-------
PESTICIDE ASSESSMENT GUIDELINES
SUBDIVISION F
HAZARD EVALUATION
HUMAN AND DOMESTIC ANIMALS
Series 81-1
Acute Oral Toxicity
ADDENDUM ON DATA REPORTING
Prepared by:
Robert P. Zendzian Ph.D.
for
Health Effects Division
Office of Pesticide Programs
US Environmental Protection Agency
-------
Subdivision F - Acute Oral Toxicity Studies
Table of Contents of Addendum
Page
Introduction 1
Response to Public Canments 1
Acute Oral Toxicity
Guideline
Cover Page 1
Table of Contents 2
Body of Report 2
I. Summary 2
II. Introduction 2
III. Materials/Methods 2
IV. Results 3
V. Discussion 3
VI. Bibliography 3
VII. Verification 3
VIII. Archives , 4
IX. Tables/Figures 4
X. Appendix(es) 4
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PESTICIDE ASSESSMENT GUIDELINES
HUMAN AND DOMESTIC ANIMALS
Acute Oral Toxicity
Subdivision F, Series 81-1
DATA REPORTING
INTRODUCTION
A. Purpose
The purpose of this addendum to the guideline is to provide a data reporting
format for acute oral toxicity studies. Subpart F, Series 81-1 provides
guidelines for carrying cut the acute oral toxicity study.
B. Objective
The objective of the Data Reporting Guidelines (DRGs) is to provide an
example of an acceptable general reporting format that can be reviewed
effectively and expeditiously in accordance with the Agency's review
process. The recommendations are intended to be consistant with the
guidelines for data reporting as described in Sections 8O4 and 81-1 and
those described in the Agency's Good Laboratory Practice Standards (40
CFR Part 160).
While following this Guideline is not mandatory, data submitters are
encouraged to submit complete reports which can be effectively reviewed
by the Agency. 40 CFR Part 158 pertains to the physical formatting of
reports (which are referred to as "studies") and submitted packages. Some
of its requirements are mandatory.
RESPONSE TO PUBLIC COMMENTS
The Agency received one response to its request for public conments on these
documents published in the FEDERAL REGISTER of March 8, 1989 (54 FR 9886).
These conments were considered in the final form of this DR.G.
GUIDELINE
The Final Report should contain the following items.
COVER PAGE
The title page and additional documentation (i.e., requirements for data
submission, Good Laboratory Practices statement and statement of data
confidentiality claims), if relevant to the study report, must precede
the content of the study format below. These requirements are described
in 40 CFR Part 158.
-------
TABLE OF CONTENTS
The table of contents should include a listing of the elements of the
final report such as the Summary, an Introduction, the Materials and
Methods, Results, Discussion Bibliography, Tables, Figures, Appendices
and key subsections as appropriate.
BODY OF THE REPORT
This item shall include all information required in §80-4(b)(2), 80-4(c) and
81-1 (h) of this subdivision and should be provided in such detail that the
reviewer can assess the quality of the study and conformity to the acute ora
toxicity Guideline. It should contain the following sections.
I. SUMMARY
As per §80-4(b)(l) of this subdivision, the test report shall contain a
summary including a brief description of the study protocol, chemical
used, the animals tested and the highlights of the results of the study
relative to the positive findings. Any deviations from the intended
protocols should be noted.
II. INTRODUCTION .
Include the objectives of the study and the Guideline reference. The
overall experimental design should be explained.
III. MATERIALS AND METHODS
A. Test substance
Test material (chemical name)
Form (technical, manufacturing-use product or formulation)
source
lot number
CAS No.
purity
state
lonization constant (if applicable)
pH (if applicable)
octanol/water partition coefficient (if Known)
vehicles used
B Test Animals
species and strain
sex
source
body weight
number of test animals
number of control animals
-------
pre-test condition
housing conditions
test environment
C. Experimental Design
Doses used
Number and sex of animals per dose group
Observations, type and time
Termination
D. Evaluation Procedures
The final report should describe the types of observations of the animals
for toxic signs, body weights, the frequency of observations, the gross
pathological examination procedures and any other observations performed.
The statistical methods used for calculation of the LDso the 95 percent
confidence limits and the slope of the dose-response curve should be
specified and referenced.
E. Deviation from protocol
Deviations fron the protocol recommended in § 81-1 should be described
along with the rational for the changes.
IV. RESULTS
This sections should provide a summary of the toxic signs, mortality,
body weight changes, gross pathology and other observations performed
associated with the doses tested. Typically, these results are summarized
in tabular form.
V. DISCUSSION
This section should provide an assessment of the results of the study, an
interpretation of the toxicity observed, the consistency of the dose-response
effects and should try to explain unexpected findings. The impact of any
deviation from the Guideline protocol should be discussed.
VI. BIBLIOGRAPHY
This itan should contain a list of references cited in the body of the
report.
VII. VERIFICATION
This item shall contain information required by §80-4(b)(2). Each test
report shall be:
A. Signed by each of the senior scientific personnel, including the
laboratory director responsible for performing and supervising the
testing, preparing, reviewing and approving the test report and
-------
B. Certified by the applicant or an authorized agent of the applicant
as a complete and unaltered copy of the report provided by the
testing laboratory whether independent or owned, operated or
controlled by the applicant.
VII. ARCHIVES
This section of the final report shall contain all information required
in 40 CFR 160.185.
IX. TABLES/FIGURES
X. APPENDIX(ES)
These should include individual animal data, historical control data,
pathology report, analytical methods and results of analysis of the test
substance, details of statistical analysis, protocol and other information
as appropriate.
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PESTICIDE ASSESSMENT GUIDELINES
SUBDIVISION F
HAZAFD EVALUATION
HUMAN AND DOMESTIC ANIMALS
Series 81-2
Acute Dermal Toxicity
ADDENDUM ON DATA REPORTING
Prepared by:
Robert P. Zendzian Ph.D.
for
Health Effects Division
Office of Pesticide Programs
US Environmental Protection Agency
7
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Subdivision F - Acute Dermal Toxicity Studies
Table of Contents of Addendum
Page
Introduction 1
Response to Public Comments 1
Acute Dermal Toxicity
Guideline 1
Cover Page 1
Table of Contents 2
Body of Report 2
I. Summary 2
II. Introduction 2
III. Materials/Methods 2
IV. Results . 3
V. Discussion 3
VI. Bibliography 3
VII. Verification 3
VIII. Archives 4
IX. Tables/Figures . 4
X. Appendix 4
8
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PESTICIDE ASSESSMENT GUIDELINES
HUMAN AND DOMESTIC ANIMALS
Acute Dermal Toxicity
Subdivision F, Series 81-2
DATA REPORTING
INTRODUCTION
A. Purpose
The purpose of this addendum to the guideline is to provide a data
reporting format for acute dermal toxicity studies. Sufcpart F, Series
81-2 provides guidelines for carrying cut the acute dermal toxicity
study.
B. Objective
The objective of the Data Reporting Guidelines (DRGs) is to provide an
example of an acceptable general reporting format that can be reviewed
effectively and expediticusly in accordance with the Agency' s review
process. The recommendations are intended to be consistent with the
guidelines for data reporting as described in Sections 80-4 and 81-2 and
those described in the Agency's Good Laboratory Practice Standards (40
CFR Part 160).
