1978 

-$6-6 75' 

OFFICIAL NTP
 

FILE COPY
 

National Cancer Institute 

CARCINOGENESIS 

DO NOT REMOVE FROM THIS OFFICE
 Technical Report Series 

No. 75
 

BIOASSAY OF 

CHLOROBENZILATE 

FOR POSSIBLE CARCINOGENICITY 

CAS No. 510-15-6 

NCI-CG-TR-75 

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

Public Health Service 

National Institutes of Health 

\ 

BIOASSAY OF
 

CHLOROBENZILATE
 

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) 78-1325
 

REPORT ON THE BIOASSAY OF CHLOROBENZILATE
 

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 chlorobenzilate 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 chemicals
 

have the capacity to produce cancer in animals. Negative results, in
 

which the test animals do not have a significantly 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 chlorobenzilate 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 Dr. B. M. Ulland (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. R. L. Schueler (7).
 

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

statistical analysis was performed by Mr. W. W. Belew (6,9) using
 

ill
 
methods selected for the Carcinogenesis Testing Program by Dr. J.
 

J. Gart (10).
 

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), task leader Dr. M. R. Kornreich (6,11), senior biologist Ms. P.
 

Walker (6), biochemist Dr. B. Fuller (6), and technical editor Ms.
 

P. A. Miller (6). The final report was reviewed by members of the
 

participating organizations.
 

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 (1,11), Dr.
 

R. A. Griesemer (1), Dr. M. H. Levitt (1), Dr. H. A. Milman (1), Dr.
 

T. W. Orme (1), Dr. R. A. Squire (1,12), Dr. S. F. Stinson (1), Dr.
 

J. M. Ward (1), and Dr. C. E. Whitmire (1).
 

1.	 Carcinogenesis Testing Program, Division of Cancer Cause and
 

Prevention, National Cancer Institute, National Institutes of
 

Health, Bethesda, Maryland.
 

2.	 No wit w h the Naylor Dana Institute for Disease Prevention, American Health Foundation, Hammon House Road, Valhalla, New York.
 

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

Vienna, Virginia.
 

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

Reston, Virginia.
 

5.	 Now with Rhodia, Inc., 23 Belmont Drive, Somerset, New Jersey.
 

6.	 The MITRE Corporation, METREK Division, 1820 Dolley Madison
 

Boulevard, McLean, Virginia.
 

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

Maryland.
 

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

Rockville, Maryland.
 

9.	 Now with the Solar Energy Research Institute, Cole Boulevard,
 

Golden, Colorado.
 

IV
 
10.	 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.
 

11.	 Now with Clement Associates, Inc., 1010 Wisconsin Avenue, N.W.,
 

Washington, D.C.
 

12.	 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 chlorobenzilate for possible carcinogenicity was conducted using Osborne-Mendel rats and B6C3F1 mice.
 

Chlorobenzilate was administered in the feed, at either of two concentrations, to groups of 50 male and 50 female animals of each species. Chlorobenzilate was administered for 78 weeks followed by an
 

observation period of 12 or 13 additional weeks in mice and 32 or 33
 

additional weeks in rats.
 

The time-weighted average dietary concentrations of chlorobenzilate were 2995 and 1600 ppm for high and low dose male rats, respectively, and 2229 and 1175 ppm for high and low dose female rats.
 

Mice received time-weighted average high and low dietary concentrations of 7846 and 4231 ppm, respectively, for males and 5908 and
 

3200 ppm, respectively, for females.
 

Survival in both species was high (over 68 percent of the high
 

dose rats and over 82 percent of the high dose mice survived on test
 

until the end of the study). Dose-related mean body weight depression,
 

observed in both species, indicated that the maximum dose for optimal
 

bioassay sensitivity was used in the high dose groups.
 

An increased incidence of hepatocellular carcinomas was observed
 

in dosed mice, i.e., 4/19 (21 percent) in control males, 32/48 (67
 

percent) in low dose males, 22/45 (49 percent) in high dose males,
 

0/20 in control females, 11/49 (22 percent) in low dose females, and
 

13/50 (26 percent) in high dose females.
 

There was a statistically significant positive association between the administration of chlorobenzilate and the appearance of
 

cortical adenoma of the adrenal gland in low dose male and high dose
 

female rats. Although suggestive, the findings of a low incidence of
 

benign adrenal tumors was not considered sufficient evidence to establish the carcinogenicity of chlorobenzilate for the Osborne-Mendel rat.
 

Under the conditions of this bioassay, orally administered chlorobenzilate was carcinogenic in male and female B6C3F1 mice, causing
 

an increased incidence of hepatocellular carcinomas. The results do
 

not, however, provide sufficient evidence for the carcinogenicity of
 

chlorobenzilate in Osborne-Mendel rats.
 

vii
 

TABLE OF CONTENTS
 

Page
 

I. INTRODUCTION	 1
 

II. MATERIALS AND METHODS	 4
 

A. Chemicals	 4
 

B. Dietary Preparation	 5
 

C. Animals	 5
 

D. Animal Maintenance	 6
 

E. Selection	 of Initial Concentrations 7
 

F. Experimental Design	 9
 

G. Clinical and Histopathologic Examinations	 12
 

H. Data Recording and Statistical Analyses	 14
 

III. CHRONIC	 TESTING RESULTS: RATS 19
 

A. Body Weights and Clinical Observations	 19
 

B. Survival	 21
 

C. Pathology	 21
 

D. Statistical Analyses of Results	 23
 

IV. CHRONIC TESTING RESULTS: MICE	 32
 

A. Body Weights and Clinical Observations	 32
 

B. Survival	 34
 

C. Pathology	 34
 

D. Statistical Analyses of Results	 36
 

V. DISCUSSION	 40
 

VI. BIBLIOGRAPHY	 42
 

APPENDIX A SUMMARY OF THE INCIDENCE OF NEOPLASMS
 

IN RATS TREATED WITH CHLOROBENZILATE A-l
 

APPENDIX B SUMMARY OF THE INCIDENCE OF NEOPLASMS
 

IN MICE TREATED WITH CHLOROBENZILATE B-l
 

APPENDIX C SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN RATS TREATED WITH CHLOROBENZILATE C-l
 

APPENDIX	 D SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN MICE TREATED WITH CHLOROBENZILATE D-l
 

ix
 
LIST OF ILLUSTRATIONS 

Figure Number Page 

1 CHEMICAL STRUCTURE OF CHLOROBENZILATE 2 

2 GROWTH CURVES FOR CHLOROBENZILATE CHRONIC 

STUDY RATS 20 

3 SURVIVAL COMPARISONS OF CHLOROBENZILATE 

CHRONIC STUDY RATS 22 

4 GROWTH CURVES FOR CHLOROBENZILATE CHRONIC 

STUDY MICE 33 

5 SURVIVAL COMPARISONS OF CHLOROBENZILATE 

CHRONIC STUDY MICE 35 

LIST OF TABLES 

Table Number Page 

DESIGN SUMMARY FOR OSBORNE-MENDEL RATS— 

CHLOROBENZILATE FEEDING EXPERIMENT 10 

DESIGN SUMMARY FOR B6C3F1 MICE-­

CHLOROBENZILATE FEEDING EXPERIMENT 11 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN MALE RATS TREATED WITH 

CHLOROBENZILATE 24 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN FEMALE RATS TREATED 

WITH CHLOROBENZILATE 27 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN MALE MICE TREATED WITH 

CHLOROBENZILATE 37 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN FEMALE MICE TREATED 

WITH CHLOROBENZILATE 38 

x
 
LIST OF TABLES (Concluded) 

Table Number Page 

Al SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

MALE RATS TREATED WITH CHLOROBENZILATE A-3 

A2 SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

FEMALE RATS TREATED WITH CHLOROBENZILATE A-7 

Bl SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

MALE MICE TREATED WITH CHLOROBENZILATE B-3 

B2 SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

FEMALE MICE TREATED WITH CHLOROBENZILATE B-6 

Cl SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN MALE RATS TREATED WITH CHLOROBENZILATE C-3 

C2 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN FEMALE RATS TREATED WITH CHLOROBENZILATE C-9 

Dl SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN MALE MICE TREATED WITH CHLOROBENZILATE D-3 

D2 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN FEMALE MICE TREATED WITH CHLOROBENZILATE D-6 

xi
 

I. INTRODUCTION
 

Chlorobenzilate (Figure 1) (NCI No. C00408) was one of a group
 

of agricultural pesticides that scientists at the National Cancer
 

Institute (NCI) selected for inclusion in the Carcinogenesis Testing
 

Program. In a study by Innes et al. (1969) evidence emerged suggesting that the incidence of hepatomas was significantly elevated in
 

male mice upon oral administration of chlorobenzilate. In addition,
 

the widespread use of chlorobenzilate and the resulting human exposure
 

emphasized the need for carcinogenicity testing.
 

The Chemical Abstracts Service (CAS) Ninth Collective Index
 

(1977) name for this compound is 4,4"-dichlorobenzilic acid, ethyl
 

*
 

ester. It is a carbinol compound and is also called ethyl 4,4'­

dichlorobenzilate.
 

Introduced in 1952 for use as a selective acaricide, chlorobenzilate had an annual domestic production of approximately 36 million
 

pounds in 1974 (Fowler and Mahan, 1976). Essentially most of the
 

chlorobenzilate that enters the environment is through its dispersion
 

as a pesticide. Ninety percent of chlorobenzilate usage in 1971 was
 

for mite control on citrus crops (Stanford Research Institute, 1975).
 

Additionally, it is effective against mites in orchards, vineyards,
 

tea plantations, field crops, and ornamental plants. Since bees are
 

not severely affected by chlorobenzilate, it is also used to control
 

*
 

The CAS registry number is 510-15-6.
 
O=C—O—CH2—CH3
 

FIGURE 1
 

CHEMICAL STRUCTURE OF CHLOROBENZILATE
 
the tracheal mite of this insect (Bartsch et al., 1971). The greatest
 

potential for exposure to chlorobenzilate appears to be among persons
 

associated with its production, formulation, and agricultural application. No chlorobenzilate was detected in food in the United States
 

in total diet studies carried out in 1966 by the U.S. Food and Drug
 

Administration (Bartsch et al. , 1971) or reported in recent analyses
 

of food composites (Johnson and Manske, 1976). Residue levels found
 

on fruits, citrus, grapes, tea, and vegetables were all below the
 

tolerance levels set by the U.S. Environmental Protection Agency
 

(Bartsch ec al., 1971).
 
II. MATERIALS AND METHODS
 

A. Chemicals
 

A single batch of technical-grade chlorobenzilate was purchased
 

by Hazleton Laboratories America, Inc., Vienna, Virginia, from Geigy
 

Agricultural Chemicals. The compound was tested by Hazleton Laboratories for purity at the start of the bioassay, at an intermediate
 

stage, and again during the final year of the feeding study. The
 

final analysis was conducted prior to termination of the bioassay.
 

The technical-grade chlorobenzilate was analyzed twice within
 

the first 12 months of the bioassay by gas-liquid chromatography
 

using both total-area analysis and the internal standard method.
 

An FDA manufacturer's standard for chlorobenzilate (99.9 percent)
 

was used as the internal standard. Total-area analysis results
 

showed a major peak of 97 and 99 percent of chlorobenzilate for the
 

first and second assays, respectively. Comparisons of the technical-


grade chlorobenzilate with the internal standard revealed that the
 

technical-grade compound contained 95 and 98 percent chlorobenzilate
 

based on the first and second assays, respectively.
 

The chemical purity of chlorobenzilate was assayed for a third
 

time 22 months after the feeding study was initiated. Total-area
 

analysis indicated a peak area of 97 percent. The internal standard
 

assay, performed to corroborate this analysis indicated a relative
 

peak area of 90 percent.
 
Throughout this report the term chlorobenzilate is used to
 

represent this technical-grade material.
 

B. Dietary Preparation
 

The basal laboratory diet for both dosed and control animals
 

consisted of Wayne Lab-Blox meal (Allied Mills, Inc., Chicago,
 

Illinois) plus 2 percent Duke's corn oil (S. F. Sauer Company,
 

Richmond, Virginia) by weight. Fresh mixtures of chlorobenzilate
 

in corn oil were prepared each week and stored in the dark. The
 

chlorobenzilate mixtures were incorporated into the appropriate
 

amount of laboratory diet in a twin-shell blender fitted with an
 

accelerator bar.
 

C. Animals
 

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.
 

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 to 15 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, while 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 mice. 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, while fresh heat-


sterilized glass water bottles and sipper tubes were provided three
 

times a week. Food (Wayne Lab-Blox®meal) and water were available
 

ad libitum.
 

Dosed	 rats were housed in the same room with other rats receiving
 

*
 

diets containing sulfallate (95-06-7); DDT (50-29-3); and TDE (72-54-8),
 

*
 

CAS registry numbers are given in paretheses,
 
Control rats were housed in a room with other rats receiving diets
 

containing trifluralin (1582-09-8); dioxathion (78-34-2); dicofol
 

(115-32-2); nitrofen (1836-75-5); endosulfan (115-29-7); and mexacarbate (315-18-4).
 

