Food

Template For Subchronic Toxicity Study in Dogs

Introduction to the Template for a Subchronic Toxicity Study in Dogs

March 2004

Return to Toxicology Template Introduction


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 (Terminal)
    14. Gross Pathology Observations
    15. Histopathology Observations
    16. 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 Comment 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. Was There A Target Organ?
    4. NOEL
  8. References

Subchronic 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. Date of Study Report:
E. Dates Study Conducted:
F. Study Objective:
G. 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

IV. Materials and Methods

A. Test Substance [4] 

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

B. Test Substance As Administered [6] 

  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?
  7. Problems with storage?

C. Animal Diet

  1. Feed
    1. Type:
    2. Name:
    3. Availability:
    4. Analysis for contaminants:
    5. Comments:
  2. Water
    1. Source:
    2. Availability:
    3. Analysis for contaminants:
    4. Comments:

D. Test Animals [7] 

  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 times:
  7. Number per cage:
  8. Environmental conditions:
  9. Comments:

E. Experimental Design [8] 

1. Targeted dose levels:

Table # [Heading]

test group conc. in diet
(ppm or mg/kg)
dose to animals
(mg/kg body-weight/day)
number of males number of females
Control        

Low

       

Mid

       

High

       

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:

F. Body Weight And Feed Intake [9] 

Parameter examined:

Table # [Heading]

Examined

Not Examined

  Feed Intake*
Water Intake*
Body Weight*
Body Weight Changes*

*These parameters are recommended in REDBOOK for subchronic toxicity studies.

2. Comments: (e.g., list frequency)

G. Cage-Side Observations [10] 

1. Parameter examined:

Table # [Heading]

Examined

Not Examined

  Appearance*
Abnormal Stool*
Morbidity*
Mortality*
Neurotoxicity Screening (Specify parameters)* [,]** 

*These parameters are recommended in REDBOOK for subchronic 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,
  • Occurrence of secretions/excretions or other evidence of autonomic activity such as lacrimation, piloerection, pupil size change, unusual respiratory pattern,
  • Presence of clonic or tonic seizure,
  • Stereotype behaviors such as excessive grooming and repetitive circling,
  • Bizarre behavior such as self-mutilating and walking backwards,
  • Gross tumor development.

2. Comments:

H.  Opthalmological Examination

  1. Parameter examined:
  2. Comments:

I. Hematology [11] 

1. Fasting duration prior to blood collection:

2. When 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]

Measurement
Related To

Examined

Not Examined

Red Blood Cells

  Hematocrit (Hct)*
Hemoglobin Conc. (Hb)*
Mean Corp. Hb. (MCH)*
Mean Corp. Hb. Conc. (MCHC)* Mean Corp. Volume (MCV)*
Total Erythrocyte Count (RBC)*

White Blood Cells

  Basophils*
Eosinophils*
Lymphocytes*
Macrophage/Monocytes*
Neutrophils*
Total Leukocytes (WBC)*

Clotting Potential

  Activated Partial-Thromboplastin Time*
Clotting Time*
Platelet Count*
Prothrombin Time*

Others

  Bone Marrow Cytology*
Reticulocyte Counts*

* These parameters are recommended in REDBOOK for subchronic toxicity studies.

6. Comments:

J. Clinical Chemistry [12] 

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]

Measurement Related To

Examined

Not Examined

Electrolyte Balance   Calcium* Chloride* [,]**  Phosphorus* Potassium* [,]** Sodium* [,]**
Carbohydrate Metabolism   Glucose* [,]**

Liver Function:
. A) Hepatocellular
(Recommend At
Least 3 Out Of 5)

B) Hepatobiliary
(Recommend At
Least 3 Out Of 5)

  Alanine Aminotransferase
(ALT or SGPT)* [,]**
Aspartate Aminotransferase
(AST or SGOT)*
Glutamate Dehydrogenase*,
Sorbitol Dehydrogenase*,
Total Bile Acids*
  Alkaline Phosphatase (ALP)* [,]**,
Gamma-Glutamyl Transferase (GGT)* [,]**
Total Bile Acids*
Total Bilirubin*
5' Nucleotidase*
Kidney Function   Creatinine* [,]**,
Urea Nitrogen* [,]**
Others
(Acid/Base Balance, Cholinesterases, Hormones, Lipids, Methemoglobin, And Proteins)
  Albumin (A)*
Globulin (G, calculated) or A/G Ratio*
Total Cholesterol*
Cholinesterase*
Total Protein* [,]**
Fasting Triglycerides*

* These parameters are recommended in REDBOOK for subchronic toxicity studies.

