1978 

National Cancer Institute 

CARCINOGENESIS 

Technical Report Series 

No. 65 

BIOASSAY OF 

CHLOROPICRIN 

FOR POSSIBLE CARCINOGENICITY 

CAS No. 76-06-2 

NCI-CG-TR-65 

U.S. DEPARTMENT OF HEALTH, EDUCATION, AND WELFARE 

Public Health Service 

National Institutes of Health 
BIOASSAY OF
 

CHLOROPICRIN
 

FOR POSSIBLE CARCINOGENICITY
 

Carcinogenesis Testing Program
 

Division of Cancer Cause and Prevention
 

National Cancer Institute
 

National Institutes of Health
 

Bethesda, Maryland 20014
 

U.S. DEPARTMENT OF HEALTH, EDUCATION, AND WELFARE
 

Public Health Service
 

National Institutes of Health
 

DHEW Publication No. (NIH) 78-1315
 

REPORT ON THE BIOASSAY OF CHLOROPICRIN
 

FOR POSSIBLE CARCINOGENICITY
 

CARCINOGENESIS TESTING PROGRAM
 

DIVISION OF CANCER CAUSE AND PREVENTION
 

NATIONAL CANCER INSTITUTE, NATIONAL INSTITUTES OF HEALTH
 

FOREWORD: This report presents the results of the bioassay of chloropicrin conducted for the Carcinogenesis Testing Program, Division of
 

Cancer Cause and Prevention, National Cancer Institute (NCI), National
 

Institutes of Health, Bethesda, Maryland. This is one of a series
 

of experiments designed to determine whether selected environmental
 

chemicals have the capacity to produce cancer in animals. Negative
 

results, in which the test animals do not have a greater incidence of
 

cancer than control animals, do not necessarily mean the test chemical
 

is not a carcinogen because the experiments are conducted under a
 

limited set of circumstances. Positive results demonstrate that the
 

test chemical is carcinogenic for animals under the conditions of the
 

test and indicate a potential risk to man. The actual determination
 

of the risk to man from animal carcinogens requires a wider analysis.
 

CONTRIBUTORS: This bioassay of chloropicrin was conducted by Hazleton
 

Laboratories America, Inc., Vienna, Virginia, initially under direct
 

contract to the NCI and currently under a subcontract to Tracer Jitco,
 

Inc., prime contractor for the NCI Carcinogenesis Testing Program.
 

The experimental design was determined by the NCI Project Officers, Dr. J. H. Weisburger (1,2) and Dr. E. K. Weisburger (1). The
 

principal investigators for the contract were Dr. M. B. Powers (3),
 

Dr. R. W. Voelker (3), Dr. W. A. Olson (3,4) and Dr. W. M. Weatherholtz (3). Chemical analysis was performed by Dr. C. L. Guyton
 

(3,5) and the analytical results were reviewed by Dr. N. Zimmerman
 

(6); the technical supervisor of animal treatment and observation
 

was Ms. K. J. Petrovics (3).
 

Histopathologic examinations were performed by Dr. R. H. Habermann (3) and reviewed by Dr. R. W. Voelker (3) at the Hazleton Laboratories America, Inc., and the diagnoses included in this report
 

represent the interpretation of these pathologists. Histopathology
 

findings and reports were reviewed by Dr. P. L. Schueler (7).
 

Compilation of individual animal survival, pathology, and summary tables was performed by FG&G Mason Research Institute (8); the
 

statistical analysis was performed by Mr. W. W. Belew (6) and Dr. J.
 

R. Joiner (7), using methods selected for the Bioassay Program by Dr.
 

J. J. Gart (9).
 

i ii
 
This report was prepared at METREK, a Division of The MITRE Corporation (6) under the direction of the NCI. Those responsible for
 

this report at METREK are the project coordinator, Dr. L. W. Thomas
 

(6), the task leader, Dr. M. R. Kornreich (6), the senior biologist,
 

Ms. P. Walker (6), and the technical editor, Ms. P. A. Miller (6).
 

The final report was reviewed by members of the participating organizat ions.
 

The statistical analysis was reviewed by members of the Mathematical Statistics and Applied Mathematics Section of the NCI: Dr.
 

J. J. Gart (9), Mr. J. Nam (9), Dr. H. M. Pettigrew (9), and Dr. R.
 

E. Tarone (9).
 

The following other scientists at the National Cancer Institute
 

were responsible for evaluating the bioassay experiment, interpreting
 

the results, and reporting the findings: Dr. K. C. Chu (1), Dr. C.
 

Cueto, Jr. (1), Dr. J. F. Douglas (1), Dr. D. G. Goodman (1), Dr. R.
 

A. Griesemer (1), Dr. H. A. Milman (1), Dr. T. W. Orme (1), Dr. R. A.
 

Squire (1,10), and Dr. J. M. Ward (1).
 

1.	 Carcinogenesis Testing Program, Division of Cancer Cause and
 

Prevention, National Cancer Institute, National Institutes of
 

Health, Bethesda, Maryland.
 

2.	 Now with the Naylor Dana Institute for Disease Prevention, American Health Foundation, Hammon House Road, Valhalla, Mew York.
 

3.	 Hazleton Laboratories America, Inc., 9200 Leesburg Turnpike,
 

Vienna, Virginia.
 

4.	 Mow with the Center for Regulatory Services, 2347 Paddock Lane,
 

Reston, Virginia.
 

5.	 No wit w h Rhodia, Inc., 23 Belmont Drive, Somerset, New Jersey.
 

6.	 Th MITR e E Corporation, METREK Division, 1820 Dolley Madison
 

Boulevard, McLean, Virginia.
 

7.	 Tracer Jitco, Inc., 1776 East Jefferson Street, Rockville,
 

Maryland.
 

8.	 EG&G Mason Research Institute, 1530 East Jefferson Street,
 

Rockville, Maryland.
 

iv 
9.	 Mathematical Statistics and Applied Mathematics Section, Biometry
 

Branch, Field Studies and Statistics Program, Division of Cancer
 

Cause and Prevention, National Cancer Institute, National Institutes of Health, Bethesda, Maryland.
 

10.	 No wit w h the Division of Comparative Medicine, Johns Hopkins
 

University, School of Medicine, Traylor Building, Baltimore,
 

Maryland.
 

V 

SUMMARY
 

A bioassay of technical-grade chloropicrin for possible carcinogenicity was conducted using Osborne-Mendel rats and B6C3F1 mice.
 

Chloropicrin in corn oil was administered 5 days a week by gavage, at
 

either of two dosages, to groups of 50 male and 50 female animals of
 

each species. Time-weighted average dosages of 25 mg/kg/day for low
 

dose male rats and 20 mg/kg/day for low dose female rats were administered during weeks 1 through 33, then administered cyclically (1 dose-


free week followed by 4 weeks of administration) from weeks 34 through
 

78. Time-weighted average dosages of 26 mg/kg/day for high dose male
 

rats and 22 mg/kg/day for high dose female rats were administered from
 

weeks 1 through 17, weeks 31 through 33, and cyclically (1 dose-free
 

week followed by 4 weeks of administration) during weeks 34 through 78.
 

Time-weighted average dosages of 66 and 33 mg/kg/day, respectively,
 

for male and female mice were administered for 78 weeks. These dosing
 

regimens were followed by observation periods of 32 weeks for rats and
 

13 weeks for mice.
 

For each species, 20 animals of each sex were placed on test as
 

vehicle controls. These animals were gavaged with corn oil. Twenty
 

animals of each sex were placed on test as untreated controls for
 

each species. These animals were not gavaged.
 

A high incidence of early death was observed among chloropicrindosed rats. Deaths among dosed rats occurred as early as week 1 for
 

females and week 6 for males. Median survival was week 48 for high
 

dose males, week 54 for low dose males, week 70 for high dose females
 

and week 59 for low dose females. Statistical tests indicate a
 

positive association between chloropicrin dosage and mortality of
 

rats.
 

No neoplasms were observed at higher incidences in dosed than
 

control rats. In rats of both sexes, incidences of adenoma of the
 

pituitary and of adenocarcinoma or fibroadenoma of the mammary gland
 

were higher in control groups than dosed groups. It is likely that
 

most dosed rats did not survive long enough to be at risk from late-


appearing tumors.
 

A rapid decrease in survival after the first year of the study
 

was observed among high dose mice of both sexes. Survival of high
 

dose male mice decreased from 80 percent in week 54 to 26 percent in
 

week 90. Survival of high dose female mice decreased from 82 percent
 

in week 54 to 36 percent in week 90. Statistical tests indicated a
 

positive association between chloropicrin dosage and mortality of
 

mice .
 

vii
 
In chloropicrin-dosed mice, proliferative lesions of the squamous
 

epithelium of the forestomach included two carcinomas and a papilloma.
 

Although these tumors were uncommon in control animals, statistical
 

analysis did not demonstrate that they were related to administration
 

of chloropicrin. Other proliterative lesions of the forestomach
 

occurring at an increased incidence in dosed mice were acanthosis and
 

hyperkeratosis. No statistically significant increase of tumor incidence was observed in mice.
 

The bioassay of chloropicrin using Osborne-Mendel rats did not
 

permit an evaluation of carcinogenicity because of the short survival
 

time of dosed animals. The bioassay of chloropicrin using B6C3F1
 

mice did not provide conclusive statistical evidence for the carcinogenicity of this compound.
 

viii
 
TABLE OF CONTENTS
 

Page
 

I. INTRODUCTION 1
 

II. MATERIALS AND METHODS 3
 

A. Chemicals 3
 

B. Dosage Preparation 4
 

C. Animals 4
 

D. Animal Maintenance 5
 

E. Gastric Intubation 6
 

F. Selection of Initial Dose Levels 6
 

G. Experimental Design 7
 

H. Clinical and Histopathologic Examinations 11
 

I. Data Recording and Statistical Analyses 13
 

III. CHRONIC TESTING RESULTS: RATS 18
 

A. Body Weights and Clinical Observations 18
 

B. Survival 20
 

C. Pathology 22
 

D. Statistical Analyses of Results 22
 

IV. CHRONIC TESTING RESULTS: MICE 26
 

A. Body Weights and Clinical Observations 26
 

B. Survival 26
 

C. Pathology 29
 

D. Statistical Analyses of Results 30
 

V. DISCUSSION 35
 

VI. BIBLIOGRAPHY 37
 

APPENDIX A SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

RATS TREATED WITH CHLOROPICRIN A-l
 

APPENDIX B SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

MICE TREATED WITH CHLOROPICRIN B-l
 

APPENDIX C SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN RATS TREATED WITH CHLOROPICRIN C-l
 

APPENDIX D SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN MICE TREATED WITH CHLOROPICRIN D-l
 

ix
 
LIST OF ILLUSTRATIONS 

figure Number Page 

1 GROWTH CURVES FOR CHLOROPICRIN CHRONIC 

STUDY RATS 19 

2 SURVIVAL COMPARISONS OF CHLOROPICRIN 

CHRONIC STUDY RATS 21 

3 GROWTH CURVES FOR CHLOROPICRIN CHRONIC 

STUDY MICE 27 

4 SURVIVAL COMPARISONS OF CHLOROPICRIN 

CHRONIC STUDY MICE 28 

LIST OF TABLES 

Table Number Page 

1 DESIGN SUMMARY FOR OSBORNE-MENDEL RATS— 

CHLOROPICRIN GAVAGE EXPERIMENT 9 

2 DESIGN SUMMARY FOR B6C3F1 MICE—CHLOROPICRIN 

GAVAGE EXPERIMENT 10 

3 ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN MALE RATS TREATED WITH 

CHLOROPICRIN 23 

4 ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN FEMALE RATS TREATED 

WITH CHLOROPICRIN 24 

5 ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN MALE MICE TREATED WITH 

CHLOROPICRIN 31 

6 ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN FEMALE MICE TREATED 

WITH CHLOROPICRIN 32 

Al SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

MALE RATS TREATED WITH CHLOROPICRIN A-3 

x
 
LIST OF TABLES (Concluded) 

Table Number Page 

A2 SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

FEMALE RATS TREATED WITH CHLOROPICRIN A-6 

Bl SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

MALE MICE TREATED WITH CHLOROPICRIN B-3 

B2 SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

FEMALE MICE TREATED WITH CHLOROPICRIN B-6 

Cl SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN MALE RATS TREATED WITH CHLOROPICRIN C-3 

C2 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN FEMALE RATS TREATED WITH 

CHLOROPICRIN C-7 

Dl SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN MALE MICE TREATED WITH CHLOROPICRIN D-3 

D2 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN FEMALE MICE TREATED WITH 

CHLOROPICRIN D-6 

XI
 

I. INTRODUCTION
 

Chloropicrin (NCI No. C00533) is an agricultural fumigant, once
 

widely used but now being phased out (Feldmesser, 1977). In the late
 

1960s, scientists at the National Cancer Institute noted that a group
 

of pesticides used extensively in agriculture had not been adequately
 

tested for carcinogenicity. In 1969 the Report of the Secretary's
 

Commission on Pesticides and Their Relationship to Environmental
 

Health (U.S. Department of Health, Education and Welfare, 1969)
 

further emphasized the need for chronic toxicity studies of certain
 

specific pesticides.
 

The Chemical Abstracts Service (CAS) Ninth Collective Index
 

.t,
 

(1977) name for chloropicrin is trichloronitromethane. It is also
 

called nitrochloroform.
 

Domestic production of chloropicrin in 1974 was approximately
 

4.8 million pounds (U.S. International Trade Commission, 1976). It
 

was developed as a tear gas, but was found to be useful as a fumigant
 

in 1918 (Matsumura, 1975). The primary use of chloropicrin as a
 

fumigant was in the treatment of stored grain (Rogers, 1966). It
 

also functions as a nematicide, fungicide, and insecticide when used
 

as a soil fumigant prior to planting. Chloropicrin is lethal to most
 

plants, necessitating the aeration of soil after treatment for one to
 

two weeks before planting (Rogers, 1966). Soil persistence of the
 

*
 

The CAS registry number is 76-06-2.
 