All mice used in the chlorobenzilate study, including controls,
 

were housed in the same room as other mice receiving diets containing
 

trifluralin (1582-09-8); dioxathion (78-34-2); sulfallate (95-06-7);
 

DDT (50-29-3); methoxychlor (72-43-5); DDE (72-55-9); IDE (72-54-8);
 

dicofol (115-32-2); pentachloronitrobenzene (82-68-8); clonitralid
 

(1420-04-8); acetylaminofluorene (53-96-3); nitrofen (1836-75-5);
 

endosulfan (115-29-7); mexacarbate (315-18-4); amitrole (61-82-5);
 

and safrole (94-59-7).
 

E. Selection of Initial Concentrations
 

In order to establish the maximum tolerated concentrations of
 

chlorobenzilate for administration to dosed animals in the chronic
 

studies, subchronic toxicity tests were conducted with both rats and
 

mice. Animals of each species were distributed among six groups, each
 

consisting of five males and five females. Chlorobenzilate was premixed with a small amount of corn oil. This mixture was then incorporated into the laboratory diet and fed ad libitum to five of the six
 

rat groups and five of the six mouse groups in concentrations of 1780,
 

3160, 5620, 10,000, and 17,800 ppm. The sixth group of each species
 

served as a control group, receiving only the mixture of corn oil and
 

laboratory meal. The dosed dietary preparations were administered
 
for a period of 6 weeks, followed by a 2-week observation period
 

during which all animals were fed the basal laboratory diet.
 

A dosage inducing no mortality and resulting in a depression in
 

mean body weight of approximately 20 percent relative to controls was
 

to be selected as the high dose for administration in the chronic bioassay. When weight gain criteria were not applicable, mortality data
 

alone were utilized.
 

All male and female rats receiving 10,000 ppm chlorobenzilate or
 

less survived the entire 8-week study. The depressions in mean body
 

weight in males receiving concentrations of 3160 and 5620 ppm were 14
 

and 38 percent, respectively. In females receiving concentrations of
 

1780 and 3160 ppm the depressions in mean body weight were 18 and 22
 

percent, respectively. The high concentrations selected for administration to rats in the chronic study were 3200 and 2350 ppm for males
 

and females, respectively.
 

In the male mice four deaths were observed in the group receiving
 

3160 ppm but no other deaths, except one male at 17,800 ppm, were recorded. All female mice survived the 8-week study, except two in the
 

group receiving 10,000 ppm. Mean body weight gain, expressed as a percentage of the weight gained by the controls, was 108 and 50 percent
 

in the males dosed with 10,000 and 17,800 ppm, respectively. In the
 

females, the body weight gains were 82 and 45 percent at concentrations of 5620 and 10,000 ppm, respectively. The high concentrations
 

selected for administration to mice in the chronic study were 12,000
 

and 6400 ppm for males and females, respectively.
 

8
 
F.	 Experimental Design
 

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

treated and untreated observation periods, and the time-weighted
 

average concentrations) are summarized in Tables 1 and 2.
 

The dosed and control rats were all approximately 6 weeks old
 

at the time they were placed on test. However, the control rats had
 

a median birth date approximately 2 weeks earlier than the dosed
 

rats and were placed on test approximately 2 weeks before the dosed
 

rats. The concentrations of chlorobenzilate initially administered
 

to males were 3200 and 1600 ppm. Throughout this report those male
 

rats receiving the former concentration are referred to as the high
 

dose group and those receiving the latter concentration are referred
 

to as the low dose group. For females, the initial concentrations of
 

chlorobenzilate were 2350 and 1175 ppm. Throughout this report those
 

female rats receiving the former concentration are referred to as the
 

high dose group and those receiving the latter concentration are referred to as the low dose group. The basal diet for all groups contained 2 percent corn oil. In week 58 of the study, administration
 

of chlorobenzilate to the high dose male rats ceased for 1 week, due
 

to toxicity of the compound, and was then followed by 4 weeks of
 

feeding at the previous concentration of 3200 ppm. This pattern of
 

cyclic administration continued for the remainder of the dosing period.
 

This same method of total intake reduction and the same rationale
 

were	 employed for the high dose female rats beginning with week 63.
 
TABLE 1
 

DESIGN SUMMARY FOR OSBORNE-MENDEL RATS
 

CHLOROBENZILATE FEEDING EXPERIMENT
 

TIME-WEIGHTED 

INITIAL CHLORO­ OBSERVATION PERIOD AVERAGE CONCENGROUP BENZILATE TREATED UNTREATED TRATION OVER A 

SIZE CONCENTRATION3 (WEEKS) (WEEKS) 78-WEEK PERIODb 

MALE
 

CONTROL 50 0 111 0
 

LOW DOSE 50 1600 78 1600
 

0 32
 

HIGH DOSE 50 3200 57 2995
 

3200° 16 5
 

0 32
 

FEMALE
 

CONTROL 50 0 111 0
 

LOW DOSE 50 1175 78 1175
 

0 33
 

HIGH DOSE 50 2350 62 2229
 

2350° 12 4
 

0 33
 

a
 

Concentrations in parts per million. »
 

b 2 (concentration X weeks received)
 

78 weeks
 

"These doses were cyclically administered with a pattern of 1 dose-free
 

week followed by 4 weeks of dosing at the levels indicated.
 

10
 
TABLE 2
 

DESIGN SUMMARY FOR B6C3F1 MICE
 

CHLOROBENZILATE FEEDING EXPERIMENT
 

TIME-WEIGHTED 

INITIAL CHLORO­ OBSERVATION PERIOD AVERAGE CONCENGROUP BENZILATE TREATED UNTREATED TRATION OVER A 

SIZE CONCENTRATION3

 (WEEKS) (WEEKS) 78-WEEK PERIODb 

MALE
 

CONTROL 20 0 90 0
 

LOW DOSE 50 6,000 9 4,231
 

4,000 69
 

0 12
 

HIGH DOSE 50 12,000 9 7,846
 

8,000 43
 

8,000C 20 6
 

0 12
 

FEMALE
 

CONTROL 20 0 90 0
 

LOW DOSE 50 3,200 78 3,200
 

0 13
 

HIGH DOSE 50 6,400 52 5,908
 

6,400C 20 6
 

0 13
 

a
 Concentrations in parts per million.
 

b . 2 (concentration X weeks received)
 Time-weighted avei rage concentration =-5 :
 

'These doses were cyclically administered with a pattern of 1 dose-free
 

week followed by 4 weeks of dosing at the levels indicated.
 

11
 
The control and dosed mice were all approximately 6 weeks old
 

on the first day of the test and they all shared the same median
 

birth date. The initial concentrations administered to male mice
 

were 12,000 and 6000 ppm. Throughout this report those male mice
 

receiving the former concentration are referred to as the high dose
 

group and those receiving the latter concentration are referred to
 

as the low dose group. In week 10 of the experiment, when the male
 

mice were 16 weeks old, the high and low concentrations were decreased to 8000 and 4000 ppm, respectively, because a hunched posture was observed in the high dose males. Female mice received
 

initial concentrations of 6400 and 3200 ppm and were maintained at
 

these levels until termination of the experiment. Throughout this
 

report those female mice receiving the former concentration are
 

referred to as the high dose group and those receiving the latter
 

concentration are referred to as the low dose group. In week 53
 

of the study, administration of chlorobenzilate to the high dose
 

male and female mice ceased for 1 week and was then followed by
 

4 weeks of dietary administration at the previous concentrations
 

of 8000 and 6400 ppm. The concentrations administered were decreased due to the reappearance of a hunched posture in the dosed
 

animals. This pattern of cyclic administration was continued for
 

the remainder of the dosing period.
 

G. Clinical and Histopathologic Examinations
 

Animals were weighed immediately prior to initiation of the
 

experiment. Body weights, food consumption, and data concerning
 

12
 
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. From the
 

first day, all animals were inspected daily for mortality. The
 

presence of 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, and gross lesions taken from sacrificed animals and, whenever possible, from animals found dead.
 

Tissues 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.
 

Slides were prepared from the following tissues: skin, subcutaneous tissue, lungs and bronchi, trachea, bone marrow, spleen, lymph
 

nodes, thymus, heart, salivary gland, liver, gallbladder (mice), pancreas, esophagus, stomach, small intestine, large intestine, kidney,
 

urinary bladder, pituitary, adrenal, thyroid, parathyroid, testis,
 

prostate, seminal vesicle, brain, muscle, uterus, mammary gland, and
 

ovary.
 

13
 
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
 

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.
 

H.	 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
 

14
 
missing; animals dying from natural causes were not statistically
 

censored. Statistical analyses for a possible dose-related effect
 

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

equality and used Tarone's (1975) extensions of Cox's methods when
 

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
 

15
 
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
 

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 when
 

appropriate. 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.
 

16
 
When appropriate, life-table methods were used to analyze the
 

incidence of tumors. Curves of the proportions surviving without an
 

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
 

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
 

17
 
percent of a large number of identical experiments, the true ratio
 

of the 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.
 

18
 
III. CHRONIC TESTING RESULTS: RATS
 

A. Body Weights and Clinical Observations
 

Dose-related mean body weight depression was observed in both
 

male and female rats beginning in week 10. This continued until
 

approximately week 90 at which time the control male and female rats
 

exhibited a slight decrease in mean body weight while the dosed
 

groups generally maintained their relative mean body weights (Figure 2). Fluctuations in the growth curve may be due to mortality;
 

as the size of the group diminishes, the mean body weight may be
 

subject to wide variations.
 

During the first year of the study the appearance and behavior
 

patterns observed among dosed rats were generally comparable to
 

those of the controls, except that by week 8, urine staining of the
 

abdominal fur was noted in a few high dose males. As the study progressed (weeks 9 to 78), abdominal urine stains and a hunched appearance were observed at a slightly greater frequency in the dosed groups
 

than in the controls. Thereafter, these signs were noted at a comparable rate in dosed and control animals. Undersized gonads were
 

observed during the last 6 months of the study in several dosed male
 

rats, an observation subsequently confirmed at necropsy as compound-


related testicular atrophy.
 

Respiratory involvement, characterized by labored respiration,
 

wheezing, and/or nasal discharge was observed at a low incidence in
 

all groups during the study. Incidental signs associated with aging
 

19
 
750 

TIME ON TEST (WEEKS) 

750­

-r 600­

oc 

a 

450­

Q 

o 

m 

< 

LU 

300­

150­

CONTROL 

LOW DOSE 

FEMALE RATS 

I 

15 30 

I 

45 60 

I 

75 

\ 

90 

HIGH DOSE 

T 

105 120 

TIME ON TEST (WEEKS) 

FIGURE 2 

GROWTH CURVES FOR CHLOROBENZILATE CHRONIC STUDY RATS 

20 
were observed in comparable numbers of dosed and control rats
 

during the last 6 months. These signs included rough fur, eyes that
 

were pale, squinted, cloudy, or showing reddish discharge, sores on
 

the tail or other parts of the body, localized alopecia, swollen
 

areas of the body, palpable nodules, and/or tissue masses. Isolated,
 

apparently incidental, observations noted sporadically during the
 

study in one to five dosed rats included ataxia, tremors, head tilt,
 

circling or loss of equilibrium, and hind-limb paralysis.
 

B. Survival
 

The estimated probabilities of survival for male and female rats
 

in the control and chlorobenzilate-dosed groups are shown in Figure 3.
 

In both sexes, the Tarone test indicated no positive association between dose and mortality.
 

In both sexes there were adequate numbers of racs at risk from
 

late-developing tumors, with 68 percent (34/50) of the high dose males
 

and 72 percent (36/50) of the high dose females surviving on test
 

until the end of the study.
 

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 was observed among both dosed and control
 

rats. The types of tumors observed have been encountered previously
 

as spontaneous lesions ,in the Osborne-Mendel rat, and the incidence
 

21
 
1.0­

b_ 

> 0.8­

cc 

CO 

LL 

o 

?­ 0.6­

03 

< 

CQ 

O 

DC 0.4­ Q_ 

COMTROL 

0.2­ LOW DOSE 

HIGH DOSE 

MALE RATS 

0.0­ I I I I \ 

15 30 45 60 75 90 105 120 

TIME ON TEST (WEEKS) 

1.0­

oc 

CO 

u_ 

O 

> 0.6­

Zi 

3 

00 

O 

cc 0.4 ­ Q. 

_ CONTROL 

0.2 .., LOW DOSE ­

. _ HIGH DOSE 

FEMALE RATS 0.0- \ I i ' r i r 

15 30 45 60 75 90 105 120 

TIME ON TEST (WEEKS) 

FIGURES 

SURVIVAL COMPARISONS OF CHLOROBENZILATE CHRONIC STUDY RATS 

22 
of tumor types, with the exceptions of malignant lymphomas and adrenal
 

neoplasms, appeared to be unrelated to group or sex.
 

Malignant lymphomas (histiocytic or lymphocytic types) occurred
 

with somewhat greater frequency in dosed than in control animals.
 

This slightly greater frequency in dosed rats when compared to controls
 

is of doubtful significance, as these tumors do not represent unusual
 

types and have been known to occur spontaneously at these incidences
 

in the Osborne-Mendel rat.
 

Cortical adenomas and pheochromocytomas of the adrenal gland
 

occurred only in dosed rats. Cortical adenomas were observed in
 

6/49 (12 percent) low dose males, 3/49 (6 percent) high dose males,
 

2/47 (4 percent) low dose females, and 5/47 (11 percent) high dose
 

females. Pheochromocytomas were observed in 1/49 (2 percent) low
 

dose males, 1/49 (2 percent) high dose males, 2/47 (4 percent) low
 

dose females and 0/47 high dose females.
 

