1978 

National Cancer Institute 

CARCINOGENESIS 

Technical Report Series 

No. 92 

BIOASSAY OF 

HYDRAZOBENZENE 

FOR POSSIBLE CARCINOGENICITY 

CAS No. 122-66-7 

NCI-CG-TR-92 

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

Public Health Service 

National Institutes of Health 

BIOASSAY OF
 

HYDRAZOBENZENE
 

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) 78-1342
 
REPORT ON THE BIOASSAY OF HYDRAZOBENZENE
 

FOR POSSIBLE CARCINOGEN1CITY
 

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 hydrazobenzene 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 hydrazobenzene was conducted by Mason
 

Research Institute, Worcester, Massachusetts, initially under direct
 

contract to the NCI and currently under a subcontract to Tracor 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. E. Smith (3) and
 

Dr. A. Handler (3). Animal treatment and observation were supervised
 

by Mr. G. Wade (3) and Ms. E. Zepp (3).
 

Histopathologic examinations were performed by Dr. Freclerickson
 

(3), Dr. D. W. Hayden (3), Dr. A. S. Krishna Murthy (3), and Dr. A.
 

Russfield (3) at the Mason Research Institute, the pathology narratives were written by Dr. A. S. Krishna Murthy (3), and the diagnoses
 

included in this report represent the interpretation of these pathologists. Histopathology findings and reports were reviewed by Dr. R.
 

L. Schueler (4).
 

Compilation of individual animal survival, pathology, and summary tables was performed by EG&G Mason Research Institute (5); the
 

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

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

Gart (7).
 

111
 
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), senior biologist Ms. P.
 

Walker (6), biochemist Dr. B. Fuller (6), chemist Dr. N. Zimmerman
 

(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), Dr. R.
 

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

Squire (1,8), 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.	 Now with the Naylor Dana Institute for Disease Prevention, American Health Foundation, Hammon House Road, Valhalla, New York.
 

3.	 Mason Research Institute, 57 Union Street, Worcester, Massachusetts.
 

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

Maryland.
 

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

Rockville, Maryland.
 

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

Boulevard, McLean, Virginia.
 

7.	 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.
 

3.	 Now with the Division of Comparative Medicine, Johns Hopkins
 

University, School of Medicine, Traylor Building, Baltimore,
 

Maryland.
 

iv
 
SUMMARY
 

A bioassay of technical-grade hydrazobenzene for possible carcinogenicity was conducted using Fischer 344 rats and B6C3F1 mice.
 

Hydrazobenzene was administered in the feed, at either of two concentrations, to groups of 50 male and 47 to 50 female animals of each
 

species. The time-weighted average dietary concentrations used in
 

the rat bioassay were 0.008, 0.03, 0.004, and 0.01 percent for low
 

dose males, high dose males, low dose females, and high dose females,
 

respectively. The time-weighted average dietary concentrations used
 

in the mouse bioassay were 0.008, 0.04, 0.004, and 0.04 percent for
 

low dose males, high dose males, low dose females, and high dose females, respectively. After a 78-week period of compound administration, observation of the rats continued for an additional 28 to 30
 

weeks and observation of the mice continued for an additional 17 or
 

18 weeks. For each species, 47 to 50 animals of each sex were placed
 

on test as controls.
 

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

The incidence of hepatocellular carcinomas was significantly
 

increased in dosed male rats and the incidence of neoplastic nodules
 

of the liver was significantly increased in dosed female rats. A
 

significant increase in the combined incidence of squamous-cell carcinomas or squamous-cell papillomas of the Zymbal's gland, the ear
 

canal, or the skin of the ear was observed among high dose male rats.
 

A significant increase in mammary adenocarcinomas was observed among
 

dosed female rats.
 

The incidence of hepatocellular carcinomas was significantly
 

increased among female mice, but no significant increase in liver
 

tumors was observed among male mice.
 