While following this Guideline is not mandatory, data submitters are
encouraged to submit complete reports which can be effectively reviewed
by the Agency. 40 CFR Part 158 pertains to the physical formatting of
reports (which are referred to as "studies") and submitted packages. Some
of its requirements are mandatory.
RESPONSE TO PUBLIC COMMENTS
The Agency received one response to its request for public comments on these
documents published in the FEDERAL REGISTER of March 8, 1989 (54 FR 9886).
These comments were considered in the final form of this DRG.
GUIDELINE
The Final Report should contain the following items.
COVER PAGE
The title page and additional documentation (i.e., requirements for data
submission, Good Laboratory Practices statement and statement of data
confidentiality claims), if relevant to the study report, must precede
the content of the study format below. These requirements are described
in 40 CFR Part 158.
-------
TABLE OF CONTENTS
The table of contents should include a listing of the elements of the
final report such as the Summary, an Introduction, the Materials and
Methods, Results, Discussion Bibliography, Tables, Figures, Appendices
and key subsections as appropriate.
BODY OF THE REPORT
This item shall include all information required in §80-4(b)(2), 80-4(c) and
81-2 (h) of this subdivision and should be provided in such detail that the
reviewer can assess the quality of the study and conformity to the acute dermal
toxicity Guideline. It should contain the following sections.
I. SUMMARY
As per §80-4(b)(l) of this subdivision, the test report shall contain a
summary including a brief description of the study protocol, chemical
used, the animals tested and the highlights of the results of the study
relative to the positive findings. Any deviations from the intended
protocols should be noted.
II. INTRODUCTION
Include the objectives of the study and the Guideline reference. The
overall experimental design should be explained.
III. MATERIALS AND METHODS
A. Test substance
Test material (chemical name)
Form (technical, manufacturing-use product or formulation)
source
lot number
CAS No.
purity
state
lonization constant (if applicable)
pH (if applicable)
octanol/water partition coefficient (if known)
vapor pressure at 20° C (if known)
vehicles used
B Test Animals
species and strain
source
sex
body weight
number of test animals
-------
number of control animals
pre-test condition
housing conditions
test environment
C. Experimental Design
Doses and duration of exposure
Number and sex of animals per dose group
Application site, preparation and method(s) of protection
Observations, type and time
Termination
D. Evaluation Procedures
The final report should describe the types of observations of the animals
for toxic signs, body weights, the frequency of observations, the gross
pathological examination procedures and any other observations performed.
The statistical methods used for calculation of the LDso the 95 percent
confidence limits and the slope of the dose-response curve should be
specified and referenced.
E. Deviation from protocol
Deviations frcm the protocol recommended in § 81-2 should be' described
along with the rational for the changes.
IV. RESULTS
This sections should provide a summary of the toxic signs, mortality,
body weight changes, gross pathology and other observations performed
associated with the doses tested. Typically, these results are surmBrized
in tabular form.
V. DISCUSSION
This section should provide an assessment of the results of the study, an
interpretation of the toxicity observed, the consistancy of the dose-response
effects and should try to explain unexpected findings. The impact of any
deviation from the Guideline protocol should be discussed.
VI. BIBLIOGRAPHY
This item should contain a list of references cited in the body of the
report.
VII. VERIFICATION
This item shall contain information required by §80-4(b)(2). Each test
report shall be:
A. Signed by each of the senior scientific personnel, including the
laboratory director responsible for performing and supervising the
testing, preparing, reviewing and approving the test report and
1 1
-------
B. Certified by the applicant or an authorized agent of the applicant
as a complete and unaltered copy of the report provided by the
testing laboratory vhether independent or owned, operated or
controlled by the applicant.
VII. ARCHIVES
This section of the final report shall contain all information required
in 40 CFR 160.185.
IX. TABLES/FIGURES
X. APPENDIX(ES)
These should include individual animal data, historical control data,
pathology report, analytical methods and results of analysis of the test
substance, details of statistical analysis, protocol and other information
as appropriate.
12
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PESTICIDE ASSESSMENT GUIDELINES
SUBDIVISION F
HAZAFD EVALUATION:
HUMANS AND DOMESTIC ANIMALS
Series 82-1 and 83-1
Subchronic and Chronic Dog
ADDENDUM ON DATA REPORTING
Prepared by:
Mission Support Section
Toxicology Branch
Hazard Evaluation Division
Project Manager:
Elizabeth M.K. Leovey, PhD
Hazard Evaluation Division
Office of Pesticide Programs
U.S. Environmental Protection Agency
13
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Subdivision F - Subchronic and Chronic Dog
Table of Content of Addendum
Page
Introduction 2
Response to Public Comments 2
Guideline
Title/Cover Page 3
Table of Contents 3
Body of Report 3
I. Summary 3
II. Introduction 3
III. Materials and Methods 3
IV. Results 5
V. Discussion/Conclusions 5
VI. Bibliography 5
VII. Verification 5
VIII. Archives 6
IX. Tables 6
X. Individual Animal Data 6
XI. Tables/Figures (other) 6
XII. Appendix(ces) 6
Examples of Tables for Report 7
14
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PESTICIDE ASSESSMENT GUIDELINES
HUMANS AND DOMESTIC ANIMALS
Subchronic and Chronic Dog
Subdivision F, Series 82-1 and 83-1
DATA REPORTING
INTRODUCTION
A. Purpose
This Data Reporting Guideline (DRG) is designed to aid the petitioner/
registrant in generating reports which are compatible with the
Agency's review process. While following this Guideline is not
mandatory, data submitters are strongly encouraged to submit
complete reports which can be efficiently reviewed by the Agency.
This DRG pertains to organizing and presenting the substance of the
data report. 40 CFR Part 158 refers to the physical formatting
of reports (which are referred to as "studies") and submittal packages.
Some of its requirements are mandatory.
B. Objective
The objective of the Guideline is to provide an example of an
acceptable format for reporting materials, methods, and results of
chronic and subchronic dog studies. These recommendations are
intended to show how the requirements of §80-4, §82-1, and §83-1,
the Agency's Good Laboratory Practice Standards (40 CFR, Part 160),
and the guidance offered by the Office of Pesticide Programs in
its Standard Evaluation Procedures, available from the National
Technical Information Center may be organized in a final report.
RESPONSE TO PUBLIC COMMENTS
The purpose of this section is to acknowledge and address the concerns
expressed in the letters of comment received by the Agency in response to
the public notice in the FEDERAL REGISTER (51 FR 18660) of May 21, 1986.
Most of the comments concerned the organization of the different sections
and of the DRG itself. The Agency has considered these comments and revised
this DRG to be consistent with other DRGs for Subdivision F and with how the
Agency reviews these studies. The latter is particularly the reason for the
inclusion of summary tables in the RESULTS sections. Individual animal data
or data for several parameters tabulated by animal is needed to allow the
Agency's toxicologists to readily recognize the interrelationships of toxic
effects. This is especially true in the case of gross-and histopathology
observations were tthe Agency's toxicologists could easily check that gross
observations haave been evaluated aand addressed by histopathology.