A wide variety of nonproliferative lesions of spontaneous disease
 

occurred in both dosed and control rats (see Appendix C). Sections
 

of testicle, however, indicated compound-related testicular atrophy in
 

the dosed male groups. Thirty-one of 49 (63 percent) high dose males
 

and 26/49 (53 percent) low dose males showed significant degrees of
 

testicular atrophy, while in the male control group only 9/44 (20 percent) of the rats were observed with testicular atrophy.
 

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
 

23
 
TABLE 3
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN MALE RATS TREATED WITH CHLOROBENZILATE5
 

TOPOGRAPHY : MORPHOLOGY
 

Subcutaneous Tissue: Fibroma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Circulatory System: Hemangiosarcoma
 N>
 

-O- P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pituitary: Chromophobe Adenoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

CONTROL
 

1/49(0.02)
 

N.S.
 

Ill
 

12/49(0.24)
 

P = O.OOl(N)
 

68
 

4/41(0.10)
 

N.S.
 

108
 

LOW
 

DOSE
 

1/50(0.02)
 

N.S.
 

0.980
 

0.013
 

75.404
 

99
 

4/50(0.10)
 

P = 0.049(N)
 

0.327
 

0.082
 

1.993
 

106
 

7/40(0.18)
 

N.S.
 

1.794
 

0.499
 

7.745
 

92
 

HIGH
 

DOSE
 

4/50(0.08)
 

N.S.
 

3.920
 

0.405
 

188.989
 

110
 

1/50(0.02)
 

P = O.OOl(N)
 

0.082
 

0.002
 

0.518
 

110
 

4/46(0.09)
 

N.S.
 

0.891
 

0.177
 

4.499
 

89
 
TABLE 3

TOPOGRAPHY : MORPHOLOGY
 

Adrenal: Cortical Adenoma
 

P Values0
 

Departure from Linear Trend
 

Relative Risk (Control)d
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Thyroid: Follicular-Cell Adenoma
 

or Follicular-Cell Carcinoma*3
 

P Values0
 

Relative Risk (Control)d
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Hematopoietic System: Malignant Lymphoma
 

P Values0
 

Relative Risk (Control)d
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

 (CONTINUED) 

CONTROL
 

0/46(0.00)
 

N.S.
 

P = 0.031
 

5/48(0.15)
 

N.S.
 

106
 

1/49(0.02)
 

N.S.
 

107
 

LOW
 

DOSE
 

6/49(0.12)
 

P = 0.016
 

Infinite
 

1.506
 

Infinite
 

97
 

3/49(0.06)
 

N.S.
 

0.588
 

0.096
 

2.846
 

106
 

5/50(0.10)
 

N.S.
 

4.900
 

0.578
 

6.748
 

95
 

HIGH
 

DOSE
 

3/49(0.06)
 

N.S.
 

Infinite
 

0.566
 

Infinite
 

110
 

6/49(0.12)
 

N.S.
 

1.176
 

0.321
 

4.557
 

110
 

2/50(0.04)
 

N.S.
 

1.960
 

0.105
 

113.312
 

107
 
TABLE 3 (CONCLUDED)
 

o
 

Treated groups received time-weighted average doses of 1600 or 2995 ppm in feed.
 

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

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

the control group when P is less than 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 control group is given beneath the incidence of tumors in the treated group when P is less than
 

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.
 Q
 

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

group when P is less than 0.05.
 
TABLE 4
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN FEMALE RATS TREATED WITH CHLOROBENZILATE3
 

TOPOGRAPHY : MORPHOLOGY
 

Circulatory System: Hemangiosarcoma
 

P Values0
 

Relative Risk (Control)d
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pituitary: Chromophobe Adenoma
 

P Values0
 

S3
 

Relative Risk (Control)d
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Adrenal: Cortical Adenoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

CONTROL
 

4/50(0.08)
 

P = 0.026(N)
 

89
 

15/50(0.30)
 

N.S.
 

100
 

0/50(0.00)
 

P = 0.014
 

LOW
 

DOSE
 

1/49(0.02)
 

N.S.
 

0.255
 

0.005
 

2.459
 

111
 

11/45(0.24)
 

N.S.
 

0.815
 

0.378
 

1.688
 

101
 

2/47(0.04)
 

N.S.
 

Infinite
 

0.315
 

Infinite
 

103
 

HIGH
 

DOSE
 

0/50(0.00)
 

N.S.
 

0.000
 

0.000
 

1.079
 

—
 

11/45(0.24)
 

N.S.
 

0.815
 

0.378
 

1.688
 

90
 

5/47(0.11)
 

P = 0.024
 

Infinite
 

1.347
 

Infinite
 

98
 
TABLE 4 (CONTINUED) 

TOPOGRAPHY : MORPHOLOGY CONTROL
 

Thyroid: C-Cell Adenoma or C-Cell Carcinomab 5/50(0.10)
 

P Values0 N.S.
 

Relative Risk (Control)d
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor Ill
 

Thyroid: Follicular-Cell Adenoma or
 

Follicular-Cell Carcinomab 1/50(0.02)
 

P Values0 N.S.
 

oo Relative Risk (Control)d
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor Ill
 

Mammary Gland: Fibroadenoma 15/50(0.30)
 

P Values0 N.S.
 

Relative Risk (Control)d
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor 87
 

LOW
 

DOSE
 

3/47(0.06)
 

N.S.
 

0.638
 

0.104
 

3.088
 

111
 

2/47(0.04)
 

N.S.
 

2.128
 

0.114
 

2.810
 

110
 

14/49(0.29)
 

N.S.
 

0.952
 

0.479
 

1.879
 

99
 

HIGH
 

DOSE
 

2/47(0. 04)
 

N.S.
 

0.426
 

0.042
 

2.454
 

111
 

4/47(0. 09)
 

N.S.
 

4.255
 

0.444
 

204.823
 

90
 

16/50(0 .32)
 

N.S.
 

1.067
 

0.558
 

2.049
 

60
 
___ 

___ 

TABLE 4 (CONCLUDED) 

LOW HIGH
 

TOPOGRAPHY : MORPHOLOGY CONTROL DOSE DOSE
 

Uterus: Endometrial Stromal Polyp 4/49(0.08) 1/47(0.02) 4/46(0.09)
 

P Values0 N.S. N.S. N.S.
 

Relative Risk (Control) 0.266 1.087
 

Lower Limit 0.005 0.215
 

Upper Limit 2.559 5.505
 

Weeks to First Observed Tumor Ill 111 94
 

Hematopoietic System: Malignant Lymphoma 1/50(0.02) 0/49(0.00) 3/50(0.06)
 

P Values0 N.S. N.S. N.S.
 

Relative Risk (Control) 0.000 3.000
 N3 Lower Limit 0.000 0.250
 VO
 

Upper Limit 9.032 154.270
 

Weeks to First Observed Tumor Ill 106
 

Treated groups received time-weighted average doses of 1175 or 2229 ppm in feed.
 

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

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

the control group when P is less than 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 control group is given beneath the incidence of tumors in the treated group when P is less than
 

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

chlorobenzilate-dosed groups of either sex is included.
 

The incidence of cortical adenomas of the adrenal gland was
 

noted in dosed rats of both sexes. For males the Fisher exact test
 

indicated a significantly (P = 0.016) higher incidence of these
 

tumors in the low dose group than in the control group. The comparison between the control and the high dose group, however, was not
 

significant. For female rats the Fisher exact test indicated a significantly higher (P = 0.024) proportion of cortical adenoma of the
 

adrenal gland for the high dose group than for the control group.
 

Additionally, the Cochran-Armitage test for positive dose-related
 

trend was statistically significant (P = 0.014), indicating a positive association between dosage and tumor incidence. In historical
 

control data compiled by this laboratory for the NCI Carcinogenesis
 

Testing Program 3/160 (2 percent) of the male and 2/160 (1 percent)
 

of the female untreated Osborne-Mendel rats had a cortical carcinoma
 

or a cortical adenoma of the adrenal gland; additionally, 2/160 (1
 

percent) of the female historical untreated controls had an adrenal
 

pheochromocytoma.
 

Based upon these results there was an association between the
 

administration of chlorobenzilate and the increased incidence of
 

cortical adenoma of the adrenal gland in both male and female rats.
 

The possibility of a negative association between compound administration and incidence was noted for hemangiosarcoma in both male
 

and female rats.
 

30
 
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 many 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
 

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

indicating the theoretical possibility of tumor induction in rats by
 

chlorobenzilate that could not be established under the conditions of
 

this test.
 

31
 
IV. CHRONIC TESTING RESULTS: MICE
 

A. Body Weights and Clinical Observations
 

Dose-related mean body weight depression was observed in both
 

male and female mice. This effect was relatively consistent throughout the study in females and until week 80 in males. At this time
 

the high dose males gained weight rapidly, and their weight equaled
 

that of the low dose group at termination of the study (Figure 4).
 

Clinical signs were restricted to a hunched appearance, observed
 

in the high dose males during the first 10 weeks of the study. Following a decrease in concentrations administered to the dosed males
 

in week 11, the incidence decreased markedly until week 54 when approximately 50 percent of all the dosed mice displayed a hunched posture.
 

By week 66 and for the remainder of the study the animals appeared to
 

have recovered, and less than 10 percent were exhibiting this clinical
 

sign.
 

Signs commonly observed in group-housed laboratory mice were
 

observed in comparable numbers of control and dosed mice with the
 

frequency of observation increasing during the latter part of the
 

study. These signs included sores on the body (particularly in the
 

males), penile, anal, or vulvar irritation with occasional prolapse
 

and/or discharge, bloated appearance or abdominal distension,
 

swollen areas of the body, and alopecia.
 

32
 
TIME ON TEST (WEEKS) 

TIME ON TEST (WEEKS) 

FIGURE 4 

GROWTH CURVES FOR CHLOROBENZILATE CHRONIC STUDY MICE 

33 

120 
B. Survival
 

The estimated probabilities of survival for male and female mice
 

in the control and chlorobenzilate-dosed groups are shown in Figure 5.
 

For both sexes, the Tarone test did not show a positive association
 

between dosage and mortality.
 

In both sexes there were adequate numbers of mice at risk from
 

late-developing tumors with 82 percent (41/50) of the high dose males
 

and 88 percent (44/50) of the high dose females surviving on test until
 

the end of the study.
 

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).
 

Hepatocellular carcinomas occurred in 4/19 (21 percent) control
 

males, 32/48 (67 percent) low dose males, 22/45 (49 percent) high
 

dose males, 0/20 control females, 11/49 (22 percent) low dose females,
 

and 13/50 (26 percent) high dose females.
 

The hepatocellular carcinomas varied greatly in appearance.
 

Some lesions contained well-differentiated hepatic cells that had a
 

relatively uniform arrangement of the cords, whereas others had very
 

anaplastic liver cells with large hyperchromatic nuclei, often with
 

inclusion bodies and with vacuolated, pale cytoplasm. Arrangement of
 

the neoplastic liver cells varied from short stubby cords to nests of
 

hepatic cells and occasionally acinar formation. Mitotic figures
 

34
 
PROBABILITY OF SURVIVAL PROBABILITY OF 

30 

< 

o 

O 

I 

33 

O 

-n 

O 

oo 

30C O 3) CD m m ,_ 2w 

N 

m 

O 

X 

o 

z 

o 

O 

o 

m 

p p o p p p o b en bo O ro *• Oi 

I I I I I 

I I

I 

H 01­

m m 

O 

H 

m 

m tn X 

en en *

4: 

s- s­
were often present. Some of the tumors were characterized by discrete
 

areas of highly anaplastic cells. The hepatic neoplasms occurring in
 

the control mice were not different in histologic appearance from
 

those noted in the dosed mice.
 

Inflammatory, degenerative, and proliferative lesions in control
 

and dosed animals occurred in numbers and kinds similar to those
 

naturally occurring lesions seen in aged laboratory mice. Focal or
 

nodular hyperplasia of the hepatocytes occurred in 1/19 (5 percent)
 

control males, 3/48 (6 percent) low dose males, 2/45 (4 percent) high
 

dose males, 0/20 control females, 5/49 (10 percent) low dose females,
 

and 4/50 (8 percent) high dose females.
 

Based on the results of this pathologic examination, evidence
 

was provided for the carcinogenicity of chlorobenzilate under the
 

conditions of this bioassay (i.e., it caused an increased incidence
 

of hepatocellular carcinoma) in both dosed male and female mice.
 

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
 

chlorobenzilate-dosed groups of either sex is included.
 

For male mice the Fisher exact test indicated a significantly
 

larger proportion of hepatocellular carcinoma in the low dose group
 

(P = 0.001) than in the control. The comparison of high dose to control had a probability level of P = 0.034, a marginal result which
 

36
 
TABLE 5
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN MALE MICE TREATED WITH CHLOROBENZILATE*
 

TOP OGRAPHY : MORP HOLOGY
 

Liver: Hepatocellular Carcinoma
 

P Values0
 

Departure from Linear Trend
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Hematopoietic System: Malignant Lymphoma
 

u> P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

LOW 

CONTROL DOSE 

4/19(0.21) 32/48(0.67) 

N.S. P = 0.001 

P = 0.002 

3.167 

1.377 

10.454 

84 68 

1/19(0.05) 1/49(0.02) 

N.S. N.S. 