1 
chemical ranges from one to three weeks (Pesticide Information Manual,
 

1966). Because of its odor and lachrymatory properties, chloropicrin
 

has been used increasingly since the 1950s as a warning agent in
 

fumigant formulations, such as 2 percent chloropicrin and 98 percent
 

methyl bromide (Pesticide Chemicals Official Compendium, 1966;
 

Feldmesser, 1977).
 

2 
II. MATERIALS AND METHODS
 

A. Chemicals
 

A single batch of technical-grade chloropicrin was purchased by
 

Hazleton Laboratories America, Inc., Vienna, Virginia, from Morton
 

International, Inc. The purity of the compound was initially determined by Hazleton Laboratories using gas-liquid chromatography (GLC)
 

and infrared spectrophotometry. The GLC total area analysis revealed
 

six peaks; the fourth peak eluted from the column, presumed to be
 

chloropicrin, accounted for 98 percent of the total area, while none
 

of the other individual peaks accounted for more than 1 percent of
 

the total area. This indication of purity in the range of 98 percent
 

was consistent with the supplier's Volhard assay determination of 98
 

percent chloropicrin. The infrared spectrum of the tested chloropicrin was consistent with that expected from the structure of the compound .
 

A second purity determination using GLC and infrared spectrophotometry was performed by Hazleton Laboratories 20 months after the
 

initial analysis to establish the stability of the chloropicrin under
 

storage conditions. The GLC total area analysis again showed six
 

peaks. The major peak accounted for approximately 99 percent of the
 

total area, with the other five observed peaks totaling less than 1
 

percent of the total area. The infrared spectrum was consistent with
 

that of the previous analysis.
 

3 
Throughout this report the term chloropicrin is used to represent
 

this technical-grade material.
 

B. Dosage Preparation
 

(R)
 Fresh solutions of chloropicrin in Duke's corn oil (S. F. Sauer
 

Company, Richmond, Virginia) were prepared weekly, sealed, and stored
 

in dark bottles at 1°C. Concentrations of chloropicrin in corn oil
 

of 2 and 5 percent (on a weight/volume basis) were utilized for rats
 

and mice, respectively. When the dose administered to mice was
 

increased during the chronic test, the chloropicrin concentration in
 

corn oil was increased to 7 percent.
 

C. Anima1s
 

Two animal species, rats and mice, were used in the carcinogenicity bioassay. The Osborne-Mendel rat was selected on the basis of a
 

comparative study of the tumorigenic responsiveness to carbon tetrachloride of five different strains of rats (Reuber and Glover, 1970).
 

The B6C3F1 mouse was selected because it has been used by the NCI for
 

carcinogenesis bioassays and has proved satisfactory in this capacity.
 

Rats and mice of both sexes were obtained through contracts of
 

the Division of Cancer Treatment, National Cancer Institute. The
 

Osborne-Mendel rats were procured from the Battelle Memorial Institute, Columbus, Ohio, and the B6C3F1 mice were obtained from the
 

Charles River Breeding Laboratories, Inc., Wilmington, Massachusetts.
 

Upon receipt, animals were quarantined for at least 10 days, observed
 

for visible signs of disease or parasites, and assigned to the various
 

dosed and control groups.
 

4 
D. Animal Maintenance
 

All animals were housed by species in temperature- and humidity-


controlled rooms. The temperature range was 20° to 24°C and the
 

relative humidity was maintained between 45 and 55 percent. The air
 

conditioning system in the laboratory provided filtered air at a rate
 

of 12 complete changes of room air per hour. Fluorescent lighting
 

was provided on a 12-hour-daily cycle. The rats were individually
 

housed in suspended galvanized-steel wire-mesh cages with perforated
 

floors. Mice were housed by sex in groups of ten in solid-bottom
 

polypropylene cages equipped with filter tops. Sanitized cages with
 

fresh bedding (Sanichips® , Pinewood Sawdust Company, Moonachie, New
 

Jersey) were provided once each week for IT ice. Rats received sanitized cages with no bedding with the same frequency. Food hoppers
 

were changed and heat-sterilized once a week for the first 10 weeks
 

and once a month thereafter. Fresh heat-sterilized glass water botfR^!
 

ties were provided three times a week. Food (Wayne Lab-Blox~'meal,
 

Allied Mills, Inc., Chicago, Illinois) and water were available ad
 

libitum.
 

The rats dosed with chloropicrin and the vehicle controls were
 

*
 housed in the same room with other rats intubated with methylchloroform (71-55-6) and trichlorofluoromethane (75-69~4). The untreated
 

control rats were housed with other rats intubated with 1 ,1 ,2 ,2-tetrachloroethane (79-34-5), allyl chloride (107-05-1), 1,2-dibromoethane
 

*
 CAS registry numbers are given in parentheses.
 

5 
(106-93-4), carbon tetrachloride (56-23-5), and chloroform (67-66-3).
 

All mice used in the chloropicrin study, including controls, were
 

housed in the same room as other mice intubated with 1,1,2,2-tetrachloroethane (79-34-5), chloroform (67-66-3), dibromochloropropane
 

(96-12-8), allyl chloride (107-05-1), 1,2-dibromoethane (106-93-4),
 

1,2-dichloroethane (107-06-2), 1,1-dichloroethane (75-34-3), trichloroethylene (79-01-6), 3-sulfolene (77-79-2), iodoform (75-47-8),
 

methylchloroform (71-55-6), 1,1,2-trichloroethane (79-00-5), tetrachloroethylene (127-18-4), hexachloroethane (67-72-1), carbon disulfide
 

(75-15-0), trichlorofluoromethane (75-69-4), and carbon tetrachloride
 

(56-23-5).
 

E« Gastric Intubation
 

Intubation was performed for five consecutive days per week on
 

a mg/kg body weight basis utilizing the most recently observed group
 

mean body weight as a guide for determining the dose. Mean body
 

weights for each group were recorded at weekly intervals for the
 

first 10 weeks and at monthly intervals thereafter. All animals of
 

one sex within a dosed group received the same dosage. Gavage of
 

dosed animals was performed under a hood to minimize extraneous
 

exposure of other animals and laboratory personnel to the chemical.
 

F. Selection of Initial Dose Levels
 

In order to establish the maximum tolerated doses of chloropicrin for administration to treated animals in the chronic studies,
 

subchronic toxicity tests were conducted with both rats and mice.
 

6 
Animals of each species were distributed among six groups, each
 

consisting of five males and five females. Chloropicrin in corn oil
 

was introduced by gavage to five of the six rat groups at dosages of
 

16, 25, 40, 63, and 100 mg/kg/day and five of the six mouse groups at
 

dosages of 10, 16, 25, 40, and 63 mg/kg/day. The sixth group of each
 

species served as a control group, receiving only corn oil. Intubation occurred 5 days per week for 6 weeks, followed by a 2-week observation period to detect any delayed toxicity.
 

None of the male rats died at chloropicrin dosages of 40 mg/kg/
 

day or less. Mean body weight depression in the males at 40 and 63
 

mg/kg/day was 11 and 38 percent, respectively. Except for one female
 

rat treated with 25 mg/kg/day, all females receiving 40 mg/kg/day or
 

less survived the 8-week study. Mean body weight depression at 40
 

and 63 mg/kg/day for female rats was 17 and 30 percent, respectively.
 

The initial high dosage for male and female rats in the chronic study
 

was set at 46 mg/kg/day.
 

No male or female mice died during the 8-week study. At levels
 

of 40 and 63 mg/kg/day, mean body weight depression in males was 12
 

and 20 percent, respectively, and in females was 3 and 6 percent,
 

respectively. The initial high dosage for mice of both sexes in the
 

chronic study was set at 50 mg/kg/day.
 

G. Experimental Design
 

The experimental design parameters for the chronic study (species, sex, group size, dosages administered, duration of treated and
 

7 
untreated observation periods, and the time-weighted average dosages)
 

are summarized in Tables 1 and 2.
 

The treated and vehicle control rats were all approximately 6
 

weeks old at the time the experiment began. The untreated rats were
 

placed on the study when they were approximately 6 weeks old, but 11
 

weeks after the initial gavage regimen was begun for dosed and vehicle control rats. Gavage was performed five consecutive days per
 

week. The doses initially utilized for both males and females were
 

46 and 23 mg/kg/day, respectively. Throughout this report the groups
 

receiving the former dosage are referred to as the high dose groups,
 

while those receiving the latter dosage are referred to as the low
 

dose groups. When the rats were 11 weeks old (week 5 of the experiment), the high and low doses for males were increased to 56 and 28
 

mg/kg/day, respectively, as the males appeared to be tolerating the
 

chemical, while the dosages for the female treated groups remained
 

unchanged. After week 17, intubation ceased for 13 weeks for the
 

high dose males and females because of apparent compound toxicity.
 

Gavage continued for the low dose groups for both sexes. Effective
 

week 31, the high dose males and females began receiving the same dose
 

as the respective low dose groups. In week 34, intubation ceased for
 

1 week for all treated groups, followed by 4 weeks of dose administration. This cyclic pattern of dose administration continued for the
 

remainder of the 78-week study period. The vehicle control rats were
 

gavaged with corn oil for 78 weeks. Observation of all rats continued
 

for an additional 32 weeks.
 

8 
TABLE 1
 

DESIGN SUMMARY FOR OSBORNE-MENDEL RATS
 

CHLOROPICRIN GAVAGE EXPERIMENT
 

TIME-WEIGHTED 

INITIAL CHLORO­ OBSERVATION PERIOD AVERAGE DOSAGE 

GROUP PICRIN TREATED UNTREATED OVER A 78-WEEK 

SIZE DOSAGE3 (WEEKS) (WEEKS) PERIODb 

MALE
 

UNTREATED CONTROL 20 0 109 0
 

VEHICLE CONTROL 20 0 78 32 0
 

LOW DOSE 50 23 4 25
 

28 29
 

28C 36 9
 

0 32
 

HIGH DOSE 50 46 4 26
 

56 13
 

0 13
 

28 3
 

28° 36 9
 

0 32
 

FEMALE
 

UNTREATED CONTROL 20 0 109 0
 

VEHICLE CONTROL 20 0 78 32 0
 

LOW DOSE 50 23 33 20
 

23° 36 9
 

0 32
 

HIGH DOSE 50 46 17 22
 

0 13
 

23 3
 

23C 36 9
 

0 32
 

Dosage, given in mg/kg body weight, was administered by gavage five
 

consecutive days per week.
 

b . . 2 (dosage. X weeks received)
 Time-weighte & d average dosage = — :


 78 weeks
 

These dosages were cyclically administered with a pattern of 1 dose-


free week followed by 4 weeks (5 days per week) of dosage at the
 

level indicated.
 

9 

c 
MALE
 

UNTREATED CONTROL


VEHICLE CONTROL


LOW DOSE


HIGH DOSE


FEMALE
 

UNTREATED CONTROL


VEHICLE CONTROL


LOW DOSE


HIGH DOSE


TABLE 2
 

DESIGN SUMMARY FOR B6C3F1 MICE
 

CHLOROPICRIN GAVAGE EXPERIMENT
 

INITIAL CHLOROGROUP PICRIN 

SIZE DOSAGE3 

 20 0 

 20 0 

 50 25 

35 

0 

 50 50 

70 

0 

 20 0 

 20 0 

 50 25 

35 

0 

 50 50 

70 

0 

OBSERVATION PERIOD
 

TREATED UNTREATED
 

(WEEKS) (WEEKS)
 

90
 

78 13
 

13
 

65
 

13
 

13
 

65
 

30
 

90
 

78 13
 

13
 

65
 

13
 

13
 

65
 

13
 

TIME-WEIGHTED
 

AVERAGE DOSAGE
 

OVER A 78-WEEK
 

PERIODb
 

0
 

0
 

33
 

66
 

0
 

0
 

33
 

66
 

Dosages, given in mg/kg body weight, .were administered by gavage five
 

consecutive days per week.
 

b . . Z(dosage X weeks received)
 Time-weighted average dosage = 2——
 

78 weeks
 

10 
The vehicle control and treated mice were all approximately 5
 

weeks old on the day their first dose was administered. The vehicle
 

control mice were born approximately 2 weeks earlier than the low and
 

high dose groups. Therefore, administration of corn oil to the vehicle controls began correspondingly earlier than did chloropicrin
 

administration to the dosed mice. The dosed mice received initial
 

dosages of 50 and 25 mg/kg/day, respectively. Throughout this report
 

the groups receiving the former dosage are referred to as the high
 

dose groups, while those receiving the latter are referred to as the
 

low dose groups. In week 14 the high and low dosages were increased
 

to 70 and 35 mg/kg/day, respectively. The entire period of chemical
 

administration for mice was 78 weeks and this was followed by a 13­

week observation period. The vehicle control mice received corn oil
 

by gavage for 78 weeks. The untreated controls, having the same
 

median birth date as the low and high dose chloropicrin groups, were
 

included in the test for the same period of time as the dosed groups.
 

H. Clinical and Histopathologic Examinations
 

Animals were weighed immediately prior to initiation of the experiment. From the first day, all animals were inspected daily for
 

mortality. Body weights, food consumption, and data concerning appearance, behavior, signs of toxic effects, and incidence, size, and
 

location of tissue masses were recorded at weekly intervals for the
 

first 10 weeks and at monthly intervals thereafter. The presence of
 

11
 
tissue masses was determined by observation and palpation of each
 

animal.
 

A necropsy was performed on each animal regardless of whether it
 

died, was killed when moribund, or was sacrificed at the end of the
 

bioassay. The animals were euthanized by exsanguination under sodium
 

pentobarbital anesthesia, and were immediately necropsied. The histopathologic examination consisted of gross and microscopic examination
 

of major tissues, organs, or gross lesions taken from sacrificed animals and, whenever possible, from animals found dead.
 

Slides were prepared from the following tissues: subcutaneous
 

tissue, lungs and bronchi, trachea, bone marrow, spleen, lymph nodes,
 

thymus, heart, salivary gland, liver, gallbladder and bil e duct
 

(mice), pancreas, esophagus, stomach, small intestine, large intestine, kidney, urinary bladder, pituitary, adrenal, thyroid, parathyroid, testis, prostate, brain, uterus, mammary gland, and ovary.
 