15
-------
GUI PET .THE
TITLE/COVER PAGE
Cover page and additional documentation requirements (i.e., requirements
for data submission, Good Laboratory Practice statement, and statement
of data confidentiality claims), if relevant to the study report, must
precede the content of the study formatted below. These requirements
are described in CFR 40 Parts 158 and 160.
TABLE OF CONTENTS
A concise listing, preceding the body of the report, should be contained
in this item of all essential elements of the study, and the page or
table number where that element is located in the report. Essential
elements include a Summary, Introduction, Materials and Methods section,
Results, Discussion, Bibliography, Tables, Figures, Appendices, and key
subsections as appropriate.
BODY OF REPORT
The report should include all information required in §80-4(b)(2), §80-4(c),
§82-1, and §83-1.
I. SUMMARY
As per §80-4(b)(l) of this Subdivision, this section of the test report
shall contain a summary and analysis of the data, and a statement of
the conclusions drawn fron the analysis. The summary should highlight
any and all positive data or observation, and any deviation from control
data which may be indicative of toxic effects. The summary shuld be
presented in sufficient detail to permit independent evaluation of the
results.
II. INTRODUCTION
Description of the objectives of the study, name, and properties of the
chemical and the rational for the type of study conducted. The rationale
for any deviations fron the protocol reccmmended in Subdivision F
should be provided.
III. MATERIALS MD METHODS
A. Test Substance
1. Chemical name and chemical abstract number or code number;
2. Source of procurement, batch or lot number of each procurement;
3. Purity and identity analysis on bulk chemical;
4. Stability of test material in feed and control subtances;
5. Assay - description of method and percent of target concentration;
16
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6. Storage/stability of test material/diet; and
7. Formulation for administration to animals.
B. Animals
1. Description:
a. Species, strain and substrain;
b. Age,
c. Body waight range, and
d. Source of supply;
2. Isolation procedures;
3. Health assessment;
4. Caging;
a. Type of cages and
b. Excreta collection;
5. Housing - number of animals per cage;
6. Diet - vendor or composition, if appropriate;
7. Water;
a. Purity or source, and
b. System; and
8. Room assignment.
C. Methods
1. Treatment groups;
a. Dosage levels;
b. Number of animals per group;
c. Statistical consideration employed in selecting the number
of groups, dosage levels and number of animals in each
group.
2. Treatment regimen;
a. Route of administration;
17
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b. Frequency;
c. Volume and concentration;
d. Carrier/soluant
e. Daily records and clinical observations
3. Experimental measurements
Describe measurements and frequency with which they are made
and procedures employed.
4. Animal assignment
a. Procedure used for identification of each animal.
b. Randomization procedure for assignment to treatment
groups. Housing, food, and water during experiment
if different from that described in subsection
"aniTBls."
5. Statistical analysis
In presenting the statistical methods used to analyze the study
data, show the method used; including computer programs and
give sufficient information in order that an independent reviewer
can re-evaluate and reconstruct the analysis.
IV. RESULTS
Pertinent summary table with appropriate statistical evaluations clearly
indicated may be placed in the text of this section - see tables included
in this package. Each parameter studied shall be addressed sequentially
regardless of whether compound related effects were noted.
V. DISCUSSION/CONCLUSION
A. Discussion of incidences of effects of biological/statistical
significance, relationship of related events.
b. Discussion of observations and adequacy of the tests for the intended
purpose.
c. Discussion of observed toxic effects, target organ(s), dose related
compound induced effects, and no observed effect levels (NOEL).
VI. BIBLIOGRAPHY
VII. VERIFICATION
This item shall contain information required by §8O4(b)(2). Each test
report shall be:
1 °
I o
-------
A. Signed by each of the senior scientific personnel, including the
laboratory director, responsible for performing and supervising
the testing, preparing, reviewing, and approval of the test report;
and
B. Certified by the applicant or an authorized agent of the applicant
as a complete and unaltered copy of the report provided by the
testing laboratory whether independent or owned, operated, or
controlled by the applicant.
VIII. ARCHIVES
The section of the final report should contain all information required
in 40 CFR 160.185.
IX. TABLES (Means or Lesions/Dose/Sex)
X. INDIVIDUAL ANIMAL DATA
by Parameter(s)/Dose/Sex
XI. TABLES/FIGURES (other)
XII. APPENDICES (Optional)
e.g. historical control data or any other information that may facilitate
the review of a report by the Agency.
19
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Tabular Data Format
(EXAMPLE)
A Table showing means or number of lesions should be presented for all
measured parameters i.e., clinical chemistry, hematology, food consumption,
etc. It is also recommended that the changes in clinical parameters over the
course of the study be presented in graphical form.
Mean Summary Body Wt Gain
Groups
Control
xx ppm
xx ppm
xx ppn
No. of animals
in group
4
4
4
4
Initial
body weight
Mean
Mean
Mean
Mean
Body Weight
after
52 weeks
Mean
Mean
Mean
Mean*
Weight
gain
Mean
Mean
Mean
Mean*
* Statistical Significance
20
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The following are examples of individual animal reporting formats for the
registrants convenience.
Clinical Observation
Test Material
Animal #1 Treatment Group; Control Female Term Sacrifice
Moribund Sacrifice
Found Dead
Date of
Observation Clinical Observations
-------
Body/Organ Weights
(EXAMPLE)
Test Compound
Animal #1 Control Group Sex; Female
Date of
Determination Body Weight (Kg) Body/organ wt. Ratio
Organ Wt (gm) Ratio
brain
heart
liver
kidney
other organs
22
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10
Food (Water) Consumption
(EXAMPLE)
Test Compound
Animal #1 Control Group Sex;
Average Daily Diet Consumption in Grams
Week 1
23
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11
(EXAMPLE)
Hematology
Test Compound
Animal #1 Control Group Sex;
Hematology
Pre-test
Months on test
12
erythrocytes
million/mm^
Henoglobin
gms/100 ml
Hematocrit
Percent
Other Parameters*
* Other determinations vhich may
be necessary for an adequate
Toxicological evaluation
24
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12
(EXAMPLE)
Clincial Chemistry
Test Compound
Animal #1 Control Group Sex;
Clinical Chemistry
Pre-test Months on test
1 3 6 12
Glucose
mg %
Other*
Parameters
* Other determinations vhich may
be necessary for an adequate
Toxicological evaluation
25
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13
(EXAMPLE)
Gross/Histopathology
Animal #1 Control Group Sex;
Organ Gross Pathology Histopathology
Brain Edema Enter description of
Pituitary Normal lesions or appropriate
Lungs Hemorrhagic code
Heart Normal
Liver Fatty infiltration
Spleen Enlarged
Kidney Normal
Adrenals Nodules
Pancreas Normal
etc.
26
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PESTICIDE ASSESSMENT GUIDELINES
SUBDIVISION F
HAZARD EVALUATION:
HUMANS AND DOMESTIC ANIMALS
Series 82-1, 83-1, 83-2 and 83-5
Subchronic Oral Toxicity (90-day Study),
Chronic Toxicity Studies,
Oncogenicity Study
and Combined Chronic Toxicity/Oncogenicity Studies
(Rodent Studies)
ADDENDUM ON DATA REPORTING
Prepared by:
Orville E. Paynter, Ph.D., DABT
Project Manager:
Elizabeth M.K. Leovey, Ph.D.