0.388 

0.005 

29.845 

90 77 

HIGH
 

DOSE
 

22/45(0.49)
 

P = 0.034
 

2.322
 

0.956
 

8.149
 

90
 

1/48(0.02)
 

N.S.
 

0.396
 

0.005
 

30.454
 

90
 

Treated groups received time-weighted average doses of 4231 or 7846 ppm in feed.
 

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

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

the control group when P is less than 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 control group is given beneath the incidence of tumors in the treated group when P is less than
 

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.
 

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

group when P is less than 0.05.
 
___ 

TABLE 6
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN FEMALE MICE TREATED WITH CHLOROBENZILATE'
 

TOPOGRAPHY rMORPHOLOGY
 

Liver: Hepatocellular Carcinoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Hematopoietic System: Malignant Lymphoma
 

u> P Values0
 

00
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

CONTROL
 

0/20(0.00)
 

P = 0.021
 

3/20(0.15)
 

P = O.Oll(N)
 

90
 

LOW
 

DOSE
 

11/49(0.22)
 

P = 0.016
 

Infinite
 

1.411
 

Infinite
 

90
 

7/50(0.14)
 

N.S.
 

0.933
 

0.246
 

5.215
 

73
 

HIGH
 

DOSE
 

13/50(0.26)
 

P = 0.007
 

Infinite
 

1.670
 

Infinite
 

86
 

0/50(0.00)
 

P = 0.021(N)
 

0.000
 

0.000
 

0.659
 

Treated groups received time-weighted average doses of 3200 or 5908 ppm in feed.
 

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

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

the control group when P is less than 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 control group is given beneath the incidence of tumors in the treated group when P is less than
 

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.
 
was not significant under the Bonferroni criterion. The results for
 

hepatocellular carcinoma were similar in female mice, as both the low
 

dose group (P = 0.016) and the high dose group (P = 0.007) had significantly higher proportions than the control groups. Additionally,
 

the Cochran-Armitage test indicated a significant (P = 0.021) positive association between dosage and tumor incidence.
 

Based upon these results there was an association between the
 

chlorobenzilate concentrations administered and the occurrence of
 

hepatocellular carcinomas in both male and female mice. There are no
 

other neoplasms for which the statistical tests were significant.
 

The possibility of a negative association between compound
 

administration and incidence was noted for malignant lymphomas in
 

the female mice.
 

39
 
V. DISCUSSION
 

In both species, adequate numbers of animals in all groups survived sufficiently long to be at risk from late-developing tumors.
 

In all groups of dosed mice, hepatocellular carcinomas occurred
 

at significantly higher incidences than in control groups. A significant positive dose-related trend for tumor incidence was observed in
 

female mice, but in male mice hepatocellular carcinomas appeared in a
 

higher proportion of the low dose group. This departure from a positive linear trend in male mice does not necessarily reduce the significance of these findings.
 

The association of liver tumors in B6C3F1 mice with oral administration of chlorobenzilate in this bioassay is in agreement with
 

results reported by Innes et al. (1969) for male B6C3F1 and B6AKF1
 

mice. The Innes study, however, reported the incidence of hepatomas,
 

a general term applied to a broad range of liver tumors. In that
 

study, the proportion of chlorobenzilate-dosed male mice of both
 

strains developing hepatomas approached the proportion found in positive controls; hepatomas were not observed, however, in female mice
 

(Innes et al. , 1969).
 

There was a statistically significant association between administration of chlorobenzilate and the appearance of cortical adenomas
 

of the adrenal gland in low dose male and high dose female rats.
 

Cortical adenomas were observed in 0/46 control males, 6/49 (12 percent) low dose males, 3/49 (6 percent) high dose males, 0/50 control
 

40
 
females, 2/47 (4 percent) low dose females, and 5/47 (11 percent) high
 

dose females. Although the incidence of cortical adenomas in control
 

rats in this bioassay was lower than the incidence in historical controls at the same laboratory (3/160 [2 percent] of male Osborne-Mendel
 

rats and 2/160 [1 percent] of female Osborne-Mendel rats), the incidence
 

in rats dosed with chlorobenzilate was elevated relative to the historical controls.
 

The lack of clearly observable carcinogenic effects in dosed
 

rats does not appear to be due to inadequate dose levels. Dose-


related mean body weight depression was observed during the chronic
 

bioassay in both male and female rats. Compound-related testicular
 

atrophy was observed in male rats.
 

Under the conditions of this bioassay, orally administered
 

chlorobenzilate was carcinogenic in male and female B6C3F1 mice,
 

causing an increased incidence of hepatocellular carcinomas. The
 

results do not, however, provide sufficient evidence for carcinogenicity of chlorobenzilate in Osborne-Mendel rats.
 

41
 
VI. BIBLIOGRAPHY
 

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

J. Wiley & Sons, New York, 1971.
 

Bartsch, E., D. Eberle, K. Ramsteiner, A. Tomann, and M. Spinder,
 

"The Carbinole Acaricides: Chlorobenzilate and Chloropropylate."
 

Residue Review 39. Springer-Verlag, New York, 1971.
 

Berenblum, I., editor, Carcinogenicity Testing. International Union
 

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

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, D.C., 1977.
 

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

Co., Ltd., London, 1970.
 

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

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

Fowler, D.L. and J.N. Mahan, The Pesticide Review, 1975. Agricultural
 

Stabilization and Conservation Service, U.S. Department of Agriculture, Washington, D.C., 1976.
 

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.
 

Innes, J.R.M., B.M. Ulland, M.G. Valerio, L. Petrucelli, L. Fishbein,
 

E.R. Hart, A.J. Pallotta, R.R. Bates, H.L. Falk, J.J. Gart, M.
 

Klein, I. Mitchell, and J. Peters, "Bioassay of Pesticides and
 

Industrial Chemicals for Tumorigenicity in Mice: A Preliminary
 

Note." Journal of the National Cancer Institute 42(6):1111-1114,
 

1969.
 

Johnson, R.D. and D.D. Manske, "Residues in Food and Feed." Pesticides
 

Monitoring Journal 9(4) : 157-169 , 1976.
 

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

Observations." Journal of the American Statistical Association
 

53:457-481, 1958.
 

42
 
Linhart, M.S., J.A. Cooper, R.L. Martin, N.P. Page, and J.A. Peters,
 

"Carcinogenesis Bioassay Data System." Computers and Biomedical
 

Research 7:230-248, 1974.
 

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

Co., New York, 1966.
 

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.
 

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.
 

Stanford Research Institute, "Stage 1A Chemical Dossier, Chlorobenzilate." Menlo Park, California, 1975.
 

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

62:679-682, 1975.
 

43
 

Review of the Bioassay of Chlorobenzilate* for Carcinogenicity
 

by the Data Evaluation/Risk Assessment Subgroup
 

of the Clearinghouse on Environmental Carcinogens
 

June 29, 1978
 

The Clearinghouse on Environmental Carcinogens was
 

established in May, 19?6, in compliance with DHEW Committee
 

Regulations and the Provisions of the Federal Advisory
 

Committee Act. The purpose of the Clearinghouse is to
 

advise the Director of the National Cancer Institute (NCI)
 

on its bioassay program to identify and to evaluate chemical
 

carcinogens 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 chemistry, biochemistry,
 biostatistics, toxicology, pathology, and epidemiology.
 Representatives of various Governmental agencies participate
 

as ad hoc members. The Data Evaluation/Risk Assessment
 

Subgroup of the Clearinghouse is charged with the responsi­ bility of providing a peer review of reports prepared on
 

NCI-sponsored bioassays of chemicals studied for carcinogenicity. It is in this context that the below critique is given
 

on the bioassay of Chlorobenzilate for carcinogenicity.
 

The reviewer agreed with the conclusion in the report
 

that Chlorobenzilate was carcinogenic in treated mice. The
 

evidence in treated rats was only "suggestive" of a carcino­ genic effect. The reviewer was critical of the high dosages
 

administered, which necessitated the intermittant treatment
 

of the animals, and of the small number of control mice.
 

Despite the shortcomings and the demonstration of a
 

carcinogenic response In only one species, he moved that
 

the report on the bioassay of Chlorobenzilate be accepted as
 

written. The motion was approved without objection.
 

Clearinghouse Members present:
 

Arnold L. Brown (Chairman), Mayo Clinic
 

Paul Nettesheim, National Institute of Environmental
 

Health Sciences
 

Verne Ray, Pfizer Medical Research Laboratory
 

Verald K. Rowe, Dow Chemical U.S.A.
 Michael B. Shimkin, University of California at San Diego
 

Louise Strong, University of Texas Health Sciences Center
 

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.
 

45
 

JJU . S . GOVERNMEN T PRINTIN G OFFICE : 1978-260-899/3200
 
APPENDIX A
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS
 

IN RATS TREATED WITH CHLOROBENZILATE
 

TABLE Al

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS TREATED WITH CHLOROBENZILATE 

(rONTKO L (VEH ) 

01-M00 1 

ANIMAL S INITIALL Y I N STUD Y 50 

ANIMAL S NECROPSIE D 49 

ANIMAL S EXAMINE D HI STOPATHOLOG I CALL Y*¥ 49 

INTEGUMENTAR Y SYSTEM 

*SKIN (It9) 

S^UAMOU S CELL CARCINOM A 

HEMAN3IOPERICYTOMA , MALIGNAN T 1 <2X ) 

*SUBCUT TISSU E 

CARCINOMA , NOS 

S^UAMOU S CELL CARCINOM A 

FIBROM A 

FIBROSARCOW A 

LIPOM A 

HEMANGIOSARCOM A 

ANGIOSARCOM A 

RESPIHATOH Y SYSTE M 

tLUNlj 

MIXE D TUMOR , MALIGNAN T 

HEMANGIOSABCOMA , METASTATI C 

HEMATOPOIETIC SYSTEM 

• MULTIPL E ORGAN S

MALIGNAN T IYMPHOMA , NOS 

MALIG.LYMPHOMA , LYHPHOCYTI C

MALIG.LYMPHOMA , HISTIOCYTIC

•SUBCUT TISSUF/GHOIN

MALIG.LYMPHOMA , HISTIOCYTIC

tSPLEEN

HEMANGIOSARCOM A

(19) 

1 (2%) 

1 (2X ) 

4 (8%) 

(19) 

1 (2%) 

2 (14%) 

 (49 ) 

 TYPE 1 (2% ) 

 TYPE 

 (49) 

 TYPE 

 (47 ) 

4 (9X ) 

LOB DOSE 

01-M00 4 

50 

50 

50 

(50) 

1 (2X) 

(50) 

1 (2*) 2 (4% ) 

3 (6*) 

1 (2%) 

(50) 

(50) 

1 (2%) 

1 (2%) 

2 (UX ) 

(50) 

(49 ) 

2 (4%) 

MALIG i LYMPHOMA £ ._HISTigCYTIC_TYP E 

* NUMriE R O F ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALL Y 

» NUMBE R OF ANIMAL S NECROPSIE D 

**EXCLUDES PARTIALLY AUTOLYZED ANIMALS
 

A-3 

HIGH DOSE 

01-M00 5 

50 

50 

50 

(50) 

(50) 

1 (2X ) 

1 (2X ) 

U (8X) 

2 (4X ) 

(50) 

(50) 

1 (2%) 

(50) 

1 (2)5) 

<H9 ) 

1 (2%) 
TABLE Al (CONTINUED) 

CONTROL (VEH ) LOS DOSE HIG H DOSE 

01-M00 1 01-M004 01-M005 

CIRCULATOR Y SYSTE M 

*HEA6 T (47) (50) (50) 

HiMANGIOSAPCOM A 1 (2X) 

HEMANGIOSAECOMA , METASTATI C 1 (2% ) 

DIGESTIV E SYSTE M 

• LIVER (49 ) (49) (49) 

NEOPLASTIC NODUL E 1 (2X) 1 (2X ) 

HEMAN3IOSARCOMA , METASTATIC 1 (2«) 

URINArt Y SYSTE M 

• KIDNEY (47) (U9) (50)
 

LIPOMA 2 (4X)
 

HEMAN3IOSARCOMA 1 (2%)
 

HAMARTOMA + 1 (2X)
 

• URINAR Y BLADDER (46) (49) (46)
 

PAPILLOMA, NOS 3 (7%)
 

ENDOCRINE SYSTEM
 

• PITUITARY (41) (40) (46)
 

CHSOMOPHOBE ADENOMA 4 (10X) 7 (18X) 4 (9%)
 

*ADPlJNA L (46) (49) (49) 

CORTICA L ADENOM A 6 (12X) 3 (6X) 

PhEOCHROMOCYTOM A 1 (2X) 1 (2% ) 

• THYROI D (48) (49) (49) 

FOLLICULAP-CEL L ADENOM A 3 (6X) 2 (4X) 4 (8X)

 

FOLLICULAR-CEL L CARCINOM A 4 (8X) 1 (2X) 2 (4X) 

C-CELL CARCINOM A 1 (2%) 1 (2X) 

• PARATHYROI D (46) (30) (31) 

ADENOMA , NOS 1 (2% ) 1 (3X) 

• PANCREATI C ISLETS (46 ) (49) (49) 

1_J2S1_ 

* NUMBE R OF ANIMAL S WIT H TISSU E EXAMINE D MICROSCOPICALLY 

* NUMBE E OF ANIMAL S NECROPSIE D 

+ THIS IS CONSIDERED TO BE A BENIGN FORM OF THE MALIGNANT MIXED TUMOR OF THE KIDNEY AND CONSISTS OF PROLIFERATIVE 

LIPOCYTES, TUBULAR STRUCTURES, FIBROBLASTS, AND VASCULAR SPACES IN VARYING PROPORTIONS. 