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

6. Comments:

K. Urinalysis [13] 

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

Table # [Heading]

Examined

Not Examined

 

Glucose*, Microscopic evaluation for sediment and presence of blood/blood cells*, pH*, Protein*, Specific Gravity*, Volume*

* These parameters are recommended in REDBOOK for subchronic toxicity studies.

4. Comments:

L. Other Tests

  1. Observations:
  2. Comments:

M.  Necropsy (Terminal)

1. Organs weighed:

Table # [Heading]

Examined

Not Examined

 

Adrenals*, Brain*, Epididymides*, Heart*,
Kidneys*, Liver*, Spleen*, Testes*, Thyroid/parathyroid*, Thymus*, Ovaries*, Uterus*

* These parameters are recommended in REDBOOK for subchronic toxicity studies.

2. Comments:

N. Gross Pathology Observations

  1. Organs/Tissues examined:
  2. Comments:

O. 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]

System

Examined

Not Examined

Digestive

  Salivary Gland*, Esophagus*, Stomach*, Duodenum*, Jejunum*, Ileum*, Cecum*, Colon*, Rectum*, Gall Bladder*, Liver (middle, left and triangular lobes)*, Pancreas*

Respiratory

  Nasal Turbinates*, Trachea*,
Lung (with main-stem bronchi)*

Cardiovascular

  Aorta*, Heart*

Reticulo- Endothelial/
Hematopoietic

  Bone Marrow (sternum)*,
Lymph Nodes (1 related to route of administration, and 1 from a distant location)*,
Spleen*, Thymus*

Urogenital

  Kidneys*, Ovaries*, Urinary Bladder*;  As applicable: fallopian tubes*, Corpus Uteri*, Cervix Uteri*, Vagina*, Prostate*, Seminal Vesicle*, Testes*

Neurologic

  Brain (at least 3 different levels)*, Spinal-Cervical*,
Spinal-Lumbar*,
Spinal-Midthoracic*,
Sciatic Nerve*,
Harderian Gland (if present)*

Glandular

  Adrenals*, Mammary Glands*, Pituitary Glands*, Thyroid/Parathyroid Glands*, Thymus*

Other

  Bone (Femur)*, Eyes*, Skeletal Muscle*, Skin*, Epididymis*

* These parameters are recommended in REDBOOK for subchronic toxicity studies.

5. Comments:

P. Statistical Methods

1. Methods of statistical analysis:

Table # [Heading]

Methods Of Statistical Analysis

Parameters Tested

   
   
   
   

2. Comments:

V. Results

A. Dose Verification [14] 

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]

Dose Groups

Targeted Daily Dose
(Mg/Kg Body-Weight/Day)

Targeted Concentration In Feed
(Ppm Or Mg/Kg)

Concentrations Found In Feed
(Ppm Or Mg/Kg)

Standarddeviation

N*

Low

 

       

Mid

 

       

High

 

       

* Number of measurements (N)

2. Verified by:

3. Comments:

B. Feed Consumption [15] 

1. Observations:

Table # [Heading]

 

Daily Feed Consumption (gram of feed consumed/day)

Sex

Males

Females

Targeted Daily Dose
(mg/kg body-weight/day)

0
Control

     

0
Control

     

Number of Animals

               

 

X

SD

X

SD

X

SD

X

SD

X

SD

X

SD

X

SD

X

SD

0 week

                               

1 [st] week

                               

2 [nd] week

                               

3 [rd] week

                               

4 [th] week

                               

5 [th] week

                               

6 [th] week

                               

7 [th] week

                               

8 [th] week

                               

9 [th] week

                               

10 [th] week

                               

11 [th] week

                               

12 [th] week

                               

13 [th] week

                               

X: Mean, SD: Standard Deviation

2. Comments:

C. Body Weight [16] 

1. Observations:

Table # [Heading]

 

Body Weights (kg body weight/day)

Sex

Males

Females

Targeted Daily Dose
(mg/kg body-weight/day)

