this Andean
Cooperation Plan.

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453454 Annex 138

COSMO-FLUX® 411-F TECHNICAL DATA SHEET

(Cosmoagro, June 2002)

COSMO-FLUX® 411-F
Adjuvant for Agrochemicals Application

Specifications

Classification Additive for spraying
Chemical Mixture of Mineral oil and non-ionic specialized
description surfactants with coupling agents.

Active Mixture of Exitane esters: 17% EPA regulation:
Ingredient Linear alcohol + aryl Tolerance
ethoxylate exempt under 40
Mixture of non-ionic CFR 180.1001
tensoactive stereospecific, (c), (e)

linear alcohol ethoxylate regulation
propoxylate with small
quantities of aryl
ethoxylate
Additive Liquid isoparafins 83% EPA regulation:
ingredients isoparafin oil of high Tolerance

purity, with very low exempt under 40
toxicity, very low aromatic CFR 180.1001
content and low superficial (c), (e)
tension that improves regulation
humidibility which

reinforces efficacy of
active ingredients.

General Characteristics

o
Appearance at 25 C Amber liquid
Flash point Above 149 Co
Specific gravity 0.84

455Annex 138

Viscosity at 25 C 40 cP
Product pH at 10% 6.0-8.0

Superficial tension 25-28 dynes/cm

Solubility

Mineral Oils Soluble

Vegetable oils Soluble
Organic solvents Soluble
Water Dispersible, forming a rapid
emulsion

Characteristics of the Mixtures Created with COSMO-FLUX®411-F

Homogeneity Excellent
Persistence Over 24 hours
Compatibility with active Excellent (See ANNEX
ingredients “Evaluation of physical

compatibility …”
Mixing speed Rapid
Adherence Excellent

Toxicological Summary

Classification as Poison Not classified
Irritation in rabbit’s eyes Practically non irritating
(According to the Kay D.
Calandra Application)

Irritation of rabbit’s skin After applying it for 24 hours
undiluted produced slight
irritation
Sensitivity on human skin It does not cause irritation, nor is
it a skin sensitizer of human skin
Biodegradability (OECD method Non-ionic components of

for Non-ionic tensoactives) COSMO-FLUX® 411-F have a
biodegradability above 98%

First Aid

456Viscosity at 25 C 40 cP ¡In any case see a physician!
Product pH at 10% 6.0-8.0

Superficial tension 25-28 dynes/cm

Solubility

Mineral Oils Soluble

Vegetable oils Soluble
Organic solvents Soluble
Water Dispersible, forming a rapid
emulsion

Characteristics of the Mixtures Created with COSMO-FLUX® 411-F

Homogeneity Excellent
Persistence Over 24 hours
Compatibility with active Excellent (See ANNEX
ingredients “Evaluation of physical

compatibility …”
Mixing speed Rapid
Adherence Excellent

Toxicological Summary

Classification as Poison Not classified
Irritation in rabbit’s eyes Practically non irritating
(According to the Kay D.
Calandra Application)

Irritation of rabbit’s skin After applying it for 24 hours
undiluted produced slight
irritation
Sensitivity on human skin It does not cause irritation, nor is Disclaimer
it a skin sensitizer of human skin
Biodegradability (OECD method Non-ionic components of The information in this safety data sheet is provided in good faith and represents what is
known about the product at the time of publishing.
for Non-ionic tensoactives) COSMO-FLUX® 411-F have a Recommendations for its use and application are based on tests conducted by
biodegradability above 98%
COSMOAGRO, but its use and application in each specific case must be evaluated to
First Aid determine the appropriateness of its use.Annex 138

COSMOAGRO guarantees that the physical-chemical characteristics of the product
correspond to those listed on the label, and that through opinion on efficacy No. 2186 of
19 April 1993, issued by ICA (Colombian Agriculture and Livestock Institute), it was

verified that it is suitable for the purposes recommended therein, in accordance with the
specifications for use, but assumes no liability for how it may be used, since its
handling is beyond its control.
This product must be used with the recommendation of an Agricultural Engineer or a
Technical Assistant.

COSMOAGRO and COSMO-FLUX 411F are registered trademarks of
COSMOAGRO.
COSMO-FLUX is a product with the technological support of ICI – Specialty
Chemicals

458 Annex 139

PRESS ITEM : “ECUADOR ACCEPTS SPRAYINGS TO CONTINUE ”,EL C OMERCIO
(ECUADORIAN NEWSPAPER ), UITO ,4 AUGUST 2004

(El Comercio, Ecuador acepta que las fumigaciones continúen,available at:
http://www.elcomercio.com/solo_texto_search.asp?id_noticia=100416&anio=…
visited 19 February 2010))

[…]

The diplomatic authorities from Ecuador are in question.

This time because they accepted Colombia to continue with its antidrug fumigations in
the border line “taking the necessary precautions to avoid effects among the Ecuadorian
population”.

Thus, the Ecuadorian Foreign Ministry abandoned the thesis that the Government of
Bogotá was to refrain from carrying out sprayings with glyphosate in a strip of 10-km
from the border line. The agreement was reached yesterday, during the closing of the
fourth meeting of the Technical Scientific Commission in Quito.

The announcement was made after Vice-Minister Edwin Johnson announced he would
request Colombia to carry out the fumigations “at a low altitude”. And, also, because he
anticipated that if they agree to that request “there is no need to ask them to stop

spraying in the area of 10-km”.

[...]

459460 Annex 140

USD EPARTMENT OF STATE, UREAU FORINTERNATIONALN ARCOTICS ANDLAW
ENFORCEMENT A FFAIR, MEMORANDUM OF JUSTIFICATIOC ONCERNING THE
SECRETARY OF STATES 2002 ERTIFICATION OC ONDITIONSRELATED TO THE
A ERIALERADICATION OFLLICITCOCA INCOLOMBIA ,W ASHINGTOND.C.,2002

(Available at: http://www.state.gov/p/inl/rls/rpt/aeicc/13232.htm (last visited 8 March 2010), p. 1)

461 Annex 140

 





 






 
 


 

















 


 

 





















 




 
 

 




















 

462 Annex 141

U NITEDSTATESENVIRONMENTAL PROTECTION AGENCY (EPA), DDENDUM TO
MEMORANDUM . UBJECT: ESCRIPTION OGLYPHOSATE USED IN TU.S.AS ABASIS
FORCOMPARISON TOG LYPHOSATE USED COLOMBIA FORCOCA ERADICATIO,

FROM : VIRGINIW ERLING ANDTIMOTHY KIELY (EPA, BEAD BIOLOGICAL AND
ECONOMIC A NALYSID IVISIO)TOJAY ELLENBERG (EPA, BEAD BIOLOGICAL AND
ECONOMIC ANALYSISDIVISIO), 21 UGUST2002

(Available at: www.epa.gov/opp00001/foia/reviews/103601/103601-2002-08-
21a.pdf (last visited 2 October 2009))

463Annex 141

464 Annex 141

465466 Annex 142

U NITEDSTATES ENVIRONMENTAL PROTECTION A GENCY (EPA), ETAILS OF THE
C ONSULTATION FOR THD EPARTMENT OFSTATE: USE OFPESTICIDES FOC OCA AND
POPPYE RADICATIONP ROGRAM IN COLOMBIA , UGUST 2002

(In: United States Department of State, Bureau of International Narcotics and Law Enforcement Affairs,
Report on issues Related to the Aerial Eradication of Illicit Coca in Colombia,December 2003 (partially
included as EM, Vol. III, Annex 143). Available at: http://www.state.gov/p/inl/rls/rpt/aeicc/26991.htm
(last visited 27 February 2010), pp. 2, 4, 5, 8, 9, 10, 13, 23, 39)

467Annex 142

 

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

 





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


 



 


 
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




 









 




 

468 Annex 142

 















 






 

 


 



 



 

 

 

 




 
 
 

 


 

 

469Annex 142

 






 


 




 

 




 







 






 


 
















 

470 Annex 142

 






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












 

 


 











 


 


 



 


 



 

 

471Annex 142

 

 


 

  
   
    

    
 

     
 
     

    

 







 









 











 




 


 

472 Annex 142

 

 


 

   
     
  

      



 

      


 

      
 






 

 









 

 




 



 





 

473Annex 142

 









 





 


 

     
     

    
     

     

    
    
 









 











 




 

474 Annex 142

 


















 














 




 





 
 

 

 







 

475Annex 142

 



 






 


 


 




 

 



 






 









 
 

 








 

476 Annex 142

 





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
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 
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 


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 


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 
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 


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
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

 

477478 Annex 143

USD EPARTMENT OF STATE, UREAU FOR INTERNATIONAL NARCOTICS ANDL AW
E NFORCEMENT A FFAIR, MEMORANDUM OF JUSTIFICATIOC ONCERNING THE
SECRETARY OFSTATE’S2003 CERTIFICATION OCONDITIONSRELATED TO THEAERIAL
ERADICATION OFILLICICOCA INCOLOMBIA ,WASHINGTON D.C.,2003

(Available at: http://www.state.gov/p/inl/rls/rpt/aeicc/27484.htm (last visited 8
March 2010), p. 4)

479 Annex 143

  






 

















 

 









 









 












































 

480 Annex 143




 





 




 
 

 
 



 


 




 



 










 




 
 








  








 



 

 
 











 






 

 

481482 Annex 144

USD EPARTMENT OF STATE, UREAU FOR INTERNATIONAL NARCOTICS ANDL AW
E NFORCEMENT A FFAIR, MEMORANDUM OF JUSTIFICATIOC ONCERNING THE
SECRETARY OFSTATE’S2004 CERTIFICATION OCONDITIONSRELATED TO THEAERIAL
ERADICATION OFILLICICOCA INCOLOMBIA ,WASHINGTON D.C.,2004

(Available at: http://www.state.gov/p/inl/rls/rpt/aeicc/57012.htm (last visited 8
March 2010), pp. 3, 4)

483 Annex 144

  






 

















 


 












 




 


 


 






 

 
 

 

 










 
 






 






 

484 Annex 144

 
 





 






 






 

 

 

 


























 


 



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 








 

485  

 

  



 









 







 









  

 


















 


 


 










 



 











 



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
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

 

486 Annex 145

ANDEAN R EGULATION FOR THECONTROL OF CHEMICAL SUBSTANCES USED IN THE
ILLEGAL MANUFACTURE OF NARCOTIC DRUGS AND PSYCHOTROPIC SUBSTANCES
ANDEAN COMMUNITY , DECISIO602 OF2004

(Andean Community Decision 602 of 2004)

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Decision 602

Andean Regulation for the Control of chemical substances used in the
illegal manufacture of narcotic drugs and psychotropic substances

THE ANDEAN COUNCIL OF MINISTERS OF FOREIGN AFFAIRS, IN AN EXTENDED

MEETING WITH THE PRINCIPAL REPRESENTATIVES TO THE ANDEAN
COMMUNITY COMMISSION,

HAVING REVIEWED: The first paragraph b) in Article 3 and Article 16 of the
Cartagena Agreement; Articles 6 and 12 of the Regulations of the Andean
Council of Foreign Affairs Ministers; and Decisions 477 (International Customs

Traffic, in place of Decision 327), 478 (Mutual Assistance and Cooperation
between Customs Authorities of Andean Community Member Countries), 505
(Andean Cooperation Plan for Fighting Against Illegal Drugs and Associated
Crimes), 562 (Directives for the preparation, adoption and application of

Technical Regulations in Andean Community Member Countries and at
Community level) and 574 (Andean Customs Control System); and,

WHEREAS:

The Andean Community is determined to reinforce the application of the control
and surveillance procedures currently being used by its Member Countries,

concerning the traffic of chemical substances which are likely to be used for the
production of illicit drugs, cocaine and heroin in particular;

In an effort to protect the Community’s customs territory against eventual
diversion of chemical substance imports or exports for use in the manufacture of
illicit drugs, it is essential to establish in the Community an early warning

mechanism on the export of such chemical substances between Member
Countries, as a complement to any mechanisms in place between each Member
Country and other countries;

While the efforts individually made by the Member Countries to control and
monitor the chemical substances included in Tables I and II of the 1988 United
Nations Convention Against the Illicit Traffic in Narcotic Drugs and Psychotropic

Substances, are recognized, there is a need to prepare a basic harmonized list of
controlled chemical substances for Community use, which can be gradually
updated based on the experience gathered in the Andean Community and the

possibilities of effective international surveillance;

The Member Countries that have signed the 1988 United Nations Convention
Against the Illicit Traffic in Narcotic Drugs and Psychotropic Substances, abide by

the recommendations of the Model Regulations for the Control of Chemical
Substances Used in the Illicit Manufacture of Narcotic Drugs and Psychotropic
Substances issued by the Organization of American States (CICAD/OEA); and

the United Nations Manual for the Transport of Hazardous Goods;

The Member Countries, in the Third Meeting of the Executive Committee of the

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Crimes, resolved to carry out mutual cooperation actions to enhance the

parameters applicable to the control and surveillance of import, export, transport
and any other type of transactions at the Andean level and from other countries,
of chemical substances which could be possibly used in the production of cocaine

and heroin;

Abiding by the recommendation of the Third Meeting of the Executive Committee
of the Andean Cooperation Plan for Fighting against Illegal Drugs and Associated

Crimes, the General Secretariat has submitted Proposal 125/Rev. 1 on the
adoption of “Andean Regulations for the Control of Chemical Substances Used in
the Illicit Manufacture of Narcotic Drugs and Psychotropic Substances”;

The representatives to the Andean Community Commission have reviewed the
said Proposal and have issued a favorable opinion for adoption thereof on the
terms set forth in Proposal 125/Rev. 1;

DECIDES :

CHAPTER I

PURPOSE AND SCOPE OF APPLICATION

Article 1.- The purpose of this Regulation is to enhance control and surveillance
of import, export, transport and any other type of transactions at the Andean

level and from other countries, of the chemical substances included in the Basic
Harmonized List for Community Use, identified in

annex Annex I hereof, which are frequently used in the illicit production,
manufacture, preparation or extraction of narcotic drugs and psychotropic
substances, cocaine and heroin in particular.

Article 2.- This Regulation shall be applicable in the entire territory of the
Member Countries. Under no circumstances shall the rules and procedures set
forth in this Regulation imply:

a) The creation of unnecessary restrictions on free trade or free border
movement, pursuant to the provisions of the Cartagena Agreement or of
bilateral or multilateral Agreements or Treaties;

b) A limitation on the application in each Member Country of the provisions
of Article 12 of the 1988 United Nations Convention Against the Illicit
Traffic in Narcotic Drugs and Psychotropic Substances;

c) An attempt to ignore the authority of each Member Country to regulate
surveillance of all stages of trading, including production, storage and
distribution, among others, in accordance with the stipulations of this

Regulation.

Any cases not prescribed by this Regulation shall be governed by the internal

regulations of each Member Country with respect to matters related to the
monitoring of the chemical substances listed in Annex I hereof, and by the 1988
United Nations Convention Against the Illicit Traffic in Narcotic Drugs and
Psychotropic Substances.

CHAPTER II
DEFINITIONS

Article 3.- For the purposes of this Regulation, the following definitions shall
apply:

Competent Administrative Authorities: They are the national entities listed
in Annex V hereof, with jurisdiction to deal with matters related to licenses,

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registration, permits and authorizations, or to issue early warning on the import,
export, customs traffic or transport, at the Andean level and from other
countries, of the controlled chemical substances identified in Annex I hereof.

CAS: Chemical Abstract Service.

Andean Community: Comprised of the sovereign States of Bolivia, Colombia,

Ecuador, Peru and Venezuela, and by the organizations and institutions forming
part of the Andean Integration System established in the Cartagena Agreement.

Andean Council of Foreign Affairs Ministers: Comprised of the Foreign

Affairs Ministers of Andean Community Member Countries, with authority to
adopt Decisions which are binding on the Member Countries.

Concentration: Physical magnitude expressing the quantity of a controlled
substance, by volume unit.

Dilution: Lowering of the concentration of a controlled substance in water.

Import and Export: Respectively, the legal entry or exit of goods to or from
customs facilities, including special customs regimes and free trade zones.

Mix: This is the product into which one or more controlled substances are
combined, and which may be used in full or in part, in the extraction and/or
refining or synthesis of natural or man-made drugs.

NANDINA: Common Nomenclature of the Andean Community Member
Countries.

Controlled Chemical Substances:Chemical substances listed in Annex I and in
Tables I and II of the 1988 United Nations Convention Against Illicit Traffic in
Narcotic Drugs and Psychotropic Substances, included in Annex II hereof.

Transfer: Transfer of controlled chemical substances, performed under the
control of a single customs facility, from one transport or freight unit to another,
or to the same unit in a different journey, including unloading to land, with the

purpose of continuing to the place of destination.

International Customs Traffic: Customs regime, according to which, the goods
are carried under customs control, from a departure customs facility to an arrival

customs facility, in the same operation and in the course of which, one or more
borders are crossed.

Harmonized System: World Customs Organization’s Basic Harmonized

Commodity Description and Coding System .

CHAPTER III

BASIC COMMON LIST OF CONTROLLED CHEMICAL SUBSTANCES

Article 4.- For the purposes of effective monitoring at the Community level, the
controlled chemical substances listed in Annex I shall be identified by their

generic and chemical names, the CAS Code and their respective numerical codes
assigned in the Harmonized System – NANDINA classification.