Hazard Evaluation Division
Office of Pesticide Prograns
U.S. Environmental Protection Agency
27
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Subdivision F - Subchronic Oral Tbxicity (90-day Study), Chronic Toxicity
Studies, Oncogenicity Study and Combined Chronic Toxicity/Oncogenicity
Studies (Rodent Studies)
Table of Contents of Addendum
Page
Introduction 1
Response to Public Garment 1
Guideline 1
Cover Page 1
Table of Contents 2
Body of Report 2
I. Summary 2
II. Introduction 2
III. Materials/Methods 2
IV. Results 4
V. Discussion 4
VI. Bibliography 5
VII. Verification 5
VIII. Archives 5
IX. Tables/Figures 5
X. Appendix 5
28
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PESTICIDE ASSESSMENT GUIDELINES
HUMANS AND DOMESTIC ANIMALS
Subchronic Oral Toxicity (90-day Study),
Chronic Toxicity Studies,
Oncogenicity Study
and Conibined Chronic Toxicity/Oncogenicity Studies
(Rodent Studies)
Subdivision F, Series 82-1, 83-1, 83-2 and 83-5
DATA REPORTING
INTRODUCTION
A. Purpose
This Data Reporting Guideline (DRG) is designed to aid the
petitioner/registrant in generating reports which are compatible
with the Agency's review process. Vhile following this Guideline
is not mandatory, data submitters are encouraged to submit
complete reports which can be efficiently reviewed by the Agency.
This DRG pertains to organizing and presenting the substance of
the data report. 40 CFR Part 158 pertains to the physical
formatting of reports (which are referred to as "studies") and
submittal packages. Some of its requirements are mandatory.
B. Objective
The objective of the Guideline is to provide an example of an
acceptable format for reporting materials, methods, and results
of toxicity and rodent oncogenicity studies. These recommendations
are intended to show how the requirements of §80-4, 82-1, 83-1,
83-2, and 83-5, the Agency's Good Laboratory Practice Standards
(40 CFR, Part 160), and the guidance offered by the Hazard
Evaluation Division's SEP Toxicity Potential (EPA 540/9-85-020)
and Oncogenicity Potential (EPA 540/9-85-019), available from
the National Technical Information Center may be organized in a
Final Report.
RESPONSE TO PUBLIC COMMENT
This document incorporates the public comments received by the Agency
in reposnse to publication in the FEDERAL REGISTER (54 FR 3136, 1/23/89).
GUIDELINE
The Final Report should contain the following items.
COVER PAGE
Cover page and additional documentation requirements (i.e.,
requirements for data submission, Good Laboratory Practice statement,
and statement of data confidentiality claims), if relevant to the
29
-------
-2-
study report, must precede the content of the study formatted below.
These requirements are described in PR Notice 86-5.
TABLE OF CONTENTS
This item should be a concise listing of the essential elements of
the final Report including the page numbers for each. Essential
elements should include a Summary, Introduction, Materials and
Methods section, Results, Discussion,. Bibliography, Tables, Figures,
Appendices, and key subsections as appropriate.
.BODY OF THE REPORT
This item shall include all information required in §80-4(b)(2), §80-4(c),
§82-l(l)(3), §83-1(3), §83-2(f)(l)(i)-(ii), and §83-2(3). If the study
is a combined chronic/ oncogenicity study, the report shall meet the
requirement of §83-5(c)(1)(i)(ii) and §83-5(3) in addition to the §80-4
requirements.
I. SIM1ARY
As per §80-4(b)(l) of this Subdivision, this section of the test
report shall contain a summary and analysis of the data, and a
statement of the conclusions drawn frcm the analysis. The summary
should highlight any and all positive data or observations, and
any deviations frcm control data vfriich" may be indicative of toxic
effects.
II. INTRODUCTION
Includes the objective of the study and any deviations from the
protocol recommended in §83-2 or §83-5. The rationale for the
deviations should be provided.
III. MATERIALS AND METHODS
A. Test Substance
1. Chemical name and chemical abstract number or code number;
2. Source of procurement, batch or lot number of each
procurement;
3. Purity and identity analysis on bulk chemical;
4. Stability of test material in feed and control substances;
5. Assay—description of method and percent of target
concentration;
6. Storage/stability of test material/diet; and
30
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-3-
7. Formulation for administration to animals.
B. Animals
1. Description:
a. Species, strain, and substrain,
b. Age,
c. Body weight range, and
d. Source of supply;
2. Isolation procedures;
3. Health assessment;
4. Caging—type of cages;
5. Housing—number of animals per cage;
6. Diet;
a. Vendor and
b. Composition;
7. Water;
a. Purity or source, and
b. Delivery System to Animals;
8. Rocm assignment.
C. Methods
1. Treatment groups:
a. Dosage levels, and
b. Number of animals per group;
2. Treatment regimen:
a. Route of administration;
b. Frequency;
c. Volume and concentration;
d. Carrier/solusnt; and
31
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-4-
e. Daily records and clinical observations.
3. Experimental measurements:
Describe measurements and frequency with which they are
made and cite procedures employed.
4. Animal assignment:
a. Procedure used for identification of each animal.
b. Randomization procedure for assignment to treatment
groups. Housing, food, and water during experiment if
different from that described in subsection "Animals."
,.5. In presenting the statistical methods used to analyze the
study, cite the method used, and give sufficient information
in order that an independent reviewer can reevaluate and
reconstruct the analysis.
IV. RESULTS
Pertinent summary table with appropriate statistical evaluations
clearly indicated may be placed in the text of this section. Each
parameter studied shall be addressed sequentially. It is sufficient
to state that there were no significant inter-group differences in
certain parameters vfrere appropriate.
V. DISCUSSION/CONCLUSION
A. Discussion of observations and adequacy of the tests for the
intended purpose.
B. Discussion of observed toxic effects, target organ(s), dose-
related compound-induced effects, and no-observed-effect levels
(NOEL).
C. The following major considerations for analysis and evaluation
of oncogenicity (see SEP - Oncogenicity Potential for further
discussion), as relating to the species and strain of rodents
used, should be specifically addressed in this part.
1. Spontaneous neoplasm/incidence in untreated animals
(concurrent and historical controls);
2. Presence and incidence of neoplasms not usually observed
(rare tumors);
3. Increased incidence of benign and/or malignant neoplasms
that are usually found;
4. Dose-response relationships; and
5. Decrease in latency (time to tumor discovery) of neoplasms
-------
-5-
that are usually found, in so far as can be assessed.
VI. SUMMARY TABLE
See tables 1 and 2 for examples for presenting historical control
data summaries and table 3 for summarizing tumor incidence data.
VII. INDIVIDUAL ANIMAL TJMDR DATA
See table 4. In addition to aninel number, the time to tumor
discovery or animal death (in days) should be contained in the
column heading for each animal.
VI11. VERIFICATION
This item shall contain information required by §80-4(b)(2). Each
test report shall be:
A. Signed by each of the senior scientific personnel, including
the laboratory director, responsible for performing and
supervising the testing, preparing, reviewing, and approval of
the test report; and
B. Certified by the applicant or an authorized agent of- the
applicant as a complete and unaltered copy of the report provided
by the testing laboratory whether independent or owned, operated,
or controlled by the applicant.
IX. ARCHIVES
This section of the Final Report shall contain all information
required in 40 CFR 160.185.