Tissues for which slides were prepared were preserved in 10 percent buffered formalin, embedded in paraffin, sectioned, and stained
 

with hematoxylin and eosin prior to microscopic examination. An
 

occasional section was subjected to special staining techniques for
 

more definitive diagnosis.
 

A few tissues were not examined for some animals, particularly
 

for those that died early. Also, some animals were missing, cannibalized, or judged to be in such an advanced state of autolysis as to
 

12
 
preclude histopathologic interpretation. Thus, the number of animals
 

for which particular organs, tissues, or lesions were examined microscopically varies and does not necessarily represent the number of
 

animals that were placed on experiment in each group.
 

I. Data Recording and Statistical Analyses
 

Pertinent data on this experiment have been recorded in an automatic data processing system, the Carcinogenesis Bioassay Data System
 

(Linhart et al., 1974). The data elements include descriptive information on the chemicals, animals, experimental design, clinical observations, survival, body weight, and individual pathologic results, as
 

recommended by the International Union Against Cancer (Berenblum,
 

1969). Data tables were generated for verification of data transcription and for statistical review.
 

These data were analyzed using the statistical techniques
 

described in this section. Those analyses of the experimental results
 

that bear on the possibility of carcinogenicity are discussed in the
 

statistical narrative sections.
 

Probabilities of survival were estimated by the product-limit
 

procedure of Kaplan and Meier (1958) and are presented in this report
 

in the form of graphs. Animals were statistically censored as of the
 

time that they died of other than natural causes or were found to be
 

missing; animals dying from natural causes were riot statistically
 

censored. Statistical analyses for a possible dose-related effect on
 

survival used the method of Cox (1972) for testing two groups for
 

1
1.33 
equality and used Tarone's (1975) extensions of Cox's methods for
 

testing a dose-related trend. One-tailed P-values have been reported
 

for all tests except the departure from linearity test, which is only
 

reported when its two-tailed P-value is less than 0.05.
 

The incidence of neoplastic or nonneoplastic lesions has been
 

given as the ratio of the number of animals bearing such lesions at a
 

specific anatomic site (numerator) to the number of animals in which
 

that site was examined (denominator). In most instances, the denominators included only those animals for which that site was examined
 

histologically. However, when macroscopic examination was required
 

to detect lesions prior to histologic sampling (e.g., skin or mammary
 

tumors), or when lesions could have appeared at multiple sites (e.g.,
 

lymphomas), the denominators consist of the numbers of animals necropsied.
 

The purpose of the statistical analyses of tumor incidence is to
 

determine whether animals receiving the test chemical developed a significantly higher proportion of tumors than did the control animals.
 

As a part of these analyses, the one-tailed Fisher exact test (Cox,
 

1970, pp. 48-52) was used to compare the tumor incidence of a control
 

group to that of a group of treated animals at each dose level. When
 

results for a number of treated groups, k, are compared simultaneously
 

with those for a control group, a correction to ensure an overall
 

significance level of 0.05 may be made. The Bonferroni inequality
 

(Miller, 1966, pp. 6-10) requires that the P-value for any comparison
 

14 4
be less than or equal to 0.05/k. In cases where this correction was
 

used, it is discussed in the narrative section. It is not, however,
 

presented in the tables , where the Fisher exact P-values are shown.
 

The Cochran-Armitage test for linear trend in proportions, with
 

continuity correction (Armitage, 1971, pp. 362-365), was also used.
 

Under the assumption of a linear trend, this test determined if the
 

slope of the dose-response curve is different from zero at the one-


tailed 0.05 level of significance. Unless otherwise noted, the direction of the significant trend was a positive dose relationship.
 

This method also provides a two-tailed test of departure from linear
 

trend.
 

A time-adjusted analysis was applied when numerous early deaths
 

resulted from causes that were not associated with the formation of
 

tumors. In this analysis, deaths that occurred before the first
 

tumor was observed were excluded by basing the statistical tests on
 

animals that survived at least 52 weeks, unless a tumor was found at
 

the anatomic site of interest before week 52. When such an early
 

tumor was found, comparisons were based exclusively on animals that
 

survived at least as long as the animal in which the first tumor was
 

found. Once this reduced set of data was obtained, the standard procedures for analyses of the incidence of tumors (Fisher exact tests,
 

Cochran-Armitage tests, etc.) were followed.
 

When appropriate, life-table methods were used to analyze the
 

incidence of tumors. Curves of the proportions surviving without an
 

15 5
observed tumor were computed as in Saffiotti et al. (1972). The week
 

during which animals died naturally or were sacrificed was entered as
 

the time point of tumor observation. Cox's methods of comparing
 

these curves were used for two groups; Tarone's extension to testing
 

for linear trend was used for three groups. The statistical tests for
 

the incidence of tumors which used life-table methods were one-tailed
 

and, unless otherwise noted, in the direction of a positive dose
 

relationship. Significant departures from linearity (P < 0.05, two-


tailed test) were also noted.
 

The approximate 95 percent confidence interval for the relative
 

risk of each dosed group compared to its control was calculated from
 

the exact interval on the odds ratio (Gart, 1971). The relative risk
 

is defined as p /p where p is the true binomial probability of the
 

incidence of a specific type of tumor in a treated group of animals
 

and p is the true probability of the spontaneous incidence of the
 

c
 

same type of tumor in a control group. The hypothesis of equality
 

between the true proportion of a specific tumor in a treated group
 

and the proportion in a control group corresponds to a relative risk
 

of unity. Values in excess of unity represent the condition of a
 

larger proportion in the treated group than in the control.
 

The lower and upper limits of the confidence interval of the relative risk have been included in the tables of statistical analyses.
 

The interpretation of the limits is that in approximately 95 percent
 

of a large number of identical experiments, the true ratio of the
 

16
 
risk in a treated group of animals to that in a control group would
 

be within the interval calculated from the experiment. When the
 

lower limit of the confidence interval is greater than one, it can be
 

inferred that a statistically significant result (a P < 0.025 one-


tailed test when the control incidence is not zero, P < 0.050 when
 

the control incidence is zero) has occurred. When the lower limit is
 

less than unity but the upper limit is greater than unity, the lower
 

limit indicates the absence of a significant result while the upper
 

limit indicates that there is a theoretical possibility of the induction of tumors by the test chemical which could not be detected under
 

the conditions of this test.
 

17
 
III. CHRONIC TESTING RESULTS: RATS
 

A. Body Weights and Clinical Observations
 

Mean body weight depression became apparent earlier for male rats
 

than for female rats. Dosed rats continued to gain less weight than
 

control rats for the duration of the bioassay (Figure 1).
 

Clinical signs were observed in the chloropicrin-dosed groups
 

early in the study. Beginning in week 3, a few rats in all dosed
 

groups started to show the following adverse clinical signs: a
 

hunched or thin appearance, squinted or reddened eyes (often with
 

a brown crust or discharge), reddened ears (through week 5 of the
 

study), and occasional abdominal urine stains. These signs were observed with greater frequency in the dosed groups than in the vehicle
 

controls from week 3 through week 50, but occurred at essentially
 

comparable rates in dosed and vehicle control rats during the second
 

year. Respiratory signs, characterized by wheezing, nasal discharge,
 

and/or labored respiration, were observed at a low to moderate incidence in all dosed and control groups during the last 10 months of
 

the study. Clinical signs associated with aging in the laboratory
 

rat were observed at a similar rate in vehicle control and dosed rats
 

during the second year of the study. These signs included localized
 

alopecia, sores on the tail and other parts of the body, fur discoloration, and palpable nodules or tissue masses.
 

18
 
FIGURE 1

 

GROWTH CURVES FOR CHLOROPICRIN CHRONIC STUDY RATS 

19 
B. Survival
 

The estimated probabilities of survival for male and female rats
 

in the control and chloropicrin-treated groups are shown in Figure 2.
 

High mortality was observed in dosed rats of both sexes.
 

For male rats the Tarone tests indicated a significant (P <
 

0.001) association between increased dosage and accelerated mortality
 

when comparing to the vehicle control. By week 16, 38 percent (19/50)
 

of the high dose male rats were dead. Fifty percent of the high dose
 

male rats had died by week 48 and 50 percent of the low dose male rats
 

by week 54. Survival to the end of the study was 6 percent (3/50) for
 

high dose male rats and 8 percent (4/50) for low dose male rats. For
 

both vehicle and untreated control groups at least 50 percent of the
 

animals survived past week 89, despite the sacrifice of five males
 

from each in week 61.
 

Female rats also experienced accelerated, dose-related mortality,
 

as indicated by the significance (P < 0.001) of the Tarone test when
 

comparing to the vehicle control. Fatalities among treated female
 

rats were observed as early as week 2. Fifty percent of the female
 

rats were dead by week 70 in the high dose group and by week 59 in
 

the low dose group. Survival at the end of the study was 20 percent
 

(10/50) for high dose females and 22 percent (11/50) for low dose
 

females. For both vehicle and untreated control groups, at least 50
 

percent of the animals survived over 108 weeks, despite the the sacrifice of five rats from the untreated control in week 61.
 

20
 
FIGURE 2

 

SURVIVAL COMPARISONS OF CHLOROPICRIN CHRONIC STUDY RATS 

21 
C. Pathology
 

Histopathologic findings on neoplasms in rats are tabulated in
 

Appendix A (Tables Al and A2); findings on nonneoplastic lesions are
 

tabulated in Appendix C (Tables Cl and C2) ..
 

A variety of neoplasms were represented among both dosed and
 

control rats. Each of the types of tuners represented has been
 

encountered previously as a spontaneous lesion in the Osborne-Mendel
 

rat. No appreciable difference in the incidence of neoplasms was
 

noted in the control and treated rats in this study.
 

Inflammatory, degenerative, and proliferative lesions seen in
 

control and treated animals were similar in number and kind to those
 

naturally occurring lesions found in aged rats.
 

This study provided no his topat'nolce ic evidence for the carcinogenicity of chloropicrin in rats.
 

D. Statistical Analyses of Results
 

The results of the statistical analyses of tumor incidence in
 

rats are summarized in Tables 3 and 4. The analysis for every type
 

of tumor that was observed in more than 5 percent of any of the
 

chloropicrin-dosed groups of either sex is included. The untreated
 

controls were not used in these analyses since the comparison to the
 

corn oil-gavaged vehicle control was the comparison of choice. Because of the differences between dosing regimens for the low and
 

high dose groups, the Cochran-Armitage test for dose-response trends
 

was not used in these analyses.
 

22 
TABLE 3
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN MALE RATS TREATED WITH CHLOROPICRIN*
 

UNTREATED VEHICLE LOW HIGH 

TOPOGRAPHY '.MORPHOLOGY CONTROL CONTROL DOSE DOSE 

Pituitary: Chromophobe Adenoma 2/20(0.10) 1/20(0.05) 1/50(0.02) 0/49(0.00) 

P Values0 N.S. N.S. ___ 

Relative Risk (Vehicle Control) _— 0.400 0.000 

Lower Limit 0.005 0.000 

Upper Limit 30.802 7.624 

Weeks to First Observed Tumor 90 97 110 

Mammary Gland: Adenocarcinoma or 

Fibroadenoma 2/20(0.10) 2/20(0.10) 0/50(0.00) 0/50(0.00) 

P Values0 N.S. N.S. 

Relative Risk (Vehicle Control)" _— 0.000 0.000 

Lower Limit 0.000 0.000 

Upper Limit 1.345 1.345 

Weeks to First Observed Tumor 90 61 

Treated groups received time-weighted average doses of 25 or 26 mg/kg by gavage.
 

Number of tumor-bearing animals/number of animals examined at site (proportion).
 

°The probability level for the Fisher exact test for the comparison of a treated group with
 

the vehicle control group is given beneath the incidence of tumors in that treated group when
 

P < 0.05; otherwise, not significant (N.S.) is indicated. A negative designation (N) indicates
 

a lower incidence in the treated group(s) than in the control group.
 

The 95% confidence interval on the relative risk of the treated group to the control group.
 
TABLE 4
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN FEMALE RATS TREATED WITH CHLOROPICRIN2
 

TOPOGRAPHY: MORPHOLOGY 

UNTREATED 

CONTROL 

VEHICLE 

CONTROL 

LOW 

DOSE 

HIGH 

DOSE 

Pituitary:

P Values0 

 Chromophobe Adenoma 6/19(0.32) ___ 3/20(0.15) 

—_ 

4/48(0.08) 

N.S. 

3/50(0.06) 

N.S. 

Relative Risk (Vehicle Control) 

Lower Limit 

Upper Limit 

0.556 

0.106 

3.546 

0.400 

0.060 

2.801 

Weeks to First Observed Tumor 109 89 76 72 

Mammary Gland:

Fibroadenoma 

 Adenocarcinoma or 

4/20(0.20) 7/20(0.35) 7/50(0.14) 6/50(0.12) 

P Values N.S. N.S. 

Relative Risk (Vehicle Control)d 

Lower Limit 

Upper Limit 

___ ___ 

0.400 

0.144 

1.187 

0.343 

0.114 

1.061 

Weeks to First Observed Tumor 60 66 95 78 

Treated groups received time-weighted average doses of 20 or 22 mg/kg by gavage.
 

Number of tumor-bearing animals/number of animals examined at site (proportion).
 

£

The probability level for the Fisher exact test for the comparison of a treated group with the
 

vehicle control group is given beneath the incidence of tumors in that treated group when
 

P < 0.05; otherwise, not significant (N.S.) is indicated. A negative designation (N) indicates
 

a lower incidence in the treated group(s) than in the control group.
 

The 95% confidence interval on the relative risk of the treated group to the control group.
 
For both male and female rats none of the Fisher exact tests
 

indicated an association between chemical administration and incidence for any type of tumor. It must be noted, however, that the
 

extreme early mortality observed in dosed animals of both sexes
 

severely restricted the usefulness of these tests for the analysis
 

of tumors.
 

To provide additional insight into the possible carcinogenicity
 

of this compound, 95 percent confidence intervals on the relative
 

risk have been estimated and entered in the tables based upon the
 

observed tumor incidence rates. In all of the intervals shown in
 

Tables 3 and 4, the value one is included; this indicates the absence of statistically significant results. It should also be noted
 

that all of the confidence intervals have an upper limit greater
 

than one, indicating the theoretical possibility of tumor induction
 

in rats by chloropicrin that could not be established under the conditions of this test.
 