Article 5.- The Chemical Substances Subcommittee shall study and recommend

the application of additional measures required for greater effectiveness of the
control exercised over the substances included in Annex I, as well as the physical
and chemical parameters required to establish control of any mix, concentration

or dilution.

CHAPTER IV

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IMPORT, EXPORT, TRAFFIC AND TRANSFER REQUIREMENTS

Article 6.- Anyone importing or exporting controlled chemical substances must

present, notwithstanding compliance with requirements pertaining to
registration, qualification, license and filing, pursuant to the internal regulations
of each Member Country (as well as with the relevant foreign trade regime), any
authorizations or permits issued by the Competent Administrative Authorities to

import or export and for international traffic under the International Customs
Traffic Declaration defined in Decision 477.

Article 7.- Anyone importing or exporting the chemical substances included in
Annex I hereof and those listed in the corresponding legislation of each Member
Country, shall be subject to a control system consisting of authorizations,
licenses and similar procedures to be followed pursuant to the internal

regulations of each Member Country.

Article 8.- Applications for authorization and permits must be filed by the

importer or exporter with the competent national authority, within the terms
stipulated in the internal regulations of each Member Country in the case of
imports. An authorization or permit may only be used once and it may not cover
future imports of substances of a different nature.

Said authorizations and permits shall expire 180 calendar days from their date of
issue. If this term has elapsed without the respective import or export having

been performed, a new authorization or permit must be sought.

Applications for a permit or authorization must include the following information:

a) Name, address, classification, license or registration number, telephone
and fax number and/or electronic mail of importer and exporter;

b) Name, address, telephone and fax number and/or electronic mail of

import or export agent and of the shipping agent, as the case may be;

c) Names, NANDINA subheading and CAS name for each chemical
substance listed in Annex I of this Regulation, as well as the description

found on the label of each piece, package and/or container;

d) Net weight or volume of the product, in kilograms or liters, with their

respective fractions;

e) Amount and gross weight of pieces or packages;

f) Number e identification of containers, if applicable;

g) Proposed date for shipping and import or export. Place of origin,
shipping points, stopovers, ports of entry and destination; and

h) Transport means and identification of carrier.

Article 9.- The Competent Administrative Authorities may deny authorization or

permit, or suspend the operation whenever they have well grounded reasons to
believe that the substances may be used in the illicit production, manufacture,
extraction or preparation of narcotic drugs and psychotropic substances.

Article 10.- International transport of controlled chemical substances within the
Andean Community may only be performed using the methods and under the
terms set forth by the competent authorities on transport matters, pursuant to

the provisions of Article 28 of Decision 477 on Customs Traffic.

CHAPTER V

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ADMINISTRATIVE MEASURES FOR REGISTRATION AND INFORMATION

Article 11.- For the purposes hereof, notwithstanding the provisions of domestic
legislation, the Competent Administrative Authorities shall keep a record of

individuals or legal entities authorized to import or export controlled chemical
substances, including the consignees, as well as authorizations granted, rejected
or revoked. The Competent Administrative Authorities must keep the

confidentiality of any information classified as industrial, business, trade or
professional secret.

Article 12.- The records must include the following information about the

importer or exporter; and when applicable, about the consignee:

a) Name and address, telephone, telex and fax number and/or electronic

mail address;

b) License or registration type and number, including issue and expiry
dates; and,

c) The main industrial activity and the chemical substances listed in Annex
I used in the industrial process.

Importers and exporters must report to the Competent Administrative
Authorities any changes in the information supplied, within thirty (30) calendar
days following the date when the change takes place.

Taking into consideration the recommendations made by the Technical
Subcommittee on Chemical Substances, the Member Countries shall centralize

the information included in the aforementioned records, on the Andean
Community General Secretariat’s web site. The General Secretariat shall set
forth the most adequate security procedures for such information, as well as the
most suitable mechanisms to facilitate information inquiries by the competent

national entities.

The information must be forwarded to the General Secretariat at least once a

year, preferably during the month of February.

Article 13.- Anyone importing or exporting controlled chemical substances must
keep and maintain records for a period of no less than two (2) years. Such

records must be complete, accurate and up to date, concerning each operation
associated with those substances, including the following information:

a) Amounts imported and exported, with specific details on:

Transaction date;

Name, address, telephone, fax, electronic mail, and license or registration
number for each and every one of the parties involved in the operation
and the ultimate consignee, if other than one of the parties involved in the

operation;

Name, NANDINA subheading, amount, unit of measurement, form of
presentation and type of package containing the chemical substance; and

Transportation means and carrier identification.

b) Amounts sold internally;

c) Quantity in stock; and

d) Quantities lost or destroyed and drops caused by shrinkage or other causes

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such as accidents and theft. The competent authorities must be informed of any
losses or unusual or significant amounts missing with respect to chemical
substances under their control. Once the information is confirmed, the

competent authorities must notify the authorities of the country of origin,
destination or transit, as soon as possible, providing them with as much detail as
may be available.

Article 14.- Importers and exporters shall keep individual files containing
records for each authorized transaction and a record of stock balances of the
chemical substances included in the respective permit, for a period equal to that
indicated in the preceding article.

Article 15.- The Competent Administrative Authorities may propose to the
General Secretariat, the inclusion or exclusion of controlled chemical substances

in Annex I of this Regulation. To this effect, they shall forward a request to the
General Secretariat using the Technical Sheet found in Annex III hereof,
including the reasons supporting their proposal.

Article 16.- The following is the procedure to be used for inclusion or exclusion
of controlled chemical substances:

a) The request submitted by the interested Member Country shall be

forwarded to the rest of the Member Countries through the General
Secretariat;

b) The General Secretariat shall forward the request to the national
representatives of the rest of Member Countries accredited to the Chemical
Substances Subcommittee, within five (5) business days from the receipt
of the said request;

c) The members of the Chemical Substances Subcommittee shall issue a
response to the request within a term of no greater than thirty (30)

business days counted as from the date of the notice forwarded by the
General Secretariat, unless a Member Country asks for a term extension;

d) The extension referred to in paragraph c) above shall be granted only

once and shall be authorized for a maximum term of fifteen (15) business
days; and

e) The General Secretariat shall issue a Resolution incorporating or

excluding the substance into or from Annex I hereof, provided the
consensus of the Member Countries is obtained; otherwise, Annex I shall
not be modified.

CHAPTER VI
INTERNATIONAL SURVEILLANCE AND COOPERATION

Article 17.- The Member Country from which territory any of the substances
listed in Annex I hereof is exported, prior to the export being carried out and
through its competent authorities, shall give prior notice of such export to the

competent authority of the importing Member Country, using the form prepared
by the International Narcotics Control Board (INCB).

Article 18.- Once the prior notice is given, the importing Member Country must

acknowledge receipt thereof and within fifteen (15) business days, it must
respond to the competent authority of the exporting Member Country stating
conformity or otherwise with respect to the transaction. If the exporting Member
Country has not received a response from the competent authority of the

importing Member Country within the aforementioned term, it shall mean that
the transaction has been accepted.

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The Member Countries agree to timely give each other every details concerning
the follow up on the information supplied and to cooperate to provide each other
with all the information relative to any presumed illicit operations.

Article 19.- The imports shall be suspended whenever, in the opinion of the
importing Member Country, there are reasonable signs to assume that the
controlled chemical substances may be subject of diversion for the illicit

manufacture of narcotic drugs or of psychotropic substances, or whenever the
exporting Member Country requests it.

The Member Countries shall cooperate to afford each other any and all
information relative to the presumed illicit operations.

Article 20.- A Member Country receiving the information subject matter hereof,

must maintain confidentiality with regard to any information classified as
industrial, business, trade or professional secret, as well as any other additional
details, in accordance with its internal regulations and international

commitments in place.

Article 21.- Any individuals and legal entities importing, exporting, trading or
transporting controlled chemical substances, must immediately report to the

Competent Administrative Authorities the transactions or proposed transactions
they are involved in, whenever there are reasonable signs to believe that such
substances could be used in the illicit production, manufacture, extraction or

preparation of narcotic drugs, psychotropic or other substances with similar
effects.

Among other cases, reasonable signs shall be considered to exist whenever the
traded amount of the chemical substances contained in Annex I, the form of
payment, or the characteristics of the buyer, are extraordinary, unusual or do
not correspond to the company’s business or industrial endeavor.

Article 22.- Whenever an inspection is to be made during the transport of
controlled chemical substances in an international customs traffic operation, as

well as whenever a fault, infraction or a crime occurs during such traffic, the
procedures established in Decision 477 concerning International Customs Traffic
shall be followed.

Article 23.- The Member Countries shall endeavor to adopt measures for
cooperation with any private sector entities undertaking activities related to the
scope of application of this Regulation, particularly with regard to the supply of

information and records to the competent authorities, prior notification
procedures and timely information with regard to suspicious and unusual
operations.

All information supplied shall be treated confidentially and shall not be disclosed,
except in the case of a court order.

CHAPTER VII
MARKING AND LABELING

Article 24.- In order to enhance surveillance over international trade between

Member Countries, each shipment of controlled chemical substances must bear
the “Standard Andean Label” in a visible place on the original packages,
including details of their designation as “controlled chemical substances”. The

operators shall make sure that the Standard Andean Label is placed before
shipping.

The aforementioned label shall be designed by the Technical Subcommittee for
Chemical Substances Control, in accordance with technological progress and
based on the enclosed model found in Annex IV of this Regulation, with the

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following features:

a) It must be easily visible and legible;

b) It must be capable of remaining on the wind and weather without its
information being notably deteriorated;

c) It must be placed on the surface of the piece, package or container; and

d) It must allow for the marking of information identifying the exporter or
the consignee, or both.

CHAPTER VIII
ADMINISTRATIVE DEFAULT

Article 25.- Notwithstanding the provisions of the Cartagena Agreement and of
the Treaty for the Creation of the Andean Community Court of Justice,
non-compliance with the following provisions of this Regulation shall be
considered faults or violations subject to administrative sanctions applied in

accordance with the internal legislation of each Member Country:

a) Individuals or legal entities not obtaining, updating or renewing

registration;

b) Iiul orlgaltiis nottigi port or export
authorization within the required time;

c) The information contained in the special records is not updated, or it is
not accurate; and

d) Transactions are carried out with companies which have not been duly
registered.

CHAPTER IX

TECHNICAL SUBCOMMITTEE FOR CHEMICAL SUBSTANCES

Article 26.- The Technical Subcommittee for Chemical Substances is the body

responsible for issuing non-binding technical opinions on matters related to
controlled chemical substances. Its set up and organization shall be determined
by the Executive Committee of the Andean Cooperation Plan for Fighting Against
Illegal Drugs and Associated Crimes, and it shall be responsible for the following

duties:

a) Adopt a mechanism for permanent evaluation and follow up on

compliance with the provisions contained herein, in order to propose the
adjustments required in view of the constant changes occurring in the
diversion methods used, in such a way that the effectiveness hereof is not
lost;

b) Conduct periodic specialized studies to provide advice to national
administrative authorities and the General Secretariat, to identify any

trends and methods observed in the Andean Region in matters pertaining
to:

i. Production, manufacture, preparation, transformation, storage,

import, export, customs traffic, trade and transport of controlled
chemical substances;

ii. Final disposal of controlled chemical substances, taking into

account environmental protection measures, whenever such disposal
implies the technical destruction of the seized substances;

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iii. Transfer of controlled chemical substances, including the
permanent re-export of the seized substances;

iv. Domestic and international diversion of controlled chemical
substances for illicit purposes;

v. Determination of acceptable levels of variation in the weight or

measurement of the imported controlled chemical substances,
produced during the period involving their transport and storage, in
order to recommend the competent national authorities on the

adoption of corrective action and investigation of any diversion of
partial quantities of the said substances;

vi. Introduction of new substances into the illicit production chain, as

well as trends on illicit drug production; and

c) Prepare the essential technical studies that will make it possible to

determine the control of the mixtures, concentrations and dilutions.

The Technical Subcommittee for Chemical Substances shall keep permanent
contact to study the problem relating to the identification, investigation and

verification of the existence of crimes, so as to recommend the most appropriate
course of action.

Article 27.- This Regulation shall come into force as from the date of its
publication in the Official Gazette of the Cartagena Agreement.

FINAL PROVISIONS

FIRST.- For the purpose of compliance with the provisions hereof, a close work
relationship must be established between the Competent Administrative

Authorities and the Andean Committee on Customs Affairs, and with the
Committee Against Fraud.

At the national level, each Member Country must design and implement an

inter-institutional coordination mechanism.

TWO.- The Member Countries must adapt their national laws so as to define as a

crime, any activity associated with the diversion of chemical substances likely to
be used in the production of illicit drugs.

THREE.- To resolve on the qualification, registration, filing and granting of

licenses, authorizations or similar permits, the Member Countries shall establish
minimum requirements such as: verification of criminal record and police record
with regard to illicit drug traffic and associated crimes of the shareholders and/or

legal representatives, and a physical and legal verification of the existence of the
company. Such circumstances may also be taken into account for revoking or
suspending any permits and authorizations granted, all of the above in

accordance with their internal legal system.

TEMPORARY PROVISIONS

ONE.- The Technical Subcommittee for Chemical Substances shall propose to the
Executive Committee of the Andean Cooperation Plan for Fighting Against Illegal
Drugs and Associated Crimes, the model and contents of the Standard Andean

Label referred to in Article 25 hereof. Once the Standard Andean Label has been
approved by the Executive Committee, it shall be forwarded to the General
Secretariat for publication by way of Resolution.

TWO.- The Member Countries hereby agree to report to the General Secretariat,
through their respective Ministries of Foreign Affairs, any changes to the list of

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Competent Organizations mentioned in Annex V hereof, within a term of no
more than thirty (30) calendar days counted as from the date when such
modification is decided at the national level. Within the following five (5)

business days, the General Secretariat shall notify the Member Countries of the
corresponding modifications.

Given in the City of Cusco, Peru, on December 6, 2004.

ANNEX I

Basic Common List of the chemical substances that are subject to
additional specific control measures within the Andean Community territory

1 CHEMICAL
NANDINA CAS NUMBER GENERIC NAME NAME

Dimethyil ketone

2914.11.00 67-64-1 Acetone / *
2-propanone

2806.10.00 7647-01-0 Hydrochloric Acid Muriatic Acid
*

2807.00.10 7664-93-9 Sulfuric Acid Sulfuric Acid *

2814.10.00 7664-41-7 Anhydrous Anhydrous
Ammonia Ammonia

Ammonium
2814.20.00 1336-21-6 Aqueous Ammonia Hydroxide

2915.24.00 108-24-7 Acetic Anhydride Acetic Anhydride
*

Sodium
2836.20.00 497-19-8 Sodium Carbonate Carbonate

2909.11.00 60-29-7 Ethyl Ether Diethyl Oxide
*

Ethyl Methyl
2914.12.00 78-93-3 Ketone Butanone *

2841.61.00 7722-64-7 Potasium Potasium
Permanganate Permanganate *

Toluene (with no

2707.20.00 108-88-3 defined chemical Toluene *
structure)

Toluene (HC
derivative, with
2902.30.00 Toluene
defined chemical
structure)

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* Substances found in Tables I and II of the 1988 United Nations Convention
Against the Illicit Traffic in Narcotic Drugs and Psychotropic Substances, which
are included by the Andean Community in Annex I due to their importance in the

region in matters relating to industrial development, and particularly with regard
to their diversion for use in the illicit production of natural origin drugs.

[1]
Chemical Abstract Substance.

ANNEX II

Tables I and II
of the 1988 United Nations Convention Against
Illicit Traffic in Narcotic Drugs and Psychotropic Substances

TABLE I S.A. CAS TABLE II S.A. CAS

N-acetyilanthranylic
2924.29 89-52-1 Acetone
acid 2914.11 67-64-1
Anthranilic
Lysergic acid 2939.63 82-58-6
acid 2922.43 118-92-3

Acetic anhydride 2915.24 108-24-7 Hydrochloric
acid 2806.10 118-92-3

Ephedrine 2939.41 299-42-3 Phenylacetic
acid 2916.34 103-82-2

Ergometrine 2939.61 60-79-7 Sulfuric acid 2807.00 7664-93-9

Ergotamine 2939.62 113-15-5 Ethyl ether 2909.11 60-29-7

1-phenyl- Ethyl methyl
2-propanone 2914.31 103-79-7 ketone 2914.12 78-93-3

Isosafrole 2932.91 2932.91 Piperidine 2933.32 110-89-4

3,4-methylene

dioxyphenyl- 2932.99 2932.92 Toluene 2902.30 108-88-3
2-propanone

Potasium 2841.61 2841.61
Permanganate

Piperonal 2932.93 2932.93

Safrole 2932.94 2932.94

Seudoephedrine 2939.42 2939.42

ANNEX III

Technical Sheet

1. Name or corporate name of the institution

2. Justification

— Legal framework applied

— Type of companies or industries in which such diversion has been
confirmed

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— Reports submitted by at least two laboratories on the product’s
chemical analysis.

— Statistics supporting product diversion.