X. TABLES/FIGURES
XI. APPENDIX(CES)
These should include individual animal data, historical control
data, analytical method and results of analyses on the test substance,
and test diet (if the test material is administered via the diet),
and other appropriate information.
33
-------
-6-
EXAMPLES TO ILLUSTRATE TABLES MDST USEFUL IN REPORTING CWCOGENICITY STUDIES
Table 1. Incidence (Percent) of Female Control Rats Bearing Thyroid
1.
2.
3.
4.
C-Cell
Study*
Group A
Group B
Group A
Group B
Group A
Group B
Group A
Group B
Tumors Among Animals
(Same
Adenoma or
Carcinoma
10/58 (17.2)
7/59 (11.9)
5/59 (8.5)
6/58 (10.3)
9/57 (15.8)
6/55 (10.9)
2/58 (3.4)
0/55 (0)
Sacrificed Post
Lab)
Adenoma
10/58 (17.2)
6/59 (10.2)
5/59 (8.5)
6/58 (10.3)
6/57 (10.5)
5/55 (9.0)
2/58 (3.4)
0/55 (0)
12-Months
Carcinoma
0/58 (0)
1/59 (2)
0/59 (0)
0/58 (0)
3/57 (5)
1/55 (2)
0/58 (0)
0/55 (0)
Total 45/459(9.8) 40/459(8.7) 5/459(1.1)
*Each listed study had two control groups, identified as Group A or B.
The rat strain is Sprague-Dawley.
Table 2. Historical Control Incidence of Lung Tumors in
Male B6C3Fi Mice Receiving Corn Oil by Gavage
(Different Labs)
Alveolar/
Alveolar/ Alveolar/ Bronchiolar
Bronchiolar Bronchiolar Adenoma or
Laboratory Adenoma Carcinoma Carcinoma
A 8/100(8.0%) 6/100(6.0%) 14/100(14.0%)
"' B 12/235(5.1%) 17/235(7.2%) 29/235(12.3%)
C 5/120(4.2%) 3/120(2.5%) 8/120(6.7%)
D 19/150(12.7%) 4/150(2.7%) 22/150(14.7%)
E 4/49(8.2%) 3/49(6.1%) 7/49(14.3%)
F 32/248(12.9%) 11/248(4.4%) 43/248(17.3%)
Total 80/902(8.9%) 44/902(4.9%) 123/902(13.6%)
34
-------
GROUP I
TABLE
GROUP
3^feCAMPLE)
GROUP III
GROUP IV
Liver
(No. Examined)
Hepatocel lular
Carcinoma
Hepatocel Lular
Adenoma*
Malignant
Lymphoma
Granulocytic
Leukemia
Angiosarcoma
Car cinema,
Metastatic
Sarcoma,
Metastatic
Reticulum Cell
Sarcoma
Hepatocho lang io-
Ogfeatcinoma
C/iMfiLti focal
Hepatocel lular
Degeneration
Basophilic Foci
Mononuclear Cell
Infiltration
Foci of Mononu-
clear Cells
Angiectasis
Focus of
Cellular Change
Multi focal
Hepatitis
Multi focal
Necrosis
Scheduled
Sacrifice
(22)
2/2
3
6
1
16
3
Moribund
Sacrifice
& Deaths
(52)
4
1/1
9
1
1
1
9
10
Total
(74)
4
3/3
12
1
1
1
15
1
26
8
Scheduled
Sacrifice
(34)
2
1/1
2
1
9
1
2
3
23
3
Moribund
Sacrifice
& Deaths
(42)
1
4/3
7
1
1
1
4
8
Total
(76)
3
5/4
9
1
1
1
10
1
3
3
27
11
Scheduled
Sacrifice
(24)
2
30/12
1
1
9
13
1
Moribund
Sacrifice
& Deaths
(52)
1
24/11
7
2
1
2
2
1
7
6
Total
(76)
3
54/23
8
2
1
3
11
1
20
7
Scheduled
Sacrifice
(22)
1
54/17
1
8
2
1
14
2
Moribund
Sacrifice
& Deaths
(53)
1
21/12
6
1
1
1
3
2
8
1
3
9
6
Total
(75)
2
75/29
6
1
1
1
3
3
16
1
5
1
23
8
-------
TABLE 4 (EXAMPLE)
ON
Liver (Animal No.)
Hepatocel lular
Carcinoma
Hepatocel lu la r
Adencma
(No. Present)
Malignant
Lymphomas
Granulocytic
Leukemia
Angiosarcoma
Carcincma,
Metastatic
Sarccma,
Mestastatic
Reticulum Cell
Sarcoma
Hepatochol-
angiocarcinoma
Mult i focal
Hepabocel lular
Degeneration
Basophilic Foci
Mononuclear Cell
Infiltration
Foci of Mono-
nuclear Cells
Angiectasis
Focus of Cellular
Change
Mult i focal
Hepatitis
Mult i focal
Necrosis
1
1
2
6
2
2
2
4
5
1
3
4
2
2
3
5
3
1
4
3
3
1
•\
5
1
2
2
1
3
2
2
3
2
7
2
1
1
3
N
1
1
2
1
4
P
4
4
2
Key: p = Present N = No Section A = Auto lysis X = Not Remarkable 1 - incomplete section
1 = Minimal 2 = Slight 3 = Moderate 4 = Moderately Severe High 5 = Severe/High
CO
-------
PESTICIDE ASSESSMENT GUIDELINES
SUBDIVISION F
HAZARD EVALUATION
HUMAN AND DOMESTIC ANIMALS
Series 82-2 and 82-3
Repeated Dose Dermal Toxicity; 21-Day Study
and
Subchronic Toxicity Study; 90-Day Study
ADDENDUM ON DATA REPORTING
Prepared by:
Robert P. Zendzian Ph.D.
for
Health Effects Division
Office of Pesticide Programs
US Environmental Protection Agency
37
-------
Subdivision F - Repeated Dose Dermal Toxicity Studies
(21-day and 90-day)
Table of Contents of Addendum
Page
Introduction 1
Response to Public Comments 1
Repeated Dose Dermal Toxicity
Guideline 1
Cover Page 1
Table of Contents 2
Body of Report 2
I. Summary 2
II. Introduction 2
III. Materials/Methods 2
IV. Results 3
V. Discussion 3
VI. Bibliography 3
VII. Verification 3
VIII. Archives 4
IX. Tables/Figures 4
X. Appendix 4
38
-------
PESTICIDE ASSESSMENT GUIDELINES
HUMAN AND DOMESTIC ANIMALS
Repeated Dose Dermal Toxicity (21-days and 90-days)
Subdivision F, Series 82-2 & 82-3
DATA REPORTING
INTRODUCTION
A. Purpose
The purpose of this addendum to the guideline is to provide a data
reporting fbrrat for repeated dose dermal toxicity studies. Sutpart F,
Series 82-2 and 82-3 provide guidelines for carrying out the repeated
dose dermal toxicity studies.
B. Objective
The objective of the Data Reporting Guidelines (DRGs) is to provide an
example of an acceptable general reporting format that can be reviewed
effectively and expeditiously in accordance with the Agency's review
process. The recommendations are intended to be consistent with the
guidelines for data reporting as described in Sections 80-4, 82-2 and
82-3 and those described in the Agency's Good laboratory Practive Standards
(40 CFR Part 160).