0
Control

     

0
Control

     

Number Of Animals

               

 

X

SD

X

SD

X

SD

X

SD

X

SD

X

SD

X

SD

X

SD

0 week

                               

1 [st] week

                               

2 [nd] week

                               

3 [rd] week

                               

4 [th] week

                               

5 [th] week

                               

6 [th] week

                               

7 [th] week

                               

8 [th] week

                               

9 [th] week

                               

10 [th] week

                               

11 [th] week

                               

12 [th] week

                               

13 [th] week

                               

X: Mean, SD: Standard Deviation

2. Comments:

D. Intake Of Test Substance [17] 

1. Observations:

  Table # [Heading]

 

Daily Intake Of Test Material (mg/kg body-weight/day))

Sex

Males

Females

Targeted Daily Dose
(mg/kg body-weight/day)

0
CONTROL

     

0
CONTROL

     

Number Of Animals

               

 

X

SD

X

SD

X

SD

X

SD

X

SD

X

SD

X

SD

X

SD

0 week

                               

1 [st] week

                               

2 [nd] week

                               

3 [rd] week

                               

4 [th] week

                               

5 [th] week

                               

6 [th] week

                               

7 [th] week

                               

8 [th] week

                               

9 [th] week

                               

10 [th] week

                               

11 [th] week

                               

12 [th] week

                               

13 [th] week

                               

X: Mean, SD: Standard Deviation

2. Comments:

E. Feed Efficiency [18] 

  1. Was feed efficiency calculated?
  2. Comments:

F. Cage-Side Observations [19] 

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

G. Mortality [20] 

  1. Observations:
  2. Comments:

H.  Ophthalmological Examination

  1. Observations:
  2. Comments:

I. Hematology [21] 

1. Observations:

Table # [Heading]

Sex

Males

Females

Daily Dose
mg/kg body-weight/day)

0
Control

 

 

 

0
Control

 

 

 

Number Of Animals

               

Red Blood Cells

               

Hematocrit (Hct)

%

               

Hemoglobin Conc. (Hb)

g/L

               

Mean Corp. Hb. (MCH)

                 

Mean Corp. Hb. Conc. (MCHC)

                 

Mean Corp. Volume (MCV)

L/L

               

Total Erythrocyte Count (RBC)

10 [12]/L

               

White Blood Cells

               

Basophils

                 

Eosinophils

                 

Lymphocytes

10 [9]/L

               

Macrophage/
Monocytes

                 

Neutrophils

10 [9]/L

               

Total Leukocytes (WBC)

10 [9]/L

               

Clotting Potential

               

Activated Partial-Thromboplastin Time

                 

Clotting Time

                 

Platelet Count

10 [9]/L

               

Prothrombin Time

                 

Others

               

Bone marrow cytology

                 

Reticulocyte counts

10 [12]/L

               

(Specify a method of statistical analysis): * p<0.05, ** p<0.01

2. Comments:

J. Clinical Chemistry [22] 

1. Observations:

Table # [Heading]

Sex

Males

Females

Daily Dose
.(mg/kg body-weight/day)

0
Control

 

 

 

0
Control

 

 

 

Number Of Animals

               
Electrolyte Balance                

Calcium

mmol/L

               

Chloride

mmol/L

               

Phosphorus

mmol/L

               

Potassium

mmol/L

               

Sodium

mmol/L

               
Carbohydrate Metabolism                

Glucose

mmol/L

               
Liver Function:
A) Hepatocellular
               

Alanine Amino-transferase
.(ALT or SGPT)

U/L

               

Aspartate Amino-transferase
(AST or SGOT)

U/L

               

Glutamate Dehydrogenase

U/L

               

Sorbitol Dehydrogenase

U/L

               
Liver Function:
B) Hepatobiliary
               

Alkaline Phosphatase (ALP)

U/L

               

Gamma-Glutamyl Transferase (GGT)

U/L

               

Total Bile Acids

mmol/L

               

Total Bilirubin

mmol/L

               

5' Nucleotidase

U/L

               
Kidney Function                

Creatinine

mmol/L

               

Urea Nitrogen

Mg/dL

               
Others                

Albumin (A)

g/L

               

Globulin

(G, calculated)

g/L

               

A/G Ratio

                 

Total protein

g/L

               