3. Date of shipment and date of receipt of the request at the General
Secretariat.

4. Signature of person responsible at the competent national authority.

ANNEX IV

Standard Andean Label Model

General provisions

General format

Color

Symbols

Ecuador proposes the following suggested model label:

ANDEAN COMMUNITY

CONTROLLED SUBSTANCE

SUBSTANCE NAME

SHIPPING COUNTRY / ORIGIN / EXPORTER

DESTINATION COUNTRY / IMPORTER

ANNEX V

Competent Organizations

Bolivia

Dirección General de Sustancias Controladas
(General Bureau on Controlled Substances)
Viceministerio de Defensa Social

(Office of the Vice-Minister of Social Defense)
Ministry of Government

Colombia

Dirección Nacional de Estupefacientes
(National Narcotic Drugs Bureau) , entity attached to the Ministry of the Interior

and Justice
Fondo Nacional de Estupefacientes del Ministerio de la Protección Social
(National Narcotic Drugs Fund) , entity forming part of the Ministry of Social

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Protection
Dirección de Impuestos y Aduanas Nacionales (DIAN)
(National Tax and Customs Bureau)

Ecuador

Dirección Técnica Nacional de Control y Fiscalización

(National Control and Monitoring Technical Bureau)
CONSEP

Peru

Dirección de Insumos Químicos y Productos Fiscalizados
(Controlled Chemical Inputs and Products Bureau)
Ministry of Production

Bolivarian Republic of Venezuela

Dirección de Control y Fiscalización de Sustancias Químicas

(Chemical Substances Control and Monitoring Bureau)
CONACUID

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USD EPARTMENT OF STATE, UREAU FOR INTERNATIONAL NARCOTICS ANDL AW
E NFORCEMENT A FFAIR, MEMORANDUM OF JUSTIFICATIOC ONCERNING THE
SECRETARY OFSTATE’S2005 CERTIFICATION OCONDITIONSRELATED TO THEAERIAL
ERADICATION OFILLICICOCA INCOLOMBIA ,WASHINGTON D.C.,2005

(Available at: http://www.state.gov/p/inl/rls/rpt/aeicc/52411.htm (last visited 8
March 2010) pp. 1, 2)

501 Annex 146

  






 




 





 








 

 











 

























 

















 

502 Annex 146

 

 






 












 

















 



   

 


 


 










  




 











 














 

503504 Annex 147

USD EPARTMENT OF STATE, UREAU FORINTERNATIONAL NARCOTICS AND LAW
ENFORCEMENT AFFAIRS, EMORANDUM OF JUSTIFICATIOCONCERNING THE
SECRETARY OFSTATE’S2006 ERTIFICATION OC ONDITIONSRELATED TO THA ERIAL
ERADICATION OFLLICITCOCA INCOLOMBIA,W ASHINGTON D.C.,2006

(Available at: http://www.state.gov/p/inl/rls/rpt/aeicc/70974.htm (last visited 8 March 2010) pp. 1, 2)

505 Annex 147

  





 







 









 



 








 
  
























 


  













 






  

 

506 Annex 147



 

  

 


 






 

 



 









 







 


  




 
 

 


 



 

 





 
 


 






 

  

 


 
 

 






 


 

 

507508 Annex 148

PRESS ITEM: “DEFENCE M INISTER WELCOMES THE NEW SUPER T UCANO AIRPLANES

OF THE AIRF ORCE”, 14DECEMBER 2006

(Web page of the Colombian Ministry of National Defence, Mindefensa da la bienvenida a los nuevos
aviones Super Tucano de la FAC, available at:
http://alpha.mindefensa.gov.co/index.php?page=181&id=4875&PHPSESSID=f60…
aa49969c7b , (last visited 20 February 2010), pp. 1,2)

[…]

“We explained to the Ecuadorian authorities, President Uribe spoke to President
Palacio, I spoke to the Minister of Defence of Ecuador, and we explained to them why

we are taking that step. We cannot allow the production of coca plants to keep growing,
or the proliferation of processing laboratories, or the presence of the guerrillas and the
increase in violence in that zone of the country”.

[…]

509510 Annex 149

US Department of State, Bureau for International Narcotics and Law
Enforcement Affairs, Memorandum of Justification Concerning the Secretary of
State’s 2007 Certification of Conditions Related to the Aerial Eradication of Illicit

Coca in Colombia, Washington D.C., 2007

(Available at: http://www.state.gov/p/inl/rls/rpt/aeicc/111210.htm (last visited 8
March 2010), p. 2)

511 Annex 149

  









 


  
 

 
 

 
 

 








  
 








  


 

  




















 













 


  

 

512 Annex 149







 


   



 


 

 

 


 


 
 






 
 


   

 
 








 

   
 



 



 

 



 







 


 


  




 

 

513514 Annex 150

PRESS ITEM : “ECUADOR WILL SUE C OLOMBIA OVER SPRAYINGS IN THE BORDER ”,
R EVISTAC AMINOS ,2 JULY 2007

(Revista Caminos, Demandará Ecuador a Colombia por fumigaciones en la frontera, available at:
http://ecaminos.org/leer.php/4745visited 20 February 2010))

President Correa and foreign minister Espinosa received the report of the Ecuadorian
scientific commission about the effects on the Ecuadorian border of Plan Colombia’s
fumigations

[…]

“We have also implemented a legal way, preparing a case that Ecuador will bring
against Colombia before the International Tribunal at The Hague” insisted the Minister
of Foreign Affairs.

Maria Fernanda Espinosa vetted the report, after stating that the government’s policy
has been supplemented “by the scientific way”.

[...]

President Correa reiterates that the report is basis for compensations sought by Ecuador
for those affected.

[…]

515516 Annex 151

C OLOMBIAN ASSOCIATION OFA UTONOMOUS REGIONAL CORPORATIONS

(ASOCARS),“S HARED RESPONSIBILITY: HE W ORLD D RUG PROBLEM FROM A
GREEN P ERSPECTIVE”, ERIÓDICO VIRTUAL, SSUEN°14,2008

(Asociación colombiana de Corporaciones Autónomas Regionales (ASOCARS), “Responsabilidad
Compartida: El Problemas Mundial de las Drogas A Través de Una Perspectiva Verde,Periódico
Virtual, Edición No. 14, 2008. Available at:
http://www.asocars.org.co/periodico/14edicion/responsabilidad/RESPONSAB…
socars_2.pdf (last visited 25 February 2010))

[Page 1]

[…]

The Silent Catastrophe

Colombia, located in the Northeast of South America, has two coasts and is the final
point of the great Andes. This creates a wide range of climates in the country. Its

territory has one of the highest levels of rain in the world and encompasses a land
equivalent to France and Spain together.

Less than 6% of the Earth’s total surface is covered by tropical jungle, and three fourths

of the planet’s biodiversity are to be found in this region. The tropical jungle of the
Amazons, responsible for 15% of the world’s oxygen, covers 40% of South America
and 35% of Colombia, one of the ten nations with more primary forests in the world.

Colombia is, after Brazil, the country with more biodiversity in the planet. It holds 18%
of the world’s bird species and is the richest country in amphibians.

[Page 2]

Moreover, Colombia has a higher potential for water production than countries of the
size of the continental United States or India.

Today there are 257 protected natural zones, a number which is likely to increase in the
near future, with a total area larger that Holland, Belgium and Denmark together.

Unfortunately, cocaine production geared by consumption threatens the conservation of

this environment and places unique species of fauna and flora at risk. The majority of

517Annex 151

the 78,000 hectares of coca in Colombia are located in or near the Amazon’s tropical
jungle.

As an aggravating factor, the Colombian Anti-Narcotics Police estimates that for each
hectare of coca cultivated, three hectares of forest are cut down. Taking into account the
frequency with which coca plantations are relocated to avoid detection, 200,000
hectares of Colombian tropical jungle are burned and cut down each year to grow coca.
This burning process is the main cause for pollution in the Colombian jungle zones.

In total, it is estimated that over the last 20 years 2.2 million hectares of Colombian
jungle have been deforested.

[…]

518 Annex 152

EMBASSY OF THEUNITEDSTATES INBOGOTÁ ,FACT SHEET2008,“COCAINE
PRODUCTION AND CULTIVATION: OLOMBIA ”,6N OVEMBER 2009,ENCLOSURE TO
PRESSITEM : “FFICIALU.S.COLOMBIA SURVEY SHOWS SHARP DROP INC OCA

CULTIVATION ANDCOCAINE PRODUCTION ”

(Press item is available at: http://bogota.usembassy.gov/pr_75_061109.html and fact sheet is available at:
http://bogota.usembassy.gov/root/pdfs/factsheet2008cocaineproductioncul… (last visited 25
February 2010))

519Annex 152

FACT SHEET

2008 COCAINE PRODUCTION AND CULTIVATION: COLOMBIA

 According to the most recent crop estimate from the CNC , potential cocaine production in
Colombia dropped fully 39 percent between 2007 and 2008.

 The potential production of pure cocaine fell from an estimated 485 metric tons in 2007 to 295
metric tons in 2008.

 Further, the area under cultivation dropped 29 pcent during the same time frame.
 The coca crop declined from an estimated 167,000 ha in 2007 to only 119,000 ha in 2008.

 The current estimate provides a genuinely comparable number to the previous year, given that the
“search area” for the estimate was su bstantially equivalent to that of the previous year.

FACTORS AFFECTING THE DECLINE

 The cumulative effect of steadfast eradication pressure delivered against the primary Colombian
growing areas diminished not only the size of the coca fields, but further the ability of remaining

fields to produce normal amounts of coca leaf.
 Increased government presence and the deployment of security forces in select growing regions

were instrumental in preventing coca cultivation and production.
 Successful operations against drug trafficking organizations kept them under constant pressure

and reduced their control over the cocaine industry.

ERADICATIONFACTORS

 Combined aerial spraying and manual eradication for 2008 was 227,605 ha.

2001 2002 2003 2004 2005 2006 2007 2008
Cultivation 169,800 144,450 113,850 114,100 144,000 157,200 167,000 119,000
Spray (Air)* 84,251 122,695 127,112 131,824 134,474 164,119 148,435 129,876
Erad (Man) 1,745 2,762 4,220 6,232 37,540 42,110 64,979 95,731

2008 COCAINE PRODUCTION AND CULTIVATION IN THE ANDES: PERU, BOLIVIA, AND

COLOMBIA

 Peru and Bolivia were found to have increased their potential cocaine production for 2008, Peru by a

slight 2 percent (to 215 metric tons), while Bolivian production was re -calculated due to increased
efficiency converting coca leaf to cocaine, providing a surge of 50 percent increased productive

potential in 2008 to 195 metric tons pure cocaine.
 All told, cocaine potential production in the three Andean nations in 2008 was no morean 705

metric tons pure, a figure last seen this low in 1997, andrepresents a decline from the peak year of
production (2001, with an estimated potential production of 1055 metric tons pure) of fully 33
percent.

 Importantly, with the steep drop in Colobia, there has not been an appreciable upsurge of coca
cultivation in either Peru or Bolivia.

 Peruvian cultivation experienced a small uptick of 14 percent (from 36,000 ha in 2007 to 41,000 ha
in 2008). Even with that uptick Peruvian cultivation remained below where it stood in 2006.

520 Annex 152

 Bolivia experienced no more than an 8 percent increase in cultivation (from 29,500 ha in 2007 to

32,000 ha in 2008).
 These modest increases in cultivation were more than offset by the steep decline of 48,000 ha in

Colombian cultivation.

DOMESTIC INDICATORS SHOW IMPACT ON U.S. STREETS DURING ATIME PERIOD
THAT EXPERIENCED COCAINE DISRUPTION

 The most recent National Survey on Drug Use and Health data showed that 18 -25 year old past-
month cocaine prevalence dropped 32 percent between 2006 and 2008. Past year initiates for

cocaine among persons aged 12 and older dropped 26% over the same period.
 Workplace positive rates, as measured by Quest Diagnostics, for cocaine dropped 42% percent

from 0.72% in 2006 to 0.42% in 2008..
 DEA STRIDE data on cocaine price and purity show that f romthe first quarter of 2007 through
nd
2 quarter of2009, the price per pure gram of cocaine increased 77.1%, from $99.48 to $176.14,
while the purity decreased 27.4%, from 67% to 4% 9.
 DEA Cocaine Signature Program data on arriva -lone seized cocaine show a drop in purity of

cocaine bricks arriving in the U.S. ports -of-entry, dropping from 83% to 85% purity (over the-5
year period mid-2003 to mid-2008) to 75% by mid-2009.

 Cocaine seizures along the Southwest Border dropped 37% from a quarterly average of 7,300
kilograms in the 1 quarter of 2007 to 4,600 kilograms in the 3quarter of 2009.

521 Annex 152

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










 

 

 

522 Annex 153

ARIAS V. DYNCORP, PLAINTIFFS’ MOTION TO DISMISS THREE INDIVIDUAL PLAINTIFFS,
23 DECEMBER 2009

(United States District Court for the District of Columbia, Case No. 1:01-cv-01908-
RWR-DAR, Document 171)

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530 Annex 154

ARIAS . DYNCORP , DEFENDANTS ’ RESPONSE ,6JANUARY 2010

(United States District Court for the District of Columbia, Case No. 1:01-cv-01908-
RWR-DAR, Document 172)

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545546 Annex 155

ARIAS V. DYNCORP , --- FUPP . D ---,2010WL94563 (DDC,2010,R OBERTS J)

(United States District Court for the District of Columbia, Case No. 1:01-cv-01908-
RWR-DAR, 2010 WL 94563)

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548 Annex 155

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550 Annex 156

A RIAS . YNCORP, EFENDANTS ’ MOTION FORSANCTIONS AGAINST THE
ARIAS/QUINTEROS PLAINTIFFS FOV IOLATIONS OD ISCOVERYO RDERS,26
JANUARY 2010

(United States District Court for the District of Columbia, Case No. 1:01-cv-01908-
RWR-DAR, Document 176)

551Annex 156

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IN THE UNITED STATES DISTRICT COURT
FOR THE DISTRICT OF COLUMBIA

)
Venancio Aguasanta Arias, et al., )
)
Plaintiffs, ) Case Number: 1:01cv01908 (RWR-DAR)
v. )

)
DynCorp, et al. )
Defendants. )
)

)
Nestor Ermogenes Arroyo Quinteros, et al., ) Case Number: 1:07cv01042 (RWR-DAR)
)
Plaintiffs, ) (Cases Consolidated for Case
v. ) Management and Discovery)

)
DynCorp, et al. )
Defendants. )

DEFENDANTS’ MOTION FOR SANCTIONS AGAINST
THE ARIAS/QUINTEROS PLAINTIFFS FOR VIOLATIONS
OF DISCOVERY ORDERS, WITH STATEMENT OF
SUPPORTING POINTS AND AUTHORITIES INCLUDED HEREIN

Filed: January 26, 2010 Joe G. Hollingsworth (D.C. Bar # 203273)
Eric G. Lasker (D.C. Bar # 430180)

Rosemary Stewart (D.C. Bar # 204438)
HOLLINGSWORTH LLP
1350 I Street, NW
Washington, D.C. 20005

Phone: (202) 898-5800
Fax: (202) 682-1639

Counsel for the Defendants

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TABLE OF CONTENTS

Page

I. BACKGROUND FACTS....................................................................................................3

A. The Court’s Prolonged Effort To Secure Prima Facie Disclosures from the

Individual Plaintiffs. ................................................................................................3

B. Plaintiffs’ Most Recent Violation of the Court’s Order to Provide
Individualized Causation Statements.......................................................................9

C. The Deposition Testimony of the 20 Test Plaintiffs Establishes Numerous
Further Violations of The Court’s Orders..............................................................11

1. The Calero Family.....................................................................................13

2. The Salas Family........................................................................................15

3. The Quevedo Family..................................................................................18

4. The Mestanza Family.................................................................................20

5. The Sandoval Family.................................................................................24

6. Elvia Alvarez .............................................................................................26

7. Edgar Balcazar...........................................................................................27

D. The Test Plaintiffs’ History of Changing Stories Regarding Their Alleged
Exposures Began Long Before This Litigation. ....................................................28

II. THE TEST PLAINTIFFS SHOULD BE SANCTIONED FOR THEIR FALSE AND

FRAUDULENT DISCOVERY RESPONSES AND REPEATED VIOLATIONS OF
COURT ORDERS.............................................................................................................32

A. Dismissal Is Warranted As Sanction For The Test Plaintiffs’ Submission Of
False and/or Fraudulent Discovery Responses......................................................32

1. The Test Plaintiffs’ Failure to Provide Accurate Information
Regarding The Alleged Factual Bases Of Their Claims Has

Significantly Prejudiced Defendants..........................................................34

2. The Test Plaintiffs’ Failure to Provide Accurate Information
Regarding The Alleged Factual Bases Of Their Claims Has
Significantly Prejudiced The Judicial Process...........................................37

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3. The Test Plaintiffs Have Shown Disrespect to the Court and
Dismissal Is Necessary to Deter Such Misconduct By These and
Other Litigants.........................................................................▯..................38

B. In the Alternative, Defendants Request That The Test Plaintiffs Be Precluded

From Defending Their False Questionnaire Responses and That The Jury Be
Instructed Regarding the Test Plaintiffs’ Misconduct...........................................39

III. THE REMAINING 2,001 INDIVIDUAL PLAINTIFFS SHOULD BE BROUGHT
INTO COMPLIANCE WITH THE COURT’S PRIOR DISCOVERY ORDERS...........40

IV. THE COURT SHOULD ORDER PLAINTIFFS’ COUNSEL TO PAY THE
DEFENDANTS THEIR EXPENSES AND FEES IN BRINGING THIS MOTION.......44

CONCLUSION........................................................................▯......................................................45

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In October and November 2009, the DynCorp defendants took the depositions of the 20

test plaintiffs (from seven plaintiff families) who were selected by the plaintiffs to serve as their

first phase trial group. During these depositions, each of the test plaintiffs repeatedly departed

from their earlier sworn Questionnaire responses regarding the purported factual bases for their

claims (i.e., the who, what, when, and where of their alleged exposures and injuries) and relied

instead on new allegations of different purported exposures and damages. The test plaintiffs’

disavowals of the most basic elements of their prior sworn disclosures – as detailed herein –

makes clear that the test plaintiffs have willfully and repeatedly violated Court Orders over the

past two years in which the Court required “verified, factual and complete” disclosures, with the

explicit warning of dismissal for noncompliance. 1 Equally troubling, given that each of the 20

test plaintiffs disavowed a significant portion of the predicate facts that they had provided in

response to the Plaintiffs’ Questionnaire, neither the defendants nor the Court can have any

confidence in the accuracy of the Questionnaire responses of the 2,001 other individual plaintiffs

who remain as plaintiffs in these cases. Indeed, some of the test plaintiffs’ testimony makes

clear that Questionnaire responses provided by other non-test-plaintiff family members are

entirely fraudulent.