While following this Guideline is not mandatory, data submitters are
encouraged to submit complete reports vhich can be effectively reviewed
by the Agency. 40 CFR Part 158 pertains to the physical formatting of
reports (vhich are referred to as "studies") and submitted packages. Some
of its requirements are mandatory.
RESPONSE TO PUBLIC COMMENTS
The Agency received one response to its request for public ccmments on these
documents published in the FEDERAL REGISTER of March 8, 1989 (54 FR 9886).
These ccmments were considered in the final form of this DRG.
GUIDELINE
The Final Report should contain the following items.
COVER PAGE
The title page and additional documentation (i.e., requirements for data
submission, Good Laboratory Practices statement and statement of data
confidentiality claims), if relevant to the study report, must precede
the content of the study format below. These requirements are described
in 40 CFR Part 158.
-------
TABLE OF CONTENTS
The table of contents should include a listing of the elements of the
final report such as the Summary, an Introduction, the Materials and
Methods, Results, Discussion Bibliography, Tables, Figures, Appendices
and key subsections as appropriate.
BODY OF THE REPORT
This item shall include all information required in §80-4(b)(2), 80-4(c),
82-2(h) and 82-3(h) of this subdivision and should be provided in such detail
that the reviewer can assess the quality of the study and conformity to the
repeated dose dermal toxicity Guideline. It should contain the following
sections.
I. SUMMARY
As per §80-4(b)(l) of this subdivision, the test report shall contain a
summary including a brief description of the study protocol, chemical
used, the animals tested and the highlights of the results of the study
relative to the positive findings. Any deviations from the intended
protocols should be noted.
II. INTRODUCTION
Include the objectives of the study and the Guideline reference. The
overall experimental design should be explained.
III. MATERIALS AND METHODS
A. Test substance
Test material (chemical name)
source
lot runtoer
CAS No.
purity
state
lonization constant (if applicable)
pH (if applicable)
octanol/water partition coefficient (if known)
vapor pressure at 20° C (if known)
vehicles used
B Test Animals
species and strain
source
sex
body weight
number of test animals
number of control animals
40
-------
pre-test condition
housing conditions
test environment
C. Experimental Design
Doses and duration of exposure
Number and sex of animals per dose group
Application site, preparation and msthod(s) of protection
Observations, type and time
Termination
D. Evaluation Procedures
The final report should describe the types of observations of the animals
for toxic signs, body weights, the frequency of observations, the gross
pathological examination procedures, histopathology and any other
observations performed.
E. Deviation frcm protocol
Deviations frcm the protocol recommended in § 82-2 or 82-3 should be
described along with the rational for the changes.
IV. RESULTS
This sections should provide a summary of the toxic signs, mortality,
body weight changes, gross pathology and other observations performed
associated with the doses.tested. Typically, these results are summarized
in tabular form.
V. DISCUSSION
This section should provide an assessment of the results of the study, an
interpretation of the toxicity observed, any the dose-response effects
and should try to explain unexpected findings. The impact of any deviation
from the Guideline protocol should be discussed.
VI. BIBLIOGRAPHY
This item should contain a list of references cited in the body of the
report.
VII. VERIFICATION
This item shall contain information required by §80-4(b)(2). Each test
report shall be:
A. Signed by each of the senior scientific personnel, including the
laboratory director responsible for performing and supervising the
testing, preparing, reviewing and approving the test report and
B. Certified by the applicant or an authorized agent of the applicant
as a ccmplete and unaltered copy of the report provided by the
41
-------
testing laboratory v*i ether independent or owned, operated or
controlled by the applicant.
VII. ARCHIVES
This section of the final report shall contain all information required
in 40 CFR 160.185.
IX. TABLES/FICURES
X. APPENDIX(ES)
These should include individual animal data, historical control data,
pathology report, analytical methods and results of analysis of the test
substance, details of statistical analysis, protocol and other information
as appropriate.
42
-------
PESTICIDE ASSESSMENT GUIDELINES
SUBDIVISION F
HAZARD EVALUATION
HUMAN AND DOMESTIC ANIMALS
Series 83-4
Reproductive and Fertility Effects
ADDENDUM ON DATA REPORTING
Prepared by:
Roger Gardner
for
Health Effects Division
Office of Pesticide Programs
US Environmental Protection Agency
43
-------
Subdivision F - Reproductive and Fertility Effects
Table of Contents of Addendum
Page
Introduction 1
Response to Public Comments 1
Reproductive and Fertility Effects
Guideline 1
Cover Pages 1
Table of Contents 2
Body of Report 2
I. Summary 2
II. Introduction 2
III. Materials/Methods 2
IV. Results 3
V. Discussion 3
VI. Bibliography 3
VII. Verification 3
VIII. Archives 4
IX. Tables/Figures 4
X. Appendix 4
Examples of Tabular Formats 5
44
-------
PESTICIDE ASSESSMENT GUIDELINES
HUMAN AND DOMESTIC ANIMALS
Reproductive and Fertility Effects
Subdivision F, Series 83-4
DATA REPORTING
INTRODUCTION
A. Purpose
The purpose of this addendum to the guideline is to provide a data reporting
forrtat for the rat multigeneration reproduction study. Sutpart F, Series
83-4 provides guidelines for carrying out this study.
B. Objective
The objective of the Data Reporting Guidelines (DRGs) is to provide an
example of an acceptable general reporting format that can be reviewed
effectively and expediticusly in accordance with the Agency's review
process. The recommendations are intended to be consistent with the
guidelines for data reporting as described in Sections 80-4 and 83-4 and
those described in the Agency's Good Laboratory Practice Standards (40
CFR Part 160).
While following this Guideline is not mandatory, data submitters are
encouraged to submit complete reports vhich can be effectively reviewed
by the Agency. 40 CFR Part 158 pertains to the physical formatting of
reports (vhich are referred to as "studies") and submitted packages. Some
of its requirements are mandatory.
RESPONSE TO PUBLIC COMMENTS
The Agency received one response to its request for public conments on these
documents published in the FEDERAL REGISTER of March 8, 1989 (54 FR 9886).
These ccmments were considered in the final form of this DRG.
GUIDELINE
The Final Report should contain the following items.
COVER PAGE
The title page and additional documentation (i.e., requirements for data
submission, Good Laboratory Practices statement and statement of data
confidentiality claims), if relevant to the study report, must precede
the content of the study format below. These requirements are described
in PR Notice 86-5.
45
-------
TABLE OF CONTENTS
The table of contents should include a listing of the elements of the
final report such as the Sunmary, an Introduction, the Materials and
Methods, Results, Discussion Bibliography, Tables, Figures, Appendices
and key subsections as appropriate.
BODY OF THE REPORT
This item shall include all information required in §8CM(b)(2), 80-4(c) and
83-4(f) of this subdivision and should be provided in such detail that the
reviewer can assess the quality of the study and conformity to the Reproductive
and Fertility Effects Guideline. It should contain the following sections.
I. SUMMARY
As per §8O4(b)(l) of this subdivision, the test report shall contain a
summary including a brief description of the experimental design, chemical
used, the animals tested and the highlights of the results of the study
relative to the positive findings. Any deviations from the intended
protocols should be noted.