25
 
IV. CHRONIC TESTING RESULTS: MICE
 

A. Body Weights and Clinical Observations
 

Progressive mean body weight depression was apparent for female
 

mice treated with chloropicrin. The weight gain patterns for the male
 

mice did not show a consistent weight differential related to chemical
 

administration until week 50 (Figure 3).
 

During the first 6 months of the study the mice in all dosed
 

groups displayed patterns of appearance and behavior that were comparable with those of the controls. Clinical signs often observed in
 

laboratory mice were observed with similar frequency among the dosed
 

and control groups. These signs included sores on parts of the body;
 

localized alopecia; penile, anal, or vulvar irritation with occasional
 

prolapse of these organs; a hunched posture; a bloated appearance;
 

abdominal urine stains; eyes showing cloudiness, redness, and/or
 

discharge; and palpable nodules and/or swollen areas on the body or
 

legs. From week 26 to cessation of chemical administration in week
 

78, observations of a hunched or bloated appearance were noted at a
 

slightly greater frequency in the dosed groups than in the controls.
 

During the last 12 weeks of the study the incidence of these signs
 

increased at a comparable rate in the surviving treated and control
 

animals.
 

B. Survival
 

The estimated probabilities of survival for male and female mice
 

in the control and chloropicrin-treated groups are shown in Figure 4.
 

26
 
FIGURES

 

GROWTH CURVES FOR CHLOROPICRIN CHRONIC STUDY MICE 

27 
FIGURE 4

 

SURVIVAL COMPARISONS OF CHLOROPICRIN CHRONIC STUDY MICE 

28 
In male mice the Tarone test for association between increased
 

dosage and accelerated mortality was significant when comparing to
 

the vehicle control. The departure from linear trend was significant
 

since the mortality in the high dose and untreated control groups was
 

greater than in the low dose and vehicle control groups. In the high
 

dose group, where 13 mice died between weeks 56 and 58, there was no
 

indication that these deaths were from a common cause. Fifty percent
 

of the male mice survived over 60, 91, and 85 weeks for the high dose,
 

low dose, and untreated control groups, respectively. The usefulness
 

of the vehicle control group was greatly restricted due to the sacrifice of 10 male mice in week 56.
 

In female mice the Tarone test for association between increased
 

dosage and accelerated mortality was significant when comparing to
 

the vehicle control. The departure from linear trend was also significant, since the accelerated mortality was primarily in the high dose
 

group starting in approximately week 55. Fifty percent of the female
 

tnice survived over 73, 91, 91, and 90 weeks in the high dose, low
 

dose, vehicle control, and untreated control groups, respectively.
 

There was no evidence that early death was associated with tumor
 

incidence.
 

C. Pathology
 

Histopathologic findings on neoplasms in mice are tabulated in
 

Appendix B (Tables Bl and B2); findings on nonneoplastic lesions are
 

tabulated in Appendix D (Tables Dl and D2).
 

299 
Squamous-cel1 carcinomas of the stomach occurred in two high
 

dose male mice. Within these tumors were both well-differentiated
 

and anaplastic areas. Microscopic invasion through the stomach wall
 

was evident. Both of these tumors metastasized to the pancreas and
 

liver, and one also metastasized to the spleen and a mesenteric lymph
 

node. A squamous-cel1 papilloma of the stomach occurred in one low
 

dose female mouse.
 

In addition to the tumor-bearing animals, acanthosis and hyperkeratosis of the forestomach occurred in 3/46 low dose males, 1/48
 

high dose males, 10/48 low dose females, and 9/48 high dose females.
 

These lesions were not observed in any of the control mice.
 

Other proliferative, inflammatory, and degenerative lesions
 

appeared consistent with lesions that occur spontaneously in this
 

mouse strain, and were observed with similar frequency in the
 

control and treated mice.
 

Proliferative lesions of the stomach were observed, including
 

two carcinomas and a papilloma. These lesions are uncommon in control animals and are considered to be related to the administration
 

of chloropicrin.
 

D. Statistical Analyses of Results
 

The results of the statistical analyses of tumor incidence in
 

mice are summarized in Tables 5 and 6. The analysis for every type
 

of tumor that was observed in more than 5 percent of any of the
 

chloropicrin-dosed groups of either sex is included. The untreated
 

30
 
TABLE 5
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN MALE MICE TREATED WITH CHLOROPICRIN*
 

TOPOGRAPHY : MORPHOLOGY
 

Lung: Alveolar /Bronchiolar
 

Adenoma^
 

P Values0
 

e
 

Departure from Linear Trend
 

Relative Risk (Vehicle Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Liver: Hepatocellular Carcinoma
 

P Values0
 

Relative Risk (Vehicle Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

UNTREATED
 

CONTROL
 

0/15(0.00)
 

1/15(0.07)
 

90
 

VEHICLE
 

CONTROL
 

3/20(0.15)
 

N.S.
 

P = 0.049
 

56
 

2/20(0.10)
 

N.S.
 

56
 

LOW
 

DOSE
 

1/46(0.02)
 

N.S.
 

0.145
 

0.003
 

1.700
 

91
 

4/46(0.09)
 

N.S.
 

0.870
 

0 = 139
 

9.144
 

73
 

HIGH
 

DOSE
 

4/48(0.10)
 

N.S.
 

0.556
 

0.106
 

3.546
 

57
 

2/48(0.04)
 

N.S.
 

0.417
 

0.033
 

5.490
 

91
 

Treated groups received time-weighted average doses of 33 or 66 mg/kg by gavage.
 

Number of tumor-bearing animals/number of animals examined at site (proportion).
 

C

The probability level for the Cochran-Armitage test is given beneath the incidence of tumors
 

in the vehicle control group when P < 0.05; otherwise, not significant (N.S.) is indicated.
 

The probability level for the Fisher exact test for the comparison of a treated group with the
 

vehicle control group is given beneath the incidence of tumors in that treated group when
 

P < 0.05; otherwise, not significant (N.S.) is indicated. For both Cochran-Armitage and Fisher
 

exact tests a negative designation (N) indicates a lower incidence in the treated group(s) than
 

in the control group.
 

The 95% confidence interval on the relative risk of the treated group to the control group.
 

e

The probability level of the test for departure from linear trend is given beneath the control
 

group when P < 0.05.
 
TABLE 6
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN FEMALE MICE TREATED WITH CHLOROPICRIN*
 

TOPOGRAPHY : MORPHOLOGY 

UNTREATED 

CONTROL 

VEHICLE 

CONTROL 

LOW 

DOSE 

HIGH 

DOSE 

Lung: Alveolar/Bronchiolar Adenoma 

P Values0 

1/20(0.05) 1/19(0.05) 

N.S. 

3/49(0.06) 

N.S. 

6/48(0.13) 

N.S. 

Relative Risk (Vehicle Control) 

Lower Limit 

Upper Limit 

1.163 

0.104 

59.809 

2.375 

0.325 

106.788 

Weeks to First Observed Tumor 90 91 91 73 

Treated groups received time-weighted average doses of 33 or 66 mg/kg by gavage.
 

Number of tumor-bearing animals/number of animals examined at site (proportion).
 

"The probability level for the Cochran-Armitage test is given beneath the incidence of tumors
 

in the vehicle control group when P < 0.05; otherwise, not significant (N.S.) is indicated.
 

The probability level for the Fisher exact test for the comparison of a treated group with the
 

vehicle control group is given beneath the incidence of tumors in that treated group when
 

P < 0.05; otherwise, not significant (N.S.) is indicated. For both Cochran-Armitage and Fisher
 

exact tests a negative designation (N) indicates a lower incidence in the treated group(s) than
 

in the control group.
 

The 95% confidence interval on the relative risk of the treated group to the control group.
 
controls were not used in these analyses since the comparison to the
 

corn oil-gavaged vehicle controls was the comparison of choice.
 

None of these statistical tests indicated an association between
 

chemical administration and tumor incidence for either male or female
 

mice. Additional analyses were performed based exclusively on mice
 

which survived at least 52 weeks, but none of the additional time-


adjusted statistical tests were significant.
 

In males an unusual tumor type--squamous-cell carcinoma of the
 

forestomach—was observed in 2/48 high dose mice but in none of the
 

other male mice. In historical data collected by Hazleton Laboratories for the NCI Bioassay Program 1/180 vehicle control male B6C3F1
 

mice had either a squamous-cell papilloma or a squamous-cell carcinoma of the stomach. Under the assumption of a binomial distribution,
 

with a probability of spontaneous incidence of 1/180, the probability
 

that 2 or more of 48 mice would have that tumor was P = 0.029. This
 

marginal result was not significant when the Bonferroni criterion was
 

applied.
 

To provide additional insight into the possible carcinogenicity
 

of this compound, 95 percent confidence intervals on the relative
 

risk have been estimated and entered in the tables based upon the
 

observed tumor incidence rates. In all of the intervals shown in
 

Tables 5 and 6, the value one is included; this indicates the absence
 

of statistically significant results. It should also be noted that
 

all of the confidence intervals have an upper limit greater than one,
 

333 
indicating the theoretical possibility of tumor induction in mice by
 

chloropicrin that could not be established under the conditions of
 

this test.
 

34
 
V. DISCUSSION
 

Because of elevated mortality, it is unlikely that an adequate
 

number of rats lived long enough to be at risk from late-appearing
 

tumors. No statistically significant increase in tumor incidence
 

was observed among mice dosed with chloropicrin, but a few unusual
 

neoplasms were noted.
 

In mice, proliferative lesions of the squamous epithelium of the
 

forestomach were observed in dosed animals but not in controls. In
 

addition to hyperplasias (acanthosis and hyperkeratosis), two carcinomas were observed in male mice and a papilloma was observed in a
 

female mouse. Both carcinomas metastasized to the pancreas and liver
 

and one carcinoma also metastasized to a mesenteric lymph node. For
 

a binomial distribution with a probability of spontaneous incidence
 

of these neoplasms of 1/180, the probability that 2 or more of 48
 

mice would have one of these neoplasms was P = 0.029; this marginal
 

result was not significant under the Bonferroni criterion.
 

The method of chloropicrin administration might have played a
 

role in inducing these neoplasms. Chloropicrin is a mucous membrane
 

irritant (Pesticide Information Manual, 1968) and proliferative
 

lesions of the forestomach have been noted in other bioassays where
 

chemical irritants and carcinogens have been administered by gavage.
 

A variety of neoplasms were observed in rats, but no significant
 

differences in the incidence of these tumors were detected between
 

the dosed and the control groups. However,, because of high early
 

35
 
mortality, the number of animals that survived long enough to be at
 

risk from late-appearing tumors does not provide enough data for the
 

results to be considered as conclusive.
 

The bioassay of chloropicrin using Osborne-Mendel rats did not
 

permit an evaluation of careinogenicity because of the short survival
 

time of dosed animals. The bioassay of chloropicrin using B6C3F1
 

mice did not provide conclusive statistical evidence for the carcinogenicity of this compound.
 

36
 
VI. BIBLIOGRAPHY
 

Armitage, P., Statistical Methods in Medical Research, Chapter 14.
 

J. Wiley & Sons, New York, 1971.
 

Berenblum, I., editor, C a r c i no ge n i c i ty T e sting. International Union
 

Against Cancer, Technical Report Series, Vol. 2. Internatioral
 

Union Against Cancer, Geneva, 1969.
 

Chemical Abstracts Service, The Chemical Abstracts Service (CAS)
 

Ninth Collective Index, Volumes 76-85, 1972-1976. American
 

Chemical Society, Washington, B.C., 1977.
 

Cox, D.R., Analysis of Binary Data, Chapters 4 and 5. Methuen and
 

Co., Ltd., London, 1970.
 

Cox, D.P., "Regression Models and Life-Tables." Journal of the Royal
 

Statistical Society, Series "B" 34:187-220, 1972.
 

Feldmesser, J.. Technical Advisor, Nematicides. Agricultural Research
 

Service, U.S. Department of Agriculture, Beltsville, Maryland.
 

Personal communication, January 25, 1972.
 

Gart , J.J., "The Comparison of Proportions: A Review of Significance
 

Tests, Confidence Limits, and Adjustments for Stratification."
 

International Statistical Institute Review 39:148-169, 1971.
 

Kaplan, E.L., and P. Meier, "Nonparametric Estimation from Incomplete
 

Observations." Journal of the American Statistical Association
 

5_3 :45 7-481 , 1958~
 

Linhart, M.S., J.A. Cooper, R.L. Martin, N.P. Page, and J.A. Peters,
 

"Carcinogenes is Bioassay Data System.." Computers and Biomedical
 

Research 7_: 2 30-248, 1974.
 

Matsumura, F., Toxicology of Jlnsecticides ., Plenum Press, New York,
 1975>
 

Miller, R.G., Simultaneous Statistical Inference. McGraw-Hill Book
 

Co., New York. 1966." ~
 

Pesticide Chemicals Official Compendium. Association of the American
 

Pesticide Control Officials , Inc., Topeka, Kansas, 1966.
 

Pesticide Information Manual. Miscellaneous Publication No. 8,
 

Virginia Cooperative Extension Service, Virginia Polytechnic
 

Institute , Biacksburg, Virginia, 1968.
 

37 
Reuber, M.D., and E.L. Glover, "Cirrhosis and Carcinoma of the Liver
 

in Male Rats Given Subcutaneous Carbon Tetrachloride." Journal
 

of the National Cancer Institute 44:419-423, 1970.
 

Rogers, J.P., Agricultural Pesticide Chemicals Handbook, 1966.
 

University of Missouri Extension Division, Columbia, Missouri,
 

1966.
 

Saffiotti, U., R. Montesano, A.R. Sellakumar, F. Cefis, and D.G.
 

Kaufman, "Respiratory Tract Carcinogenesis in Hamsters Induced
 

by Different Numbers of Administration of Benzo (a) Pyrene and
 

Ferric Oxide." Cancer Research 32:1073-1079, 1972.
 