A-4 
TABLE A1 (CONTINUED) 

CONTRO L (VEH ) LOli DOSE HIG H DOSE 

01-H001 01-M00 4 01-M00 5 

REPRODUCTIV E SYSTF H 

*NAMMAH Y GLAN D (49 ) (50) (50)
 

ADENOCARCINCKA , NO S 2 (4%)

 

FIBBOADENOH A 1 (2%) <2X)
 

tPROSTAT E (34) (43) (35)
 

HEMANGIOSARCOMA , (1ETASTATIC 1 (3%)
 

*SEM1NAL VESICLE (49) (50) (50)
 

HtfMANGIOSAECOMA, BETASTATIC 1 (2X)
 

NERVOU S SYSTEM
 

#BRAIN (47) (50) (50)
 

GLIOMA, NOS I (29!)
 

EPENDYHOHA 1 (2%)
 

SPECIAL SFNSE ORGANS
 

NONE
 

MUSCULOSKELETAL SYSTEM
 

"SKELETAL MUSCLE (49) (50) (50)
 

FIBBOSABCGMA 1 (2%)
 

*KUSCLE OF THORAX (49) (50) (50)
 

HEMANGIOSARCOMA 1 (2%)
 

BODY CAVITIES
 

"MEDIASTINUM (49) (50) (50)
 

FIBROSARCOBA 1 (2%)
 

"ABDOMINAL CAVITY (49) (50) (50)
 

LIPOBA 1 (2%)
 

"TUNICA VAGINALIS (49) (50) (50)
 

* NUMBEP OF ANIMALS WITH TISSUE FXAHINED fllCBOSCOPICA LLY
 

* NUMBER OF ANIMALS NECROPSIED
 

A-5
 
TABLE Al (CONCLUDED) 

CONTROL (VEH) LOW DOSE HIG H DOSE 

01-M001 01-MOO t 01-M005 

A LL OTHE R SYSTEM S 

*MULIIPL E ORGAN S (U9) (50) (50)
 

HEMANGIOSAECOM A 1 (2*)
 

DISPOSITION SUMMAR Y
 

ANIMALS INITIALLY IN STUDY 50 50 50
 NATURAL DEATHS 24 20 15
 MORIBUND SACRIFICE 2 1
 

SCHEDULED SACRIFICE
 

ACCIDENTALLY KILLED
 

TERMINAL SACRIFICE 30
 A1.IMAL MISSING
 

INCLUDES AUTOLYZED ANIMALS
 

TUMOR SUMMAR Y
 

TOTAL ANIMAL S WITH PRIMAR Y TUMORS* 28 31 23
 

TOTAL PRIMARY TUMORS 39 30
 

TOTA L ANIMAL S WIT H BENIG N TUMOR S 1 3 20 TOTA L BENIG N TUMOR S 1 7 21 19 

TOTAL ANIMAL S WIT H MALIGNAN T TUMORS 1 8 15 9

 

TOTA L MALIGNAN T TUMOR S 2 <• 17
 

TOTAL ANIMALS WITH SECONDARY TUMORS* 2
 TOTAL SECONDARY TUMORS 6
 

TOTAL ANIMALS WITH TUM3RS UNCERTAINBENIGN OR MALIGNANT 1 2
 

TOTAL UNCERTAIN TUMORS 1 2
 

TOTAL ANIMAL S WITH TUMOPS UNCERTAIN-
 PRIrlARY OE MfTASTATIC
 

TOTAL UNCERTAIN TUMORS
 

PRIilARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS
 SECuNDARY TUMORS: METASTATIC TUMORS OR TUMORS INVASIVE INTO AN ADJACENT ORGAN
 

A-6 
TABLE A2

 

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

ANIMAL S INITIALL Y IN STUD Y

ANIMAL S MISSIN G 

ANIMAL S NECROPSIE D

ANIMAL S EXAMINE D HI STOPATHOLOGICALL Y **

INTEGUMENTAR Y SYSTEM 

*SUBLUT TISSUE 

BASAL-CEL l CARCINOM A 

FIBROM A 

"IBROSARCOM A 

LIPOMA 

HEMANGIOSARCOM A 

RESPIRATOR Y SYSTE M 

tLUNG 

ALVEOLAR/BRONCHIOLAR ADENOMA
 

HEMATOPOIETIC SYSTEM
 

*MULTIPLE ORGANS
 

MALIG.LYMPHOMA, LYMPHOCYTIC TYPE
 

MALIG. LYMFHCMA, HISTIOCYTIC TYPE
 

ISPLEEN
 

HEMANGIOSAHCOMA
 

ICERVICAL LYMPH NODE
 

MALIG.LYMPHOMA, LYMPHOCYTIC TYPE
 

•SMALL	 INTESTINE
 

MALIG.LYMPHOMA, HISTIOCYTIC TYPE
 

CIRCULATORY SYSTEM
 

IENDOCARDIOM
 

SARCCMAt_fiQS___
 

CONTRO L (VEH ) 

01-F001 

 5 0 

 5 0 

 50 

(50) 

1 (2% ) 

2 (UX ) 

1 (2% ) 

3 <6X ) 

(50)
 

(50)
 

(50)
 

(48)
 

1 (2%)
 

(50)
 

(50) 

L OW DOSE HIG H DOSE 

01-F006 C1-F007 

50 50 

1 

49 50 

48 148 

(49) (50 ) 

2 (4X ) 

1 (2%) 

(18)	 (48)
 

1 (2%)
 

(49)	 (50)
 

1 (2X)
 

1 (2X)
 

(47)	 (48)
 

1 (2%)
 

(46) (43)
 

(48)	 (48)
 

1 (2%)
 

(48) (48) 

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

* NUMBE R OF ANIMAL S NECROPSIE D 

**EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

A-7 
TABLE A2 (CONTINUED) 

CONTROL (VEH ) L OW DOSE HIG H DOSE 

01-F001 01-F006 01-F007 

DIGESTIV E SYSTE M 

*LIVZR (50) (48) (48)
 NEOPLASTIC NODULE 1 (2%)
 HdPATOCELLULAR CARCINOMA 1 (2%)
 

tDUODENUM (50) (48) (48)
 HLMANGIOSAFCONA 1 (2%)
 

URINARY SYSTFM
 

•KIDNEY	 (50) (48) (48)
 

LIPOMA 1 (2X>
 FIXED TUMOR, MALIGNANT 1 (2X)
 HAMARTOHA + 1 (2%) 1 (2*)
 

ENDOC3INE SYSTEM
 

•PITUITARY	 (50) (45) (45)
 

CHRONOPHOEE ADENOMA 15 (30*) 11 (24S) 11 (24X)
 

•ADRENAL	 (50) (47) (47)
 

CORTICAL ADENOMA 2 (U*) 5 (11*)
 

PHEOCHHCMCCYTOMA 2 (4X)
 

• THYtiOID (50) (47) (47)
 

FOLLICULAR-CELL ADENOMA 2 («X) 3 (6*)
 FOLLICULAR-CELL CARCIN3MA 1 (2*) 1 (2*)
 C-CELL ADENOMA 5 (10*) 2 (4X) 2 (4X)
 C-C^LL CARCINOMA	 1 (2*)
 

•PANCREATIC	 ISLETS (50) (48) (48)
 

ISLET-CELL ADENOMA 2 (4X) 1 (2X)
 

REPRODUCTIVE SYSTEM
 

*MAMMAR Y GLAND (50) (49) (50)

 

CARCINOMA,NOS 1 (2*)

 ADENOMA, NOS 1 (2*) 1 (2X)

 ADENOCARCINOMA, NOS 1 (2X)

 FIBROADENCMA 15 (30*) 14 (29*) 16 (32X)

 

fUTERU S (49) (47) (46)

 

Lfc 10 MYOM A J-J2J1 

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

' NUMBE R OF ANIMAL S NECROPSIE D 

t- THIS IS CONSIDERED TO BE A BENIGN FORM OF THE MALIGNANT MIXED TUMOR OF THE KIDNEY AND CONSISTS OF PROLIFESATIVE 

LIPOCYTES, TUBULAR STRUCTURES, FIBROBLASTS, AND VASCULAR SPACES IN VARYING PROPORTIONS. 

A-8 

I
TABLE A2 (CONTINUED) 

CONTROL (VEH ) LOW DOSE HIG H DOSE 

01-F001 01-F006 01-F007 

ENDOMETRIA L STROMA L POLYP 4 (8») 1 (2«) 4 (9X) 

HUTERUS/ENDOHETRIO M (49) (47) (46) 

ENDOMETHIA L STROMA L POLY P 1 (2X) 

• OVAR Y (H9) (U8) (46) 

CAECINOMA.NO S 1 (2%) 

PAPILLAB Y CYSTADENOMA . NOS 1 (2X) 

GHANULOSA-CEL L TUMO R 1 (2% ) 

OSTEOSARCOM A 1 (2X) 

NERVOU S SYSTE M 

NONE 

SPECIAL SENS'; ORGAN S 

NON E 

MUSCULOSKELETA L SYSTEM 

NON E 

BODY CAVITIES 

*ABDOMINAL CAVITY (50) (49) (50)
 

LIPOMA 1 (2%)
 

*ABDOMINA L VISCERA (50) (49) (50) 

HEMANGIOSARCOM A 1 (2*) 

A LL OTHER SYSTEM S 

* HUHBE R OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBE R OF ANIMAL S NECROPSIED 

A-9 
TABLE A2 (CONCLUDED)
 

CONTROL (VEH) LOW DOSE HIGH DOSE
 

01-F001 01-F006 C1-F007
 

ANIMAL DISPOSITION SUMMARY
 

ANIMALS INITIALLY IN STUDY 50 50 50 

NATURAL DEATHS 11* 11

 

MORIBUND SACRIFICE 2 

SCHEDULED SACRIFICE
 

ACCIDENTALLY KILLED 1

 TERMINAL SACRIFICE 36 38 36

 ANIHAL MISSING 1

 

INCLUDES AUTCLYZE D ANIMAL S 

TUMO R SUMMAR Y 

TOTAL ANIMAL S WIT H PRIMAR Y TUBORS* 36 38 3U 

TOTAL PRIMAR Y TUMORS 56 U9 51 

TOTAL ANIMAL S KITH BENIGN TUMORS 31 31 32 

TOTA L BISIGN TUMORS <t U U3 45 

TOTAL ANIMAL S WITH MALIGNAN 9 T TUMORS 6 6 

TOTAL MALIGNAN T TUMORS 1 0 6 6 

TOTAL ANIMAL S WITH SECONDARY TUUORSI 

TOTAL SECONDAR Y TUMORS 

TOTAL ANIMAL S WIT H TUMORS UNCERTAIN ­

BENIG N OR MAIIGNAN T 2 

TOTAL UNCERTAI N TUMORS 2 

TOTAL ANIMAL S WITH TUMORS UNCERTAIN ­

PRIMAR Y OR METASTATIC 

TOTAL UNCERTAI N TUMORS 

* PRIMAR Y TUMORS : ALL TUMORS EXCEPT SECONDAR Y TUMORS 

* SECONDAR Y TUMORS : METASTATIC TUMORS OR TUMOR S INVASIV E INTO AN ADJACEN T ORGA N 

A-10 
APPENDIX B
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS
 

IN MICE TREATED WITH CHLOROBENZILATE
 

TABLE Bl

 

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

CONTROL (VEH )

02-M006 LOW DOSE 02-H007

 HIGH DOSE 

 02-B008 

ANIMALS INITIALLY IN STUDY 20 50 50 

ANIMAL S MISSING 1 1 

ANIMAL S NECROESIED 19 49 48 

ANIMAL S EXAMINE D HISTOPATHOLOGICALLY** 17 47 45 

INTEGUMENTAR Y SYSTEM 

*SUBCUT TISSUE (19) (19) (48) 

FIBHOSARCCHA 1 (2X) 

RESPIRATOR Y SYSTEM 

ILUNG (19) (48) (4<l) 

ALVEOLAR/BRONCHIOLA R ADENOM A 1 (5X) 1 (2%) 1 (2X) 

HEMATOPOIETIC SYSTEM 

*MULTIPLE ORGAN S (19 ) (49) (48) 

HALIG.LYMPHOMA , HISTIOCYTIC TYPE 1 (2*) 1 (2X) 

*SPLEEN (18) (47) (114)
 

HEHANGIOSARCOMA 1 (2«)
 

IKIDNEY (19) (47) (45)
 

MALIG.I.YMPHOMA, LYMPHOCYTIC TYPE 1 (5X)
 

CIRCULATORY SYSTEM
 

NONE
 

DIGESTIVE SYSTEM
 

tLIVER (19) (48) (45)
 