Total Cholesterol

mmol/L

               

Fasting Triglycerides

mmol/L

               

Cholinesterase

U/L

               

(Specify statistical method of analysis): * p<0.05, ** p<0.01

2. Comments:

K. Urinalysis [23] 

1. Observations:

Table # [Heading]

Sex

Males

Females

DAILY DOSE
.(mg/kg body-weight/day)

0
CONTROL

 

 

 

0
CONTROL

 

 

 

NUMBER OF ANIMALS

               

Glucose

mmol/L

               

Microscopic evaluation for sediment and presence of blood/blood cells

                 

pH

                 

Protein

g/L

               

Specific Gravity

                 

Volume

L/time

               

2. Comments:

L. Neurotoxicity (If Indicated) [24] 

1. Observations:

Table # [Heading]

 

Number Of Animals

Sex

Males

Females

Daily Dose
.(Mg/Kg Body-Weight/Day)

0
Control

 

 

 

0
.Control

 

 

 

Number Of Animals Examined

               
Observations Of Nervous System Toxicity [+]                

Observation

               

Observation

               

Observation

               

 

               
Gross- And Histo-Pathology Changes In The Neurologic System [# ]                
Organ/Tissue                

Gross Lesion

               

Non-Neoplastic Lesion

               

Non-Neoplastic Lesion

               

Non-Neoplastic Lesion

               

 

               

Neoplastic Lesions

               

Neoplastic Lesions

               

Neoplastic Lesions

               

+ 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 end of dosing period can be shown; however, if there were additional noteworthy findings at earlier timepoints, 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 [25]

  1. Observations:
  2. Comments:

N. Organ Weights [26] 

1. Observations:

Table # [Heading]

Sex

Males

Females

Daily Dose
.(Mg/Kg Body-Weight/Day)

0
Control

 

 

 

0
Control

 

 

 

Number Of Animals

               
Body Weight (gram) [a]                
Brain                
Absolute Weight [ a]

gram

               
Per Body Weight [ a]

%

               
Adrenals                
Absolute Weight [ a]

gram

               
Per Body Weight [ a]

%

               
Per Brain Weight [a]

%

               
Epididymides          
Absolute Weight [ a]

gram

       
Per Body Weight [ a]

%

       
Per Brain Weight [a]

%

       
Heart                
Absolute Weight [ a]

gram

               
Per Body Weight [ a]

%

               
Per Brain Weight [a]

%

               
Kidneys                
Absolute Weight [ a]

gram

               
Per Body Weight [ a]

%

               
Per Brain Weight [a]

%

               
Liver                
Absolute Weight [ a]

gram

               
Per Body Weight [ a]

%

               
Per Brain Weight [a]

%

               
Spleen                
Absolute Weight [ a]

gram

               
Per Body Weight [ a]

%

               
Per Brain Weight [a]

%

               
Testes          
Absolute Weight [ a]

gram

       
Per Body Weight [ a]

%

       
Per Brain Weight [a]

%

       
Thyroid and Parathyroid                
Absolute Weight [ a]

gram

               
Per Body Weight [ a]

%

               
Per Brain Weight [a]

%

               
Thymus                
Absolute Weight [ a]

gram

               
Per Body Weight [ a]

%

               
Per Brain Weight [a]

%

               
Ovaries          
Absolute Weight [ A]

gram

       
Per Body Weight [ a]

%

       
Per Brain Weight [a]

%

       
Uterus          
Absolute Weight [ a]

gram

       
Per Body Weight [ a]

%

       
Per Brain Weight [a]

%

       

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

(Specify methods of statistical analysis): * p<0.05, ** p<0.01

2. Comments:

O. Gross Pathology Changes Observed [27] 

  1. Observations:
  2. Comments:

P. Histopathology Changes Observed [28] 

1. Observations:

Table # [Heading)

 

Number Of Animals
With, Gross, Non-Neoplastic, Or Neoplastic Lesions

Sex

Males

Females

Daily Dose
.(Mg/Kg Body-Weight/Day)

0
Control

 

 

 

0
Control

 

 

 

Number Of Animals Examined

               
Digestive System                
Organ/Tissue [#]                

Gross Lesion

               

Non-Neoplastic Lesion

               

Non-Neoplastic Lesion

               