By Orders of the Magistrate Judge and the District Judge dated November 27, 2007,

October 21, 2008, December 1, 2008, May 5, 2009, July 7, 2009, and July 17, 2009, the Court

set forth clear directions to the individual plaintiffs requiring them to provide both (1) their best

1
Oct. 21, 2008 Order (Ex. A); see also the Nov. 27, 2007 Hr’g Tr. at 11-14 (Ex. B); the Nov. 25,
2008 Hr’g Tr. at 21-22, 52 (Ex. C); the Dec 1, 2008 Order; the May 5, 2009 Order; the July 17,
2009 Hr’g Tr. at 13-15, 50 (Ex. D); and the Sept. 19, 2009 order. (All discussed in detail below.)

2 Following the Court’s dismissal of 681 plaintiffs in its Order of September 16, 2009 and its
dismissal of 590 plaintiffs in its Order of January 12, 2010, there are 2021 remaining individual
plaintiffs in the Arias and Quinteros cases.

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information as to the dates and circumstances of their alleged exposures to Plan Colombia

spraying and the specific nature of their alleged personal injuries and property damages, and (2)

individualized scientific causation statements by qualified experts linking each plaintiff’s alleged

exposures to his or her alleged personal injuries and property damages. Plaintiffs’ repeated

failures to provide the ordered information required defendants to engage in extensive motions

practice and required the Court to conduct five separate motions hearings, each of which resulted

in Court findings that plaintiffs had violated the Court’s prior Orders. Despite this extraordinary

expenditure of time and resources, defendants and the Court find themselves today in the same

place now as they were when this case first began: with no credible information from any

3
individual plaintiff setting forth a plausible factual basis for his or her claim.

The question now before the Court is what – if anything – can be done to bring the

individual plaintiffs into compliance with the Court’s long-standing discovery Orders and move

this case on to a meaningful path towards trial. The DynCorp defendants submit that the 20 test

plaintiffs’ unambiguous violations of the Court’s Orders – and the resulting hopelessly-muddled

nature of their factual allegations – require that each of the test plaintiffs’ claims be dismissed

with prejudice. In the alternative, defendants request that if the test plaintiffs are allowed to

proceed to trial, they be precluded from offering any argument to explain away their misconduct

and that the jury be instructed by the Court that the plaintiff provided false answers under oath in

their 2008 Questionnaire responses.

The DynCorp defendants also submit that the claims of the remaining 2,001 individual

plaintiffs should not be allowed to proceed until each of those plaintiffs have provided

3 See Ashcroft v. Iqbal, 129 S. Ct. 1937, 1949 (2009) (“To survive a motion to dismiss, a
complaint must contain sufficient factual matter, accepted as true, to ‘state a claim to relief that is
plausible on its face.’”), citing Bell Atlantic Corp. v. Twombly, 550 U.S. 544, 570 (2007).

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meaningful assurance that they have provided accurate factual disclosures in their Questionnaire

responses and that each plaintiff be required, as previously ordered, to provide individualized

expert causation statements in support of their claims. Further, in light of numerous test

plaintiffs’ testimony that their children had been put forward as plaintiffs without their

knowledge, approval or authorization, defendants request that plaintiffs be required to provide a

parental authorization for each minor plaintiff within the group of remaining individual

plaintiffs. Defendants also seek recovery of their reasonable attorneys’ fees and costs in

preparing this motion. A proposed Order including these requested remedies is being filed

4
simultaneously with this motion.

I. BACKGROUND FACTS

A. The Court’s Prolonged Effort To Secure Prima Facie Disclosures from the

Individual Plaintiffs.

This case involves 2,021 individual plaintiffs in Ecuador who – notwithstanding the

repeated findings of the safety of the Plan Colombia spraying operations by the U.S. Department

of State, the U.S. Environmental Protection Agency, the U.S. Department of Agriculture, and an

5
expert panel assembled by the Organization of American States – allege physical harm and

property damage stemming from the defendants’ contract with the United States government to

spray herbicides in order to eradicate Colombian cocaine and heroin farms. At the initial

4The DynCorp defendants certify that before filing this motion, they conferred in good faith with
the Arias/Quinteros plaintiffs about the relief to be sought in this motion but were not able to
reach agreement (or to narrow the areas of disagreement) about the subjects and requests for

relief set out herein. Plaintiffs’ counsel have indicated that they will oppose this motion.
5
See, e.g., Keith R. Solomon, et al., Human Health and Environmental Risks from the Use of
Glyphosate Formulations to Control the Production of Coca in Colombia: Overview and
Conclusions, 72 J. Toxicology and Envtl. Health, Part A, 914-920 (2009) (Ex. E); United States
Department of State, Memorandum of Justification Concerning the Secretary of State’s 2007
Certification of Conditions Related to the Aerial Eradication of Illicit Coca in Colombia
(submitted to Congress on Aug. 10, 2007) (Ex. F).

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scheduling conference on November 27, 2007, the Court recognized that the science behind

plaintiffs’ claims was “skimpy” and thus required each of the individual plaintiffs to provide the

prima facie “who, what, when, and where” of their alleged individual exposures and injuries. 6

The Court explained:

There ought to be the kind of detail that would be useful by way of
identification of dates or times, if that’s possible, airplanes,
descriptions, what they saw, colors of fumes, any kind of details
like that, and added into temporal connections. Plus, if they did

have any doctors that they visited after the onset, assuming the
onset was temporally connected in some way, what the doctors
might have told them. . . . [T]o move the case forward to get some
of these issues teed up, it would be required to put in some basis of

that type.

(11/27/07 Hr’g. Tr. at 13:9-14:5 (emphasis added) (Ex. B)). Plaintiffs’ counsel agreed that these

facts were of the type that should be gathered before the filing of a lawsuit and assured the Court

that providing this information for each individual plaintiff “would be very doable.” (id. at

12:10).

Over the next 18 months, however, it became increasingly clear that what counsel had

represented was “very doable” was not, in fact, getting done. What followed instead was a series

of delays, obfuscations, and violations of multiple Court Orders:

• Plaintiffs’ first attempt to provide the ordered disclosures by the Court’s January

28, 2008 deadline was so grossly deficient that plaintiffs did not even seek to defend it. Instead,

6See 11/25/08 Hr’g Tr. 60:1-4 (Court discussion of bases for its earlier November 2007 ruling)
(Ex. C).

7
At the same hearing, the Court clarified that the disclosures about when the alleged exposures
occurred and what the plaintiffs saw would be required in addition to eight categories of
information promised by the plaintiffs, including: “(1) name; (2) address; (3) exposure address
(if different than current address … (6) claimed physical injuries (if any); (7) claimed property
damages (if any) …” Id. at 11 (referencing plaintiffs’ promise to produce all such data in the
Joint Rule 16.3 Statement (ECF No. 62 in Arias, ECF No. 19 in Quinteros, Nov. 19, 2007) at
16).

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following a meet-and-confer with defendants, the individual plaintiffs agreed to provide the

ordered disclosures as part of their responses to the Plaintiffs’ Questionnaire, the text of which

had been negotiated and agreed to in advance by their counsel and would by agreement of the

parties be treated as supplemental initial disclosures and interrogatory responses. Because the

plaintiffs also represented that they would be unable to meet the Court’s April 25, 2008 deadline

to submit the first 800 Plaintiffs’ Questionnaire responses, defendants agreed to extend this first

Questionnaire “due-date” to June 25, 2008. (Consent Notice (ECF No. 68 in Arias; ECF No. 25

in Quinteros), April 17, 2008 at 1).

• Plaintiffs’ initial responses to the Plaintiffs’ Questionnaires, however, were

equally deficient, requiring defendants to file a motion to compel with the Court. On October

21, 2008, the Magistrate Judge granted defendants’ motion in full, and ordered each of the

individual plaintiffs to “provide verified, factual and complete responses to the Plaintiffs’

Questionnaire, using the previously agreed-upon text of that Questionnaire.” Ex. A ¶1. The

October 21, 2008 Order also listed specific requirements for completing the Questionnaire,

including:

1. Providing the month, day and year of each alleged exposure

to the “Plan Colombia” herbicide, and then responding to
all follow-up questions with respect to each alleged
exposure event;

2. The marking of the maps appended to the Questionnaire to
show where the plaintiffs, their affected farms, and/or their

affected farm animals were located when each alleged
exposure event occurred;

3. The submission of medical records in accordance with the
instructions in the Questionnaire by the plaintiffs who

claim health injuries from their alleged exposure events …;
and

4. Providing responses to Section VII.A of the Plaintiffs’
Questionnaire to provide each individual plaintiff’s

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monetary computation of each category of damages

claimed.

Id. The same Order required the parties to discuss the “voluntary dismissal of plaintiffs who

have failed to provide adequate responses to the Questionnaire” and stated that “[a]ny resulting

dismissals shall be with prejudice and the plaintiffs are not entitled to any further opportunity to

supplement their Questionnaire responses in support of their responses to the defendants’ motion

to dismiss.” Id. ¶ 6. Moreover, in light of plaintiffs’ inability (or unwillingness) to provide the

prima facie disclosures that had been ordered nearly a year earlier in November 2007, the

Magistrate Judge further required that “[i]n addition, the Questionnaire responses provided by

the individual plaintiff shall be accompanied by sworn statements by qualified medical or

scientific witnesses that specifically link each plaintiff’s alleged personal injuries or property

damages to the effects of the ‘Plan Colombia’ spraying.” Id. ¶ 5.

The plaintiffs filed Objections to the October 21, 2008 Order, accusing the Magistrate

Judge of “cultural bias and lack of complete sensitivity to the reality of these Plaintiffs’ lives.”

Plaintiffs acknowledged their obligation “to provide information within their knowledge to the

best of their recollection” but represented that “[t]hey have done this, and this is all they can do.”

Id. at 6. The District Judge rejected plaintiffs’ objections and affirmed the October 21, 2008

Order in full. The Court did provide plaintiffs a further extension of time to produce “all of the

completed questionnaires and supplements to make them complete” until January 21, 2009 and

an extension of time until March 23, 2009 to submit the individualized causation statements by

qualified medical or scientific witnesses. See 11/25/08 Hr’g Tr. (Ex. C) at 24; Minute Entry

entered on Nov. 28, 2008 in the Arias and Quinteros dockets; and the Court’s Dec. 1, 2008 Order

8 Plaintiffs Objections to Magistrate Judge’s Oct. 21, 2008 Order on Defs.’ Mot. to Compel
Disc., ECF No. 77 in Arias; ECF No. 40 in Quinteros, Nov. 3, 2008, at 2.

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(ECF No. 84 in Arias; ECF No. 47 in Quinteros).

In response to plaintiffs’ counsel’s claimed confusion as to the causation statement

requirement, the Court explained that plaintiffs were required to produce for each individual

plaintiff an expert causation statement that “at minimum, … mak[es] some connection based

upon some scientific assessment between the allegation that spraying happened and that spraying

caused these symptoms.” 11/25/08 Hr’g. Tr. (Ex. C) at 60; see also id. at 53 (“each one of the

2500 or so questionnaires that’s returned will have to have appended to it a separate sworn

statement from some expert opining about causation with regard to that individual”).

• Despite being provided yet another opportunity to make sure that they had really

done their best to truthfully answer the Questionnaire, plaintiffs again failed to provide

defendants with the “verified, factual and complete responses” explicitly required by the Court’s

Orders. To the contrary, the individual plaintiffs made no changes whatsoever to their previous

9
Questionnaire responses. When confronted with this fact in defendants’ subsequent motion to

dismiss, plaintiffs’ counsel again assured the Court that “all 2,463 Plaintiffs who submitted a

completed Questionnaire responded as fully as they could based on personal knowledge,” and

argued that the plaintiffs could not be required to produce information that they did not have.

Joint Mot. to Dismiss Nonresponsive Plts. and Plts. Without Claims, ECF No. 86 in Arias, ECF

No. 48 in Quinteros, Feb. 19, 2009, at 10. Based upon this representation, the District Court

granted in part and denied in part the defendants’ motion to dismiss but allowed the majority of

individual plaintiffs who had completed the Questionnaire to proceed with their claims. 7/17/09

9This is precisely what the Court lamented in its January 12, 2010 Memorandum Opinion
dismissing 590 plaintiffs with prejudice: “Multiple orders have directed the plaintiffs to respond
in full to the questionnaires, and the plaintiffs received three extensions of time in which to do
so,” and yet, “[d]espite the plaintiffs’ ample opportunity to fill in the information gaps,” they did

not do so. 1/12/10 Mem. Op., at 5-6.

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Hr’g Tr. (Ex. D) at 13-15 . In a subsequent dismissal order addressing two categories of

disclosure failures, however, the Court made clear that individual plaintiffs who failed “to

plainly state data to which they have access” or who had “produce[d] discovery responses only

selectively” would be dismissed with prejudice. 1/12/10 Mem. Op., ECF No. 173 in Arias , ECF

No. 133 in Quinteros, at 4, 7; see also 7/17/09 Hr’g Tr. (Ex. D) at 15 (explaining reasoning for

dismissal categories).

In response to a separate defendants’ motion relating to the causation statements, the

Court held that the plaintiffs’ submission of aggregate general causation statements (one for

personal injuries and one for property damages) violated the Court’s order that they submit

individualized causation assessments. In granting the defendants’ request for sanctions, the

Magistrate Judge explained:

The Court finds that there is nothing in this Court’s order, or in the
order of the assigned District Judge, which permits the plaintiffs to
substitute “an aggregate review” for the paragraph 5 requirement
. . . of this Court’s October 21, 2008 order, . . . that the plaintiffs

provide a sworn statement of a qualified medical or scientific
witness regarding “each plaintiff’s alleged personal injuries or
property damages.” The plaintiffs simply have not done so. The
plaintiffs concede they haven’t done so, and there’s nothing
ambiguous about the order directing that they do so.

5/5/09 Hr’g Tr. (Ex. G) at 43:16-44:5; see also the Court’s May 5, 2009 Minute Order. After

rejecting plaintiffs’ subsequent objections to the Magistrate Judge’s ruling, the District Court

ordered plaintiffs to produce the “individualized assessments with regard to each individual

plaintiff and each individual plaintiff’s complaints about harm or damage” by October 2, 2009.

7/17/09 Hr’g Tr. (Ex. D) at 39; see also id. at 50 (“the [submitted] reports . . . that had been

provided did not comply with what I had asked for from the beginning. The Magistrate Judge

was quite right that they didn’t and that the reports did not comply, and I find no error in their

conclusion that the individualized expert assessments are required.”); see also 7/22/09 Minute

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Order entered in the Arias and Quinteros without ECF numbers and the Court’s amended

scheduling Order (ECF No. 147 in Arias; ECF No. 109 in Quinteros, Sept. 17, 2009).

B. Plaintiffs’ Most Recent Violation of the Court’s Order to Provide
Individualized Causation Statements.

Despite the explicit and repeated direction from both the Magistrate Judge and the

District Judge, the individual plaintiffs on October 2, 2009, again failed to provide individualized

expert assessments supporting their claims. Instead, plaintiffs simply repackaged the general

causation statements previously rejected by the Court by merging them into a single causation

statement that they then produced to defendants over 2,000 times (one for each individual

plaintiff). The only change in the language of the previous aggregate causation statements was

the addition of a single paragraph in each statement that summarizes each individual plaintiff’s

alleged personal injuries and property damages (taken directly from his/her Plaintiffs’

Questionnaire response), followed by an identical statement, repeated for each of the 2,000-plus

plaintiffs that the injuries “could have been caused by Plan Colombia spraying.” See the sample

Campana causation statement attached at Ex. H ¶ 16.