HEPATOCELLOLAR CARCINOMA 4 (21*) 32 (67*) 22 (49X)
 

HEMANGIOMA 1 (2*)
 

HEMANGIOSAECOMA, NETASTATIC 1 (2*)
 

URINARY SYSTEM
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS N1CROPSIED
 

**EXCLUDES PARTIALLY AUTOLYZED ANIMALS
 

B-3
 
ENDOCRIN E SYSTE M 

NONE 

REPRODUCTIVE SYSTEM
 

NONE
 

NERVOUS SYSTEM
 

NONE
 

SPECIAL SENSE ORGANS
 

*HARDERIAN GLAND


ADENOMA, NOS


KUSCOLOSKELETAL SYSTSM
 

NONE
 

BODY CAVITIES
 

NONE
 

ALL OIHFR SYSTEMS
 

NONE
 

ANIMAL DISPOSITION SUMMARY
 

ANIMALS INITIALLY IN STUDY
 NATURAL DEATHS


MORIBUND SACRIFICE


SCHEDULED SACRIFICE
 

ACCIDENTALLY KILLED


TERMINAL SACRIFICE


ANIMAL MISSING


TABLE Bl (CONTINUED) 

CONTROL(VEH ) LOB DOSE HIGH DOSE 

02-H006 02-M007 02-H008 

 (19)	 (19) («8)
 

1 (2*)
 

 20 50 50
 7 15 6
 

1
 

2
 

 12 3U U1
 

1 1
 

* NUMBER OF ANIMALS BITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NEC80PSIED
 

B-4
 
TABLE Bl (CONCLUDED)
 

CONTROL (VEH) LOi DOSE HIGH DOSE
 

02-(1006 02-N007 02-M008
 

TUMOK SUMMAR Y
 

TOTAL ANIMALS WITH PRIMARY TUMORS* 3U 25
 TOTAL PRIMARY TUMORS 37 25
 

TOTAL ANIMALS WITH BENIGN TUMORS
 TOTAL BENIGN TUMORS
 

TOTAL ANIMALS WITH MALIGNANT TUMORS 33 2U
 TOTAL MALIGNANT TUMORS 3H 2«
 

TOTAL ANIMALS WITH SECONDARY TUMORSt 1
 TOTAL SECONDARY TUHORS 1
 

TOTAL ANIMALS WITH TUHORS UNCERTAIN­ BENIGN OR MALIGNANT
 

TOTAL UNCERTAIN TUMORS
 

TOTAL ANIMALS WITH TUMORS UNCERTAIN­ PRIMARY OR METASTATIC
 

TOTAL UNCERTAIN TUMORS
 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMOES
 * SECONDARY TUMORS: METASTATIC TUMORS OR TUMORS INVASIVE INTO AN ADJACENT ORGAN
 

B-5
 
TABLE B2

 

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

CONTROL (VE8) LOW DOSE HIG H DOSE 

02-F006 02-P009 02-F010 

ANIMAL S INITIALL Y IN STUD Y 20 50 50 ANIMAL S NECROPSIE D 20 50 50 ANIMAL S EXAMINE D HISTOPATHOLOGICALLY ** 20 49 50 

INTEGUMENTAR Y SYSTEM 

NON E 

RESPIRATOR Y SYSTE M 

#LUNG (19) C*9) (50) ALVEOLAR/ERONCHIOLAR ADENOMA 2 (11X) 1 (2*) 2 (4X) ALVEOLAR/BRONCHIOLAR CARCINOMA 1 (2*) 

HEMATOPOIETIC SYSTEM 

*BULTIPIE ORGAN S (20 ) (50) (50) 

MALIG.LYMPHOMA , LYMPHOCYTIC TYPE 1 (5X) 

MALIG.LYMPHOMA , HISTIOCYTIC TYPE 6 (12X) 

MALIGNAN T LYMPHOMA , MIXE D TYPE 2 (10*) 

tSPLEEN (20) (49) (49)
 

MALIGNANT LYMPHOMA, MIXED TYPE 1 (2%)
 

CIRCULATORY SYSTEM
 

NONE
 

DIGESTIVE SYSTEM
 

iLIVER (20) (49) (50) 

HEPATOCELIULAR CARCINOMA 11 (22X) 13 (26*) 

»STOMACH (19) (49) (50) S^UAMOUS CELL CARCINOMA 1 (2%) 

URINAB Y SYSTEM 

* NUMBE R OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBE R OF ASIMAL S NECROPSIED 

**EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

B-6 
ENDOCRINE SYSTEM
 

NONE
 

REPRODUCTIVE SYSTEM
 

fUTERUS


S^UAMOHS CELL PAPILLOMA


ENDOMETRIAL STROMAL POLYP


NERVOUS SYSTEM
 

NONE
 

SPECIAL SENSE ORGANS
 

NONE
 

KUSCULOSKELETAL SYSTEM
 

NONE
 

BODY LAVITIES
 

NONE
 

ALL OTHER SYSTEMS
 

NONE
 

ANIMAL DISPOSITION SUMMARY
 

ANIMALS INITIALLY IN STUDY


NATURAL DEATHS


MORIBUND SACRIFICE


SCHEDULED SACRIFICE
 

ACCIDENTALLY KILLED


TERMINAL SACRIFICE


ANIMAL MISSING
 

2_IH£1.11DES_AU1QLYZED_ANIMALS
 

TABLE B2 (CONTINUED) 

CONTROL (VEH ) LOW DCSE HIGH DOSE 

02-F006 02-F009 02-F010 

 (20)	 (U9) (U8)
 

1 (2S)
 

1 (2%)
 

 20 50 50
 

6 U
 

1
 

1
 

 20 44 It
 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIHALS NECROPSIED
 

B-7
 
TABLE B2 (CONCLUDED) 

CONTROL (VEH) LO U DOSE HIG H DOSE 

02-F006 02-F009 02-F010 

TUMOR SUMMAR Y
 

TOTAL ANIMALS WITH PRIMARY TUMORS* 5 18 15
 

TOTAL PRIMAR Y TUMORS 5 22 16
 

TOTAL ANIMALS HITH 2 BENIGN TUMORS 3 2
 

TOTAL BENIGN TUMORS 2 3 2
 

TOTAL ANIMALS HITH MALIGNANT TUMORS 3 16 11
 

TOTAL MALIGNANT TUMORS 3 19 1U
 

TOTAL ANIMALS WITH SECONDARY TUMORS*
 

TOTAL SECONDARY TUMORS
 

TOTAL ANIMALS WITH TUMORS UNCERTAINBENIGN OR MALIGNANT
 

TOTAL UNCERTAIN TUMORS
 

TOTAL ANIMALS WITH TUMORS UNCERTAINPRIMARY OR METASTATIC
 

TOTAL UNCERTAIN TUMORS
 

* PRIHARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS
 # SECONDARY TUMORS: METASTATIC TUMORS OR TUMORS INVASIVE INTO AN ADJACENT ORGAN
 

B-8
 
APPENDIX C
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN RATS TREATED WITH CHLOROBENZILATE
 

TABLE Cl

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE 

RATS TREATED WITH CHLOROBENZILATE 

CONTROL (VEH ) LOU DOSE HIGH DOSE 01-M001 01-M004 C1-M005 

ANIMAL S INITIALL Y III STUDY 50 50 50 

ANIMAL S NECBOFSIED 49 50 50 

ANIMAL S EXAMINE D HISTOPATHOLOGICA LLY *» 49 50 50 

INTEGUMENTAR Y SYSTEM 

• SKIN EPIDERMAL INCLUSION INFLAMMATION, CHRON CYST IC 

(49) 

1 (2*) 

(50) 

1 (2X) 

(50) 

*SUBCUT TISSUE 

ABSCESS, NOS 

NECROSIS, FAT 1 (2X) 

(50) (50) 

1 (2X) 

RESPIRATORY SYSTEM
 

*TRACHEA (50) (50) 

INFLAMMATION, NOS 4 (100*) 

INFLAMMATION, CHRONIC 6 (12%) 1 (2X) 

HYPEHPLASIA, LYBPHOID 2 (ttX) 

tLUNG/BHONCHIOLE (49) (50) (50) HYPEBPLASIA, LYMPHOID 18 (36X) 6 (12*) 

fLHNG (49) (50) (50)
 HEMORRHAGE 1 <2X)
 LOBAE PNEUMONIA, NOS 1 (2X)
 INFLAMMATION, NOS 2 (4%) 9 (18X)
 INFLAMMATION, FOCAL 5 (10X) 7 (14X)
 PNEUMONIA, ASPIRATION 1 (2%) 1 (2X)
 LOBAR PNEUMONIA NECHOTIZING 1 (2%)
 ABSCESS, NCS 1 (2X>
 PNEUMONIA, CHRONIC MURINE 20 (41X) 12 (24X) 1 (2X)
 CALCIFICATION, NOS 1 (2*)
 HYPERPLASIA, ALVEOLAR EPITHELIUM 1 (2X)
 

HEMATOPOIETIC SYSTEM 

tSPLLEN (47 ) (49) (49) 

ElfiROSIS 

* NUMBE R OF ANIMAL S KITH TISSUE EXAMINE D MICPOSCOPICALLY 

* NUMBE R OF ANIMAL S NECHOPSIED 

**EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

C-3 
TABLE Cl (CONTINUED) 

CONTROL (VEH) L OW DOSE HIG H DOSE 

01-M001 01-H004 01-N005 

HEMOSIDESOSIS 3 (6%) 1 (2»)

 HYPEEPLASIA , LYMPH3I D 1 (2*)

 

HEHATOPOIESI S 1 (2X) 3 (6*) 1 (2*)

 

»MESENTERI C L. NODE (15) (18) (17) CYST, NOS 1 (2%)

 

FIBROSIS 1 <2X )

 

CIRCULATORY SYSTEM 

*HEART/ATRIU M (17) (50) (50)

 

EBBOLUS , SEPTIC 1 (2«)

 

*HEAET/VENTRICL E (17) (50) (50) THPOMBOSIS , NOS 1 (2X ) 

*MYOCARDIU M (17) (50) (50) INFLAMMATICN , NO S 11 (30%) INFLAMMATION , FOCAL 1 (2X) INFLAMMATION , INTERSTITIA L 1 (2%) INFLAMMATION , HEMORRHASI C 1 (2«) INFLAMMATION , CHRONIC 1 (2%) 

INFLAMMATION , CHRONIC FOCAL 1 (2X) FIBROSIS 1 (2%) 5 (10X) 7 (14*) FIBROSIS, FOCAL 8 (15X) DEGENERATION , SOS 1 (2*) 2 (IX) CALCIFICATION , FOCAL 1 (2%) 1 (2*) 

(ENDOCARDIU M (17) (50) (50) 

HYPERPLASIA , NOS 2 (1%) 2 («() 

*ARTERIOL E (19) (50) (50)

 HYPERTROPHY , NO S 1 (2*)

 

*AORTA (19) (50) (50)

 ARTERIOSCLEROSIS , NOS 1 (8%)

 MEDIA L CALCIFICATIO N 1 (2*)

 CALCIFICATION , NO S 2 (i»%)

 

• PULMONAR Y ABTERY (19) (50) (50) CALCIFICATION , NO S 5 (10%) 1 (2*) 

DIGESTIVE SYSTEM 

*LIVER (19) (19) (19)

 SXS.Ti.NOS _ _

 2-iazi 

I NUMBE R OF ANIMAL S WIT H TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBE R OF ANIMAL S NECROPSIE D 

C-4 
TABLE Cl (CONTINUED) 

CONTROL (VEH ) 

01-M001 

LOW D

01-M

OSE 

001 

HEMORRHAG E 

INFLAMMATION , NOS 

ABSCESS, NOS 

NECROSIS, NOS 

NECROSIS, FOCAL 

METAMORPHOSI S FATTY 

FOCAL CELLULA R CHANG E 

HYPERPLASIA , NOS 

ANGIECTASI S 

3

3

5

 <6X) 

 (6X) 

 (10X) 

1

2

 (2X) 

 (IX) 

*LIViH/CENTRILOBULA R 

METAMORPHOSIS FATTY 

(19) (19) 

1 (2*) 

*BILE DUCT 

INFLAMMATION , CHRONIC 

HYPERPLASIA , NO S 

(19) 

3 (6X) 

(50) 

1

13

 (2X) 

 (26X) 

#PANCHEA S 

INFLAMMATION, ACUTE 

INFLAMMATION, ACUTE/CHRONIC 

PERIARTERITIS 

ATROPHY, NOS 

ATROPHY, FOCAL 

(16) 

5 (11X) 

(19) 

1

1

3 

1 

5 

 <2X) 

 (2X) 

<6X) 

(8X) 

(10X) 

IESOPHAGUS 

INFLAMMATION, NOS (1) 

1 (1COX) 

(10) 

tSTOBACH 

MINERALIZATION 

HEMORRHAGE 

INFLAMMATION, NOS 

INFLAMMATION, CHRONIC 

ULCER, CHRCNIC 

CALCIFICATION, NOS 

HiPERKEHATCSIS 

(16) 

1 (2X) 

(50) 

1

1

2

1

1 

2 

 (2X) 

(2%) 

 (IX) 

 (2X) 

(2X) 

