Introduction to the Chronic (1 Year) Dog Toxicity Study Template

Return to Toxicology Template Introduction

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Table of Contents

1. Identification of Study
2. Good Laboratory Practice
3. Executive Summary
4. Materials and Methods
   1. TEST substance
   2. TEST substance AS ADMINISTERED
   3. ANIMAL DIET
   4. TEST ANIMALS
   5. EXPERIMENTAL DESIGN
   6. BODY WEIGHT AND FEED INTAKE
   7. CAGE-SIDE OBSERVATIONS
   8. OPTHALMOLOGICAL EXAMINATION
   9. HEMATOLOGY
   10. CLINICAL CHEMISTRY
   11. URINALYSIS
   12. OTHER TESTS
   13. NECROPSY (INTERIM, if any)
   14. NECROPSY (TERMINAL)
   15. GROSS PATHOLOGY OBSERVATIONS
   16. HISTOPATHOLOGY OBSERVATIONS
   17. STATISTICAL METHODS
5. Results
   1. DOSE VERIFICATION
   2. FEED CONSUMPTION
   3. BODY WEIGHT
   4. INTAKE OF TEST SUBSTANCE
   5. FEED EFFICIENCY
   6. CAGE-SIDE OBSERVATIONS
   7. MORTALITY
   8. OPHTHALMOLOGICAL EXAMINATION
   9. HEMATOLOGY
   10. CLINICAL CHEMISTRY
   11. URINALYSIS
   12. NEUROTOXICITY (IF INDICATED)
   13. OTHER TESTS
   14. ORGAN WEIGHTS
   15. GROSS PATHOLOGY CHANGES OBSERVED
   16. HISTOPATHOLOGY CHANGES OBSERVED
6. Evaluation and Comments on Study
7. Summary and Conclusions
   1. BRIEF SUMMARY OF MAJOR FINDINGS FROM THE STUDY
   2. RELATIONSHIP BETWEEN DOSE AND INCIDENCE/SEVERITY OF LESIONS OR ABNORMALITIES
   3. IS THERE A TARGET ORGAN?
   4. NOEL
8. References

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Chronic (1 Year) Toxicity Study in Dogs [1]

Date of Submission:

Title of Petition or Notification:

Name and Address of Petitioner or Notifier:

I. Identification of Study [2] 

A. Study File Location:
B. Study Title/Report Number:
C. Name and Address of Testing Facility:
D. Study Author(s) and/or Study Director(s):
E. Date of Study Report:
F. Dates Study Conducted:
G. Study Objective:
H. Comments:

II. Good Laboratory Practice [3] 

A. Good Laboratory Practice (GLP) Compliance?
B. Quality Assurance (QA) Statement?
C. Availability and Location of Original Data/Specimens/Test Substance:  

III. Executive Summary

KEYWORDS: [4] 

IV. Materials and Methods [5] 

A. TEST substance [6] 

1. CAS name:
2. Other name(s):
3. CAS registry number:
4. Molecular structure: [http://www.chemfinder.com] [7]
5. Purity:
6. Impurities:
7. Stability:
8. Comments:

B. TEST substance AS ADMINISTERED [8] 

1. Batch/Lot Number(s): 
2. Route:
3. Vehicle used:
4. Tested adequately for concentration ? (See Part V-A for calculation)
5. Tested for homogeneity?
6. Tested for stability? [9] 

• Before treatment period   Yes   No
• After treatment period   Yes   No
• After changes to stock   Yes   No
• (If No, list dates tested for stability  )

7. Problems with storage?  

C. ANIMAL DIET

1. Feed

1. Type:
2. Name:
3. Availability:
4. Analysis for contaminants:
5. Comments: (e.g., describe contaminants found)

2. Water

1. Source:
2. Availability:
3. Analysis for contaminants:
4. Comments: (e.g., describe contaminants found)

D. TEST ANIMALS

1. Species/strain:
2. Sex:
3. Age range at initiation of study:
4. Weight range at initiation of study:
5. Quarantine/acclimation?
6. Physical examination schedule:
7. Number per cage:
8. Environmental conditions (e.g. temperature, humidity, photoperiod, etc.)
9. Comments:

E. EXPERIMENTAL DESIGN [10] 

1. Targeted dose levels:

Table # [Heading]

2. Total number of animals:
3. Duration of study (including recovery period, if any):
4. Length of exposure to test substance:
5. Were animals randomized?
6. Recovery period:
7. Comments: (e.g. dose selection rationale, dosing differences in males and females, etc.)