Under the conditions of this bioassay, hydrazobenzene was carcinogenic to Fischer 344 rats of both sexes, causing increased incidences of hepatocellular carcinoma and Zymbal's gland squamous-cell
 

neoplasms in male rats, neoplastic nodules of the liver in female
 

rats, and mammary adenocarcinomas in female rats. Hydrazobenzene was
 

also carcinogenic to female B6C3F1 mice, causing an increased incidence of hepatocellular carcinomas; The compound was not carcinogenic
 

to male B6C3F1 mice.
 

TABLE OF CONTENTS
 

Page
 

I. INTRODUCTION 1
 

II. MATERIALS AND METHODS 3
 

A. Chemicals 3
 

B. Dietary Preparation 3
 

C. Animals 5
 

D. Animal Maintenance 5
 

E. Selection of Initial Concentrations 9
 

F. Experimental Design 11
 

G. Clinical and Histopathologic Examinations 15
 

H. Data Recording and Statistical Analyses 17
 

III. CHRONIC TESTING RESULTS: RATS 22
 

A. Body Weights and Clinical Observations 22
 

B. Survival 22
 

C. Pathology 25
 

D. Statistical Analyses of Results 27
 

IV. CHRONIC TESTING RESULTS: MICE 42
 

A. Body Weights and Clinical Observations 42
 

B. Survival 42
 

C. Pathology 45
 

D. Statistical Analyses of Results 46
 

V. DISCUSSION 53
 

VI. BIBLIOGRAPHY 55
 

APPENDIX A SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

RATS TREATED WITH HYDRAZOBENZENE A-l
 

APPENDIX B SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

MICE TREATED WITH HYDRAZOBENZENE B-l
 

APPENDIX C SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN RATS TREATED WITH HYDRAZOBENZENE C-l
 

APPENDIX D SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN MICE TREATED WITH HYDRAZOBENZENE D-l
 

Vll
 
LIST OF ILLUSTRATIONS
 

Figure Number
 

1
 

2
 

Table Number
 

Al
 

Page 

CHEMICAL STRUCTURE OF HYDRAZOBENZENE 4 

GROWTH CURVES FOR HYDRAZOBENZENE CHRONIC 

STUDY RATS 23 

SURVIVAL COMPARISONS OF HYDRAZOBENZENE 

CHRONIC STUDY RATS 24 

GROWTH CURVES FOR HYDRAZOBENZENE CHRONIC 

STUDY MICE 43 

SURVIVAL COMPARISONS OF HYDRAZOBENZENE 

CHRONIC STUDY MICE 44 

LIST OF TABLES 

DESIGN SUMMARY FOR FISCHER 344 RATS— 

HYDRAZOBENZENE FEEDING EXPERIMENT 12 

DESIGN SUMMARY FOR B6C3F1 MICE—HYDRAZOBENZENE FEEDING EXPERIMENT 13 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN MALE RATS TREATED WITH 

HYDRAZOBENZENE 28 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN FEMALE RATS TREATED 

WITH HYDRAZOBENZENE 34 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN MALE MICE TREATED WITH 

HYDRAZOBENZENE 47 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN FEMALE MICE TREATED 

WITH HYDRAZOBENZENE 50 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

MALE RATS TREATED WITH HYDRAZOBENZENE A-3 

viii
 
LIST OF TABLES (Concluded) 

Table Number Page 

A2 SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

FEMALE RATS TREATED WITH HYDRAZOBENZENE A-8 

Bl SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

MALE MICE TREATED WITH HYDRAZOBENZENE B-3 

B2 SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

FEMALE MICE TREATED WITH HYDRAZOBENZENE B-7 

Cl SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN MALE RATS TREATED WITH HYDRAZOBENZENE C-3 

C2 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN FEMALE RATS TREATED WITH HYDRAZOBENZENE C-9 

Dl SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN MALE MICE TREATED WITH HYDRAZOBENZENE D-3 

D2 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN FEMALE MICE TREATED WITH HYDRAZOBENZENE D-8 

ix
 

I. INTRODUCTION
 

Hydrazobenzene (NCI No. C01854) is a hydrazine derivative selected
 

for bioassay by the National Cancer Institute because of the documented
 

carcinogenicity of the parent compound hydrazine and of certain substituted hydrazines (International Agency for Research on Cancer, 1974) .
 