(IX) 

tGASTRIC MUCOSA 

HEMORRHAGE 

CALCIFICATION, SOS 

(16) (50) 

*LARGE INTESTINE 

NEMATODIASIS 

(16) (18) 

5 (10X) 

iCOLJN (16) (18) 

____ INFLAMSiTICNx _NOS ____________________ J_ 

# NUMBE R OF ANIMAL S SITH TISSDE EXAMINE D MICROSCOPICALL Y 

* NUMBE R OF ANIMAL S NECROPSIE D 

C-5 

HIG H DOSE 

C1-M005 

1 (2X) 

1 (2X)
 

1 (2X)
 

1 (2X)
 

(19)
 

(50)
 

7 (11X)
 

(19)
 

2 (IX)
 

3 (6X)
 

1 (2X)
 

(11)
 

(19)
 

2 (IX)
 

3 (6X)
 

(19)
 

1 (2X)
 

1 (2%)
 

(50)
 

6 (12X)
 

(50)
 
TABLE Cl (CONTINUED) 

CONTROL (VEH) L OW DOSE 

01-B001 01-MOO U 

URINAK Y SYSTE M 

tKIDHE Y (47) (K9 ) 

MINERALIZATIO N 1 (2X) 

PYELONEPHRITIS , NOS 2 (4X) 

PYONEPHROSI S 1 (2X ) 

ABSCESS, NOS 1 (2%) INFLAMMATION , CHRONI C 37 (79*) 35 (71X) 

• KIDNEY/CORTE X (47) (U9) ABSCESS, NOS 1 (2X) 

• RENA L PAPILL A (47 ) (19) CALCIFICATION , NOS 1 <2X ) 

#KIDNEY/PKLVI S (47 ) (19) CALCIUM DEPOSIT 

HYPERPLASIA , EPITHELIA L 1 (2*) 

• URINAR Y BLADDE R («6 ) (49) INFLAMMATION , NO S 1 (2*) INFLAMMATION , HEHORRHAGI C 1 (2X) 

ENDOCRINE SYSTEM 

• PITUITAR Y (41) (10) CYST , NOS 1 (2)5) 3 (8X) HYPERPLASIA , FOCAL 1 (3X) HYPERPLASIA , CHROB3PHOBE-CELL 1 (3X) 

• ADRENA L (46) (49)

 ANGIECTASI S 8 (17X )

 

tADRENA L CORTE X (46) (49) DEGENERATION , NOS 9 ( 1 8X ) NECROSIS, NOS 1 (2X) M£TAMOEPHCSI S FATTY 5 (10*) 

• ADRENA L MEDULL A <t6 ) (19) HYPERPLASIA , NOS 1 (2%) 

• THYROI D (18) (49) CYST, NOS 1 (2X) FOLLICULA R CYST.. N3S 6_ii2SL 

f NUMBE R OF ANIMAL S KIT H TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBE R OF ANIMAL S NECROPSIE D 

C-6 

HIG H DOSE 

01-M005 

(50) 

2 (4X) 

34 (68X) 

(50) 

(50) 

(50) 

1 (2«) 

(46) 

(46) 

2 (4)1) 

1 (2%) 

(49) 

1 (2%) 

(49) 

10 (20X) 

(19) 

(49) 

3_16«1 
TABLE Cl (CONTINUED) 

CONTEOL (VEH ) 

01-M00 1 

LOW DOS': 01-M00 4 

HIG H DOSE 

01-M00 5 

HYPERPLASIA , C-CELL 1 (2%) 8 (16%) 5 (10% ) 

HYPEEPLASIA , VOLLICU L AR -CELL 4 (8% ) 1 (2%) 

iPARATHYROI D 

HYPEEPLASIA, NOS 

(U6 ) 

2 (4%) 

(30 ) 

2 (7%) (J1 )3 (10%) 

HFPRODUCTIV E SYSTE M 

*MAMMAS Y GLAN D (49) (50) (50) 

GALACTOCEL F 

CYST, NO S 

1

1

 (2% ) 

 (2% ) 1 (2% ) 

*PEObTATE (34 ) (43 ) (35) 

INFLAMMATION , NO S 9 (26% ) 

INFLAMMATION , HEMORRHAGI C 1 (2% ) 

INFLAMMATION , ACUTE 6 (14%) « (11%) 

INFLAMMATION , CHRONI C 6 (14% ) 3 (9% ) 

HYPEEPLASIA , FOCAL 1 (2%) 

*SEMINA L VESICLE (49) (50) (50) 

INFLAMMATION , NO S 1 (2% ) 

ITESTIS (D4 ) (49) (49 ) 

EDEMA , NO S 7 (14%) 8 (16%) 

GrtANULOMA , SPERMATI C 

PERIAETERITI S 

DEGENERATION , NO S 

1

8

3

 (2% ) 

 (16% ) 

 (6%) 

17

2

 (35%) 

 (4% ) 

ATROPHY ,

ATROPHY ,

 NCS 

 FOCA L 

9 (20%) 26

5

 (53%) 

 (10% ) 

31 (63X) 

HYPEEPLASIA , INTERSTITIA L CELL 1 (2%) 

*EPIDIDYMI S 

NECROSIS , FAT 

(49 ) (50) (5C) 

1 (2%) 

*SCROTU H 

NECEOSIS , FA T 

(49 ) (50) (50) 

1 (2% ) 

NERVOU S SYSTE M 

*BEAIN/MENINGE S 

GRANULCMA , NO S 

(47 ) (50) 

1 (2%) 

(50) 

•CEREBRU M 

MALACI A 

(47 ) (50) (50 ) 

1 12%1 _ 

t NUMBE R O F ANIMAL S lilTH TISSU E EXAMINE D MICEOSCOPIC A LL Y 

* NUMBF 8 OF ANIMAL S NECROPSIE D 

C-7 
SPECIAL SENSE ORGANS
 

*EYE


CATARACT
 

PHTHISIS BULBI


*EYE/LACRIMAL GLAND


INFLAMMATION, FOCAL
 

*HAFDERIAN GLAND


INFLAMMATION, NOS


MUSCULOSKELETAL SYSTEM
 

*SKELETAL MUSCLE


DEGENERATION, NOS


BODY CAVITIES
 

*PLEURA


INFLAMMATION, CHRONIC
 

*PERICARDIUM


INFLAMMATION, NOS


INFLAMMATION, CHRONIC NECROTIZIN
 

*EPICARDIUM '


CALCIFICATION, NOS
 

*MESiiNTEHY


INFLAMMATION, ACUTE
 PERIARTERITIS


ALL OTHER SYSTEMS
 

NONE
 

SPECIAL MORPHOLOGY SUMMARY
 

AUTOLYSIS/NO NECROPSY
 

TABLE Cl (CONCLUDED)
 

CONTROL (VEH) LOW DOSE
 01-M001 01-M004
 

 (49) (50)
 

1 (2X)
 

1 (2X)
 

 (49)	 (50)
 

2 (4X)
 

 (49)	 (50)
 

1 (2X)
 

 (U9) (50)
 

1 (2%)
 

 (49) (50)
 

 (49)	 (50)
 

5 (10X)
 

 (49) (50)
 

 (49) (50)
 

1 (2%)
 

2 (4X) 1 (2%)
 

#	 NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

*	 NUMBER OP ANIMALS NBCROPSIZD
 

C-8
 

HIGH DOSE
 

01-H005
 

(50)
 

(50)
 

(50)
 

(50)
 

(50)
 

1 (2%)
 

(50)
 

1 (2X)
 

(50)
 

1 (2X)
 

(50)
 

1 (2X)
 
TABLE C2

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS 

IN FEMALE RATS TREATED WITH CHLOROBENZILATE 

CONTROL (VEH ) LOU DOSE HIGH DOSE 

01-F001 01-F006 01-F007 

ANIMAL S INITIALL Y IN STUDY 50 50 50 ANIMAL S MISSIN G 1 

ANIMALS NECBOPSIED 50 1*9 50 ANIMAL S EXAMINE D HISTOPATHOLOGICALLY* » 50 48 18 

INTEGUMENTARY SYSTEM
 

*SKIN (50) (K9) (50)
 INFLAMMATION, NOS 1 (2*)
 

*SUB(-UT TISSUE (50) (1*9) (50)
 

NECROSIS, FAT 1 (2X)
 

EESPIBATOBY SYSTEM
 

*TRACHEA (5) (1*7) (18)
 

INFLAMMATION, NOS 5 (100*)
 

INFLAMMATION, CHRONIC 3 (6%) 1 (8X)
 

•LUNG/BRONCHIOLE	 (50) (18) («8)
 

HYPERPLASIA, LYMPHOID 12 (25X) 9 (19*)
 

#LUNG (50) (18) (18)
 

HEMORRHAGE 1 (2%)
 

LUBAE PNEUMONIA, NOS 1 (2%)
 

INFLAMMATION, NOS 7 (15*) 5 (10*)
 

INFLAMMATION, FOCAL 11 (23*)
 

INFLAMMATION, MULTIFOCAL 1 (2%) 1 (8*)
 

INFLAMMATION, DIFFUSE 1 (2*)
 PNEUMONIA, CHRONIC MDRINE 15 (30%) 2 (151) 1 (8*)
 

#LUNG/ALVEOLI (50) (18) (18)
 

MINERALIZATION 1 (2*)
 

HEMATOPOIETIC SYSTEM
 

*BONE MARRO W (50) (17) (18) 

METAMORPHOSI S FATTY 1 (2*) 

ISPLEEN (50) (17) (48) 

HEMOBBHAGF . 

* NUMBE R OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBER OF ANIMAL S NECROPSIED 

**EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

C-9 
TABLE C2 (CONTINUED) 

CONTROL (VEH ) L O U DOSE HIG H DOSE 

01-F001 01-F006 01-F007 

HEMOSIDEROSI S 1 (9%) 

HEMATOPOIESIS 5 (10%) 1 (2X) 2 (4%)

 

CIRCULATOR! SYSTE M 

• MYOCARDIU M (50) (US ) (48) 

MINERALIZATIO N 2 (tX) FIBROSIS 3 (6%) 1 (2%) 

FIBROSIS, FOCAL 3 (6%) 

DEGENERATION , SOS 2 (4% ) 

• ENDOCARDIU M (50) (18) (48) 

HYPERPLASIA , NOS 1 (2% ) 1 (2% ) 

• AORT A (50) (U9 ) (50) MINERALIZATIO N 1 (2% ) 

ARTERIOSCLEROSIS , NO S 1 (2% ) 

DIGESTIV E SYSTE M 

•LIVE R (50) (U8 ) (48) 

INFLAMMATION , NO S 4 (8% ) METAMORPHOSI S FATTY 

FuCAL CELLULA R CHANG E 

1 (2S) 

1 (2«) 2 (4%) 

ANGIECTASI S 1 (2% ) 

*BILL DUCT (50) (149) (50) DILATATION , NO S 1 (2%) INFLAMMATION , NO S 1 (2H ) 

FIBROSIS 

HYPEEPLASIA ,

HYPERPLASIA ,

 NO S 

 CYSTIC 

2 (4X) 

3 (6%) 

9 (18%) 12

1

 (21%) 

 (2X ) 

#PANCREA S 

AISOPHY ,

ATROPHY ,

 NOS 

 FOCAL 

(50) (18) 

2 (t%) 

(48) 

2 (4%) 

1 (2X) 

•STOHAC H 

MINERALIZATIO N 

(50) (48) 

2 (U% ) 

(48) 

HEMORRHAG E 

ULCER, FOCAL 

INFLAMMATION , CHRONIC 

5 (10%) 1

1

 (2X) 

 (2X ) CALCIU M DEPOSIT 1 (2% ) 

•LARG E INTESTIN E 

NEMATODIASI S 

(19) (48) 

4 (8%) 

(46 ) 

3 (7X) 

EiSiSITISfl _ l.Uil — 

• NUMBE R OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALL Y 

* NUMBE R OF ANIMAL S NECROPSIE D 

c-io 
TABLE C2 (CONTINUED) 

CONTROL (VEH ) L OW DOSE 

01-F001 01-F006 

DBINAR Y SYSTE M 

• KIDNE Y (50) (1»8) 

PYELONEPHRITIS . NOS 1 (2*) 

INFLAMMATION , CHRONIC 23 (46X) 15 (31*) 

tRENA L PAPILL A (50) (48) 

CALCIFICATION. NOS 17 (35*) 

HYPERPLASIA , EPITHELIAL 1 (2X) 

AtiGIECTASIS 1 (2%) 

METAPLASIA . SQUAMOUS 1 (2%) 

• KIDNEY/PELVI S (50) (48) 

INFLAMMATION , NOS 

CALCIFICATION . NOS 1 (2X ) 

tUEINAH Y BLADDE R (49 ) (46)

 

METAPLASIA , SQUAMOD S 

ENDOCalN E SYSTE M 

tPITUITAR Y (50) (45) 

COLLOID CYST 3 (7%) 

DEGENERATION , NOS 1 (2*) 

HYPERPLASIA , CHBOMOPHOBE-CEL L 4 (9X) 

ANGIECTASIS 1 (2*) 1 (2%) 

tADHENA L (50) (47) 

THROMBOSIS , NOS 1 (2*) 

ANGIECTASIS 17 (34*) 14 (30*) 

tADRENA L CORTEX (50) (47 ) 

DEGENFRATION , NOS 7 (15*) 

*ADRENA L MEDULL A (50) (47) 

HYPEHPLASIA , NO S 1 (2X ) 

tTHYROID (50) (47) 

FULLICULA R CYST, NOS 3 (6*) 

HYPERPLASIA , C-CELL 2 (t%) 4 (9*) 

HYPERPLASIA , FOLLICttLAR -CELL 2 (UK ) 

• PARATHYROI D (48) (24 ) 

HYPERPLASIA. t _NOS J. 