Non-Neoplastic Lesion

               

 

               

Neoplastic Lesions

               

Neoplastic Lesions

               

Neoplastic Lesions

               

 

               
Respiratory System                
Organ/Tissue [#]                

Gross Lesion

               

Non-Neoplastic Lesion

               

Non-Neoplastic Lesion

               

Non-Neoplastic Lesion

               

 

               

Neoplastic Lesions

               

Neoplastic Lesions

               

Neoplastic Lesions

               

 

               
Cardiovascular System                
Organ/Tissue [#]                

Gross Lesion

               

Non-Neoplastic Lesion

               

Non-Neoplastic Lesion

               

Non-Neoplastic Lesion

               

 

               

Neoplastic Lesions

               

Neoplastic Lesions

               

Neoplastic Lesions

               

 

               
Reticulo-Endothelial /Hematopoietic System                
Organ/Tissue [#]                

Gross Lesion

               

Non-Neoplastic Lesion

               

Non-Neoplastic Lesion

               

Non-Neoplastic Lesion

               

 

               

Neoplastic Lesions

               

Neoplastic Lesions

               

Neoplastic Lesions

               

 

               
Urogenital System                

Organ/Tissue [#]

               

Gross Lesion

               

Non-Neoplastic Lesion

               

Non-Neoplastic Lesion

               

Non-Neoplastic Lesion

               

 

               

Neoplastic Lesions

               

Neoplastic Lesions

               

Neoplastic Lesions

               

 

               
Neurologic System [29]                 
Organ/Tissue [#]                

Gross Lesion

               

Non-Neoplastic Lesion

               

Non-Neoplastic Lesion

               

Non-Neoplastic Lesion

               

 

               

Neoplastic Lesions

               

Neoplastic Lesions

               

Neoplastic Lesions

               

 

               
Glandular System                
Organ/Tissue [#]                

Gross Lesion

               

Non-Neoplastic Lesion

               

Non-Neoplastic Lesion

               

Non-Neoplastic Lesion

               

 

               

Neoplastic Lesions

               

Neoplastic Lesions

               

Neoplastic Lesions

               

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

2. Comments:

VI. Evaluation and Comment on Study [30] 

VII. Summary and Conclusions [31] 

A. Brief Summary Of Major Findings From The Study

B. Relationship Between Dose And Incidence/Severity Of Lesions Or Abnormalities

C. Was There A Target Organ?

D. NOEL

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

VIII. References


End Notes

This section includes some comments that are only relevant when using the Word template. 

[1]This Template is set up for typical 90-Day 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 the Questions A and B.  However, you may want to elaborate on the type of GLP compliance (USFDA, OECD, etc.) or make other notations 

[4]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? 

[5]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, use other methods of depicting the molecular structure. 

[6]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. 

[7]Note anything that might have affected the study. Use the comments to indicate extremes or other factors that impacted the study. 

[8]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. 

[9]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. 

[10]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. 

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

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

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

[14]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, was there a statement in the study report verifying, for example, that a 100 ppm feed mixture was analyzed to contain 100 ± 5 ppm, or 100 ± 10 ppm of the test substance?  Was this done more than once? Were there adequate data to calculate the actual dose that was 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. 

[15]Note any statistically or biologically significant feed consumption changes. Also 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). 

[16]Note any statistically or biologically significant body weight changes. Also 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 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 'Body Weight' tab on the lower left-hand side of the file) to calculate the Means and Standard Deviations (SD). 

[17]Knowing what the 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 the 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). 

[18]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. 

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

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

[21]Note findings that were statistically or biologically treatment-related.  Also 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. 

[22]Note findings that were statistically or biologically treatment-related.  Also 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. 

[23]Note findings that were statistically or biologically treatment-related.  Also 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. 

[24] 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 was indicated, this section may be omitted or deleted. 

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

[26]Note findings that were statistically or biologically treatment-related.  Also 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]Note findings that were statistically or biologically treatment-related.  Also 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. 

[28]Note findings that were statistically or biologically treatment-related.  Also 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. 

[29]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. 

[30]Give your evaluation of the study: what was done well; what the problems were; did the study accomplish what it was supposed to do? 

[31]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.


Some of the internet links may have been changed in this document. Guidance documents can be found in the guidance area of the Food section of www.fda.gov

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