F. BODY WEIGHT AND FEED INTAKE [11] 

1. Parameter examined:

Table # [Heading]

*These parameters are recommended in Toxicological Principles for the Safety of Food Ingredients: Redbook 2000 for chronic toxicity studies.

2. Comments: (e.g., list frequency; control vs. treatment group measurements)

G. CAGE-SIDE OBSERVATIONS [12] 

1. Parameter examined:

Table # [Heading]

*These parameters are recommended in Toxicological Principles for the Safety of Food Ingredients: Redbook 2000 for chronic toxicity studies.

** The parameters for neurotoxicity screening may include, but are not limited to, the following:

• Changes in skin, fur, eyes, mucous membranes, gait, posture, and response to handling,
• autonomic activity such as lacrimation, piloerection, pupil size change, unusual respiratory pattern,
• Presence of clonic or tonic seizure,
• Stereotypic behaviors such as excessive grooming and repetitive circling,
• Bizarre behavior such as self-mutilation and walking backwards,
• Gross tumor development.

2.   Comments: (e.g., list frequency)

H.  OPTHALMOLOGICAL EXAMINATION

1. Parameter examined: 2. Comments:

I.   HEMATOLOGY [13] 

1. Fasting duration prior to blood collection:
2. When during the study (dates, times) were the blood samples collected? .
3. How were the blood samples drawn?
4. Dose groups and number of animals tested: 5. Parameter examined:

Table # [Heading]

* These parameters are recommended in Toxicological Principles for the Safety of Food Ingredients: Redbook 2000 for chronic toxicity studies.

6. Comments:

J.   CLINICAL CHEMISTRY [14] 

1. Fasting duration prior to blood collection:
2. When in the study were the blood samples collected?
3. How were the blood samples drawn?
4. Dose groups and number of animals tested: 5. Parameter examined:

Table # [Heading]

* These parameters are recommended in Toxicological Principles for the Safety of Food Ingredients: Redbook 2000 for chronic toxicity studies.

** These parameters should generally be given priority when adequate volumes of blood samples cannot be obtained from test animals.

6. Comments:

K. URINALYSIS [15] 

1. When and how were urine samples collected?
2. Dose groups and number of animals tested:
3. Parameter examined:  

Table # [Heading]

* These parameters are recommended in Toxicological Principles for the Safety of Food Ingredients: Redbook 2000 for chronic toxicity studies.

4. Comments:

L.   OTHER TESTS

1. Observations: 2. Comments:

M. NECROPSY (INTERIM, if any)

1. Organs weighed:

Table # [Heading]

* These parameters are recommended in Toxicological Principles for the Safety of Food Ingredients: Redbook 2000 for chronic toxicity studies.

2. Comments: (e.g., note scheduled or unscheduled necropsy)
 

N. NECROPSY (TERMINAL)

1. Organs weighed:

Table # [Heading]

* These parameters are recommended in Toxicological Principles for the Safety of Food Ingredients: Redbook 2000 for chronic toxicity studies.

2.   Comments:

O. GROSS PATHOLOGY OBSERVATIONS

1. Organs/Tissues examined: 2. Comments:

P.   HISTOPATHOLOGY OBSERVATIONS

1. Organs were collected from which dose groups?
2. Organs were examined from which dose groups?
3. How were the organs/tissues prepared for histopathology observation?
4. Organs/tissues collected:  

Table # [Heading]

* These parameters are recommended in Toxicological Principles for the Safety of Food Ingredients: Redbook 2000 for chronic toxicity studies.

5. Comments:  

Q. STATISTICAL METHODS

1. Methods of statistical analysis:  

Table # [Heading]

2. Comments:  

V. Results [16] 

A. DOSE VERIFICATION [17] 

1. Were doses verified?

We suggest using an Excel file to calculate the Means and Standard Deviations (SD). (Optional): A sample Excel file (calculation.xls) is provided.