The increased incidence of bladder cancer observed among workers in the
 

dye manufacturing industry (Wynder et al., 1963; Anthony and Thomas,
 

1970) was also a factor in the selection of this chemical for testing.
 

The Chemical Abstracts Service (CAS) Ninth Collective Index* (1977)
 

name for this compound is 1,2-diphenylhydrazine. It is also known as
 

symmetrical (sym-) diphenylhydrazine (Weast, 1972) .
 

Treatment of hydrazobenzene with hot mineral acid results in the
 

production of benzidine (the so-called "benzidine rearrangement") and
 

hydrazobenzene finds application in the dye manufacturing industry as a
 

precursor of this important dye intermediate and potent carcinogen. An
 

additional use of hydrazobenzene is in the preparation of phenylbutazone (via condensation with diethylbutyl malonate), a widely used
 

agent against rheumatoid arthritis and related conditions (Wenner,
 

1967) .
 

Although specific production figures are not available, the
 

listing of hydrazobenzene in the 1977 Directory of Chemical Producers,
 

U.S.A. (Stanford Research Institute, 1977) implies an annual commercial
 

*The CAS registry number is 122-66-7.
 
production in excess of 1000 pounds or $1000 in value. This production would appear to be in addition to the quantities produced at dye
 

manufacturing facilities by, for example, the reduction of nitrobenzene with zinc dust and sodium hydroxide prior to the benzidine rearrangement (Bannister, 1967).
 

The potential for exposure to hydrazobenzene is greatest for
 

workers in the dye manufacturing industry although workers in the
 

pharmaceutical industry may also experience contact with the chemical
 

Hydrazine derivatives tend- to be local irritants, convulsants,
 

hepatotoxins and hemolytic agents and are absorbed by all routes of
 

administration (Button, 1967). It has been suggested that benzidine
 

may be produced from hydrazobenzene, following ingestion, by acidity
 

in the stomach (International Agency for Research on Cancer, 1972).
 
II. MATERIALS AND METHODS 

A. Chemicals
 

Technical-grade hydrazobenzene (Figure 1) was purchased from K &
 

K Labs. Chemical analysis was performed by Mason Research Institute,
 

Worcester, Massachusetts. The experimentally determined melting point
 

of 120° to 124°C suggested the presence of impurities due to its
 

difference from the literature value of 129° to 131°C. Thin-layer
 

chromatography showed a major spot with an Rf of 0.63 and a single
 

unidentified minor spot indicating the presence of at least one
 

impurity. Infrared analysis was not inconsistent with the structure
 

of hydrazobenzene.
 

Throughout this report the term hydrazobenzene is used to represent this technical-grade material.
 

B. Dietary Preparation
 

The basal laboratory diet for both treated and control animals
 

(R)
 was Wayne Lab-Blox (Allied Mills, Inc., Chicago, Illinois). Hydrazobenzene was administered to the treated animals as a component of
 

the diet. The hydrazobenzene was first ground to a powder in a
 

Quaker City Crystal Mill and then returned to the original metal
 

container. To prepare the chemical diet mixture, the chemical was
 

removed from the stock container, weighed out in proper amounts for
 

dosage preparation, and hand-blended in an aluminum bowl with an
 

aliquot of ground feed. Once visual homogeneity was attained, the
 

mixture was placed into a 6 kg capacity Patterson-Kelley standard
 
h-N— N— I l

 

H H 

FIGURE 1 

CHEMICAL STRUCTURE OF HYDRAZOBENZENE 
model twin-shell stainless steel V-blender with the remainder of the
 

diet. After 20 minutes of blending, the mixtures were placed in
 

double plastic bags and stored in the dark at 4°C. Mixtures were
 

prepared once weekly and stored for a maximum period of one week.
 