This boilerplate paragraph does not make the required connection “based upon some

scientific assessment between the allegation that spraying happened and that spraying caused

these symptoms” in any of the plaintiffs. 11/25/08 Hr’g Tr. (Ex. C) at 60:18-21 (the District

Judge’s description of what was expected). Campana’s paragraph 16 provides no information

whatsoever as to the dates, times and frequencies of the individual plaintiffs’ alleged exposures,

the location of each plaintiff in proximity to contemporaneous Plan Colombia spraying, or the

amount of the herbicide as to which each plaintiff purportedly could have been exposed. The

paragraph also provides no scientific data linking the Plan Colombia herbicide spray to the

specific types of illnesses alleged, no test data showing even the presence of the herbicide in any

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of the plaintiffs’ blood, and no explanation why the symptoms alleged by the plaintiffs should be

linked to Plan Colombia spraying as opposed to the widespread health problems endemic to the

impoverished communities in which the plaintiffs live. 10 Instead, paragraph 16, like the rest of

the aggregate causation statement, makes clear that plaintiffs’ expert simply assumed that any

and all ailments in the region – as asserted in any one of the plaintiffs’ Questionnaire responses –

might be attributed to the spraying, no matter how outlandish the claim might be. See, e.g.,

Campana Decl. (Ex. H) ¶19 (recounting a Sucumbios resident’s claim that “since the fumigations

[my son] no longer gets good grades in school”); id. ¶20(b) (stating that the affected population

generally attribute infectious diseases like influenza, bronchitis, and pneumonia to Plan

Colombia spraying); id. ¶20(d) (stating that “women think” that the number of miscarriages in

the region must be associated with the spraying despite admission from the Provincial health

authorities that they cannot make such a link); id. ¶¶ 29-34 (identifying mosquito-borne diseases

like malaria and dengue fever as possible results of the spraying).

Moreover, Campana’s paragraph 16 largely ignores the separate requirement in the

Court’s Order that the individual plaintiffs provide “sworn statements by qualified … scientific

witnesses that specifically link each plaintiff’s alleged … property damages to the effects of the

10
For example, published epidemiologic studies in communities in Northern Ecuador identified
alarmingly high rates of bacterial and insect-born diseases of the eyes, skin, and gastrointestinal
tract. See, e.g., Simonetta Gatti., et al., Amebic Infections Due To The Entamoeba Histolytica-
Entmoeba Dispar Complex: A Study of the Incidence in a Remote Rural Area of Ecuador, 67(1)
Am. J. Trop. Med. Hyg. 123, 125-26 (2002) (176 of 178 individuals in study population (98.9%)
tested positive for intestinal parasites; “This high detection rate is clearly related to poor
sanitation, nutrition [and] use of contaminated water …”); Rodrigo X. Armijos, et al., The
Epidemiology of Cutaneous Leishmaniasis in Subtropical Ecuador, 2(2) Tropical Med. and Int’l
Health 140-152 (1997) (14% of study population tested positive for active parasitic skin disease,
leishmaniasis, and 33% had evidence of prior disease); P.J. Cooper, et al., Onchocerciasis in
Ecuador: Ocular Findings in Onchocerca Volvulus Infected Individuals, 79 Brit. J. of
Opthalmology 157-162 (1995) (insect-born disease; Onchocercal ocular lesions identified in over
33% of the study population). Copies of these studies were attached as Exhibit H to the
defendants’ April 3, 2009 motion for sanctions at ECF No. 91 in Arias; ECF No. 53 in

Quinteros, April 3, 2009.

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Plan Colombia spraying.” 10/21/08 Order (Ex. A) ¶ 5 (emphasis added). Other than passing

parenthetical references to the alleged impacts of the spraying on each plaintiff’s crops and

animals – also taken directly from each Plaintiffs’ Questionnaire response – plaintiffs’ expert

simply combines these allegations with the plaintiffs’ alleged personal injuries and concludes –

for all plaintiffs – that the alleged property damages “could have been caused by Plan Colombia

spraying.” Ex. H ¶ 16. Again, the statements contain nothing in the way of an individualized

scientific assessment: no scientific studies linking the alleged exposure scenarios to the types of

crop and animal losses alleged; no soil, air, or water testing detecting even the presence of the

herbicide on the plaintiffs’ farms; and no discussion of potential alternative causes. Plaintiffs’

expert’s scant references to property damages are not surprising because the expert, as a medical

doctor with purported specialties in the areas of mental health and public health, does not even

claim to have the expertise or experience in animal toxicology or agricultural science that he

11
would need to address causation for the plaintiffs’ property damage claims.

C. The Deposition Testimony of the 20 Test Plaintiffs Establishes Numerous
Further Violations of The Court’s Orders.

In its July 17, 2009 ruling, the Court granted plaintiffs’ request that they be allowed to

select 20 test plaintiffs who plaintiffs believed could best advance their claims as a first phase

trial group. Plaintiffs had previously represented to the Court that they would be selecting in this

initial group, “people who had the most information [about the alleged facts of their claims] so

that we could in the first trial be in a position to really prove what happened in detail.” 11/25/08

11At the November 25, 2008 hearing, the Court specifically addressed the fact that plaintiffs
would need to have an expert who could address each individual plaintiff’s claims of crop or
animal damage. See 11/25/08 Hr’g Tr. (Ex. C) at 20:13-17 (“Do you at this point … have any
expert or experts lined up, scientific or medical, who have either examined the plaintiffs or
examined their medical records or evidence of crop or animal damage so far?”). (Emphasis

added.)

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Hr’g Tr. (Ex. C) at 11:14-12:1. 12 During the depositions of these test plaintiffs in October and

November 2009, however, these self-selected test plaintiffs repeatedly demonstrated that they

had not or could not provide any accurate account of the basic facts underlying their legal claims.

To the contrary, all of the test plaintiffs repeatedly changed their stories as to their previously-

disclosed dates and circumstances of alleged exposures and as to the personal injuries and

property damages purportedly arising therefrom. Thus, either the sworn Questionnaire responses

– and expert causation statements derived solely from those responses – or the deposition

testimony provided by each of the test plaintiffs, or both, are materially false with respect to the

basic factual underpinnings of the test plaintiffs’ claims. Because the test plaintiffs have not

produced any independent documentary evidence supporting their claims (e.g., medical records

relating to the injuries allegedly caused by the Plan Colombia spraying, business or tax records

relating to the alleged property losses), the defendants have no meaningful way to choose among

the plaintiffs’ varying representations and can only guess at what story these same plaintiffs will

present at trial. Further, with this clear showing of the meaningless of the Plaintiffs’

Questionnaire responses and causation statements of the plaintiffs’ “best” individual cases, the

defendants and the Court can have no confidence whatsoever in the information provided to date

by the remaining 2,001 individual plaintiffs.

The changing stories of each of the 20 test plaintiffs, as grouped by plaintiff family,13are

discussed below:

12
In their written proposal about how to select the test plaintiffs, plaintiffs’ counsel likewise
promised that they would “select plaintiff cases that provide the parties with the most useful
information.” Pls.’ Proposal for Selection of 20 Sample Test Plaintiffs, ECF No. 88 in Arias,
ECF No. 50 in Quinteros, March 23, 2009, at 8.

13Each test plaintiff family has multiple other family members who are also plaintiffs in the
litigation but who are not serving as test plaintiffs.

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1. The Calero Family

Four of the test plaintiffs are members of the Calero family, as follows:

• Santos Calero (husband of Calixta; father of Betty; grandfather of Yuli)
• Calixta Pineda (wife of Santos; mother of Betty; grandmother of Yuli)
• Betty Calero Pineda (daughter of Santos and Calixta; mother of Yuli)

• Yuli Calero Pineda (11-year-old daughter of Betty; granddaughter of
Santos and Calixta)

In their Questionnaire responses, the Calero plaintiffs provided general and inconsistent

representations that their alleged exposures to Plan Colombia herbicide occurred sometime in

2001 (Yuli), sometime in 2002 (Betty), sometime in 2002 or 2003 (Calixta), or sometime in

2001, 2002, 2003, 2004, 2005, and/or 2006 (Santos). (App. 1, 5, 24, 37, 55.) 14 At their

depositions, however, each of the Calero plaintiffs told a different story. The adult Caleros

(Santos, Calixta, and Betty) now each testified that they were exposed only on a single occasion

in August 2003, and while Santos and Calixta did not identify the number of helicopters they

saw in their Questionnaire responses, and Betty said that she saw 3 helicopters in her

Questionnaire response, at deposition they each testified to having seen 2 helicopters at the time

of the alleged exposure. (App. 2, 11-13, 24, 32, 39-40, 45-47.) On the other hand, Yuli Calero,

who in her Questionnaire response provided specific details of her recollection of an alleged

spraying event in 2001 (e.g., that she saw two white spray planes and a green helicopter on a

clear morning at 9 am, that she saw a white cloud sprayed from the plane and both smelled the

herbicide and felt it on her body) (App. 56-57), admitted in her deposition that she had no

recollection of seeing (or smelling) any spraying event whatsoever. (App. 61, 62-63.)

14In response to the Questionnaire inquiry: “When were you or your property exposed to
herbicide?” Santos Calero responded: “2001.” However, in response to the question: “What was
the date and time of the day that your crops were exposed to the Plan Colombia herbicide?
Santos Calero responded: “2001, 2002, 2003, 2004, 2005, 2006.” (App. 1, 5.) All of the
citations to the test plaintiffs’ sworn Questionnaire responses and deposition transcripts have

been compiled in an Appendix filed as the last exhibit to this motion.

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Defendants also learned that the maps provided by the Caleros with their Questionnaire

responses were wildly inaccurate in identifying the alleged location of their exposures. (App. 10,

30, 44-B, 60.)

Further, the Caleros presented a notably different account at deposition of their alleged

personal injuries from the purported spraying event. In their Questionnaire responses, both

Calixta and Betty Calero alleged that they had incurred medical expenses of $5,000 in

connection with their alleged exposures, which they had paid in cash. (App. 28-29, 44-45.) At

their depositions, however, both women admitted that they had incurred no such expenses:

Calixta acknowledged that she did not even have access to that amount of money (App. 35-36),

and Betty testified that her medical expenses were in fact only 5,000 sucres or less than $20 (a

sucre is an Ecuadorian currency that was discontinued in 1999 – four years before Betty’s

alleged exposure – when Ecuador adopted the U.S. dollar as its currency). (App. 53.) The

Caleros also changed their stories as to the nature of their alleged personal injuries: Santos

Calero testified about bone and kidney pains that were nowhere mentioned in his Questionnaire

response (App. 3-4, 5, 17), and Betty Calero testified about previously-unmentioned mental
15
problems, stroke, pain in the kidneys, and pain in the legs (App. 39, 41-43, 48, 49, 51). Not

surprisingly, because they were prepared solely from the Questionnaire responses, the causation

statements submitted for these two plaintiffs say nothing about these newly alleged health

claims. (App. 23, 54.) Moreover, Betty Calero’s claim of medical injuries arising from Plan

Colombia spraying completely fell apart when it was pointed out at deposition that the medical

15 While Calixta Pineda’s testimony as to her alleged personal injuries from the spraying was
generally consistent with her Questionnaire response, she separately admitted that the medical

history in her Questionnaire – in which she represented that she had suffered from chagas,
respiratory illness, migraine headaches, bacterial and fungal infections, and anemia or other
blood disorders – was entirely incorrect because she suffered from none of these ailments. (App.
25-27, 33, 34.)

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record she had produced only days earlier as evidence of her injuries was dated March 2003,

some seven months prior to her alleged exposure. (App. 50, 51.) Betty Calero’s only

explanation for this obvious and fatal inconsistency was that the medical record must have been

inaccurately dated. (App. 52.)

While Santos Calero claimed in his Questionnaire response that the family had crop

losses of nearly $15,000 (App. 8-9), at his deposition, he admitted that he had not in fact

calculated any monetary losses for his purportedly damaged crops and did not know most of his

crops’ market value because he used them only for personal consumption. (App.18-20, 21-22.)

Mr. Calero also repeatedly reversed himself during his deposition on the question whether he had

planted crops in the years following the alleged crop damage from Plan Colombia spraying,

initially testifying that he did not plant crops for the following four years, then (after being

shown his Questionnaire response claiming damage to crops planted in 2004) testifying that he

could not remember whether he had planted any crops between 2003 and 2007, and then

testifying that “of course we planted because with the crops we continued to live. That’s what

we live off of.” (App. 5, 14-16.)

2. The Salas Family

Three of the test plaintiffs are members of the Salas family:

• Jorge Salas (husband of Laura; father of John)
• Laura Sanchez (wife of Jorge; mother of John)
• John Salas (15-year-old son of Jorge and Laura)

In contrast to the Calero family, Jorge Salas provided very specific information in his

Questionnaire response as to the date of his alleged exposure: October 4, 2002. (App. 67.) His

wife, Laura Sanchez gave a consistent, though more general date of exposure of “since 2002” in

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her Questionnaire Response. (App. 101.) 16 At deposition, however, the Salas’s abandoned those

dates and testified to a hodgepodge of exposure dates that do not match up with either their

Questionnaire responses or with the testimony of the other family members who were purporting

to describe the same alleged exposures. Jorge testified that the exposures occurred in December

2000, May 2001, and October 2003 (App. 76, 80-81, 85-86); Laura testified that the exposures

occurred in June 2002, January 2003, and October 2003 (App. 105, 106, 109); and their son John

admitted that he cannot remember when the exposures occurred, but that he believed he was

exposed on two occasions. (App. 118-120.) Defendants can only guess what dates of exposure

the Salas family might allege at trial.

The Salas family’s testimony as to what they saw during the alleged spraying events also

changed significantly, with the number of spray planes decreasing (from multiple planes to 1-2

planes for Jorge and from 5 planes to 2-3 planes for John) (App. 67, 77, 81, 86, 114, 118) and the

number of helicopters increasing (with Jorge and Laura now claiming to have seen a helicopter

after having not so claimed in their Questionnaire responses). (App. 67, 77, 81, 86, 101, 105,

107, 110.) The Salas family also changed their statements as to where they were allegedly

exposed, with John testifying that the exposure took place 2 kilometers from the border (rather

than the 300 meters from the border stated in his and his mother’s Questionnaire responses and

causation statements (App. 101, 113, 114, 119, 123)), and Jorge testifying that the exposure was

3 kilometers from the border (up from the 2 kilometers response in his Questionnaire and in his

causation statement). (App. 67, 75, 77, 82, 87, 98.) As with the Caleros, the marked maps of

the location of the alleged exposures provided with the Salas family’s Questionnaire responses

were inaccurate and thus do not provide any clarification. (App. 74, 104, 117.)

16 John Salas stated in his Questionnaire response that the exposure occurred in 2000 (App. 114),
but he would have only been six years old at the time.

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The Salas family plaintiffs also testified to a whole host of alleged personal injuries that

are at odds with the personal injuries identified in their Questionnaire responses. Jorge Salas

testified that the herbicide spray caused inflammation of the eyes and caused him to lose his

vision whereas he had not mentioned any problems with his eyes in his Questionnaire response.

(App. 68, 70-71, 97.) Laura Sanchez, on the other hand, abandoned the claims of respiratory

disease and stomach burning set forth in her Questionnaire response. (App. 102, 108, 111.) 17

John Salas likewise abandoned his Questionnaire response alleging respiratory problems; at his

deposition, John testified instead that the herbicide exposure caused previously-unidentified

body pain, bone pain, sore throat and a strong headache. (App. 115, 121.) Thus, once again, the

causation statements submitted for the Salas family test plaintiffs do not even correctly identify

the personal injuries alleged by the plaintiffs at deposition. (App. 98, 113, 123.)

The Salas family also changed their claims of injuries to farm animals. In his Questionnaire

response, Jorge Salas claimed monetary damages for the death of 40 chickens, 2 cows, and 2

pigs. (App. 72, 73.) At his deposition, however, Mr. Salas testified that the cows were not his,

but he added new damages claims for 2 horses, 2 dogs, and an additional 2 pigs. (App. 87-A to

92.) He produced no documentation whatsoever to support either set of numbers. Jorge Salas

also testified at deposition that the Salas family members are plaintiffs in a separate lawsuit

alleging contamination by waste from an oil well 500 meters from their home. (App. 93-96.) In

his Questionnaire response, Jorge had specifically denied being exposed to oil pollution and

denied being involved in such litigation. (App. 64-66.) 18

17
When confronted with this fact at her deposition, Laura responded that she had testified about
her sore throat, which she considered to be a respiratory disease. (App. at 112 to 112-A.)

18Test plaintiff John Salas testified that an oil spill near the family’s home “is like black crude
that comes down … and it flows along the edge of the road like water, and the spill reached the
river … where we used to wash.” (App. 122.)