Tarone, R.E., "Tests for Trend in Life-Table Analysis." Biometrika
 

^:679-682, 1975.
 

U.S.	 Departmen o t f Health, Education and Welfare, Report of the
 

Secretary's Commission on Pesticides and Their Relationship
 

to Environmental Health. Parts I and II. E. M. Mrak. NTIS
 

No. PB-251-360, 1969.
 

U.S.	 International Trade Commission, Synthetic Organic Chemicals.
 

United States Production and Sales, 1974. USITC Publication
 

776, Washington, D.C., 1976.
 

38
 
Review of the Bioassay of Chloropicrin* for Carcinogenicity
 

by the Data Evaluation/Risk Assessment Subgroup of the
 

Clearinghouse on Environmental Carcinogens
 

January 18, 1978
 

The Clearinghouse on Environmental Carcinogens was
 

established in May, 1976 under the authority of the National
 

Cancer Act of 1971 (P.L. 92-218). The purpose of the Clear­ inghouse is to advise on the National Cancer Institute's
 

bioassay program to identify and evaluate chemical carcino­ gens in the environment to which humans may be exposed. The
 

members of the Clearinghouse have been drawn from academia,
 industry, organized labor, public interest groups, State
 

health officials, and quasi-public health and research
 

organizations. Members have been selected on the basis of
 

their experience in carcinogenesis or related fields and,
 collectively, provide expertise in organic chemistry, bio­ statistics, biochemistry, toxicology, pathology, and epide­ miology. Representatives of various Governmental agencies
 

participate as ad hoc members. The Data Evaluation/Risk
 

Assessment Subgroup of the Clearinghouse is charged with
 

the responsibility of providing a peer review of NCI bioassay
 

reports on chemicals studied for carcinogenicity. In this
 

context, below is the edited excerpt from the minutes of
 

the Subgroup's meeting at which Chloropicrin was reviewed.
 

The primary reviewer said that the bioassay was
 

inadequate to allow a conclusion to be reached with respect
 

to the carcinogenicity of Chloropicrin in treated rats. In
 

the mice, there was no statistical evidence to conclude a
 

carcinogenic effect produced by the chemical. However,
 the primary reviewer expressed concern that the study may
 

represent a "false negative" based on the known biological
 

activity of Chloropicrin and the fact that it is fairly
 

persistent. He recommended that Chloropicrin be retested.
 

Due to its extreme toxicity, a Subgroup member said
 

that it is not used widely by itself. It has been employed
 

as a warning agent because of its lacrimatory properties.
 

It was moved that the report be accepted as written
 

but that Chloropicrin be referred to the Chemical Selection
 

Working Group for possible retest. The motion was seconded
 

and approved by all the Subgroup members except Dr. Rowe,
 who abstained.
 

39
 
Members Present Were:
 

Arnold Brown (Acting Chairman), Mayo Clinic
 

Lawrence Garfinkel, American Cancer Society
 

Joseph Highland, Environmental Defense Fund
 

Charles Kensler, Arthur D. Little Company
 

Verald Rowe, Dow Chemical, U.S.A.
 Sheldon Samuels, Industrial Union Department, AFL-CIO
 

Louise Strong, University of Texas Health Sciences Center
 

Sidney Wolfe, Health Research Group
 

*	 Subsequent to this review, changes may have been made
 

in the bioassay report either as a result of the
 

review or other reasons. Thus, certain comments and
 

criticisms reflected in the review may no longer be
 

appropriate.
 

40
 »U.S. GOVERNMENT PRINTING OFFICE: 1978 260-899/3059 1-3 
APPENDIX A
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS
 

IN RATS TREATED WITH CHLOROPICRIN
 

TABLE Al

 

SUMMARY OF THF, INCIDENCE OF NEOPLASMS IN MALE RATS TREATED WITH CHLOROPICRIN 

-Zll^Zll"Z.----~ 

CONTROL (UNTR ) CONTROL (VEH ) LOU DOSE 

01-031M 01-0<»1N 01-OU2M 

ANIMAL S INITIALL Y I N STUDY 20 20 50 

ANIMAL S HECROPSIED 20 20 50 

ANIMAL S EXAMINE D HISTOPATHOLOGICA LLY ** 20 20 50 

INTEGUMENTAR Y SYSTE M 

*SUBCUT TISSUE (20) (20.1 (50) 

FIflBCSARCCM A 1 (5K) 

LIPOSARCOMA 1 (5*) 

8ESEIRATCR Y SYSTE M 

NCN S 

HEMATOPCIE1I C SYSTE M 

• MULTIPL E ORGAN S (20) (20] (50) 

LYBPHCCYII C LEUKEMI A 1 (5X ) 

*SPIEE N (20) (20 | (50) 

HEMANGICSABCON A 1 (2%) 

CIRCULATOR Y SYSTEM 

NON E 

EIGESTIV E SYSTEM 

• SALIVAR Y GLAN D (It) (16 ) (20) CARCINOM A ,NOS 1 (7*) 

fSKAL L INTESTIN E (20) (20) (50) 

FIBRCSARCCM A 1 (5X ) 

URINAR Y SYSTE M 

NON E 

« NUMBE R OF ANIMAL S HITH TISSUE EXAMINE D MICROSCOPICALL Y 

« NUMBE R OF ANNAL S NECROPSIE D 

**EXCLUDES PARTIALLY AUTCLYZED ANIMALS 

HISH DOSE 

01-OU3H 

50 

50 

50 

(50) 

(50) 

(l»9) 

(11) 

(19) 

A-3 
TABLE Al (CONTINUED) 

CONTSOL(OMtB) CONTROL (VEH) LOW DCSE HIGH DOSE 

01-031N 01-011!! 01-OI)2« 01-OU3M 

ENCCCBIN E SYSTE M 

*PI'K)IIARY (20) (20) (50) C*9) 

CH80MCEHOBE ACENOilA 2 (10X) 1 <5X) 1 (2%) 

•ADRENAL 

CCBTICAL CARCINOM A 

(20) 

2 (10X) 

(20) C*9) (U9) 

•THYROID 

FCLIICULAR-CELL ADENOM A 

(19) (20) (i»9) 

1 (2X) 

(l«7) 

FOLIICULAR-CELL CARCINOMA 1 <5X) 1 (5*> 

REPRCtUCTIV E SYSTEM 

•HAKNAR Y GLAND (20) (20) (50) (50) 

ADESCCARCIHCHA , NOS 1 (5X) 2 (10S) FIBROADENOMA 1 (5X) 

NERVCU S SYSTE M 

NONE 

SPECIA L SEKS E ORGAN S 

NONE 

MUSCUtOSKELETA L SYSTE M 

NON E 

BODY CAVITIE S 

DON E 

A LL OTHER SYSTEMS 

_ flONE __ __ __ 

* NUMBE R OF ANIMAL S HUH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBE R CF ANIMAL S NEC80PSIED 

A-4 
TABLE A1 (CONCLUDED) 

CONTROL (UNta) CONTROL (VEH) LCM DCSE HIGH DOSE 

01-0 3 1M 01-041N 01-OU2 H 01-OU3M 

ANIMA L DISPOSITION SUMMAR Y 

AHIMAI S INITIALL Y I N STUD Y 20 20 50 50

 

NATURA L DEATH * I 1 18 U6 47

 

MOaiBTIKD SACRIFICE 

SCHEDULE D SACRIFICE 5

 

ACCIDENTALL Y KILLE D 

TESMINA L SACRIFICE 14 2 4 3 

ANIKA L MISSIN G 

a ! INCLUDE S MJTOLYEE D ANISAL S 

TUMC ? SUfHAR Y 

TOTA L ANIMAL S WIT H PSIMAH Y TUMORS* 6 5 3

 

TOTA L PRIMAR Y TUMOR S 11 5 3 

TOTAL ANIMAL S WIT H BENIG N TUHCR S 2 1 2

 

TOTAL BENIG N TUMOR S 3 1 2 

TOTA L ANIMAL S WIT H MALIGBAN T TUMOE S 6 it 1 

TCTAL MALIGNAN T TUNO8 S 8 i* 1

 

TOTA L ANIMAL S WIT H SECONDAR Y TUMOSS K 

TOTA L SECONDAR Y TIJMOHS 

TOTA L ANIMAL S WIT H TUMOR S UNCEHTAIN ­

BENIG N O R MALIGNAN T 

IOJA L IINCEHTAI N TU>!Oas 

TCTA L AMMAL S WII H TUMOE S UNCERIAIN ­

PSIBAE Y Ofi METASTATIC 

TOTA L UNCE3TAI B TUM03 S 

* PHIMAH Y TUMORS : AL L TUMOR S EXCEPT SECONDAR Y TUMORS 

t SECOMDAS I TUMORS : METASTATIC TUMOR S OH TUMOR S INVASIV E INTO AN ADJACENT ORGA N 

A-5 
TABLE A2

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE RATS TREATED WITH CHLOROP1CR1N 

HIG H DO SE 

01-OU5 F 

50 

50 

50 

(50) 

2 (0% ) 

CiO) 

(50) 

(50) 

<U9 ) 

(50) 

CONTRO L (UNTS ) CONTRO L (VEH ) 

0 1-031F 0 1-Otl F 

ANIMAL S INITIALL Y I N STUD Y 20 20 

ANIKAI S NECRCFSIE D 20 20 

ANIMAL S EXAMINE D HIST09ATHOLOGICALLY* * 20 20 

IHTEGUHENTAH Y SYSliM 

• SUBCU T TISSUE (20) (20)

 FIBSOM A

 

RESPIRATOR Y SYSTE M 

NONE 

HEMATOPOIE1IC SYSTE M 

NON E 

CIBCULAIORY SYSTEM 

tHEAST (20) (20)

 BIXIC IOBCR , MI1ASTAIIC 1 <5« )

 

DIGESTIVE SYSTEM 

•LIVER (20) (20) HIEATOCELLULA R CARCINOM A 1 (5%)

 CORTICAL CARCINOMA , METASTATIC

 

tPANCBEAS (20) (20)

 IIEOM A

 

MES01HELIOKA, METASTATIC 

*SMAII INTESTINE (20) (20) IEIOHYCSABCOM A

 

URINAR Y SYSTEM 

#KIDKEY (20) (20) CORTICA L CARCINOMA . METASTATI C .

* NUMBE R O F ANIMAL S WITH TISSU E EXAMINE D MICROSCOPICALL Y 

* NUMBE R CF ANIMAL S NECBOPSIED 

**EXCLUDES PARTIALLY AUTCLYZED ANIMALS. 

A-6 

LCii DCSE 

0 1-04UF 

50 

50 

50 

(50) 

(1?) 

C*9) 

1 (2X) 

(50) 

1 (2«) 

1 (2*) 

(50) 

1 (2%) 

(49) 

- 1 i»i ­
TABLE A2 (CONTINUED) 

CONTRO L (UNTR ) CONTRO L (VEH ) L CH DCSE HIG H DOSE 

01-031F 01-041F 01-04U F 01-OI45 F 

MIXE D TUMOH , MALIGNAN T 1 (5*) 

H A M A Ei TOM A + 2 (10*) 1 (5*) 

ENEOC3IK E SYSTE M 

• PITUITAR Y (19) (201 (148) (50) 

CHRCKCFH03 i ADENOM A 6 (32%) j (15X ) it (8*) 3 (6X ) 

• ADRENA L (20) (20 ) (U9 ) (50) 

fCEIICA L CABCINCK A 1 (2%) 

•THY SCID (20) (20) («9) (46) 

FCLIICULAE- C EL L CARCINOM A 1 (2% ) C-CELL ADBNOM A 2 (10% ) 1 (2%) 

C-CEL L CARCINOM A 2 (10*) 

• PANCREATI C ISLET S 

ISIIT-CEI.I. ABENCH A 

(20 ) 

1 (5*) 

(20; (50; (50 ) 

REPRODUCTIV E SYSTE M 

*MAMMAR Y GLAN D (20) (20) (50) (50) 

ADENOCARCINOMA , NO S 2 (10*) 1 (5*) 1 (2% ) 

FIBBOADENCM A 2 (10%) 7 (35*) 7 (14%) 5 (10%) 

• UTE3U S (20) (20) (19) (50) ACENCCAPCINCM A , NO S 1 (2*) ENCOM2IRIA L STBOMAL POLYP 1 <5X) HEMANGIOM A 1 (5% ) 

• UTERUS/ENDCSBiaiU M (20) (20 ) (K9 ) (50) CAECINOM A ,NCS 1 (2*) 

•CVASY/CVICUCT (20) (20) (49) (50) 

i'ESCTHELICM A , METASTATI C 1 (2% ) 

• OVAR Y (20 ) (19 ) (49 ) (50) 

ACZSCCARC I NCM A , NOS , MZ1ASTATI C 1 (2X ) 

CYSTADENOCASCINOMA , SOS 1 (5*) GSANOLOSA-CEL L TUMO R 1 (5%) 

MESOTHELIOMA , METAST'TIC 1 <2S) 

NERVOU S SYSTE M 

NON E 

* NUMBE R O F ANIMAL S WIT H TISSUE EXAMINE D MICROSCOPICALL Y 

* NUMBE R CF ANIMAL S NECEOPSIED 

+ THI S IS CONSIDERED TO BE A BENIGN FORM OF THE MALIGNANT MIXED TL'MOR OF THE KIDNEY AND CONSISTS OF PROLIFERAT1VE 

LIPOCYTFi;-;, Tl'BULAR STRUCTURES, FI8ROBLASTS, AND VASCULAR SPACES j N VARYING PROPORTIONS. 