C. Animals
 

Two animal species, rats and mice, were used in the carcinogenicity bioassay. Fischer 344 rats and B6C3F1 mice were obtained through
 

contracts of the Division of Cancer Treatment, National Cancer Institute. High dose rats and their controls and all mice were supplied by
 

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

Low dose rats and their controls were supplied by Laboratory Supply
 

Company, Inc., Indianapolis, Indiana. Animals to be treated were
 

received separately from their respective control groups, except low
 

dose treated and control rats, which were received on the same date.
 

Upon arrival, a sample of animals was examined for parasites and other
 

signs of disease. The remaining animals were quarantined by species
 

for 2 weeks prior to initiation of test. Animals were assigned to
 

groups and distributed among cages so that the average body weight
 

per cage was approximately equal for a given species and sex.
 

D. Animal Maintenance
 

All animals were housed by species in rooms having a temperature
 

®
 

range of 23° to 34°C. Incoming air was filtered through Tri-Dek
 

15/40 denier Dacron filters (Tri-Dim Filter Corp., Hawthorne, New
 
Jersey) providing six changes of room air per hour. Fluorescent
 

lighting was provided on a 12-hour-daily cycle.
 

Rats were housed five per cage by sex. During quarantine and
 

for the first 13 months of study, all rats were kept in galvanized-


or stainless-steel wire-mesh cages suspended above newspapers. Newspapers under cages were replaced daily and cages and racks washed
 

weekly. For the remainder of the study, high dose, low dose, and
 

control rats were housed in suspended polycarbonate cages equipped
 

with nonwoven fiber filter sheets. Clean bedding and cages were provided twice weekly. Corncob bedding (SAN-I-CEL , Paxton Processing
 

Company, Paxton, Illinois) was used for low dose treated and control
 

rats for the first 9 months that they were housed in polycarbonate
 

cages, and Aspen hardwood chip bedding (American Excelsior Company,
 

Baltimore, Maryland) was used for the remainder of the study. High
 

dose treated and control rats received Aspen bedding for the entire
 

time that they were housed in polycarbonate cages. Stainless steel
 

cage racks were cleaned once every two weeks, and disposable filters
 

were replaced at that time.
 

Mice were housed by sex in polycarbonate cages. During quarantine and periods of compound administration, cages were fitted with
 

perforated stainless steel lids,. During the untreated observation
 

period, stainless steel wire bar lids were used. Both types of lids
 

were from Lab Products, Inc., Garfield, New Jersey. Nonwoven fiber
 

filter bonnets were used over cage lids. Low dose mice and their
 
controls were housed ten per cage for the first 16 months of study
 

and five per cage thereafter. High dose mice and their controls were
 

housed ten per cage initially and then five per cage after 11 months.
 

Cages, lids, filters, and bedding were provided three times per week
 

when cage populations were ten and twice per week when cage popula­

®
 tions were reduced to five. Hardwood chip bedding (Ab-sorb-dri ,
 

Wilner Wood Products Company, Norway, Maine) was used for the first
 

30 weeks for low dose and low dose control mice, for the first 6
 

weeks for high dose mice, and for 2 weeks for high dose control mice.
 

®
 SAN-I-CEL corncob bedding was used for all mice for the next 12
 

months. For the remainder of the study, Aspen bedding was used.
 

Reusable filter bonnets and pipe cage racks were sanitized every 2
 

weeks throughout the study.
 

Water was available for both species from 250 ml water bottles
 

equipped with rubber stoppers and stainless steel sipper tubes.
 

Bottles were replaced twice weekly and, for rats only, water was
 

supplied as needed between changes. Food and water were available
 

ad libitum.
 