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3. The Quevedo Family

There are four test plaintiffs from the Quevedo family:

• Luciano Quevedo (husband of Rosa; father of Edith and Robinson)
• Rosa Altamirano Miranda (wife of Luciano; mother of Edith and
Robinson)
• Edith Quevedo Altamirano (14-year-old daughter of Luciano and Rosa)

• Robinson Quevedo Altamirano (9-year-old son of Luciano and Rosa)

The deposition testimony of the Quevedos follows the same winding course as that of the

other test plaintiffs. Although Luciano Quevedo stated in his Questionnaire response that he was

exposed to Plan Colombia herbicide “from 2002 til 2006,” he testified during his deposition that

he could only recall seeing a single spray plane on one occasion flying close to the San Miguel

River (which is 3 km from his farm), and that he could not recall the date. (App. 124, 129, 131-

133.) Luciano’s wife Rosa had claimed exposure “from October 2002 on” in her Questionnaire

response and stated further that she had seen 2 lead-colored planes and a green helicopter in 2002

(App. 157-158); at her deposition, however, Rosa testified that she had not seen a helicopter and

had only had heard what she believed to be a spray plane. (App. 162-163, 164-165, 166-167.)

Luciano and Rosa’s daughter Edith claimed exposure in her Questionnaire response “since April

10, 2002” and claimed in the same response that she saw 2 spray planes, but Edith admitted

during her deposition that she could not provide any date for the one time she allegedly saw a

single spray plane. (App. 173-174, 177, 178.) The Quevedos did provide testimony consistent

with their Questionnaire responses that their home was 4 km from the border, but at deposition,

Edith testified that she was exposed at school, not at home as stated in her Questionnaire

response. (App. 173, 178.)

As to personal injuries, in his Questionnaire responses, Luciano Quevedo alleged that the

spraying had caused him to suffer itchiness, headaches, aching bones and fever. (App. 125.)

The individual causation statement submitted on his behalf listed these same alleged injuries (as

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well as another ailment listed in the medical history section of Luciano’s Questionnaire). (App.

126-A, 154.) At his deposition, however, Luciano initially abandoned his claims of headaches,

aching bones, and fever and substituted in their place only a claim of vision problems.9 He then

revised his testimony (after being shown a copy of his Questionnaire response) to once again

allege that he had suffered all of the injuries. (App. 139, 140.) Finally, Luciano abandoned all

of his personal injury claims, testifying that he was not seeking any damages at all in connection

with any alleged personal injuries. (App. 149.) Meanwhile, Luciano’s wife, Rosa, who had

alleged “headache, fever, diarrhea, and spots” in her sworn Questionnaire response, came to

deposition with different complaints of a rash, headaches, and kidney and bone pains. (App.

159, 168-169.)

The Quevedos’ claims of property damages likewise changed. In his Questionnaire

response, Luciano had alleged damage to 1 hectare of plantains, 5 hectares of coffee, and 5

hectares pasture. (App. 127, 128.) At deposition he bumped those numbers up, adding another 5

hectares of pasture and 1 hectare of cacao beans. (App. 130.) Further despite having provided a

dollar value for his lost crops in his Questionnaire response, Luciano initially testified at

deposition that he had never placed a monetary value on his allegedly lost crops. (App. 134.)

When confronted with his Questionnaire response, Luciano (again) changed his testimony and

stated that he had calculated the stated dollar amounts for his crops. (App. 145-148.) But then,

when the written response was taken away, Luciano conceded once more that he had not – and

19See App. 137-138 (Q: Other than the itchiness on your skin and the eyesight problems, do you
believe you have suffered any additional personal injuries from the spray? A: No, just that).

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could not – quantify the value of his allegedly lost crops (App. 150-151), and he produced no

20
farming records to assist in making such calculations.

4. The Mestanza Family

The Mestanza family provides 4 of the test plaintiffs in the first phase trial group:

• Victor Mestanza (husband of Ercilia; father of Edy; grandfather of
Jennifer)
• Ercilia Bosquez (wife of Victor; mother of Edy; grandmother of Jennifer)
• Edy Mestanza (son of Victor and Ercilia; father of Jennifer)

• Jennifer Mestanza (14-year-old daughter of Edy; granddaughter of Victor
and Ercilia)

Although the Mestanza family demonstrated problems with changing testimony similar to

the other test plaintiffs, the misstatements in their Questionnaire responses raise even more

serious concerns. The four Mestanza test plaintiffs family all stated in their Questionnaire

responses that they were exposed to Plan Colombia herbicide at their “current home address” in

Puerto Mestanza, a small community in the Sucumbíos province of Ecuador, right beside the

river border with Colombia. Each plaintiff likewise submitted an individual causation statement

asserting that he or she had lived in Puerto Mestanza for most of their lives (App. 179-180, 221,

225-226, 241, 242-243, 273, 275, 301.) At deposition, however, all four of the test plaintiffs

admitted that their principal place of residence both now and on the date of their alleged

exposures was not in Puerto Mestanza but in Guayaquil, which is 275 miles from the Ecuador-

Colombia border (App. 189, 229-230, 247-248, 282-283). While Victor and Ercilia testified that

they had spent the majority of their time at the family farm in Puerto Mestanza during 2000-2004

and that their granddaughter Jennifer had been with them at the farm when spraying allegedly

20 Luciano likewise reversed course with regard to his alleged animal losses, first testifying that
he did not know the value of the animals, then testifying that he had calculated the values in his
Questionnaire response, and then, after the response was taken away, admitting again that he

could not state how much the animals were worth. (App. 135-136, 141-144, 152-153.)

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occurred between 2000 and 2002, no other members of their family were at the farm during the

times that Victor and Ercilia allegedly saw spray planes. (App. 190-191, 231 to 231-A, 236-A,

293-295, 283-A to 283-B.) Victor’s grown son Edy acknowledged in direct contravention to his

Questionnaire response that he was rarely in Puerto Mestanza and that he had never been

exposed to any Plan Colombia spraying. (App. 250-252, 256-257.) In his Questionnaire

response, Edy had falsely alleged exposure in 2002 and claimed personal injuries including

headaches, dizziness, vomiting, skin irritation, and sore throat, all of which were likewise falsely

attributed to Plan Colombia spraying in his individual causation statement. (App. 244-245, 273-

21
274.) Moreover, as addressed more fully in Defendants’ Response to Plaintiffs Motion to

Dismiss Three Individual Plaintiffs, ECF No. 172 in Arias, ECF No. 132 in Quinteros (Jan. 6,

2009) at 3-10, the deposition testimony of the Mestanza test plaintiffs made clear that

Questionnaire responses of four other Mestanza family members in the larger individual plaintiff

group of non-test-plaintiffs were completely fictitious. Each of these plaintiffs (two of Victor’s

other grown children and two of Edy’s minor children) live full time in Guayaquil and were not

in Puerto Mestanza during any of the alleged spraying events. (App. 190, 193, 236-236-A, 253-

255, 262-264, 267, 269.) Nonetheless, as set out in the defendants’ Jan. 6th response brief at 3-

10, each of these four Mestanza family members submitted sworn Questionnaire responses

containing numerous false representations about, e.g., having seen spray planes and helicopters

overhead and having suffered a wide variety of personal injuries. Moreover, this apparent fraud

21Edy Mestanza claimed at his deposition that his correct signature did not appear on the
Questionnaire response provided for him, although he acknowledged that certain portions of the
response were specific to him. (App. 258.) Edy also identified his own signature on other
Questionnaire responses submitted for his two minor sons and for his daughter (test plaintiff
Jennifer). (App. 265, 266, 268-269, 270-272.) Edy insisted, however, that he had not authorized
his minor sons (9-year-old Victor Manuel and 5-year-old David) to serve as plaintiffs in this

litigation and that he did not believe that they were plaintiffs. (App. 264, 269 to 269-A.)

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cannot be attributed solely (or even at all) to these other (non-test-plaintiff) Mestanza family

22
members, as two of them are minors and one is mentally retarded.

The testimony of the Mestanza test plaintiffs also includes the same changing stories that

characterize the testimony of the other test plaintiffs. For example, in their Questionnaire

responses, the four Mestanza test plaintiffs provide only broad and conflicting statements as to

the dates of their alleged exposures to Plan Colombia herbicide, with Victor alleging exposure in

2002, Ercilia alleging exposure from 2000 until October 2002, Edy alleging exposure in 2002,

and Jennifer alleging exposure in 2002-2006. (App. 181-182, 227, 244, 276.) At deposition,

Edy admitted that he had not been exposed at all, as noted above. (App. 255-256, 257.) Victor

and Ercilia, on the other hand, were now in agreement as to five specific dates of alleged

exposure (December 2000, January 2002, September 2002, and October 7 and 10, 2002). (App.

192, 196, 201-203, 205, 232, 233, 235, 237-240.) Ercilia also changed her Questionnaire

response stating that she had seen 3 planes and 1 helicopter during the alleged spraying events,

testifying at deposition -- again now consistent with her husband Victor – that she in fact had

seen 5 spray planes and 5 helicopters. (App. 227, 233-234.) Jennifer, meanwhile, changed her

prior five-year potential exposure period to just two years, 2000 and 2002, and further admitted

that she doesn’t recall anything from 2000 and is relying solely on her grandfather Victor

Mestanza’s say-so as to her alleged exposure during that year. (App. 284, 285, 286, 288, 297.)

After having provided a detailed Questionnaire response stating that she had seen 5 spray planes,

multiple helicopters marked with the Colombian flag, a white cloud in the air and residue on the

22 On December 23, 2009, without offering any explanation for the fraudulent Questionnaire
responses, plaintiffs counsel moved to dismiss the claims of Victor Mestanza’s two minor
grandsons and the claim of his mentally-challenged daughter. The DynCorp defendants’
January 6, 2009, response addresses in detail the misrepresentations made by each Mestanza
non-test-plaintiff and by test plaintiff Edy Mestanza (ECF No. 172 in Arias, ECF No. 132 in

Quinteros), to which plaintiffs’ January 13, 2010 reply brief did not respond in any detail.

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ground, Jennifer acknowledged at deposition that (other than claiming that she once saw a plane

spraying in 2002), she could not recall anything else she had seen at the times of the alleged

spraying events. (App. 276-277, 284-285, 286-290.)

The Mestanza test plaintiffs also changed course repeatedly on their claims of personal

injuries and property damages. Edy Mestanza, whose Questionnaire response had alleged that

exposures to Plan Colombia spraying had caused him to suffer from headaches, dizziness,

vomiting, skin irritation, and a throat infection, conceded at deposition that he had not suffered

any of these injuries (App. 245, 256-257), although he raised an entirely new claim that he had

suffered emotional and psychological damages due to his alleged property losses and the alleged

personal injuries of family members. (App. 256-257.) Edy’s father, Victor Mestanza, backed

away from the claims made in his Questionnaire response that his exposures had caused

headaches, stomach aches, gastric problems and vomiting, stating at deposition that he wasn’t

sure what was causing those and other new medical problems he was experiencing. (App. 183,

195, 197, 204, 206, 209-210.) Jennifer Mestanza abandoned claims of headaches and stomach

aches in her Questionnaire response and causation statement and further admitted that she had no

basis for the $20,000 in paid medical expenses she had claimed in her Questionnaire response.

(App. 278, 281, 290-291, 296, 301.) On the flip side, however, Victor Mestanza expanded at

deposition the already expansive claims of property damage set out in his Questionnaire response

and causation statement, adding new allegations of damages to 10 hectares of sugar cane,

tomatoes, green peppers, cassava and grasslands, and the alleged deaths of large numbers of

sheep, ducks, pigs and chickens. (App. 184, 187-188, 194, 198-200, 207, 208, 218, 221-222.) 23

23Mr. Mestanza’s allegation that all of his animals were dead as of the date of his last alleged
exposure to Plan Colombia spraying in October 2002 (App. 214-215), is belied by a video he
produced to defendants before his deposition, which Victor confirmed was taken at his property

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Remarkably, despite contending in his Questionnaire response that his property damages

exceeded $600,000, Victor Mestanza could produce no business or farming records to support

his claims. (App.186, 187, 188, 213-214, 217, 211-212, 214-216, 220.) Edy Mestanza testified

consistently with his Questionnaire response as to the hectares of crops and numbers of animals

allegedly killed by the Plan Colombia spraying, but testified that he was relying solely on his

father Victor for these numbers because he (Edy) had never even seen the crops or animals

allegedly at issue, and Edy further completely disavowed the monetary values set forth in his

Questionnaire response for the alleged losses. (App. 252, 259-261.)

24
5. The Sandoval Family

Three members of the Sandoval family are serving as test plaintiffs:

• Dociteo Sandoval (father of Edgar and Wilber)
• Edgar Sandoval (25-year-old son of Dociteo; brother of Wilber)

• Wilber Sandoval (14-year-old son of Dociteo; brother of Edgar)

The Sandoval test plaintiffs present the same pattern of changing alleged dates of

exposure, changing alleged personal injuries, and changing alleged property damages. In their

Questionnaire responses, the three Sandoval test plaintiffs disagreed as to the date of their

in Nov. 2002 (App. 207-208) and which is replete with pictures of lush green plants and
apparently healthy farm animals. Excerpted photos from that video are attached hereto as Ex. I.

24 Other children of Dociteo Sandoval are also named plaintiffs in this litigation, despite Mr.

Sandoval’s testimony that he did not want any of his children (except 25 year old Edgar) to be
plaintiffs:

Q: I understand that you do not want Marcelo and Roque to be plaintiffs. Do you
want Wilber to be a Plaintiff?

A: . . . I don’t want any of my children – I do not accept that they are plaintiffs. . . .
I’m surprised. I’m very surprised for having signed that document with that
explanation.

(App. 322) (referring to his surprise at seeing his son Roque’s Questionnaire response).

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alleged exposure to Plan Colombia spraying, with Dociteo claiming that the exposure occurred at

the end of 2003, Edgar claiming that the exposure occurred in 2006, and Wilber claiming that the

exposure took place in 2004. (App. 304-305, 325-326, 341-342.) At deposition, however,

Dociteo and Edgar were now in agreement that the exposure took place during three days at the

end of 2003; Wilber also testified that the exposure took place on three separate days, but he

could not recall the year in which this occurred. (App. 313, 319-320, 321, 330-331, 334, 338,

351-352.) Wilber’s memory was far more detailed at deposition, however, as to what he saw

during these purported spraying events: while his Questionnaire response states that he didn’t

see anything (App. 326), at deposition, Wilber testified that he saw 5 spray planes and 2

helicopters on 3 separate occasions, all flying in a specific formation. (App. 331-333, 335-339.)

Dociteo and Edgar also provided new answers to these questions at deposition, with Dociteo and

Edgar for the first time specifying that they had seen 2 helicopters and Edgar increasing the

number of planes he had seen from 3 to 5. (App. 305, 314, 342, 348.) As a result, at deposition,

the three Sandoval test plaintiffs were all suddenly testifying to the same story. Moreover,

whereas Edgar’s Questionnaire response had claimed exposure 4 kms from the border, at

deposition, Edgar testified (now consistent with his father) that he was exposed on the family

farm adjacent to the river border. (App. 312, 313, 342, 347.) Wilber, meanwhile, who stated in

his Questionnaire response that he was exposed at his home, testified at deposition that two of

his alleged exposures occurred while he was at school. (App. 326, 332, 336.)

As to personal injuries, Edgar and Wilber both abandoned in their depositions the claims

in their Questionnaire responses that they had incurred specified out-of-pocket expenses for

medical treatment ($20 and $300, respectively) (App. 329, 340, 346, 350 ), but Edgar and

Dociteo added new personal injury allegations to those claimed in their Questionnaire responses

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and causation statements, with Dociteo adding a new claim of vision loss (App. 306-307, 323,

324), and Edgar newly contending that his alleged exposure had caused him to suffer vomiting

and diarrhea. (App. 343-344, 349, 353-354.) Dociteo likewise added to the family’s claims for

crop damage, testifying that in addition to the alleged crop damages set forth in his Questionnaire

response and causation statement, the herbicide spray also had killed 10 coconut trees, as well as

cassava, bananas and oritos. (App. 308, 310-311, 315-318, 324.)

6. Elvia Alvarez

Ms. Alvarez is the only member of her family who has been proffered as a test plaintiff.

Her testimony is typical of the other test plaintiffs. In her Questionnaire response, Ms. Alvarez

stated that she was exposed to Plan Colombia spraying several times between 2002 and 2006.

(App. 355-356.) In her deposition, however, she testified that she witnessed only two spray

events, in April 2001 and sometime around October 2001. (App. 366, 378-379.) Whereas her

Questionnaire response did not answer the question about the number of helicopters that she had

seen, Elvia testified at deposition that she saw 6 helicopters. (App. 356, 366-367.) The location

of the alleged exposures also changed, from 3 km from the border in her Questionnaire response

and causation statement, to 7 km from the border at her deposition. (App. 356, 365, 396.) Ms.

Alvarez’s testimony regarding her alleged exposure in April 2001 became particularly

convoluted when she was confronted with the fact that her son Byron – who Ms. Alvarez had

alleged was with her during the April 2001 exposure – had stated in his Questionnaire response

(App. 401, 402) that he had not seen any spray planes. Ms. Alvarez responded that Byron had

not seen the planes because he was working on another part of the farm that was more

mountainous, a fact she claimed to recall because she had recorded that information in a

notebook along with the April 2001 date. (App. 372-373, 402.) When asked if she could

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produce the notebook, however, Ms. Alvarez claimed to have thrown it away three years ago.

(App. 373-374, 375-377.)