A-7 
TABLE A2 (CONTINUED) 

COKTHOL (UNTR) CONTROL (VEH ) 

01-031F 0 1-011F 

SPECIA L SENS E ORGAN S 

N JN E 

MUSCULCSKELETA L SYSTE M 

NON E 

BOC1 CAVITIE S 

*PEHITONEU M (20 ) (20 ) 

tESCTHELIOMA , MALIGNAN T 

A L L OTHE R SYSTEM S 

NON E 

ANIfA L DISECSI1IC N SUMMAR Y 

ANIMAL S INITIALL Y I N STUD Y 20 20

 

NATURA L DEATH S 2 9 

MORIBUN D SACRIFICE 1

 

SCHEDULE D SACRIFICE 5 ACCIDENTALL Y KILLE D

 

TERMINA L SACRIFICE 13 10 

ANICA L MISSIN G 

T> INCLUDE S AUTOLYZE D ANIMAL S 

» NUMBE R O F ANIMAL S HIT H TISSUE EXAMINE D MICROSCOPICALL Y 

* NUMBE R OE ANIMALS NEC80PSIED 

LCX DCSE 

01-OHU F 

(50) 

1 (2% ) 

50 

39 

11 

HUH DOSE 

01-1U5F 

(50 ) 

50 

«0 

1 0 

A-8 
TABLE A2 (CONCLUDED) 

CONTROL (UNTR) CONTROL (VEH) 1C* DCSS 

01-031F 01-041F 01-OUU F 

TUHOR SUMMAR Y 

TOTA L ANIMAL S HU H PRIMAR Y TUMORS * in 11 1<4 

TCTA L PRIMAR Y IOMOBS 21 lit 17 

TOTA L ANIMAL S WIT H BENIG N TUMORS 12 1 1 10

 

TOTA L EENIG N TBMOHS 11 12 12

 

TOTAL ANIMAL S WITH MALIGNAN T TUMOR S 6 1 5

 

TOTAL MALIGNAN T TUMORS 7 1 5 

TOTAL ANIMAL S WIT H SECONDAR Y TUMORS * 1 3

 TCTA L SECONDAR Y TUMOR S 1 6

 

TOTA L ANIMAL S WITH TUMOR S UNCE3TAIN ­

BEMIG N O H MALIGNAN T 1 TOTAL UNCERTAI N TUMOBS 1

 

TCTA L ANIMAL S KIT H TtlMORS UNCERTAIN ­

P5IMAE Y C8 HETASTATIC 

TOTA L UNCERTAI N TUMOR S 

* PRIMAR Y TUMORS : ALL TUMOR S EXCEPT SECONDAR Y TUMORS 

* SECONDAR Y TUMORS : SETASTATIC TUMORS OH TUMORS INVASIV E INTO AN ADJACENT ORGA N 

HIGH DOSE 

01-OK5F 

9 

13 

9 

11 

2 

2 

A-9 

APPENDIX B
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS
 

IN MICE TREATED WITH CHLOROPICRIN
 

TABLE Bl

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE TREATED WITH CHLOROPICRIN 

LOW DCSE 

02-M012 

50 

1 

46 

«6 

(U6 ) 

1 (21) 

(1*6) 

1 (2% ) 

1 (2% ) 

(i»6 ) 

(16) 

(«6 ) 

U (9*) 

(16) 

HIGH DOSE 

02-MOU 3 

50 

48 

«7 

(48) 

4 (8S) 

1 (2*) 

(48) 

1 (2X ) 

(46) 

1 (2*) 

(47) 

1 (2S) 

(48) 

2 (4X ) 

2 (4%) 

(46) 

2_i43L. 

ANIMAL S INITIALL Y I N STUD Y 

ANIHAI S KISSIN G 

ANIdBI S NECHOPSIE D 

ANIMAL S EXAMINE D HISTOPATHOLOGICAL L Y** 

INTEGUMENTAR Y SYSTE M 

NON E 

32SPI8AT03 Y SYSTE M 

(tLUN G 

ALVECLAB/BBCNCHIOLA H ADENOM A 

ALVEOI. AR/BBONCHIOLA S CAHCINOM A 

HEHATOPOIETI C SYSTE M 

• MUllIFl E OSGAN S 

"ALIG.LYMPHOHA , LYMPHOCYTI C TYPE 

MALIG.LYMPHOMA , HISTIOCYTI C TYP E 

*SPI.EE N 

SCUAMCU S CELL CARCINOMA ,

*«ESENTEBI C L . NCC E 

SGUAMCO S CELL CARCINOMA ,

CIECUIATOE Y SYSTE M 

NON E 

DIGESTIV E SJSTEM 

#LI V E R 

SCUAHCU S CELL CARCINOMA , HEPATOCELLULA R CARCINOM A 

tPAKCHEA S 

 METAST A 

 METAST A 

 MSIAST A 

SiUAMCUS_CELL_CAECINgMA i _«ElAST A 

CONTSOL (UNTR) 

02-MOH 1 

20 

18 

15 

(15) 

(18) 

(15) 

(15) 

(15) 

1 (7X ) 

(14) 

CONTROL (VEH) 

02-M03 1 

20 

20 

20 

(20 ) 

3 (15X) 

(20) 

1 (5S) 

(20 ) 

(19 ) 

(20 ) 

2 (10X ) 

(20) 

* NUMBE R O F ANI'IAL S WIT H TISSUE EXAMINE D MICROSCOPICALL Y 

* NUMBE B O E ANIMAL S NECBOPSIE D 

**EXCLUDE S PARTIALLY AUTOLYZED ANIMAL S 

B-3 
TABLE Bl (CONTINUED) 

CONTROL (UNT8) 

02-M04 1 

CONTROL (VEH) 

02- MO 31 

LCD DOSE 

02-KOU 2 

HIGH DOSE 

02-MOU 3 

*STC MACb l 

SQUAMCU S CILL CARCINOM A 

(15) (20) C*6) C*8) 

2 C*X) 

URINS3 Y S 1ST EM 

HKIDNE Y 

5C.UAMCUS CELL CARCINOMA, METAST A 

(15 ) (20) (46) («B) 

1 (2%) 

ENDOCRIN E SYSTE M 

NON E 

HclPBODUClIV E

NON 2 

 SYSTE1 

NERVOU S

NCS E 

 3YSTEK 

SPECIA L

NCli E 

 SE^S E CKGAN S 

MUSCaiCSKELEIA L

SCN E 

 SYSTE M 

BODY CAVITIE S 

NON E 

A L L CTHI B SYSTEM S 

NON E 

» NUHBE H C F ANIMAL S WIT H TISSU E

* NUHBE B O E ANIMAL S IJECBOPSIED 

 EXAMINE E MICROSCOPICALL Y 

B-4 
TABLE Bl (CONCLUDED) 

CONTROL (UNTR ) 

02-MO U1 

CONTRO L (VEH ) 

02-B031 

L C H DCSE 

C2-MOU 2 

HIG H DO SE 

02-MOt3 

ANIMA L DISPOSITIO N SUMMAR Y 

ANIMAL S INITIALL Y I N STUD Y 

NATURA L DEATH S 

MORIBUN D SACRIFIC E 

SCHEDULE D SACRIFICE 

ACCIDENTALL Y KILLJ D 

TERMINA L SACRIFICE 

ANIMA L MISSIN G 

20 

12 

8 

20 

U 

10 

6 

50 

16 

33 

1 

50 

37 

13 

S INCLUDE S AUTOLYZE D ANIMAL S 

TUBO B SUfFAR Y 

TOTA L ANIMAL S WITH PRIMAR Y

TOTA L PRIMAR Y TUMOR S 

 TUMORS * 1 

1 5 6 

7 

7 

9 

1 0 

TOTA L ANIMAL S HITH BENIG N

TOTAL BENIG N TUMOR S 

 TUMC3 S 3 

3' 

1 

1 

4 

I! 

TOTA L ASIMAL S HIT H MALIGNAN T

TCTA L HALIGNAN T TUMOB S 

 TUMOR S 1 

1 

3 

3 

6 

6 

5 

6 

TOTA L ANIMAL S HITH SECONDAR Y

TCTA L SECONDAR Y TU10RS 

 TUMORS * 2 

7 

TOTA L ANIMAL S WIT H TUMOR S

BENIG N O R MALIGNAN T 

TOIA L UNCERTAI N TUMOR S 

 UNCERTAIN ­

TCTA L AKIMAL S HIT H TUMOR S

PBIBAF Y C E HETASTATI C 

TOTA L UNCERTAI N TUMOR S 

 UNCERTAIN ­

* PHIBAB Y TUMORS : AL L TUMOR S EXCEP T SECONDAR Y TUMORS 

* SECONDAR Y TUMORS : HETASTATIC TUMOR S O H TUMOR S INVASIV E INTO A N ADJACEN T ORGA N 

3-5 
TABLE B2

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICE TREATED WITH CHLOROPICRIN 

CONTROL (UNTR) CONTROL (VEH) 

02-FOU1 02-F031 

ANIMAL S INITIALL Y I N STUD Y 20 20 

ANINAI S NECECPSIE D 20 19 ANIMAL S EXAMINE D HISTOPATHOLOGICALLY** 20 19 

INTEGUBEKTAR Y SYSTE M 

*SOBCU1 1ISSUE (20) (19) 

SQUAMOU S CELL CARCINOM A 

LEIOMYOSARCOM A 1 (5%) 

RESPIEATCB Y SYSTE M 

#LUN G (20) (19) 

ALVECLAB/BHCNCHIOLA R ADENOM A 1 (5«) 1 (5X) 

HEHATOPOIiTIC SYSTE M 

'MULTIPL E ORGAN S (20) (19) 

MAIIG. LY UPHOMA, LYMPHGCYT1C TYPE 1 (5%) 

MALIG.LYMPHOMA , HISTIOCYTIC TYP E 2 (10*) 1 (5X) 

tSPLEE N (20) (19 ) 

IEIOKYCSABCCMA , METASTATI C 1 (5%) 

tMESENTEEIC L. NODS (20) (19) 

IBIOKYCSABCCMA , METASTA1I C 1 (51) 

CIRCULATCH Y SYSTE M 

NON S 

DIGESTIV E SYSTE M 

ILIVE B (20) (19) 

HIPATOCELLULA R CARCINOM A 1 (5%) 

tSTOWACH (20) (19) 

SiUAflCU S CELL PA£ILLOMA 

# NUPIBE E OF ANIMAL S KITH TCSS'J E SiCA.IISED :i ICSOS COf ICftLI. i 

* NUMBE E OE ANIMAL S BECSOPS1ED 

**EXCLUDESPARTIALLY AUTOLYZEDANIMALS 

LCK DOSE 

02-FOU1*

50 

50 

U9 

(50) 

<U9 ) 

3 (6*) 

(50) 

1 (2%) 

3 <6S) 

(49) 

(16) 

(«9) 

(U8 ) 

1 12*1 

HIGH DOSE 

 02-FOU 5 

50 

H8 

48 

(48) 

1 <2X ) 

(48) 

6 (13%) 

(48) 

2 (4*) 

(48) 

(44 ) 

(48) 

(48) 

B-6 
TABLE B2 (CONTINUED) 

UBISAH Y SYSTE M 

NON E 

ENDCCEIN E SYSTE M 

• PITUITAR Y 

CHRCKCEHCB S ADENCN A 

HTHYRCI D FCLLICULAH-CEL L ADENOM A 

RHPHOCOCTIV E SYSTE M 

lUTEIiU S 

ENDOMETRIA L S1BCMAL EOLYP 

• OVAB Y 

CYSTACESCMA , NO S 

NEHVCU S SYSTEfl 

NJN E 

SPECIA I SENS E ORGAN S 

*KAHDERIAN GLAN D 

ADEKCKA , SOS 

MUSCULOSKELETA L SYSTEM 

NON E 

BODY CAVITIES 

NON E 

A L L OTHE R SYSTEM S 

NOHE_ _ „ _ 

# NU1BE8 OF ANIMAL S WITH TISSUE

* SDMBE B CF AHINAL S NECROPSIE D 

CONTEOLIDNTR) CONTROL (VEH) 

02-POU1 02-F031 

(17) (18) 

1 <6X) 

(16) (16) 

(20) (19) 

(20) (19) 

1 (5S) 

(20) (19) 

1 (5*) 

 EXAMINED MICROSCOPICALLY 

LCH DOSE 

02-F04U 

("»») 

C"*) 

("9)

1 (2*) 

(»9) 

(50) 

HIGH DOSE 

02-FOU5 

(32) 

(36) 

1 (3«) 

(U7) 

(17) 

(13) 

B-7 
TABLE B2 (CONCLUDED)
 

HIG H DOSE 

02-FOU5 

50 

32 

18 

10 

10 

7 

7 

3 

3 

ANIMA L DISPOSITION SUMMAR Y 

ANISALS INITIALLY IN SIUDY 

NATURA L DEATHS 

MORIBUN D SACRIFICE 

SCHEDULE D SACRIFICE 

ACCIDENTALL Y KILLE D 

TERMINA L SACRIFICE 

ANIRA L >IISSING 

3 INCLUDES AUTOLYZE D ANIMAL S 

TUfiC R SUMKAR Y 

TOTAL ANIMAL S KITH PRIMARY

TOTA L PRIMAR Y TUMOR S 

TOTA L ANIMAL S SITH BENIG N

TOTAL BENIGN TUMORS 

 TUMORS* 

 TUHCR S 

TCTA L ANIMAL S HU H MALIGNAN T TUMOR S 

TOTA L MALIGNAN T TUMOB S 

TOTA L ANIMAL S WITH SECONDAR Y TUMORS * 

TCTAL SECONDARY TUBORS 

TOTA L ANIMAL S WIT H TUMOR S UNCERTAIN ­

BENIG N O R MALIGNAN T 

TOTAL UNCERTAI N TUMOR S 

TCTA L ANIMAL S HIT H TUMOR S UNCERTAIN ­

PBIMAE Y CR METASTATIC 

TOTAL UNCERTAI N TUMORS 

CONTRO L (UNTR ) 

02-FOt 1 

20 

3 

1 

16 

7 

7 

2 

2 

5 

5 

1 

2 

CONTRO L (VEH ) LC» DOSE 

02-F031 02-FOUU 

20 50 

2 7 

1 

18 t2 

4 9 

a 9 

3 5 

3 5 

1 a 1 i» 

* PRIMAR Y TUMORS : ALL TUSOHS EXCEPT SECONDAR Y TUMOR S 

* SECONDAR Y TUMORS : METASTATIC TUMOR S 08 TUPIOE S IHVASIV E INTO AN ADJACENT ORSA I 

B-8 
APPENDIX C
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN RATS TREATED WITH CHLOROPICRIN
 

TABLE Cl

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS TREATED WITH CHLOROPICRIN 

CONTROL (HNTR) CONTBOL (VEH ) 

0 1-031M 01-041(1 

ANIMAL S INITIALL Y I N STUD Y 20 20 

ANIKAI S BECRCPSIE D 20 20 

ANIMALS EXAMINE D HI STOPATHOLOGICALLY ** 20 20 

IHTEI3UHESTAB Y SYSTE M 

*SKIK (20) (20 ) INFLAMMATION , NO S 1 (5%) 

*SUBCUT IISS'JE (20 ) (20) ABSCESS , NCS 

RESPIRATOR Y SYSTE M 

*LON G (20) (20) PHEUHCNIA , CHRCNIC MURIN E 16 (SOX) 18 (90S) 

CALCIU M DEPOSIT 1 (5%) 

HEMATCPCIETI C SYSTE M 

iSPLEEN (20 ) (20) 

HEMATCFCIESIS 1 (5») 2 (10X) 

(tCEBVICA L LYMP H UOD B (19 ) (20 ) 

INFLAPBATICN , NO S 1 (5*) 1 <5X ) 

#MESENTEEI C L . NCD E (19 ) (20 ) IliFLAPfATICN , NO S 2 (10X) 

CIRCULATOR Y SYSTE M 

tHEAR T (20) (20) 

CALCIU M DEPOSIT 1 <5X ) 

#BYOCA8DIU M (20) (20) INFIAMBATICH , NO S 2 (10X) F I EROS IS J 15X1 

* NUMBE R OF ANIMAL S WIT H TISSU E EXAMINE E MICHOSCOPICA LLI 

* NUMBE R OE ANIMAL S NECROPSIED 

**EXCLUDES PARTIALLY AUTOLYZED ANIMAL. ' 

L CU DCSE 

01-042M 

50 

50 

50 ­

(50) 

3 (65S) 

(50) 

1 (2*) 

(50) 

36 (72%) 

(50) 

1 (2X) 

(50) 

(50) 

(50)

1 (21) 

(50) 

. 1 J2X1 .... 