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

* NUMBE R OF ANIMAL S NECBOPSIED 

C-ll 

HIG H DOSE 01-F007 

(48) 

17 (35*) 

(48) 

15 (31X) 

(48) 

1 (2*) 

(42 ) 

1 (2*) 

(45) 

1 (2*) 

(47) 

11 (23X) 

(47) 

7 (15X) 

(47) 

(47) 

4 (9*) 

(24 ) 
HEPPODUCTIV F SYSTE M 

*1AMrtAR Y GLAN D 

HYPEFPLASIA , NO S 

9 IT "a as 

HYDHOMETP A 

INFLAMMATION , NO S 

ttUTEhOS/ENDOKETRIU M 

CYST , NO S 

HiPERPLASIA , CYSTIC 

*JVAH Y 

CYST , NO S 

NFBVO'J S SYSTE M 

• MEDULL A OBLCNGAT A 

HEMORRHAG E 

SP-:C:AL SENS E CEGAN S 

*E YE 

SYNCCHIA , ANTERIO R 

CATARAC T 

tCYVFTIN A 

AIPOEHY , NC S 

• EYE/CONJUNCTIV A 

INFLAMMA^lrN , CHRONI C 

• "•"YE/LACRIMA L GLAN D 

INFLAMMATCN , CHRONI C 

HYPERDLASIA , LYMPHOI D 

MU3CULOSKELETA L SYSTE M 

NON E 

ECD Y ^AVITIF S 

*ABDOBINA L CAVIT Y 

NECROSISj_FA T 

* NUMBE R O F ANIMAL S WIT H TISSU E

* NUMB" P O F ANIMAL S NFCROPSIE D 

TABLE C2 (CONTINUED) 

CONTRO L (VEH ) L OW DOSE 

01-F001 01-F006 

(50) (49) 

1 (2% ) 

(49 ) (47) 

9 (18% ) 1 (2%) 

2 (4% ) 

(49 ) (47 ) 

4 (9S) 

3 (6% ) 

(49 ) (48) 

2 (4% ) 

(50) (48) 

2 (4*) 

(5C) (49 ) 

1 (2%) 

1 (2S) 

(50) (49 ) 

1 (2t) 

(50 ) (49 ) 

(50) (49 )

1 (2X) 

1 (2%) 

(50 ) (49 ) 

 EXAMINE D MICROSCOPICALL Y 

C-12 

HIG H DOSE 

C1-F007 

(50) 

(46 ) 

(46 ) 

6 (13%) 

(46 ) 

2 (4X ) 

(48) 

(50) 

(50 ) 

(50 ) 

1 (2% ) 

(50) 

(50) 
TABLE C2 (CONCLUDED) 

:ONTROL(VEH ) LO W DOS E HIG H DOSE 

01-F001 01-F006 C1-F00 7 

*EPICARDIU M (50 ) (<49 ) (50 ) 

INFLAMMATION , ACUTE 1 (2% ) 

ALL OTHFP SYSTEMS
 

NONZ
 

SPECIAL MOREHCLCGY SUMMAR Y
 

NO LESION tEPOHTED 1
 ANIMA L MISSING/NO NECROPSY 1
 

NtCROPSY PERF/NO HISPO PEHFORHED 2
 

AUTO/NTCRCESY/NO HISTO 1
 

» NUMtJER OF ANIMAL S HITH TISSUE EXAMINE D MICROSCOPICALLY
 * NUMdPR OF ANIMAL S NFCBOPSIED
 

C-13
 

APPENDIX D
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN MICE TREATED WITH CHLOROBENZILATE
 

TABLE Dl

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS 

IN MALE MICE TREATED WITH CHLOROBENZILATE 

CONTRO L (VEH )

02-M00 6 LOW DOSE 02-H007

 HIG H DOSE 

 02-M008 

ANIMAL S INITIALL Y IN STUDY 20 50 50 

ANIMAL S MISSIN G 1 1 

ANIMAL S NECFOESIE D 19 49 48 

ANIMAL S EXAMINED HISTOPATHOLOGICALLY** 17 47 45 

INTEGUMENTAR Y SYSTEM 

*SKIN (19) (49) (48) 

INFLAMMATION , DOS 2 (4X) 

INFLAMMATION , CHRONIC 3 (16% ) 

ACANTHOSIS 2 (4X) 

*SUBuUT TISSUE (19) (49) (18) 

ABSCESS , N03 1 (2X ) 

EESPItiATOR Y SYSTE M 

ULUN G (19) (U8 ) (14) 

PNEUMONIA , CHRONIC MURIN F 3 (16)t) 1 (2%) 2 (5X) 

HEMATOPOIETIC SYSTEM 

tSPLtE N (18) («7) (11) 

AJlYLOIDOSI S 5 (28% ) 

CIRCULATOR Y SYSTE M 

*MYOLARDIU M (19 ) (17) (44) 

INFLAMMATION , NOS 1 (5X) 

«AORI A (19) (49) (18) 

INFLAMMATION , NOS 1 (5%) 

DIGESTIV E SYSTE M 

tLIVER (19 ) (18) (45) 

TjjEgMBUS^CRGANIZE D JLX2S.L 

* NUMBE R OF ANIMAL S KIT H TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBE R OF ANIMAL S HECROPSIED 

**EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

D-3 
INFLAMMATION , NO S 

FIBRCSI S 

INFABCT , NO S 

AMYLOIDOSI S 

HYPFHPLASIA , NODULA R 

HYPEHPLASIA , FOCA L 

*BILE DUCT 

HYPERPLASIA , NO S

 

URINAR Y SYSTE M 

1KIDNE Y 

PYELONEPHRITIS , NO S 

INFLAMMATION , CHRONI C 

((KIDNEY/PELVI S 

NECROSIS , NOS 

SURIHAR Y BLADDE R 

CALCULUS , NO S

 

INFLAMMATION , NOS 

EKDOCKIN E SYSTE M 

4ADBENA L 

AMYLOIDOSI S 

*THYtiOI D 

FULLICULA H CYST, NO S 

REPRODUCTIV E SYSTE M 

"tPPOSTATE 

INFLAMMATION , NO S 

»TESIIS 

ATROPHY , NCS 

*EPIDIDYMI S 

GhANULOMA , SPERMATI C 

NECROSIS , FA T 

NERVOU S SYSTE M 

NON E 

*	 NUMBER OF ANIMALS HITH TISSUE


*	 NUMBER OF ANIMAL S NECBOPSIED
 

TABLED! (CONTINUED) 

CONTROL (VEH ) L OW DOSF HIG H DOSE 

02-M00 6 02-M007 02-M00 8 

2 (4/5) 2 ("*%) 

1 (2% ) 

1 (2%) 

3 (16X ) 

3 (6JS) 2 (U% ) 

1 (5% ) 

(19 ) (U9 ) (1*8) 

1 (2% ) 

(19 ) (17) (15) 

1 (2% ) 

6 (32% ) 1 (2% ) 

(19 ) (17) (1*5) 

1 (5% ) 

(15 ) (11) (13) 1 (2%) 1 (2% ) 

2 (5%) 

(16 ) (15) (11) 

2 (13% ) 

(15) (1*1) (13) 

1 (7X ) 

(9) (28) (23) 

1 (i*%) 

(16 ) (17) (15) 

1 (2% ) 

(19 ) (19) (18) 

1 (2%) 

1 (5%) 

 EXAMINED MICROSCOPICALLY
 

D-4
 
TABLE Dl (CONCLUDED) 

CONTRO L (VEH ) LO W DOSE HIG H DOSE 

02-MOQ6 02-M007 Q2-M008 

SPECIA L SENS F CEGAN S 

NONi, 

MUSCULOSKELETA L SYST1'-! 

NON E 

BOD Y CAVITIE S 

NON E 

A L L OTHE R SYSTEM S 

NON E 

SPECIA L MORPHOLOG Y SUHP1AB Y 

NO LESIO N REPORTE D 1 5 1 <4 

AdlMA L HISSING/N O NECROPS Y 1 1 

NECROPS Y PERF/N O HISTO EEHFORME D 1 

AUTO/NECPOPSY/N O HISTO 2 2 2 

AUTOLYSIS/N O NECROPS Y 1 1 

D-5 
TABLE D2

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS 

IN FEMALE MICE TREATED WITH CHLOROBENZILATE 

ANIMAL S INITIALL Y IN STUDY 

ANIMAL S NECROPSIE D 

ANIMAL S EXAMINE D HISTOPATHOLOGICALLY* * 

INTEGUMENTAR Y SYSTEM 

NONi 

RESPIRATOR Y SYSTE M 

tiUNG 

PNEUMONIA, CHRONIC MURINE
 

HEMATOPOIETIC SYSTEM
 

tSPLEEN
 

HEMATOFOIESIS
 

*MESENTERIC L. NODE 

INFLAMMATION , NO S 

HtfPKRPLASIA , LYMPHOI D 

CIRCULATORY SYSTEM
 

NONE
 

DIGESTIV E SYSTB M 

*LIVER
 

THROMBUS, ORGANIZED
 

INFLAMMATION, NOS
 

PELIOSIS HEPATIS
 

INFARCT, NCS
 

METAMORPHOSIS FATTY
 

HYPESPLASIA, NODUHE
 

* NUMBE R O F ANIMAL S HITH TISSU E

* NUMBE H OF ANIMAL S NECROPSIED 

**EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

CONTRO L (VEH ) 

02-F006 

20 

20 

20 

(19)
 

(20) 

(18) 

(20)


L OW DOSE HIG H DOSE 02-F009 02-F010 

50 50 

50 50 

U9 50 

(50)
 

3 (6*)
 

(09) (<*9) 

2 (OX) 

(06) (140) 

1 (2%) 

1 (2%) 

 (09)	 (50)
 

1 (2X)
 1 (2*)
 

1 (2%)
 1	 (2%)
 

1 (2*)
 5 (10*) 0 <8X)
 

 EXAMINE D MICROSCOPICALL Y 

D-6
 
TABLE D2 (CONTINUED) 

CONTROL (VEH) LOW DOSE 

02-F006 02-F009 

• PANCREA S (20 ) («8) 

DILATATION/DOCTS 1 (2%) 

INFLAMMATION , NO S 1 (2% ) 

•STOMACH (19 ) (U9 ) 

INFLAMMATION , NOS 

HYPERKERATOSIS 1 (2%) 

ACANTHOSI S 1 (2% ) 

UBINARY SYSTEM
 

•KIDNEY	 (20) (49)
 

INFLAMMATICN, CHRONIC 1 (2%)
 

AHYLOIDOSIS 1 (2%)
 

ENDOCRINE SYSTE M 

NONE 

REPRODOCTIVE SYSTEM
 

tUTEROS (20) (H9)
 

HYDROSETRA 5 (10%)
 

INFLAMMATION, NOS 8 (16%)
 

ABSCESS, NOS 1 (5%)
 

•UTERUS/ENDOMETRIUM	 (20) (<*9)
 

INFLAMMATION, SUPPURATIVE 2 (10%)
 

HYPERPLASIA, CYSTIC 12 (60%) 5 (10%)
 

IOVARY/OVIDUCT (20) (U9)
 

INFLAMMATICN, NOS 3 (6%)
 

•OVAKY	 (19) (U8)
 

CYST, NOS 2 (11%) i* (8%)
 

FOLLICULAR CYST, NOS U (21%) 2 (U%)
 

INFLAMMATION, NOS 6 (1 3%)
 

ABSCESS, NOS 2 (11%)
 

INFLAMMATION, CHRONIC SUPPURATIV 1 (5%)
 

NERVOUS SYSTEM 

--M2HE
 

f NUMBER OF ANIMALS WITH TISSUE EXAMINED MICBOSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

D-7
 

HIG H DOSE 02-F010 

(U9 ) 

1 (2%) 

(50) 

1 (2%) 

(50)
 

(U8) 

3 (6%) 

1 (2% ) 

(18) 

6 (13% ) 

(18) 

|i*8)
 

2 (U% )
 
TABLt D2 (CONCLUDED)
 

CONTROL (VEH) LOW DOSE HIGH DOSE
 

02-F006 02-F009 02-F010
 

SPECIAL SENSE ORGANS
 

NONE
 

MUSCULOSKELETAL SYSTEM
 

NONE
 

BODY CAVITIES
 

*MESENTEHY (20) (50) (50)
 

CYST, NOS 1 (28)
 

INFLAMMATION, NOS 1 (2%)
 

ALL OTHEH SYSTEMS
 

NONE
 

SPECIAL MORPHOLOGY SUMMAR Y
 

NJ LESION BEPORTED 14 6 22
 

AUTO/NECROPSY/NO HISTO 1
 

» NUMBER OF ANIMAL S WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NEC80PSIED
 

D-8
 


DHE

W Publication No. (NIH) 78-1325 

i