®
 Pelleted Wayne Lab-Blox was fed to low dose rats and their
 

controls during quarantine and to all rats and mice during the untreated observation period. During the dosing period, all treated
 

®­ animals were fed Wayne Lab-Blox meal containing the appropriate
 

concentration of hydrazobenzene. Control animals had untreated meal
 

®
 available. Alpine aluminum feed cups (Curtin Matheson Scientific,
 
Inc., Woburn, Massachusetts) containing stainless steel baffles were
 

used to distribute powdered feed to all mice and to low dose rats and
 

their controls throughout the study. High dose treated and control
 

®
 rats were fed from Alpine feed cups during quarantine and for the
 

first 11 months of study. For the remainder of the study, high dose
 

treated and control rats were fed from stainless steel gangstyle feed
 

hoppers (Scientific Cages, Inc., Bryan, Texas). During the final observation period, mice were fed pellets from a wire bar hopper incorporated into the cage lid, and rats were fed pellets on the cage floor.
 

Food hoppers were changed on the same schedule as were cages. Food was
 

replenished daily in Alpine feed cups.
 (R)
 

* .
 

Low dose rats and their controls and high dose rats were in a
 

room with other rats receiving diets containing 5-nitro-o-toluidine
 

(99-55-8); 2-aminoanthraquinone (117-79-3); 3-amino-9-ethylcarbazole
 

hydrochloride; 6-nitrobenzimidazole (94-52-0); 1-nitronaphthalene
 

(86-57-7); 2,4-diaminoanisole sulfate (615-05-4); and APC (8003-03-0).
 

High dose control rats were in a room with other rats receiving diets
 

containing amitrole (61-82-5); 2-methyl-l-nitroanthraquinone (129-15­

7); and 3-nitro-p-acetophenetide (1777-84-0).
 

Low dose mice were housed in a room with other mice receiving diets
 

containing 2,5-toluenediamine sulfate (6369-59-1); 5-nitro-o-toluidine
 

(99-55-8); 3-amino-9-ethylcarbazole hydrochloride; 6-nitrobenzimidazole
 

(94-52-0); 5-nitro-o-anisidine (99-59-2); 1-nitronaphthalene (86-57-7);
 

*
 CAS registry numbers are given in parentheses,
 
and 2,4-diaminoanisole sulfate (615-05-4). High dose mice were housed
 

in a room in which other mice were receiving diets containing 2,3,5,6­

tetrachloro-4-nitroanisole (2438-88-2); tris(2,3-dibromopropyl)
 

phosphate (126-72-7); N-(l-naphthyl)ethylenediamine dihydrochloride
 

(1465-25-4); 2-chloro-p-phenylenediamine sulfate (61702-44-1); and
 

acetone (67-64-1). All control mice were in a room where other mice
 

were receiving diets containing amitrole (61-82-5); N,N-dimethylp-nitrosoaniline (138-89-6); 2,5-toluenediamine sulfate (6369-59­

1); 2,4-dinitrotoluene (121-14-2); 2-aminoanthraquinone (117-79-3);
 

3-amino-4-ethoxyacetanilide (17026-81-2); 3-amino-9-ethylcarbazole
 

hydrochloride; l-amino-2-methylanthraquinone (82-28-0); 5-nitro-oanisidine (99-59-2); 4-nitroanthranilic acid (619-17-0); 1-nitronaphthalene (86-57-7); 2,4-diaminoanisole sulfate (615-05-4); and APC
 

(8003-03-0).
 

E. Selection of Initial Concentrations
 

In order to establish the maximum tolerated concentrations of
 

hydrazobenzene for administration to treated animals in the chronic
 

studies, subchronic toxicity tests were conducted with both rats and
 

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

each consisting of five males and five females. The chemical was
 

incorporated into the laboratory diet and supplied ad libitum to the
 

rat and mouse groups for a total of 4 weeks. This dosing period was
 

followed by a 2-week observation period during which animals were
 
fed the basal diet. Eight of nine male rat groups received concentrations of 0.007, 0.014, 0.028, 0.055, 0.108, 0.214, 0.301 and 0.423
 

percent. The ninth male rat group served as a control, receiving
 

only the basal laboratory diet. Nine of ten female rat groups were
 

fed dietary concentrations of 0.00008, 0.0003, 0.0011, 0.002, 0.004,
 

0.015, 0.104, 0.731, and 5.138 percent. The tenth group served as a
 

control group.
 