Table # [Heading]

* Number of measurements (N)

2. Verified by:
3. Comments:  

B. FEED CONSUMPTION [18] 

1. Observations: We suggest using an Excel file to calculate the Means and Standard Deviations (SD).

Table # [Heading]

X: Mean, SD: Standard Deviation

2. Comments:  

C. BODY WEIGHT [19] 

1. Observations: We suggest using an Excel file to calculate the Means and Standard Deviations (SD).

Table # [Heading]

X: Mean, SD: Standard Deviation

2. Comments:  

D. INTAKE OF TEST SUBSTANCE [20] 

1. Observations: We suggest using an Excel file to calculate the Means and Standard Deviations (SD).

Table # [Heading]

X: Mean, SD: Standard Deviation

2. Comments:  

E. FEED EFFICIENCY [21] 

1. Was feed efficiency calculated?
2. Comments: (e.g., groups, time periods) 

 F. CAGE-SIDE OBSERVATIONS [22] 

1. Appearance:
2. Abnormal Stool:
3. Morbidit:
4. Signs of Neurotoxicity:
5. Comments:  

G. MORTALITY [23] 

1. Observations:
2. Comments:

H.  OPHTHALMOLOGICAL EXAMINATION

1. Observations:
2. Comments:

I.   HEMATOLOGY [24] 

1. Observations: (n.b.; include dates and times of blood sampling and alter table as necessary)  

Table # [Heading]

(Specify a method of statistical analysis): * p<0.05, **=""><>

2. Comments:  

J.   CLINICAL CHEMISTRY [25] 

1. Observations: (n.b.; include dates and times of clinical chemistry sampling and alter table as necessary)

Table # [Heading]

(Specify statistical method of analysis): * p<0.05, **=""><>

2. Comments:  

K. URINALYSIS [26] 

1. Observations: (n.b.; include dates and times of urine sampling and alter table as necessary) 

Table # [Heading]

2. Comments:  

L.   NEUROTOXICITY (IF INDICATED) [27] 

1. Observations: (n.b.; include dates and times of neurotox screening and alter table as necessary)

Table # [Heading]

+ See under  section IV.G for the types of observation for nervous system toxicity: List noteworthy findings. If additional parameters (other than those in the Template) showed noteworthy changes, these should be added to the tables. In general, data at the end of dosing period can be shown; however, if there were additional noteworthy findings at earlier time points, these should be included. Footnotes should be used as needed to provide additional information about the tests or the results. Note the severity of the abnormal observations using the following scales; + Mild, ++ Moderate, and +++ Marked or other scales, as appropriate.

# Organs/tissues listed under  Section IV.O.

M. OTHER TESTS [28]

1. Observations:
2. Comments:  

N. ORGAN WEIGHTS [29] 

1. Observations:  

Table # [Heading]

a: Group means at the end of terminal necropsy are shown.

(Specify methods of statistical analysis): * p<0.05, **=""><>

2. Comments:  

O. GROSS PATHOLOGY CHANGES OBSERVED [30] 

1. Observations:
2. Comments:  

P.   HISTOPATHOLOGY CHANGES OBSERVED [31] 

1. Observations:  

Table # [Heading]

[# ]See Section IV.O for the list of organs or tissues.

2. Comments:  

VI. Evaluation and Comment on Study [33] 

VII. Summary and Conclusions [34] 

A. BRIEF SUMMARY OF MAJOR FINDINGS FROM THE STUDY

B. RELATIONSHIP BETWEEN DOSE AND INCIDENCE/SEVERITY OF LESIONS OR ABNORMALITIES

C. IS THERE A TARGET ORGAN?

D. NOEL

1. Was there a no observed effect level?
2. Comments:  

VIII. References

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End Notes

[1]This Template is set up for submitting data from typical chronic dog studies. The sections are adjustable; if you find you need more space for a larger group of animals, you can add them to the section. If you have a smaller group of animals, you can delete the unneeded sections. The same applies to sections on feed mixtures and data for levels of test substances in feed. 