II. INTRODUCTION
Include the objectives of the study and the Guideline reference. The
overall experinental design should be explained.
III. MATERIALS AND METHODS
A. Test substance
Test material (chemical name)
Form (technical, manufacturing-use product or formulation)
source
lot number
CAS No.
purity
state
lonization constant (if applicable)
pH (if applicable)
octanol/water partition coefficient (if known)
vehicles used
B Test Animals
species and strain
sex
source
body wsight
pre-test condition
46
-------
housing conditions
test environment
C. Experimental Design
Dosing and breeding schedule
Mating procedure
Doses used
Route of administration
Number and sex of animals per dose group
Observations, type and time
Termination and tissue collection
D. Evaluation Procedures. The final report should describe:
(A) Observations and procedures used to calculate percentages, weight
changes, or mating, gestation, viability, and lactation indices.
(B) Methods for adjusting data (censoring, pooling, or transforming
results) prior to statistical analysis,
(C) Statistical procedures as recommended in §80-4(c) of this
subdivision.
E. Deviation from protocol
Deviations from the protocol recommended in § 83-4 should be described
along with the rationale for the changes.
IV. RESULTS
This section should provide a summary of the toxic signs, mortality,
body weight changes, gross pathology, histopathology and other observations
associated with the doses tested. Typically, these results are summarized
in tabular form. Example formats in this section are generalized from
those caruTDnly submitted to the Agency. They do not represent required
or preferred formats, but they are provided as a guide.
V. DISCUSSION
This section should provide an assessment of the results of the study, an
interpretation of the toxicity observed, the consistency of the dose-response
effects and should try to explain unexpected findings. The impact of any
deviation from the Guideline protocol should be discussed.
VI. BIBLIOGRAPHY
This item should contain a list of references cited in the body of the
report.
VII. VERIFICATION
This item shall contain information required by §80-4(b)(2). Each test
report shall be:
47
-------
A. Signed by each of the senior scientific personnel, including the
laboratory director responsible for performing and supervising the
testing, preparing, reviewing and approving the test report and
B. Certified by the applicant or an authorized agent of the applicant
as a complete and unaltered copy of the report provided by the
testing laboratory whether independent or owned, operated or
controlled by the applicant.
VII. ARCHIVES
This section of the final report shall contain all information required
in 40 CFR 160.185.
IX. TABLES/FIGURES
(1) Summary Tables. Paragraph 83-4(f) and §80-4 require results to be
summarized in tabular form. Example Formats 1-3 show common presentations
of surmHry data that are accepted by the Agency.
X. APPENDIX(ES)
These should include individual animal data (Example Formats.4 through 7),
historical control data, pathology report, analytical methods and results
of analysis of the test substance, details of statistical analysis,
protocol and other information as appropriate.
-------
Example Format 1
Example of frequently submitted
summary table presentations of clinical signs
Observation
Dose (mg/kg/day)
Behavioral changes; clinical signs; signs
of difficult or prolonged parturition;
clinical signs; mortalities;
Number of animals
Number of males mated; number of females
mated; number pregnant; number of males
and females that died; number aborted;
number not pregnant
Duration (days) of gestation;
number of pups; stillbirths; live pups;
pups with gross alterations; litter weights;
individual pup weights at birth as well Mean
as at Days 4, 7, 14, and 21 after birth S. D.
(Day 0); N
0
X
N
Y
X
N
5Y
X
N
10Y
X
N
Mean
S. D.
N
Mean
S. D.
N
Mean
S. D.
N
Example Format 2
Example of a frequently submitted summary table format for
maternal body weight results in a Reproduction Ibxicity Study
Observation
Dose (mg/kg/day)
Y 5Y
10Y
Body weight (g) at
Week 0
Each treatment
week when weights
are obtained.
Day of sacri-
fice
Body weight
change, food
and water con-
Mean
S. D.
N
Mean
S. D.
N
Mean
S. D.
N
Mean
S. D.
N
Mean
S. D.
N
Mean
S. D. .
N
Mean
S. D.
N
Mean
S. D.
N
Mean
S. D.
N
Mean
S. D.
N
Mean
S. D.
N
Mean
S. D.
N
Meanb
S. D.
N
Meanb
S. D.
N
Meant*
S. D.
N
Mean
S. D.
N
sumption
bMeans that are statistically significantly different from
control group means should be highlighted.
49
-------
Example Format 3
Presentation of some optional parental toxicity end points
such as liver weighta and weight ratios (% body weight)
Liver wt
Dosee Body weight (g) at
(mg/kg/day) Day 0
0 Mean
S. D.
N
Y Mean
S. D.
N
5Y Mean
S. D.
N
ICY Mean
S. D.
N
Sacrifice
Mean
S. D.
N
Mean
S. D.
N
Mean
S. D.
N
Mean
S. D.
N
parameters
Absolute
Mean
S. D.
N
Mean
S. D.
N
Mean
S. D.
N
Mean
S. D.
N
Relative*
Meant*
S. D.
N
Meant)
. S. D.
N
Meant*
S. D.
N
Meanto
S. D.
N
include other organs as appropriate. "
tMeans that are statistically significantly different from control
group means should be highlighted.
*Expressed as % body weight at sacrifice.
Example Format 4
Example of a format for reporting individual
survival and toxic signs results
Dose Animal Day of death
group no. or sacrifice
X
2
3
X
X
Toxic signs
Onset day Duration
X X
X
X
X
X
Description
Brief description of
changes in behavior,
appearance, or other
clinical signs.
50
-------
.
CHEMICAL LIBRARY
01 M ST., S.W., TS-793
Example Format 5 WASHINGTON, D.C. 20460
An example of a forrtat for reporting individual
maternal body weight data
Animal Animal
Week number »## number ###
(Premating)
1
XXX
XXX
Animal
number ###
XXX
10
(Gestation)
12
13
14
(Lactation)
15
16
17
Sacrifice
XXX
XXX
XXX
XXX
XXX
XXX
XXX
XXX
XXX
XXX
XXX
XXX
XXX
XXX
XXX
XXX
XXX
XXX
XXX
XXX
XXX
XXX
XXX
XXX
51
-------
Example Format 6
An example of the presentation of individual animal data for postnatal
observations to be reported for each mating in each generation of
the multigeneration reproduction study
Animal
number
xxxxxx
xxxxxx
xxxxxx
At birth, Day 4 (before culling of litters),
and on lactation days 7, 14, and 21
Number alive
Males Females
X X
X X
X X
Number dead
Males
X
X
X
Females
X
X
X
Per cent
alive
X
X
X
Mean pup
weight (g)
X
X
X
Sex
ratio*
X
X
X
Animal
number
xxxxxx
xxxxxx
xxxxxx
Day 4 (after culling of litters)
Number alive Mean pup Sex
Males Females weight (g) ratio*
X
X
X
X
X
X
X
X
X
X
X
X
* Expressed as a percentage for one sex or as a ratio of the number of
males to females.