Eight of the nine male mouse groups were given dietary concentrations of 0.007, 0.014, 0.028, 0.055, 0.108, 0.214, 0.301, and 0.423
 

percent. The ninth group served as a control, receiving only the
 

basal laboratory diet. Nine of the ten female mouse groups were given
 

dietary concentrations of 0.0003, 0.0008, 0.0011, 0.002, 0.004, 0.015,
 

0.104, 0.731, and 5.138 percent. The tenth group served as a control.
 

The highest concentration causing no deaths, no compound-related
 

gross abnormalities, and no mean group body weight depression in
 

excess of 11 percent relative to controls during the 6-week subchronic
 

test was selected as the high concentration utilized for the rat and
 

mouse chronic bioassays.
 

Two of the five male rats receiving concentrations of 0.108 percent died. All rats receiving higher concentrations died. One male
 

mouse receiving a concentration of 0.301 percent, two male mice receiving 0.423 percent, four female mice receiving 0.731 percent, and all
 

female mice receiving 5.138 percent died. Mean body weight depression
 

10
 
patterns were not consistent. Intestinal hemorrhage was the single
 

gross abnormality consistently observed in these mice.
 

The initial high dietary concentrations used in the chronic study
 

were 0.03 percent for male rats, 0.01 percent for female rats, and
 

0.04 percent for male and female mice.
 

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 time-weighted average
 

concentrations) are summarized in Tables 1 and 2.
 

Male rats were all approximately 6 weeks old at the time they
 

were placed on test. The initial dietary concentrations of hydrazobenzene were 0.007 and 0.0035 percent. After week 9, the higher of
 

these doses was raised to 0.008 percent. For male rats the group
 

receiving the lower dose (0.0035 percent) was sacrificed after 41
 

weeks because the dose level was considered, on the basis of weight
 

depression, to be inadequate. No histopathologic examinations of
 

these animals were performed. A new group, receiving 0.03 percent,
 

was started with its own control group. Throughout this report,
 

those male rats receiving 0.03 percent hydrazobenzene and their controls are referred to as the high dose and high dose control groups,
 

respectively. Throughout this report, those male rats initially
 

receiving a concentration of 0.007 percent and their controls are
 

referred to as the low dose and low dose control groups, respectively.
 

11
 
TABLE 1
 

DESIGN SUMMARY FOR FISCHER 344 RATS
 

HYDRAZOBENZENE FEEDING EXPERIMENT
 

MALE
 

LOW DOSE CONTROL
 

HIGH DOSE CONTROL
 

LOW DOSE
 

HIGH DOSE
 

FEMALE
 

LOW DOSE CONTROL
 

HIGH DOSE CONTROL
 

LOW DOSE
 

HIGH DOSE
 

a
 

INITIAL


GROUP
 

SIZE
 

50
 

49
 

50
 

50
 

50
 

50
 

50
 

50
 

 HYDRAZOBENZENE OBSERVATION PERIOD TIME-WEIGHTED
 

CONCENTRATION TREATED UNTREATED AVERAGE
 

(PERCENT) (WEEKS) (WEEKS) CONCENTRATION3
 

0 0 108 0
 

0 0 109 0
 

0.007 9 0.008
 

0.008 69
 

0 29
 

0.03 78 0.03
 

0 28
 

0 0 109 0
 

0 0 109 0
 

0.004 78 0.004
 

0 30
 

0.01 78 0.01
 

0 29
 

s(concentration X weeks received)
 

receiving chemical)
 

12
 
MALE
 

LOW DOSE CONTROL
 

HIGH DOSE CONTROL
 

LOW DOSE
 

HIGH DOSE
 

FEMALE
 

LOW DOSE CONTROL
 

HIGH DOSE CONTROL
 

LOW DOSE
 

HIGH DOSE
 

a .
 