HIGH DO SE 

01-OU3M 

50 

50 

50 

(50) 

1 <2X ) 

(50) 

1 (2<) 

(50) 

29 (58*) 

(U9 ) 

1 (2X) 

(47) 

(47) 

(50) 

1 (2X ) 

(50) 

j_jm_ 

C-3 
DEGENERATION , NOS 

CALCIFICATION , NOS 

IENDCCARDIU M 

HYPI5ELASIA, NOS 

*AORTA 

MEDIA L CALCIFICATIO N 

*CORONAR Y ARTER Y 

(1EDIAL CALCIFICATION 

*MESENT£BIC AETER Y 

BHDIA L CALCIFICATION 

DIGESTIV E SYSTEM 

*LIVES 

INFLAKfATICN , NOS 

CIRRHOSIS , NOS 

PELIOSIS HEPATIS 

METAMORPHOSI S FATTY 

ANGIECTASIS 

• GALLBLADDE R 

PIGMENTATION , NO S 

*3ILE EUCT 

HYPERPLASIA , NO S 

• PANCREA S 

PERIAETERITIS 

*STC«ACH 

ULCER , FCCAL 

CALCIUM DEPOSIT

 

HYPEP.KERATOSI S 

ftCANTHOSIS 

*CCLC N 

NIMATCEIASI S 

URINAR Y SYSTE M 

tKIDNE Y 

PYJLCN|PHRITISt_NOS

* NUMBE R OF ANIMAL S WIT H TISSUE

* NUM3E R CF ANIMAL S NECHOPSIE D 

TABLE Cl (CONTINUED) 

CONTROL (UNTR) CONTR OL (VE H) 

01-031H 01-0l*1« 

1 (5%) 

1 (5% ) 

(20) (20) 

1 (5%) 

(20) (20) 

2 (10%) 2 (10% ) 

(20 ) (20) 

1 (5*) 

(20) (20) 

1 (5X) 1 (5%) 

(20) (20) 

1 (5X) 

2 (10% ) 

3 (15*) 1 (5%) 

(20) (20) 

(20 ) (20) 

1 (20%) 

(20 ) (20) 

U (20*) t (20%) 

(20) (20) 

2 (10X) 2 (10S) 1 (5%) 

(19) (20) 

1 (5*1 

(20 ) (20) 

 I-J53&1 JLJ5S1

 EXAMINE D MICROSCOPICALLY 

I CW DOSE 

01-OH2 M 

(50) 

(50) 

2 (4X) 

(50) 

(50) 

1 (2%) 

(50) 

1 (2«) 

1 (2%) 

(50) 

(50) 

(50) 

3 (6%) 

(19) 

1 (2X ) 

2 (lt%) 

1 (2*) 

1 (2% ) 

(50) 

1 (2X) 

(19) 

 J_i2*L. 

HIG H DO SE 

01-OU3 M 

(50) 

1 (2%) 

I 50) 

1 (2%) 

(50) 

(50) 

(K9 ) 

1 (2%) 

(50) 

1 (2*) 

(50) 

1 (2*) 

0*8) 

1 (2%) 

l«9 ) 

2 (H* ) 

1 (2*) 

8 (16%) 3 (6X) 

(49) 

(U9 ) 

3_I6*l._ 

C-4 
TABLHC1 (CONTINUED) 

INFLAMMATION , CHRONI C 

CALCIU M DEPOSIT 

#i; BINAR Y BLADDER 

INFLACKATICN , NO S 

HYPEHPLASIA , EPITHELIA L 

POLY P 

ENEOC3IN E SYSTE M 

tPIIUITAS i 

ANGIECTA3I S 

tADSENA L 

CALCI0. 1 DEPOSI T 

tAERJNA l COBTEX 

CEGENESATICN , SOS 

ANG1ECTASI S 

#THYFOI D 

ULTIBCBRA8CHIA L CYST 

KJPEaPLASIA , C-CEL L 

HYPEEP1ASIA , FOLLICULAR-CEL L 

#FASVriiYFCI B 

HYPE? ? LA SI A, DOS 

REPRODUCTIV E SYSTE M 

#PM STATE 

INFLAKK A IICN , SO S 

'SiKINA I VESICL E 

INFLAMMATION , NO S 

* IE 3 IIS 

;;.5ANOLCHA , SFEBMATI C 

ATROPHY , NO S 

*EPIDIDYMI 3 

KECHCSIS , FA T 

ATROPHY , NO S 

NEHVOW S SYSTE M 

N 1 3 ]' 

* S'JNBEB OP ANIMAL S WITH TISSUE

* NUMBE R O B ANIMAL S NECBOPSIE D 

CONTRO L (UNTR ) 

0 1-031M 

15 (75X) 

1 (5*) 

(19) 

1 (5%) 

(20 ) 

1 <5X ) 

(20 ) 

(20} 

1 (5%) 

(19 ) 

2 (11%) 

1 (5% ) 

1 (5X ) 

(•'!)

2 (67*) 

(20 ) 

5 (25%) 

(20 ) 

1 (5%) 

(20 ) 

1 (5% ) 

11 (55% ) 

(20 ) 

1 <5* ) 

3 (15% ) 

 EXAMINE D MICROSCOPICALLY 

CONTRO L (VEH ) 

0 1-04 1N 

8 (l»0») 

2 (10X ) 

(20) 

(20) 

(20 ) 

(20) 

(20) 

1 1 5%) 

(1ft ) 

it I25X ) 

(17 ) 

1 (6% ) 

(20 ) 

(20 ) 

12 (60%) 

(20) 

I CW DOSE 

01-OU2M 

1« (29%) 

1 (2*) 

(U8 ) 

(50) 

(19) 

1 (2*) 

(l»9) 

1 (2*) 

2 (U% ) 

(49) 

(26) 

2 (8*) 

(30) 

(50) 

(50) 

10 (20%) 

(50) 

HIG H DOSE 

01-OU3M 

10 (20%) 

1 (2% ) 

("9)

1 (2% ) 

1 (2% ) 

1 (2% ) 

C*9) 

(49 ) 

(49) 

(1*7) 

(33 ) 

(21) 

1 (4%) 

(50 ) 

(49 ) 

9 (18% ) 

(50) 

1 (2%) 

>5 
TABLECl (CONCLUDED) 

- CONTEOL (UNTB ) CONTEOL (VEH ) L OK DOSE 

0 1-031M 01-OU1M 01-042M 

SPECIA L SEfcS E CHGAN 3 

*EYS/COBN£ A (20 ) (20) (50) 

IKFLAMMATION , NO S 1 (5%) 

*EYE/LACBIMAL GLAN D (20) (20) (50) 

INFLAKFATICN , NO S 1 15%) 

MUSCUICSKELETA L SYSTEM 

NCS E 

BOD Y CAVITIE S 

•PERITONEU M (20) (20 ) (50) 

INFLAMMATION , NOS 1 (5«) 

*PEBICA8DIUM 

INFLAPHATICN , NO S 

(20) 

2 (10X) 

(20) (50) 

*1ESENTEB Y 

PEEIABIEBI1IS 

(20 ) 

t (20X) 

(20)

3 (15*) 

(50) 

i» (8*) 

A L L OTHE R SYSTEM S 

NON E 

SPECIA L BCEPHOLOG Y SUMMAH Y 

N O LESIO N BEPORTE D 1 11 

* NUMBE R OF ANIMAL S WIT H TISSUE EXAMINE D MIC BOSCOPICALLY 

* NUMBE R O E ANIMAL S SECROPSIED 

HIG H DOSE 

01-OU3M 

(50) 

(50) 

(50) 

(50) 

(50) 

1 <2X) 

15 

C-6 
TABLE C2

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS TREATED WITH CHLOROPICRIN 

CONTROL (UNTR ) CONTROL (VEH ) L CM DOSE HIGH DOSE 

01-031F 01-OM1 F 01-0<*1(F 01-OU5 F 

ANIMALS INITIALLY IN STUDY 20 20 50 50 

ANIMAL S KECBCFSIE D 20 20 50 50 

ANIMAL S EXAMINE D HISTOEATHOLOGICALLY* * 20 20 50 50 

INTEGUMENTAR Y SYS1EB 

*SKI N (20) (20) (50) (50) EPIDERMA L INCLUSIO N CYST 1 (5%) INFLAMMATION , NO S 1 (5X) 

*SUBCUT TISSU E (20 ) (20) (50) (50) 

ABSCESS, NOS 1 (5X) 

EESFIKATCB Y SYSTE M 

tLUN G (20) (20 ) (50) (50) PNEUMCNIA , CHEONIC MURIN E 18 (90%) 20 (100X) 31 (62X) 30 (SOX) 

HEMATOPOIETI C SYSTE M 

iSPLEEN (20) (20) (50) (50) HEBATCFOIESI S U (20X) 2 (4*) 1 (2X) 

•CERVICA L LYMP H NOD E (20) (18) («8) <«8 ) INFLAMMATION , NO S 1 (6X) 

CIRCULATOR Y SYSTE M 

#MYOCAPDIU M (20) (20) (l»9) (50) FIBBCSIS 1 <5X) 1 (2X ) 

• ENDOCARDIU M (20) (20) (19) (50) HYPEBFLASIA , SOS 1 (2S) 1 (2X) 

*ACHT A (20) (20) (50) (50) MEDIA L CALCIFICATIO N 1 15*1. 1.12XL. 

* NUMBE R CF ANIMAL S KI1H TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBE R OE ANIMAL S NECROPSIE D 

**EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

C-7 
EIGBSTIV E SYSTE M 

#LIVER 

BETAKCBPHCSI S FATT Y 

FOCAL CELLULAR CHANGE 

tLIVER/CENTBILOBULA R KECECSIS , NOS

 

*BILE DUC T 

HYPEBPLASIA , NO S 

IPANCREA S 

PIHIABTEBITIS 

tSTCHAC H 

INFLAPHATICN , NO S 

ULCER , FOCAL 

CALCIU M DEPOSIT 

HJPERKESATOSI S 

ACANTHOSIS 

*COUN 

BIMATCDIASI S 

USINAB Y SYSTE M 

tKIDNE Y 

FYELCNEPHBITIS , NO S 

INFLAMMATION , CHRONIC 

CALCIU M DEPOSIT 

tUBINAR Y BLADDJSB 

INFLAtMATICN , NOS 

ENDOCRIN E SYSTE M 

#PITUITARY 

CYST , NO S 

• ADRENA L 

CYST , NO S 

tADRENA L CORTE X 

CEGENERATICNj , NOS

i NUMBE R Cf ANIMALS WITH TISSUE

* NUMBE R OE ANIMAL S NECROPSIED 

TABLE C2 (CONTINUED) 

CONTROL (UNTR) CONTROL (VEH ) 

01-031F 01-OU1F 

(20) (20) 

1 <5X) 

1 (5*) 

(20) (20) 

1 (5X) 

(20) (20) 

(20) (20) 

1 (5%) 

(20) (20) 

1 (5X) 

1 (5%) 

1 <5X ) 

1 (5*) 

(19) (20) 

1 (5%) 

(20) (20) 

1 (5%) 9 (US*) 5 (25*) 

1 (5X) 

(19) (20) 

(19) (20) 

1 (5X) 

(20) (20) 

(20) (20) 

_ 1_J5$1

 EXAMINE D MICROSCOPICALLY 

C-8 

LCD DOSE 

01-041F 

(U9) 

(U9) 

(50) 

1 (2*) 

(50) 

(50) 

1 (2*) 

2 (»<) 

2 (it*) 

(50) 

(U9) 

« (8X) 

1 (2X) 

(U8) 

1 (2X) 

C*8) 

1 (2X) 

(U9 ) 

1 (2X) 

(«9) 

 -LiJ*! 