Example Format 7
Example format for presentation of clinical observations
of offspring fron each mating in each generation
of a multigeneration reproduction study
Animal
number
xxxxxx
xxxxxx
xxxxxx
Number of pups
Males Females
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
X
Day
X
X
X
X
X
X
X
X
X
Observations
Group Y
Description of clinical observation 1
Description of clinical observation 2
Description of clinical observation n
Description of clinical observation 1
Description of clinical observation 2
Description of clinical observation n
Description of clinical observation 1
Description of clinical observation 2
Description of clinical observation n
52
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PESTICIDE ASSESSMENT GUIDELINES
SUBDIVISION F
HAZARD EVALUATION
HUMANS AND DOMESTIC ANIMALS
Series 84
Mutagenicitv Studies
ADDENDUM ON DATA REPORTING
Office of Pesticide Programs
U.S. Environmental Protection Agency
53
-------
Subdivision F - Mutagenicity Studies
Table of Contents of Addendum
Page
Introduction 1
Response to Public Comments 1
Guideline
Cover Page 2
Table of Contents 2
Body of Report 2
I. Summary 2
II. Introduction 2
III. Materials and Methods 2
IV. Results 4
V. Discussion/Conclusion 4
VI. Bibliography 4
VII. Verification 4
VIII. Archives 5
IX. Tables/Figures 5
X. Appendix(ces) 5
54
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PESTICIDE ASSESSMENT GUIDELINES
HUMANS AND DOMESTIC ANIMALS
Mutaqenicity Studies
Subdivision F, Series 84
DATA REPORTING
INTRODUCTION
A. Purpose
This Data Reporting Guideline (DRG) is designed to aid the
petitioner/registrant in generating reports which are compatible
with the Agency's review process. This DRG pertains to organizing
and presenting the substance of the data report. 40 CFR Part 158
pertains to the physical formatting of reports (which are referred
to as "studies") and submittal packages. Some of its requirements
are mandatory.
B. Objective
The objective of the Guideline is to provide an example of an
acceptable format for reporting materials, methods, and results of
mutagenicity studies. These recommendations are intended to show
how the requirements of Series 84 of Subdivision F, the Agency's
Good Laboratory Practice Standards (40 CFR, Part 160) , and the
guidance on mutagenicity testing offered by the Health Effects
Testing Guidelines (Subpart F - Genetic Toxicity of 40 CFR, Part
798) may be organized in a Final Report. While following this
Guideline is not mandatory, data submitters are encouraged to
submit complete reports which can be efficiently reviewed by the
Agency.
RESPONSE TO PUBLIC COMMENTS
The Agency received no response to its request for public
comments on these documents published in the FEDERAL REGISTER of
March 8, 1989 (54 FR 9886).
55
-------
GUIDELINE
The final report should contain the following items.
COVER PAGE
Cover page and additional documentation requirements (i.e.
requirements for data submission, Good Laboratory Practice
statement, Quality Assurance statement and procedures for claims
of confidentiality of data), if relevant to the study report, must
precede the content of the study formatted below. These
requirements are described in 40 CFR Part 160.
TABLE OF CONTENTS
This item should be a concise listing of the essential elements of
the Final Report including the page numbers for each. Essential
elements should include a Summary, Introduction, Materials and
Methods section, Results, Discussion/Conclusion(s), Bibliography,
Tables, Figures, Appendices, and key subsections as appropriate.
BODY OF REPORT
This item shall include all information required in Series 80-
4(b)(2), Series 80-4(c) and Series 84 of this Subdivision.
Additional guidance is provided in the Health Effects Testing
Guidelines for mutagenicity testing (40 CFR, Part 798, Subpart F
- Genetic Toxicity; hereafter referred to as the Health Effects
Testing Guidelines).
I. SUMMARY
As per Series 80-4(b)(1) of this Subdivision, this section of
the test report shall contain a summary and analysis of the data,
and a statement of the conclusions drawn from the analysis. The
summary should highlight any and all positive data or observations,
and any deviations from control data which may be indicative of
toxic effects.
II. INTRODUCTION
Includes the objective of the study and any deviations from
the protocol recommended in the Health Effects Testing Guidelines.
The rationale for the deviations should be provided.
III. MATERIALS AND METHODS
A. Test Substance; Negative and Positive Control Substances
1. Chemical name and chemical abstract number; any code
numbers.
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2. Source of procurement, batch or lot number of each
procurement; procurement date.
3. Purity and identity analysis on bulk chemical.
4. Physical description.
5. Solvent(s)/vehicle(s) used and percent of final solution.
6. Stability (e.g. shelf life) and stability in
solvent/vehicle.
7. Additional for in vivo tests: storage/stability of test
substance/diet; formulation for administration to animals; percent
of target concentration.
B. Test System
1. Description:
a. In vitro; Cell type; maintenance; source.
b. In vivo; Species, strain, substrain; age; body weight
range; sex; source; maintenance (including isolation procedures,
health assessment, caging with types of cages, housing - number of
animals per cage, diet - vendor and composition, water).
2. Metabolic activation system (if required)
a. Description of source and derivation.
b. Composition of final preparation.
C. Methods
In this item, present description of protocol, include the
following where appropriate for in vitro or in vivo situations:
1. Treatment groups
a. Concentration/dosage levels; present rationale for
selection of concentrations/doses used (e.g. preliminary toxicity
test).
b. Number of cultures/animals per test group.
c. Replicates per test group.
2. Treatment regimen
a. Method of administration/exposure.
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b. Frequency.
c. Volume and concentration.
d. Daily records and observations (e.g. cell condition;
clinical observations).
3. Experimental measurements
Describe measurements and frequency with which they are made
and cite procedures employed.
4. In presenting the statistical methods to analyze the study,
cite the method used and give sufficient information in order that
an independent reviewer can reevaluate and reconstruct the
analysis.
IV. RESULTS
A pertinent summary table(s) with appropriate statistical
evaluations clearly indicated may be placed in the text of this
section. Each parameter studied shall be addressed. Due to the
large number of mutagenicity test types, a single Example table is
not appropriate. Indicate in the table all pertinent information
from which an unambiguous conclusion can be made.
V. DISCUSSION/CONCLUSION
1. Discussion of observations and adequacy of the tests for
the intended purpose.
2. Discussion of observed effects of endpoint of interest,
toxic effects, concentration-/close-related compound-induced effects
and statistical and biological significance.
3. Discussion of necessity for repeat, if warranted.
4. Overall conclusion(s) of the study.
VI. BIBLIOGRAPHY
This item should contain a list of references cited in the
body of the report. They should be complete so that an independent
reviewer may readily identify and obtain any reference.
VII. VERIFICATION
This item shall contain information required by Series 80-
4(b)(2). Each test report shall be:
1. Signed by each of the senior scientific personnel,
including the laboratory director, responsible for performing and
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supervising the testing, preparing, reviewing, and approval of the
test report; and
2. Certified by the applicant or an authorized agent of the
applicant as a complete and unaltered copy of the report provided
by the testing laboratory whether independent or owned, operated,
or controlled by the applicant.
VIII. ARCHIVES
This section of the Final Report shall contain all information
required in 40 CFR Part 160.185.
IX. TABLES/FIGURES
If a summary table was not included in the results section of
the Final Report, include a summary table(s) here.
X. APPENDIX(CES)
These should include individual data, historical control data,
analytical method and results of analyses on the test substance and
test diet (if the test substance is administered via the diet) ,
details of statistical analyses, complete protocol (if not already
described completely in body of Final Report), and other
information as appropriate.
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