Ms. Alvarez also used her deposition testimony as an opportunity to expand upon the

broad allegations of personal injury and property damage set forth in her Questionnaire response.

As compared to her Questionnaire response and causation statement, Ms. Alvarez testified to

new personal injuries (headache, dizziness, stomach ache, diarrhea), new alleged crop damages

(2 additional hectares of pasture, 1 additional hectare of plantains, 1 hectare of peanuts and 30

assorted plants of guava, grape, and avocado), and new alleged animal deaths (2 additional cows,

1 additional horse, 1 additional dog, and 40 guinea pigs). (App. 357-359, 361-364, 368-371,

386-387, 388.) Like other test plaintiffs, Ms. Alvarez first tried to justify in her deposition the

high values placed on her crops in her Questionniare response, see e.g., App. 389-390, but her

calculations quickly fell apart upon questioning. (App. 390-393.)

7. Edgar Balcazar

Mr. Balcazar is the only test plaintiff from his family. Like the other test plaintiffs, his

deposition was replete with new and different claims as to his alleged exposures and damages.

In his Questionnaire response, Mr. Balcazar alleged that he had been exposed to Plan Colombia

spray “in the year 2001 and 2002 until the year 2007,” while at his deposition, Mr. Balcazar was

unwilling even to guess at the years of his alleged exposure. (App. 403, 413, 414, 415.) Whereas

his Questionnaire stated that he had not seen a spray plane, at deposition he testified that he had,

albeit at a distance. (App. 403, 412, 413.) While Mr. Balcazar repeated at deposition his claim

in his Questionnaire responses to a variety of non-specific personal injuries, he admitted that he

25
Like test plaintiffs Edy Mestanza and Dociteo Sandoval, Edgar Balcazar was surprised to learn
at his deposition that his son is also a plaintiff, testifying that he did not “want [his] son Diego to
be identified as a plaintiff in this litigation.” (App. 424.)

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has no basis for and cannot confirm the representation in his Questionnaire response that he

spent $3,000 out-of-pocket for medical expenses. (App. 406, 416-417.) Moreover, Mr.

Balcazar’s testimony in explaining why he initially forgot during his deposition to mention his

claimed respiratory symptoms (symptoms for which, as with all of the medical symptoms

claimed by all the test plaintiffs, there is no documentary evidence) raises obvious red flags as to

the reliability of his testimony: “You cannot be after every detail for what happened to you

because time goes by and, but that’s something we can recognize if it comes into your eyes and

your nose why shouldn’t your respiratory ways be affected as well?” (App. 422-423.)

Finally, Mr. Balcazar’s testimony as to his alleged crop damages bears virtually no

resemblance to his identification of crop damages in his Questionnaire responses: Some of the

alleged crop damages decreased (6 hectares of cacao in his Questionnaire dropped to 2 hectares

at deposition, 5 hectares of coffee dropped to 2-3 hectares, 35 hectares of pasture dropped to 20

hectares, and ½ hectare of allegedly damaged coconuts disappeared altogether), while others

increased (1 hectare of rice became 3 hectares at deposition, ½ hectare of yucca increased to 1-

1½ hectares, and 2 hectares of lost plantains and a variety of lost fruit trees, including avocado,

lime, orange, and grapefruit were identified at deposition whereas such crops had not been

identified in his Questionnaire). (App. 407, 418-420.) Further, at his deposition, Mr. Balcazar

conceded that he did not even own the two farms at which he alleged that these damages

occurred, as the title to one farm is held solely by his mother and title to the other is held by his

wife, neither of whom are plaintiffs in this litigation. (App. 408-409.)

D. The Test Plaintiffs’ History of Changing Stories Regarding Their Alleged
Exposures Began Long Before This Litigation.

In Section V.H. of the agreed-upon Plaintiffs’ Questionnaire, each of the test plaintiffs

was asked whether they had made any previous complaints about their alleged exposures to Plan

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Colombia spraying to any other party. In their responses, 19 of the 20 test plaintiffs either

26
denied having made any such complaints or failed to respond to this question, and the one

plaintiff who answered “yes” failed to provide the requested information as to whom he had

complained and what had been communicated. (App. 246.) These representations were not true.

Rather, as evidenced by documents produced by the test plaintiffs on the eve of their depositions,

as well as an Ecuadorian judicial complaint located by the DynCorp defendants through their

own investigation, at least six of the seven test plaintiff families had raised claims of alleged

exposures and damages from Plan Colombia spraying in other forums. While the test plaintiffs’

failure to disclose these complaints in their Questionnaire responses is improper in its own right

(and impeded the DynCorp defendants in their ability to effectively prepare for the test plaintiffs’

depositions), the documents now available to defendants are particularly telling in that they

demonstrate that the test plaintiffs’ history of false and changing stories regarding alleged

exposures to and injuries from Plan Colombia spraying began long before this litigation.

For example, in December 2002, various Ecuadorian plaintiffs sued the President of

Ecuador and other government officials in an Ecuadorian court for failing to protect them from

alleged personal injuries and property damages purportedly arising from exposure to the

Republic of Colombia’s spraying of herbicide during the period December 2000 through July

2002. (App. 427 et seq.) Test plaintiff Santos Calero signed this complaint despite his

acknowledgment in this litigation that he was not exposed (even allegedly) to Plan Colombia

spraying until August 2003, eight months after the Ecuadorian complaint was filed. (App. 11,

26App. 6-7, 25, 41, 59, 69, 103, 116, 126, 160-161, 172, 175-176, 185, 228, 279-280, 309, 328,
345, 360, 405.

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15, 443.)27 Similarly, in March 2002, test plaintiff Elvia Alvarez signed the same complaint

(App. 448) and also prepared two certifications with her brother, the President of her residential

commune Rivera Del Oriente, in support of her complaint that Plan Colombia spraying in

December 2000 and January 2001 had, inter alia, caused the death of her husband in February

2001. (App. 380-385, 397-398, 399-400) But in this litigation, Ms. Alvarez testified that she

was not exposed to Plan Colombia spraying until April 2001, two months after her husband’s

death, and conceded that her husband’s illness began in early 2000, well before even the dates of

alleged exposure asserted in the prior certification she prepared with her President-brother.

(App. 366, 382-383, 394-395A.) (Ms. Alvarez is not claiming damages for the death of her

husband in this litigation.)

Further, the third party complaints by other test plaintiffs set forth litanies of alleged

personal injuries and property damages that are far different from those they claim in this case –

either in their 2008 sworn Questionnaire responses or in their 2009 deposition testimony. In the

DynCorp defendants’ prior briefing in response to plaintiffs’ motion to dismiss three non-test

plaintiff Mestanza family members, defendants demonstrated that test plaintiff Victor Mestanza

had secured, for purposes of a separate litigation in Ecuador, a fraudulent medical certification

identifying alleged personal injuries to his grandson Victor Manuel, despite the now-

acknowledged fact that his grandson was not exposed to any spraying operations and was living

275 miles away in Guayaquil at the time of the alleged spraying events. See Defendants’

Response to Plaintiffs’ Motion to Dismiss Three Plaintiffs, ECF No. 172 in Arias, ECF No. 132

in Quinteros, at 9-10. In addition to this, however, Victor Mestanza also secured medical

27
A second test plaintiff, Rosa Altamirano, also signed on to the Ecuadorian complaint (App.
481), despite stating in her Questionnaire that her first exposure was not until October 2002,
three months after the end of the exposure period alleged in that complaint. (App. 163.) She
also failed to answer the Questionnaire section asking about other complaints. (App. 166-167.)

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certifications on his own behalf and on behalf of his granddaughter, test plaintiff Jennifer

Mestanza, in which he claimed personal injuries that his and Jennifer’s deposition testimony now

indicates did not occur. (App. 223, 302.) In particular, while the 2003 medical certification

Victor secured for his granddaughter Jennifer states that Plan Colombia spraying had caused her

to suffer both a 35% vision loss requiring eyeglasses and chronic gastritis, Jennifer testified at

her deposition in this case that she has never worn eyeglasses or had vision problems, and she

could not recall ever suffering from any stomach problems. (App. 292, 298-300.) The 2003

medical certification for Victor Mestanza is virtually identical to that prepared for his

granddaughter (and indeed virtually identical to the admittedly-fraudulent certification prepared

for his young grandson, Victor Manuel), similarly claiming that Plan Colombia spraying had

caused him to lose 35% of his vision and caused chronic gastritis. Victor Mestanza notably

backed away from those claims at his deposition in November 2009. (App. 209-210.)

Test plaintiff Jorge Salas signed a December 2002 certification in which he alleged that

exposure to Plan Colombia spraying caused skin infections, acute sores, inflammation of the

throat and dry cough, fungus in the intestines, dizziness, and fevers – a laundry list of alleged

maladies that only marginally intersects with his claims of nose and eye irritation, throat

inflammation, and skin itching and infection at his deposition in this case. (App. 78-79, 83-84,

99-100.) Test plaintiff Rosa Altamirano assisted in preparing a March 2002 certification signed

by the president of her cooperative setting out Rosa’s complaint that the Plan Colombia spraying

had caused her family (the Quevedos) to suffer respiratory problems – an allegation not made in

any of the Quevedo test plaintiffs’ Questionnaire responses, causation statements, or deposition

testimony – as well as Rosa’s complaints of crop damage to rice and cassava that likewise are

nowhere alleged in this litigation. (App. 155-156, 170-171.) Finally, test plaintiff Edgar

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Balcazar prepared an alleged “report of damages and harms from Plan Colombia” in May 2002

in which he made no mention of the now-alleged loss of cacao, coffee, coconuts, sugarcane, rice

and yucca, did not claim of the now alleged death of 2 or 3 horses and 30-50 hens, and alleged

only 4 pigs dying as compared to the 10 now alleged in this case. (App. 407, 421, 425-426.)

The only consistent message that emerges from the test plaintiffs’ third party complaints,

the test plaintiffs’ Questionnaire responses, and the test plaintiffs’ deposition testimony, is that

every time these test plaintiffs are asked about their alleged exposures to Plan Colombia

spraying, they come forward with a different story – a pattern that defendants can only assume

will continue if these cases ever get to trial. While this record would surely provide defendants

with a wealth of material for cross-examination, the test plaintiffs’ repeated changes in their

allegations dating back before this litigation was filed significantly prejudices defendants in their

ability to prepare their defense (to exposures allegedly occurring when? occuring where? causing

what alleged personal injuries and property damages?), and they demonstrate that the problems

in the plaintiffs’ Questionnaire responses and their violations of repeated Court orders cannot be

blamed on technical glitches or misunderstandings. The inherent unreliability of the test

plaintiffs’ factual claims cannot support the continued prosecution of their claims.

II. THE TEST PLAINTIFFS SHOULD BE SANCTIONED FOR THEIR FALSE AND
FRAUDULENT DISCOVERY RESPONSES AND REPEATED VIOLATIONS OF
COURT ORDERS

A. Dismissal Is Warranted As Sanction For The Test Plaintiffs’ Submission Of
False and/or Fraudulent Discovery Responses.

Although dismissal is “a severe sanction, and should be resorted to only to the extent

necessary to induce future compliance and preserve the integrity of the system . . . the most

severe in the spectrum of sanctions provided by statute or rule must be available to the district

court in appropriate cases.” Weisberg v. Webster, 749 F.2d 864, 869-870 (D.C. Cir. 1984). As

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the Supreme Court has explained, the Rule 37 sanction of dismissal “must be available . . . not

merely to penalize those whose conduct may be deemed to warrant such a sanction, but to deter

those who might be tempted to such conduct in the absence of such a deterrent.” Nat’l Hockey

League v. Metro. Hockey Club, Inc., 427 U.S. 639, 643 (1976). The availability of dismissal is

essential for effective case management. “[I]f district court judges are to discharge their heavy

case processing responsibilities effectively, their power to dismiss . . . must be more than

theoretical.” Bristol Petroleum Corp. v. Harris, 901 F.2d 165, 167 (D.C. Cir. 1990) (internal

citations and quotation marks omitted). As this Court noted in its January 12, 2010 Opinion

dismissing the claims of 590 other plaintiffs in this litigation with prejudice for failure to submit

accurate Questionnaire responses, dismissal is particularly appropriate here, because lesser

sanctions have been imposed previously but without success. See 1/12/10 Mem. Op., at 3-4.

The test plaintiffs’ misconduct in this case in providing false evidence under oath strikes

at the core of the judicial process and cannot be countenanced. As the United States Supreme

Court has explained, “[f]alse testimony in a formal proceeding is intolerable. We must neither

reward nor condone such a ‘flagrant affront’ to the truth-seeking function of adversary

proceedings.” ABF Freight Sys., Inc. v. N.L.R.B., 510 U.S. 317, 323 (1994) (citations omitted).

Accordingly, numerous courts faced with discovery abuses of the type here at issue have

concluded that dismissal is the appropriate sanction. See Chavez v. City of Albuquerque, 402

F.3d 1039, 1046 (10th Cir. 2005) (affirming district court’s dismissal of plaintiff’s jury verdict

where plaintiff perjured himself during discovery); Archibeque v. Atchison, Topeka and Santa Fe

Ry. Co., 70 F.3d 1172, 1175 (10th Cir. 1995) (affirming dismissal of case for plaintiff’s

submission of false and misleading discovery responses); Dotson v. Bravo, 202 F.R.D. 559, 574

(N.D. Ill. 2001) (“Dismissal is an appropriate sanction for giving false interrogatory responses.”)

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(bold and italic typeface omitted); In re Amtrak “Sunset Limited” Train Crash, 136 F. Supp. 2d

1251, 1257, 1271 (S.D. Ala. 2001) (dismissing plaintiff’s claim with prejudice for providing

knowing false interrogatory answers).

The D.C. Circuit has set forth three basic justifications that support the use of dismissal

as a sanction for misconduct:

• “First, the court may decide that the errant party’s behavior has severely

hampered the other party’s ability to present his case – in other words, that the

other party has been so prejudiced by the misconduct that it would be unfair to

require him to proceed further in the case.” Webb v. District of Columbia, 146

F.3d 964, 971 (D.C. Cir. 1998) (internal quotations and citations omitted).

• “Second, the court may take account of the prejudice caused to the judicial system

when the party’s misconduct has put an intolerable burden on a district court by

requiring the court to modify its own docket and operations in order to

accommodate the delay.” Id. (internal quotations and citations omitted).

• “And finally, the court may consider the need to sanction conduct that is

disrespectful of the court and to deter similar misconduct in the future.” Id.

(internal quotations and citations omitted).

Any one of these justifications alone is sufficient grounds for dismissal. Id. In this case, all

three justifications are present.

1. The Test Plaintiffs’ Failure to Provide Accurate Information
Regarding The Alleged Factual Bases Of Their Claims Has

Significantly Prejudiced Defendants.

The test plaintiffs’ submission under oath of the factual bases of their claims, which they

have now acknowledged are false, has significantly prejudiced defendants at every stage of the

pretrial process. As this Court noted in its January 12, 2010 Opinion, plaintiffs’ failure to

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provide accurate and complete Questionnaire responses, “impedes the defendants’ ability to

prepare their defense.” 1/12/10 Mem. Op., at 6 (“Without the requested information, the

defendants are hampered in knowing the full extent, nature and location of the plaintiffs’ alleged

damages.”) (citing In re Phenylpropanolamine (PPA) Prods. Liab. Litig., 460 F.3d 1217, 1234

th
(9 Cir. 2006). This prejudice began with defendants’ enormous expenditure of time and

resources in repeatedly seeking judicial assistance over the course of five separate motions

hearings in 2008 and 2009 in an (apparently futile) effort to secure accurate Plaintiff

Questionnaire responses and with defendants’ prodigious but wasted efforts in analyzing

Questionnaire responses that have now been shown to be wholly unreliable.

The prejudice continued with the defendants’ frustrated efforts to meaningfully prepare

for the 20 test plaintiff depositions, conducted over a four-week period in Quito, Ecuador. While

defendants were able to expose during these depositions at least some of the falsehoods in the

test plaintiffs’ Questionnaire responses, defendants did not and could not question the test

plaintiffs as to every response set forth in their Questionnaires and have no way to determine

whether the other Questionnaire responses are equally false. See In re Amtrak, 136 F. Supp. 2d

at 1260 (“A party is entitled to rely on an opposing party’s written responses to interrogatory

questions; he/she/it is not required to ask a party deponent every question in his/her deposition

that the party previously answered in the set of interrogatories. Indeed, such a practice would

render the interrogatories superfluous and unnecessarily increase the expense of a deposition.”).

Moreover, because of the false information in the Questionnaire responses, defendants prepared

their deposition questions to address now-abandoned factual allegations, and defendants were

unable to adequately prepare question regarding the new factual allegations the test plaintiffs

made for the first time at the depositions themselves. In addition, as noted above with respect to

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the test plaintiffs’ false Questionnaire responses as to complaints to third parties, the test

plaintiffs’ misconduct also prevented defendants from pursuing potential fruitful areas of

investigation in preparation for the depositions.