[2]Make note of: Study location (Volume, pages), Study title/Report #, Testing facility, publication dates and comments, if needed. 

[3]Indicate Yes or No for questions A and B. However, you may want to elaborate on the type of GLP compliance (USFDA, OECD, etc.) or make other notations 

[4]Please include keywords that may be helpful for indexing and accessing study reports from electronic archives. 

[5]Chronic studies often include treatment groups that will be terminated before completion of the study. Intermediate or interim sampling protocols should be included and noted. Scheduled or unscheduled necropsies should be clearly stated. 

[6]Description of the test substance should be given, including purity, any possible contaminants or impurities, any properties of the test substance that could affect its integrity. Are there factors that could affect the actual administered dose, as opposed to the intended dose? 

[7]After you put a blinking cursor on the http address, (CTRL +) left-click once to follow the link and wait. Enter a chemical name, CAS Number, or molecular formula inside the search window and click 'search' button. Right-click inside the molecular structure and select 'copy' submenu. Return to your document and put the cursor underneath item #4 (molecular structure). Right click to open up the menu options and select 'paste' submenu. You can drag the structure to any position you want and resize.

If preferred, the reviewer is free to use other methods of depicting the molecular structure. 

[8]Indicate how the test substance was given and whether any vehicle was used to dissolve or suspend the test substance (e.g., dissolved in corn oil and mixed into the feed). Also note if there was adequate testing for concentration and homogeneity and whether stability tests were done. Was the strength of the test material checked over the period and under the conditions that the feed mix, test article in vehicle, etc., would be stored? Note anything that was not normal or routine. 

[9]Information about methods used for testing stability. A brief description may be helpful to clarify how often stability of test articles was tested (e.g., was stability tested for each dilution made from the stock solution). 

[10]Provide adequate details. Use the comments to indicate additional information about the experimental design. To change the table, place cursor in the row or column that you wish to modify, then from the "Table" menu, use "Select Row" or "Select Column" to highlight the row or column that you wish to change, and use the "delete" or "insert" functions to make your changes. For major changes, it may be easiest to create a new table and paste it into the template (e.g. if doses differ in male and female treatments). 

[11]Indicate the measurements that were made by highlighting the parameter and dragging the term into the "Examined" side. Those parameters not examined will remain in the "Not Examined" side. Use the comments to indicate when observations were made and any other notable fact. 

[12]Indicate the measurements or observations that were made by highlighting the parameter and dragging the term into the "Examined" side. Those parameters not examined will remain in the "Not Examined" side. Use the comments to indicate when observations were made and any other notable fact. 

[13]Drag and drop to indicate the parameters that were examined. Use comments to indicate other important information. 

[14]Drag and drop to indicate parameters that were examined. Use comments to indicate other important information. 

[15]Note when urines were collected for testing, the groups that were tested, and drag and drop the parameters that were tested. Make comments on anything significant. 

[16]Intermediate or interim data points should be included and noted. Scheduled or unscheduled necropsies should be clearly noted. Footnotes may be useful in tables to describe any changes in sample size that may be due to planned or unplanned deviation in animal number. Tables can be expanded or modified to include scheduled interim data collection. 

[17]Note here whether there was a check or verification of the doses being administered (by what analytical methodology and in which laboratory). If the test substance was mixed in the feed, provide a statement in the study report verifying, for example, that a 100 ppm feed mixture was analyzed to contain 100 E 5 ppm, or 100 E 10 ppm of the test substance? Was this done more than once? Were there adequate data to permit the calculation of the actual dose administered?

We suggest using an Excel file to calculate the Means and Standard Deviations (SD). (Optional): Download the Excel file named 'calculation.xls' to your computer. This Excel file contains four sheets named: Dose verification, Body Wt, Feed Consumption, and Intake of test substance. Each sheet can be accessed by clicking the name tab that appears on the lower left-hand side of the Excel file. 

[18]Note any statistically or biologically significant feed consumption changes. You may also want to note feed consumption changes that were not statistically significant (e.g., at low or mid dose) but could indicate a treatment-related trend. You may wish to insert graphs or use a table as provided in this section or delete it if not needed. To use the optional Excel file (calculation.xls) click the 'Feed Consumption' tab on the lower left-hand side of the file) to calculate the Means and Standard Deviations (SD). 