HIGH DOSE 

01-0«5F 

(50) 

2 <UX ) 

(50) 

(50) 

(50) 

<«9) 

1 (2X) 

1 (2X) 

1 (2*) 

(50) 

(50) 

6 (12*) 

1 (2*) 

(U9) 

(50) 

(50) 

(50) 
TABLE C2 (CONTINUED) 

CONTROL (UNTR) CONTROL (VEH ) L OU DOSE 

01-031F 01-OU1F 01-04UF 

ANGIECTASI S 3 (15*) 3 (15X) « (8*) 

tTHYHCI D (20) (20) (19) 

FCILICULA R CYST, NOS 1 (5X) 

HYPE9PLASIA , C-CELL << (20%) 

#PARATHY3CI D (D (11 ) (32) 

HYPEBFLASIA , NO S 1 (100*) 

REPECDUCTIV E SYSTEM 

"VAGIN A 

INFLAMMATION , NO S 

(20 ) (20) 

1 (5X) 

(50) 

#UTERU S (20) (20) («9) 

HYCEONET3 A i* (20X) 2 (10X) 5 (10X) 

*UTERUS/ENDCHET8IU M (20) (20) (U9 ) 

INF1APKATICN , NO S 

HYPESPLASIA , CYSTIC 

1 (5X) 

1 <5X ) 

1 (2S) 

IOVAR Y (20 ) (19) (U9 ) 

CYST, NOS 1 (5%) 

INFLAMMATION , NOS 

NEHVOtI S SYSTE M 

NON E 

SPECIAL SENSE ORGAN S 

*HARI:EHIA N GLAN C INFIAMNATICH , NOS (20) (20) 

1 (5X ) 

(50) 

MUSCUIOSKE1ETA L SYSTE M 

NON J 

BOD Y CAVITIE S 

*MESENTER Y (20 ) (20) (50) 

EEBIA11BBITIS l_i5«L 

* NUTOF.fi OF ANIMAL S KITH TISSUE EXAMINE E MICROSCOPICALL Y 

* JUJUBES CF ANIMAL S NHCHOPSIE D 

HIG H DOSE 

01-045F 

6 (12X ) 

(46) 

(22) 

(50) 

1 (2X ) 

(5") 

U (8X) 

(50) 

(50) 

2 (<*X) 

1 (2») 

(50) 

(50) 

C-9 
TABLE C2 (CONCLUDED) 

CONTROL (UNT8 ] CONTROL (VEH ) L CH DCSE HIGH DOSE 

01-031F 01-OU1F 01-OI»«F 01-045F 

A L L CTHE B SYSTEM S 

NON E

S P E C I AS P E C I A L KCEPHCLOG KCEPHCLOG Y SUMMAR Y 

1C L E S I OL E S I O N BEECHlt BEECRlt D 16 188 

* NUMBE R O F ANIMAL S WIT H TISSUE EXAMINE D MICBOSCOPICALL Y 

* »U«BE R O F ANIMAL S NECBOPSIE D 

C-10 
APPENDIX D
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN MICE TREATED WITH CHLOROPICRIN
 

TABLE Dl 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE MICE TREATED WITH CHLOROPICRIN 

1MNTBOL (UNTR) CONTROL (VEH ) LOH DOSE HIG H D3 SE 

02-MO« 1 02-M031 02-MOK 2 02-M013 

ANIMAL S INITIALL Y IN STUD Y 20 20 50 50 

ANIMAI S fISSIN G 1 

ANIMAI S NECROPSIt C 18 20 46 18 

ANIMAL S EXAMINE D HISTOPATHOLOGICALLY ** 15 20 '46 47 

INTEGUMENTAR Y SYSTE M 

*SKI K (18) (20) (16) (18) 

INFLACMATICN , NO S 2 (10X) 1 (2X) 

*SUBCUT TISSUE (18) (20) (U6 ) (18) 

ABSCESS , NOS 1 <6X) 1 (5*) 5 (11*) 

RESPIBATCB Y SYSTEM 

tLUN G (15) (20) ("6) (19) 

PNEUMONIA , CHBCNIC MURIN E 6 (13X) 3 (6)5) 

HEMATOPOIETIC SYSTE M 

#SPLEEN (15) (20) (16) (16) 

ABYLCIDCSI S 7 (47X) 2 (10*) 5 (11*) 

HEMATOPOIESI S 3 (15*) 

#LY11EH NCD E (15) (19) (16) (17) 

IKFLAPHATICN , NO S 1 (2*) 

#CERVICA L LYMP H NOD E (15) (19) (16) (17) 

INFLACPATICN , NO S 1 (5*) 1 (2*) HIPERPLASIA , LXBPHOID 1 (2*) 

#MESENTESIC L. NCD E (15) (19) (16) (17) 

INFLAMMATION , NO S 1 <7X ) 7 (37*) 1 (2*) 5 (11% ) 

CIBCULATOR Y SYSTE M 

*AoaiA (18) (20) (16) (18) 

IBFLAf.BATICN t NOS 1_I2*1 _. 

t NUMBE R OF ANIMAL S WITH TISSUE EXAMINE D MICBOSCOPICALLY 

* NUMBE R CE ANIMAL S NECEOFSIE D 

*»EXCI.UUES PARTIALLY AUTOLYZED ANIMALS 

D-3 
TABLE Dl (CONTINUED) 

CONTROL (UNTR ) CONTROL (VEH ) 

02-MOU 1 02- MO 31 

EIGESTIV E SYSTE M 

ILIVEB (15) (20) 

AMYLCIECSI S 

METAMORPHOSI S FATTY 

• PANCREA S (14) (20)

 

INFLAPKATICN , NO S 1 (7*)

 

•STOMACH (15) (20) 

HYFERKERATCSI S 

ACANTHOSI S 

URINAH Y SYSTE M 

• KIDKE Y (15) (20 ) 

HYCRCNEFHRCSI S 

CYST, NOS 1 (5%) 

PYELONEPHRITIS , NOS 2 (10*) 

INFLAMMATION , CHBONIC 12 (SOX ) 2 (10X) 

AMYLOIDOSIS 7 (1*7*) 2 (10?S) 

CALCIU M DEPOSIT 

MRINAE Y BLAtEER (15) (19) 

INFLAPKATICN , NO S 2 (11%) 

ENDOCRIN E SYSTE M 

ITHYHOID (10) (20 )

 FCLIICULS K CYST , NO S 1 (10X) 1 (5%)

 

REPROIUCTIV E SYSTE M 

• PROSTATE (15) (12)

 INFLACfATICN , NO S

 

tTESTIS (15) (20)

 ATRCEHY , NQS 1 (7%)

 

HEHVOU S SYSTE M 

NON E _ 

» NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBE R CE ANIMAL S NECROPSIE D 

L OU DOSE 

02-MOU 2 

(46) 

3 (7*) 

1 <2X ) 

(46) 

(46) 

3 (7X) 

3 (7X) 

(«6) 

1 (2X) 

6 (12*) 

5 (11*) 

(46) 

1 (2X) 

(35) 

(46) 

(46) 

1 (2X) 

HIGH DOSE 

02-M04 3 

(48) 

(46) 

(48) 

1 (2X) 1 (2X) 

(48) 

1 (2*) 

1 (2*) 

(47) 

1 (2*) 

(40) 

(45) 

2 (4*) 

(46) 

D-4 
TABLE Dl (CONCLUDED) 

CONTRO L (BNTR) CONTBOL(VEH ) 

02-B011 02- MO 31 

SPECIAL SZKS E ORGAN S 

• EYE/LACRIMA L GLAN D (18) (20)

 

HYFEBELASIA , NO S 

*HAB[E8IA N GLAN D (18) (20)

 HYPESPLASIA , NOS

 

HUSCUICSKELETA L SYSTEM 

NJN E 

30DJ CAVITIE S 

• ABDOMINA L CAVIT Y (18) (20)

 KECKCSIS , FA1

 

Hll OTHE R SYSTEM S 

NJN E 

SPECIA L NCEPHOLOG Y SUI1MAB Y 

N C LESIO N BEPORTE D 3 7 

flNIMAL MISSING/SO NECEOES Y 

AUTO/NECROPSY/N O HISIO 3 

AUTCLYSIS/N C NECBOFS Y 2 

* NUMEE H O F ANIMAL S WIT H TISSUE EXAMINE D MICBOSCOPICALL1 

* NUMBE R CI ANIMAL S NECEOPSIE D 

LC» DOSE HIGH DO SE 

02-MOU2 02-M04 3 

(U6 ) (US )

 

2 («*) 

(U6 ) (18) 

1 (2*) 

C*6) (18)

 

1 (2*) 

18 31

 

1 1

 

3 2

 

D-5 
TABLE D2 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE TREATED WITH CHLOROPICRIN 

CONTROL (ONTR) CONTROL (VEB) LCW DCSE HIGH DO SE 

02-FQ141 02-F031 02-FOa« 02-FOU5 

ANIMAL S INITIALL Y I N STUD Y 20 20 50 50 

AMIRAI S KECSOFSIBD 20 19 50 H8 

ANIMAL S EXAMINE D HISTOPATHOLOGICALL Y *» 20 19 U9 i»8 

INTEGUflEKIAH Y SYS1EM 

NDN E 

EESFIBATCR Y SYSTE M 

• LUN G (20) (19) (l»9) (U8) 

PNEUKCNIA , CHBCNIC MURIN E 1 (5*) 1 (5X) 2 (!»*) 15 (31X) 

HEMATCPOIETI C SYSTEM 

tSPLEE N (20) (19) (»9) («8) 

APYICIDOSI S 1 (2%) HEMATOPOIESIS 3 (6*) 

tLYCF H NCC E (20) (19) <U6 ) (UK ) INFLAKfATICN , NO S 1 (SX) 

IC2HVICA L LYMP H NODE (20) (19) (46) (tt) INFLAP.KA1ICN . NO S 3 (7*) 

*MES£NTIBIC L. NCD E (20) (19) (t6) (UK ) 

INFLACSATICS , NO S 1 (5X) 1 (2*) ANGIECTASI S 1 (5*) 

CIRCULATOR Y SYSTEM 

NON E 

DIGESTIV E SYSTE M 

tLIVE R (20) (19) («9) (US) 

PJ LIC SIS HEPA.TIS_ _ 1 12*L . 

* NUMBE R OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBE R OF ANIMAL S NECROPSIE D 

**EXCLUDESPARTIALLY AUTOLYZEDANIMALS 

D-6 
AHYLOIDOSI S 

LEUKENOI D BEACTIO N 

*BI1E DUCT 

EILATA1ICN , NO S 

HYPHEPLASIA , NO S 

• PANCREA S 

CYST, NOS 

INFLAMMATION , NO S 

tSTCBAC H 

UICEB, NO S 

HYPEBKEEATOSI S 

ACANTHOSI S 

#PEYER S PATCH 

HYPEHPLASIA , LYMPHOI D 

ILARG E INTESTIN E 

HEMCRRHAGI C CYS T 

URINAR Y SYSTE M 

*KIDKE Y 

PYILCNEPHSIT I S, NOS 

INFLAMMATION , CHRONI C 

AMYLOIDOSI S 

ENDOCRIN E SYSTEM 

ITHYSCI D 

FCLIICULA B CYST , NO S 

HSPERPIASIA , F01LICUIAH-CELL 

SEPRCCUCTIV E SYSTE M 

*VAi3INA 

INFLAMMATION , NOS 

*UTEBUS 

HYCROMETR A 

iUTERUS/ENDCMZTRIU M 

* NUMBE B OF ANIMAL S BITH TISSU E

* NUMBE B OF ANIMALS NECBOPSIED 

TABLE D2 (CONTINUED) 

CONTROL (UNTS) CONTROL (VEH ) L CW DOSE HIGH DOSE 

02-F041 02-F031 02-F014 02-F015 

1 (2*) 1 (5*) 

(20) (19) (50) (U8) 

1 (5*) 

1 <5X ) 

(20 ) (19) (17) <U8 ) 

1 <5X ) 

1 (5*) 

1 (2%) 1 <2X ) 

(20) (19) (18) 

1

10

 (2X ) 

 (21X) 

(18) 

9 (19X) 

10 (21X) 9 (19*) 

(20) (19) (17) (45) 

1 (2*) 

(19) (19) (13) 

1 (2*) 

(46) 

(20 ) (19) (U9 ) (18) 

1 (2X ) 

1

1

 <2X ) 

 (2») 

(16 )

1 (61) 

1 <6X ) 

(16) (11) (36) 

(20) (19) (50) (48) 

1 (5%) 

(20) 

5 (25%) 

(19) 

2 (11*) 

(19) 

6 (12X) 

(17) 

5 (11*) 

(20) (19) (19) (47) 

 EXAMINE D MICROSCOPICALLY 

D-7 
TABLE D2 (CONCLUDED) 

CONTROL (UNTR) CONTROL (VEH ) LOW DOSE 

02-FOU1 02-F031 02-FOiJU 

HYPERPLASIA , CYSTIC 6 (30*) 9 (U7X ) 16 (33*) 

IOVAHY/CVIDUC T (20) (19) (1*9) INFLACfATICN , NO S 1 (5%)

 

*OVAfiY (20) (19) (49) CYST, NOS 12 (60%) « (21)1) 9 (18%) 

INFLAMMATION , NOS 5 (25X) 14 (21%) it (8%) 

NERVOU S SYSTE M 

NONI 

SPECIA L SHUS H ORGAN S 

*EYE (20) (19 ) (50) 

PHTHISIS BULBI 

*HAPCEHIAN GLAN E (20) (19) (50) 

HYPE3P1ASIA , NOS 1 (2*) 

MUSCOLOSKELETA L SYSTEM 

NON E 

BODY CAVITIES 

• PERITONEU M (20) (19) (50)

 

INFLAmilCN, NOS 1 (5*)

 

A LL OTHER SYSTEMS 

NON E 

SPECIA L MCEPHOLOG Y SUMMAR Y 

NO LESION REPORTE D 1 3 11 

NECROPS Y PERF/NO HISTO PERFORME D 1 

AOTOHSIS/8O NECSOPSY 1 

t NURSE S CF ANIMAL S HITH TISSU E EXAMINE E MICROSCOPICALLY 

* NUMBE 8 CF ANIMAL S NHCSOPSIED 

HIGH DOSE 

02-FOt»5 

7 (15%) 

(«7) 

(47) 

8 (17%) 

(48) 

1 (2%) 

(48) 

(48) 

1 (2*) 

11 

2 

D-8 
DHEW Publication No. (NIH) 78-1315