Looking forward, defendants will be significantly prejudiced in their ability to

meaningfully prepare for trial. Defendants plainly cannot rely on any of the information set forth

in the test plaintiffs’ Questionnaire responses. Nor, given the ever changing nature of the

plaintiffs’ allegations, can defendants have any confidence that the test plaintiffs will stick to the

stories told at deposition. The prejudice to defendants is particularly significant given the central

importance of the information at issue in the test plaintiffs’ false Questionnaire responses. For

example, in preparing their expert case, it is essential that defendants have accurate information

about, e.g., the times and locations of the alleged exposures (to rebut plaintiffs’ necessary

showing that any herbicide spray in the vicinity could have reached the individual plaintiff or

reached them at a sufficient dose to cause injury), the types of personal injuries allegedly caused

by the spraying (as well as an accurate history of other medical or environmental conditions that

might present alternative causes), and the types of crops and livestock allegedly impacted by the

spray and the nature of these alleged impacts. As it is, however, defendants can only proceed

based upon guesswork and speculation, must sift through plaintiffs’ varying and often mutually

exclusive discovery responses, and must prepare for any variety of different allegations that

might be made by the test plaintiffs at trial. As this Court explained in its January 12, 2010

Opinion, “draw[ing] conclusions based on incomplete information … is not the defendant’s

duty.” 1/12/10 Mem. Op. at 6.

More fundamentally, of course, defendants are prejudiced in having to respond at all to

inherently unreliable and demonstrably false factual allegations. The test plaintiffs have been

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provided numerous opportunities to present a reliable factual predicate for their claims, and they

have failed to do so. They should not be allowed to continue to impose upon defendants the

significant costs of litigating their nebulous and shifting claims.

2. The Test Plaintiffs’ Failure to Provide Accurate Information
Regarding The Alleged Factual Bases Of Their Claims Has

Significantly Prejudiced The Judicial Process.

The Court has since November 2007 devoted considerable time and resources in an effort

to secure accurate factual disclosures from the individual plaintiffs. At each turn, when

defendants were forced to file motions to compel, to seek sanctions for plaintiffs’ noncompliance

with prior orders, or to defend against plaintiffs’ objections to the Magistrate Judge’s orders, the

Court has ruled in favor of the defendants. In so doing, the Court was required to wade through

a total of 435 pages of briefing (plus over 1,150 pages of exhibits) and to partake in motions

hearings totaling some 278 transcript pages. In an effort to resolve the Court’s concerns,

plaintiffs’ counsel provides specific assurances that each of the individual plaintiffs had

answered the Questionnaire “as fully as they could based upon personal knowledge.” Jt. Mot. to

Dismiss, ECF No. 86 in Arias, ECF No. 48 in Quinteros, Feb. 19, 2009, at 10. The Court

accepted these assurances absent evidence to the contrary, but the Court made clear through its

dismissal with prejudice of two categories of plaintiffs who failed to provide information that

they clearly should have known that a plaintiff’s knowing failure to provide accurate information

in his or her Questionnaire response would not be tolerated. See 1/12/10 Mem. Op.

The testimony of the test plaintiffs demonstrates that the plaintiffs’ counsel’s

representation that plaintiffs had provided full and accurate Questionnaire responses to the best

of their knowledge was false and that all of the Court’s efforts over the past two years have been

for naught. Moreover, if the test plaintiffs are allowed to continue with their claims, the Court

undoubtedly will be required to intervene again and again in the day-to-day pretrial process in an

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effort to somehow mitigate the prejudice caused to the defendants by the test plaintiffs’

misconduct. The Court will also be required to preside over a trial where it will be “virtually

impossible for the jury to determine the truth” behind the test plaintiffs’ allegations. In re

Amtrak, 136 F. Supp. 2d at 1268.

As the D.C. Circuit explained in affirming a dismissal sanction in Weisberg, 749 F.2d at

193 (citation omitted), “if parties are allowed to flout their obligations [to respond to

interrogatories], choosing to wait to make a response until a trial court has lost patience with

them, the effect will be to embroil trial judges in day-to-day supervision of discovery, a result

directly contrary to the overall scheme of the federal discovery rules.” The test plaintiffs have

failed to abide by the Court’s orders, they have repeatedly embroiled the Court in futile efforts to

compel compliance, and they have through their false discovery responses subverted the judicial

process. Their claims should be dismissed.

3. The Test Plaintiffs Have Shown Disrespect to the Court and Dismissal
Is Necessary to Deter Such Misconduct By These and Other Litigants.

The history of the test plaintiffs’ repeated violations of the Court’s discovery orders

leaves no question that the test plaintiffs have failed to provide the Court with the proper respect.

Dismissal is necessary to sanction the test plaintiffs’ disrespectful behavior and to deter other

litigants from engaging in similar misconduct. The need for a clear sanction in this case is

particularly compelling because the Court’s response to the test plaintiffs’ misconduct likely will

determine whether the Court will be able to maintain any control whatsoever with respect to the

remaining 2,001 individual plaintiffs in this case, whose Questionnaire responses must be

assumed to contain the same glaring problems as those of the 20 test plaintiffs. Moreover,

litigants in other proceedings may be tempted to resort to similar misconduct if they believe that

the Court is unwilling to impose appropriate and meaningful sanctions to ensure compliance with

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its orders. See Nat’l Hockey League, 427 U.S. at 643 (stating that if the court of appeals’

reversal of the sanction of dismissal were to remain undisturbed, “other parties to other lawsuits

would feel freer than we think Rule 37 contemplates they should feel to flout other discovery

orders of other district courts”).

The Court has shown ample patience in its dealings with the plaintiffs. The Court has

repeatedly given the plaintiffs additional time to provide factual information they should have

had in hand before they even filed their claims, and it has repeatedly indulged plaintiff counsel’s

now-disproved assurances that the information had been accurately provided. The time for

patience is over. If the Court’s orders are to have any meaning, the 20 test plaintiffs’ claims

must be dismissed. Moreover, because the test plaintiffs’ misconduct was willful and because

the test plaintiffs have now, in any event, conclusively demonstrated their inability to present any

reliable, consistent evidence that would state a cause of action for alleged harms from purported

Plan Colombia spray exposures, their dismissals should be with prejudice. See 1/12/10 Mem.

Op.; see also Norman v. United States, 467 F.3d 773 (D.C. Cir. 2006) (affirming dismissal with

prejudice where evidence demonstrated that refiling of claim would be futile); Handy v. Shaw,

Bransford, Veilleux & Roth, No. 00-2336, 2006 WL 3791387, at *7 (D.D.C. Dec. 22, 2006)

(dismissal with prejudice appropriate as sanction for willful misconduct).

B. In the Alternative, Defendants Request That The Test Plaintiffs Be
Precluded From Defending Their False Questionnaire Responses and That

The Jury Be Instructed Regarding the Test Plaintiffs’ Misconduct.

If the Court elects to allow the test plaintiffs to proceed with their claims, defendants

request that the Court issue an alternative sanctions order that provides defendants with at least

some measure of relief from the prejudice imposed upon them by the test plaintiffs’ misconduct.

First, defendants request that the test plaintiffs be precluded from presenting any evidence or

argument at trial to defend, explain, or mitigate the fact that they provided false statements under

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oath in their Questionnaire responses. As this Court has noted, such a preclusion order is an

“unexceptional remedy that is contemplated in the federal rules” and it is plainly warranted here.

See Moore v. Napolitano, No. 00-953 (RWR-DAR), 2009 WL 2450280, at *1-2, 9 (D.D.C. Aug.

7, 2009) (precluding defendant from introducing as evidence any information responsive to an

interrogatory that was not produced in a timely manner); see also Jung v. Neschis, No. 01 Civ.

6993, 2009 WL 762835, at *24 (S.D.N.Y. Mar. 23, 2009) (adopting report and recommendation

precluding the plaintiffs from using fraudulent evidence or amending reports to take it into

account). Second , defendants request that the Court instruct the jury at any future trials that the

test plaintiffs provided false information under oath regarding the basic factual foundations of

their alleged claims and that this conduct may be considered by the jury in assessing the test

plaintiffs’ credibility. See, e.g., Jung, 2009 WL 762835, at *24 (adopting report recommending

jury instruction that jurors should consider fabricated evidence and false statements in assessing

the plaintiff’s credibility on other matters); Bernal v. All Am. Investment Realty, Inc., 479 F.

Supp. 2d 1291, 1301 (S.D. Fla. 2007) (ordering jury instruction that jurors may take into account

the defendant’s false statements and conduct when considering his credibility); Rybner v.

Cannon Design, Inc., No. 95 Civ. 0279 (SS), 1996 WL 470668, at *6 (S.D.N.Y. Aug. 20, 1996)

(“[D]efendants will be permitted to inform the jury of [plaintiff’s] dishonesty and a jury charge

will be given that any falsehood under oath should be considered seriously by jurors in assessing

[plaintiff’s] credibility.”).

III. THE REMAINING 2,001 INDIVIDUAL PLAINTIFFS SHOULD BE BROUGHT
INTO COMPLIANCE WITH THE COURT’S PRIOR DISCOVERY ORDERS.

In addition to establishing their own inexcusable misconduct, the test plaintiffs’

depositions demonstrated without question: (1) that defendants (and the Court) can have no

confidence in the factual information provided to date by the remaining 2,001 individual

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plaintiffs in this litigation and (2) that without further order from the Court to finally secure

accurate and meaningful information from these plaintiffs, this litigation cannot meaningfully

proceed. Defendants accordingly request the Court to issue a detailed order providing the 2,001

individual plaintiffs with one last opportunity to provide what the Court has long required:

verified, factual and complete responses to the Plaintiffs’ Questionnaire and individualized

expert causation statements based upon a meaningful scientific assessment that links each

individual plaintiff’s alleged exposure to his or her alleged personal injuries and/or property

damages. The Court’s order should further provide that if plaintiffs fail to provide the ordered

information and causation statements, their claims will be dismissed with prejudice.

With respect to the Plaintiffs’ Questionnaire, defendants submit that the Court need not

order the individual plaintiffs to prepare completely new Questionnaire responses. Indeed,

absent some showing that the individual plaintiffs would approach the Questionnaires

differently, such an order would not provide any meaningful assurance that the plaintiffs’ new

responses would be any more accurate than their old ones. Instead, defendants request that each

individual plaintiff be required to provide a sworn certification that confirms the accuracy of his

or her prior Questionnaire responses and/or provides corrected information to those questions

that were answered inaccurately. To ensure that the individual plaintiffs are providing accurate

information, the proposed certification should contain a number of additional safeguards:

First, the certification should state that plaintiffs are being required to prepare the

certificate because of the Court’s concerns about the reliability of the previous Questionnaire

responses. Second, the certification should state that if the certification is shown to be

knowingly inaccurate in any respect, the plaintiffs’ claims will be dismissed and that the plaintiff

(or his or her counsel) will be required to pay defendants’ costs in seeking such dismissal. Third,

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plaintiffs should be required to certify that he or she has read both the certification and his or her

prior Questionnaire responses in full (or, if the plaintiff is unable to do so, that the documents

have been read to him or her). Fourth, the plaintiffs should be required to physically sign the

certificate (as opposed to providing an electronic signature), have it witnessed, and have the

original copy of the signed certificate maintained by plaintiffs’ counsel in the United States and

available for inspection by defendants upon reasonable demand. A proposed certification

providing such assurances is attached to the proposed order being filed with this motion.

With respect to the previously-ordered causation statements, the DynCorp defendants

submit that the only possible course of action is an order requiring plaintiffs to provide new

statements. As set forth above, the aggregate causation statements provided by plaintiffs to date

(with only a one-paragraph boilerplate recitation of the individual plaintiff’s allegations) are

directly contrary to the Court’s repeated Orders that plaintiffs provide individualized causation

statements that make “some connection based upon some scientific assessment between the

allegation that spraying happened and that spraying caused these symptoms.” 11/25/08 Hr’g Tr.

(Ex. C) at 60:19-21; see also 7/17/09 Hr’g Tr. (Ex. D) at 39 (“I have made it clear that there’s

got to be some individualized assessments with regard to each individual plaintiff and each

individual plaintiff’s complaints about harm or damage.”).

Moreover, because the prior causation statements are based solely on Questionnaire

responses that have now been shown unreliable, the boilerplate paragraph 16 in each causation

statement likely does not even address the individual plaintiffs’ actual alleged exposures or

harms. Further, the failure of the test plaintiffs after three years of litigation – and after

defendants’ production of hundreds of thousands of pages of Plan Colombia documents and nine

years of electronic “spray line” data covering the entire Colombia-Ecuador border – to provide

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any reliable and consistent information as to the factual predicates of their claims makes the need

for meaningful causation statements all the more evident. See In re Vioxx Products Liability

Litigation, 557 F. Supp. 2d 741, 744 (E.D. La. 2008) (noting that while individual causation

statements “may not have been appropriate at an earlier stage before any discovery had taken

place … this case is no longer in its embryonic stage”).

In light of plaintiffs’ repeated – albeit baseless – claims that the Court’s prior orders

requiring causation statements were ambiguous, defendants request that the Court issue a

detailed order that clearly specifies what the Court is requiring by way of a scientific assessment

of each individual plaintiff’s claims: First, the causation statement must specifically address the

individual plaintiff’s allegations of exposure, including some scientific assessment of the

proximity of the plaintiff to any alleged spraying events on the dates of alleged exposure and

some scientific basis for a conclusion that the spray could have reached the plaintiff or their

property at a dose level sufficient to cause injury or damage. Second, the causation statement

must specifically address the individual plaintiff’s claims of personal injury and provide some

scientific basis for a conclusion that his or her alleged exposure to Plan Colombia spray could

have caused each of the specific types of injury alleged. Third, the causation statement must

specifically address the individual plaintiff’s claims of property damage and provide some

scientific basis for a conclusion that the alleged exposure to Plan Colombia spray could have

caused each of the specific property damages alleged.

In addition, each of the foregoing assessments in the causation statements must be

proffered by an expert who is specifically qualified to speak to the issue at question. See Acuna

v. Brown & Root, Inc., No. SA-96-CA-543-OG, 1998 WL 35283824, at *5 (W.D. Tex. Sept. 30,

1998) (That the court’s orders requiring “affidavits from qualified experts” did not “specif[y] the

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particular type of expert required . . . does not excuse Plaintiffs from the requirement that the

experts they offer actually be qualified in the subject matter area in which [they] testif[y].”),

aff’d, 200 F.3d 335 (5th Cir. 2000). Thus, for example, plaintiffs may not rely, as they have to

date, solely on Dr. Campana to prepare the causation statements because Dr. Campana does not

even purport to have the requisite expertise in exposure assessment, animal toxicology, or

agronomy to speak to the plaintiff’s exposure and property damage allegations.

Finally, in light of the testimony from three of the test plaintiff families that minor

children have been put forward as plaintiffs in this litigation without their parents’ knowledge,

approval, or authorization, see supra nn. 21, 24, and 25, defendants request that the Court require

signed parental authorizations on behalf of each of the minor plaintiffs in the remaining

individual plaintiff group. See Fed. R. Civ. P. 17(c) (addressing capacity of minors to bring

suit); Weinbaum v. City of Las Cruces, New Mexico, 465 F. Supp. 2d 1164, 1166 n.1 (dismissing

claim of minor plaintiff because she “lacks the legal capacity to sue on her own behalf”); Woods

v. Wills, 400 F. Supp. 2d 1145, 1181 (E.D. Mo. 2005) (“To maintain a suit in federal court, a

minor must be represented by a competent adult”).

IV. THE COURT SHOULD ORDER PLAINTIFFS’ COUNSEL TO PAY THE
DEFENDANTS THEIR EXPENSES AND FEES IN BRINGING THIS MOTION.

Finally, the defendants request their reasonable expenses, including attorney’s fees, in

bringing this motion. Rule 37 provides that when granting a motion for sanctions for failure to

comply with a court order, “the court must order the disobedient party, the attorney advising that

party, or both to pay the reasonable expenses, including attorney’s fees, caused by the failure,

unless the failure was substantially justified or other circumstances make an award of expenses

unjust.” Fed. R. Civ. P. 37(b)(2)(C); see also Kornagay v. AT&T, No. 05-0001, 2008 WL

4482970, at *1 (D.D.C. Sept. 29, 2008) (“This section of Rule 37 is mandatory unless ‘the

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disobedient party’ meets its burden to avoid expenses”). Plaintiffs’ failures here are not justified

and no known circumstances make an award of fees unjust. In light of defendants’

understanding that plaintiffs’ counsel have agreed to cover all of plaintiffs’ costs in this

litigation, defendants request that the Court specify in its order that the payment of defendants’

expenses be made by plaintiffs’ counsel, as the Court did in its previous order sanctioning

plaintiffs in this case. See 7/17/09 Hr’g Tr. at 50:12-51:1 (Ex. D).

CONCLUSION

For the foregoing reasons, defendants respectfully request that their motion for sanctions

be granted.

Dated: January 26, 2010 Respectfully submitted:

/s/ Eric Lasker

Joe G. Hollingsworth (D.C. Bar # 203273)
Eric G. Lasker (D.C. Bar # 430180)
Rosemary Stewart (D.C. Bar # 204438)
HOLLINGSWORTH LLP
1350 I Street, NW
Washington, D.C. 20005

Phone: (202) 898-5800
Fax: (202) 682-1639

Counsel for the Defendants
DynCorp International, et al.

658041v1

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600

Document Long Title

volume III