[19]Note any statistically or biologically significant body weight changes. You may also want to note body weight changes that were not statistically significant (e.g., at low or mid dose) but could indicate a treatment-related trend. You may wish to insert additional graphs or tables (e.g. body weight change per unit time), and/or use a table as provided in this section or delete it if not needed. To use the optional Excel file, (calculation.xls) click the 'Body Weight' tab on the lower left-hand side of the file) to calculate the Means and Standard Deviations (SD). 

[20]What was the level of intake of feed and the level of test substance in the feed was, what was the dose actually being delivered to the animals? You may wish to use a table as provided in this section or delete it if not needed. To use the optional Excel file (calculation.xls), click the 'Intake of Test Substance' tab on the lower left-hand side of the file to calculate the Means and Standard Deviations (SD). 

[21]Note in this section if feed efficiency was determined; if any changes were observed; if feed efficiency was calculated for all groups or only selected groups; and if calculations were made per group or per animal. 

[22]List significant, dose-related abnormal cage-side observations reported. Also, use a Table in Section V. L. to note any significant, dose-related abnormal neurotoxicological observations (see Section IV. G for a list of neurotox parameters). 

[23]For each animal with an unscheduled death, note the group, date of death, and any observations, including lesions observed at necropsy. 

[24]Note findings that were statistically or biologically treatment-related. You may also want to note changes that were not statistically significant (e.g., at low or mid dose) but could indicate a treatment-related trend. The comment section is available to explain data findings, if needed. 

[25]Note findings that were statistically or biologically treatment-related. You may also want to note changes that were not statistically significant (e.g., at low or mid dose) but could indicate a treatment-related trend. The comment section is available to explain data findings, if needed. 

[26]Note findings that were statistically or biologically treatment-related. You may also want to note changes that were not statistically significant (e.g., at low or mid dose) but could indicate a treatment-related trend. The comment section is available to explain data findings, if needed. 

[27] If indicated, list treatment-related findings in the nervous system that were noted under results sections V. F: Abnormal cage-side observations, V. O: Gross pathological changes, and V.P: Histopathological changes. Provide a statement whether or not the test substance presents a potential neurotoxicity hazard. If no neurotoxicity is indicated, this section may be omitted or deleted. 

[28] When other tests were conducted, make note of the tests and any significant treatment-related effects. 

[29]Note findings that were statistically or biologically treatment-related. You may also want to note changes that were not statistically significant (e.g., at low or mid dose) but could indicate a treatment-related trend. The comment section is available to explain data findings, if needed. 

[30]Note findings that were statistically or biologically treatment-related. You may also want to note changes that were not statistically significant (e.g., at low or mid dose) but could indicate a treatment-related trend. The type of lesion should be noted. The comment section is available to explain data findings, if needed. Also, use a Table at the next histopathology section to note any findings. 

[31]See section IV.O for the list of organs or tissues.

This table should serve as a summary of the comprehensive histopathology report. Footnotes (or addenda) can be added to describe critical findings or individual observations that should be noted. Complex histopathology findings may not be transferable to this summary table.

Note findings that were statistically or biologically treatment-related. You may also want to note changes that were not statistically significant (e.g., at low or mid dose) but could indicate a treatment-related trend. Note treatment-related findings in the nervous system in this section, if not already noted in Section V. L. The type of lesion should be noted. The comment section is available to explain data findings, if needed.

To change the table, place cursor in the row or column that you wish to modify, then from the "Table" menu, use "Select Row" or "Select Column" to highlight the row or column that you wish to change, and use the "delete" or "insert" functions to make your changes. 

[32]If this information has already been entered in section V.L., delete the relevant rows in this table to prevent duplication of neurologic system entries. 

[33]Give your comments on the study: what was done well; what the problems were; did the study accomplish what it was supposed to do? 

[34]Summarize the key findings from the study. Was there a dose-effect relationship? Were target organs identified? Was a NOEL achieved for each significant effect/observation? What was the NOEL? The whole study should be summarized in this section.

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