FDA Logo U.S. Food and Drug AdministrationCenter for Food Safety and Applied Nutrition
 U.S. Department of Health and Human Services

FDA Home Page | CFSAN Home | Search/Subject Index | Q & A | Help

horizontal rule

CFSAN/Office of Food Additive Safety
April 2005

horizontal rule

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

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 (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
horizontal rule

Chronic (1 Year) Toxicity Study in Dogs1

Date of Submission:

Title of Petition or Notification:

Name and Address of Petitioner or Notifier:

I. Identification of Study2 

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 Practice3 

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 Methods5 

A.  TEST substance6 

1.    CAS name:
2.    Other name(s):
3.    CAS registry number:
4.    Molecular structure: http://www.chemfinder.com7 /

 

5.    Purity:
6.    Impurities:
7.    Stability:
8.    Comments:

B.  TEST substance AS ADMINISTERED8 

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  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 DESIGN10 

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: (e.g. dose selection rationale, dosing differences in males and females, etc.)

 

F.  BODY WEIGHT AND FEED INTAKE11 

1.    Parameter examined:

 

Table # [Heading]

 

examined

not examined
  Feed Intake*,
Water Intake*,
Body Weight*,
Body Weight Changes*

*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 OBSERVATIONS12 

1.    Parameter examined:

 

Table # [Heading]

 

examined

not examined
  Appearance*,
Abnormal Stool*,
Morbidity*,
Mortality*,
Neurotoxicity Screening (Specify parameters)**

*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:

 

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

 

H.  OPTHALMOLOGICAL EXAMINATION

1.    Parameter examined:
2.    Comments:

I.      HEMATOLOGY13 

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]

 

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 Toxicological Principles for the Safety of Food Ingredients: Redbook 2000 for chronic toxicity studies.

 

6.    Comments:

 

J.    CLINICAL CHEMISTRY14 

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 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.  URINALYSIS15 

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 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]

 

examined not examined
  Adrenals*, Brain*, Epididymides*, Heart*, Kidneys*, Liver*, Spleen*, Testes*, Thyroid/parathyroid*, Thymus*, Ovaries*, Uterus*

* 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]

 

examined not examined
  Adrenals*, Brain*, Epididymides*, Heart*, Kidneys*, Liver*, Spleen*, Testes*, Thyroid/parathyroid*, Thymus*, Ovaries*, Uterus*

* 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]

 

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*  
   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 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]

 

methods of statistical analysis parameters tested
   
   
   
   

 

2.    Comments:

 

V. Results16 

A.  DOSE VERIFICATION17 

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 CONSUMPTION18 

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

 

Table # [Heading]

 

  Weekly Summary of daily feed consumption (mean gram of feed consumed/day)
SEX males females
TARGETED DAILY DOSE(mg/kg body-weight/day) 0
CONTROL		       0 CONTROL      
NUMBER OF ANIMALS                
Week X SD X SD X SD X SD X SD X SD X SD X SD
0                                
1                                
2                                
3                                
4                                
5                                
6                                
7                                
8                                
9                                
10                                
11                                
12                                
13                                
14                                
15                                
16                                
17                                
18                                
19                                
20                                
21                                
22                                
23                                
24                                
25                                
26                                
27                                
28                                
29                                
30                                
31                                
32                                
33                                
34                                
35                                
36                                
37                                
38                                
39                                
40                                
41                                
42                                
43                                
44                                
45                                
46                                
47                                
48                                
49                                
50                                
51                                
52                                

X: Mean, SD: Standard Deviation

 

2.    Comments:

 

C.  BODY WEIGHT19 

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

 

Table # [Heading]

 

  Weekly Summary of body weights (mean kg body weight/day)
SEX males females
TARGETED DAILY DOSE(mg/kg body-weight/day) 0
CONTROL		       0 CONTROL      
NUMBER OF ANIMALS                
Week X SD X SD X SD X SD X SD X SD X SD X SD
0                                
1                                
2                                
3                                
4                                
5                                
6                                
7                                
8                                
9                                
10                                
11                                
12                                
13                                
14                                
15                                
16                                
17                                
18                                
19                                
20                                
21                                
22                                
23                                
24                                
25                                
26                                
27                                
28                                
29                                
30                                
31                                
32                                
33                                
34                                
35                                
36                                
37                                
38                                
39                                
40                                
41                                
42                                
43                                
44                                
45                                
46                                
47                                
48                                
49                                
50                                
51                                
52                                

X: Mean, SD: Standard Deviation

 

2.    Comments:

 

D.  INTAKE OF TEST SUBSTANCE20 

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

 

Table # [Heading]

 

  Weekly Summary of 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                
Week X SD X SD X SD X SD X SD X SD X SD X SD
0                                
1                                
2                                
3                                
4                                
5                                
6                                
7                                
8                                
9                                
10                                
11                                
12                                
13                                
14                                
15                                
16                                
17                                
18                                
19                                
20                                
21                                
22                                
23                                
24                                
25                                
26                                
27                                
28                                
29                                
30                                
31                                
32                                
33                                
34                                
35                                
36                                
37                                
38                                
39                                
40                                
41                                
42                                
43                                
44                                
45                                
46                                
47                                
48                                
49                                
50                                
51                                
52                                

X: Mean, SD: Standard Deviation

 

2.    Comments:

 

E.  FEED EFFICIENCY21 

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

 

F.   CAGE-SIDE OBSERVATIONS22 

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

 

G.  MORTALITY23 

1.    Observations:
2.     Comments:

 

H.  OPHTHALMOLOGICAL EXAMINATION

1.    Observations:
2.     Comments:

 

I.      HEMATOLOGY24 

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

 

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)
1012/L
                
white blood cells                
Basophils
                 
Eosinophils
                  
Lymphocytes
 109/L                
Macrophage/Monocytes
                  
Neutrophils
 109/L                
Total Leukocytes (WBC)
 109/L                
clotting potential                
Activated Partial-Thromboplastin Time
                  
Clotting Time
                  
Platelet Count
 109/L                
Prothrombin Time
                  
others                
Bone marrow cytology
                  
Reticulocyte counts
 1012/L                

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

 

2.    Comments:

 

J.    CLINICAL CHEMISTRY25 

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

 

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.  URINALYSIS26 

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

 

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)27 

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

 

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 #           </