TABLE 2
 

DESIGN SUMMARY FOR B6C3F1 MICE
 

HYDRAZOBENZENE FEEDING EXPERIMENT
 

INITIAL HYDRAZOBENZENE OBSERVATION PERIOD TIME-WEIGHTED 

GROUP CONCENTRATION TREATED UNTREATED AVERAGE 

SIZE (PERCENT) (WEEKS) (WEEKS) CONCENTRATION3 

50 0 0 95 0
 

50 0 0 96 0
 

50	 0.007 9 0.008
 

0.008 69
 

0 17
 

50	 0.04 78 0.04
 

0 17
 

50 0 0 96 0
 

50 0 0 96 0
 

47	 0.004 78 0.004
 

0 17
 

50	 0.04 78 0.04
 

0 18
 

s(concentration X weeks received)
 

13
 
Female rats were all approximately 6 weeks old at the time they
 

were placed on test. The initial dietary concentrations of hydrazobenzene were 0.004 and 0.002 percent. The female rats receiving
 

0.002 percent were sacrificed after 41 weeks because the dose level
 

was considered, on the basis of weight depression, to be inadequate.
 

No histopathologic examinations of these animals were performed. A
 

new dosed group, receiving 0.01 percent, was started with a new
 

control. Throughout this report, those female rats receiving 0.01
 

percent and their controls are referred to as the high dose and high
 

dose control groups, respectively. Throughout this report those
 

female rats initially receiving a concentration of 0.004 percent and
 

their controls are referred to as the low dose and low dose control
 

groups, respectively.
 

Male mice were all approximately 6 weeks old at the time they
 

were placed on test. The initial dietary concentrations of hydrazobenzene were 0.007 and 0.0035 percent. After week 9, the higher dose
 

was increased to 0.008 percent. The male mice initially receiving
 

0.0035 percent were sacrificed after 22 weeks because the dose level
 

was considered, on the basis of weight depression, to be inadequate.
 

No histopathologic examinations of these animals were performed. A
 

new dosed group, receiving 0.04 percent, was started with a new
 

control group. Throughout this report, those male mice receiving
 

0.04 percent and their controls are referred to as the high dose and
 

high dose control groups, respectively. Throughout this report those
 

14
 
male mice initially receiving a concentration of 0.007 percent and
 

their controls are referred to as the low dose and low dose control
 

groups, respectively.
 

Female mice were all approximately 6 weeks old at the time they
 

were placed on test. The initial dietary concentrations of hydrazobenzene were 0.004 and 0.002 percent. The female mice initially
 

receiving 0.002 percent were sacrificed after 22 weeks because the
 

dose level was considered, on the basis of weight depression, to be
 

inadequate. No histopathologic examinations of these animals were
 

performed. A new dosed group, receiving 0.04 percent, was started
 

with a new control group. Throughout this report, those female mice
 

receiving a concentration of 0.04 percent and their controls are
 

referred to as the high dose and high dose control groups, respectively. Throughout this report those female mice initially receiving
 

a concentration of 0.004 percent and their controls are referred to
 

as the low dose and low dose control groups, respectively.
 

Treated rats and mice (except for groups terminated early) were
 

supplied with dosed feed for a total of 78 weeks followed by a 26-week
 

observation period for rats and a 13-week observation period for mice.
 

G. Clinical and Histopathologic Examinations
 

Animals were weighed immediately prior to initiation of the experiment. Body weights were recorded twice weekly for the first 12
 

weeks of the study and at monthly intervals thereafter. From the
 

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

15
 
Food consumption, for two cages from each group, was monitored for
 

seven consecutive days once a month for the first nine months of
 

the bioassay and for three consecutive days each month thereafter.
 

The presence of tissue masses and lesions was determined by monthly
 

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 carbon dioxide inhalation,
 

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, seminal vesicle, pituitary, adrenal, thyroid,
 

parathyroid, ear, testis, prostate, brain, Zymbal's gland, uterus,
 

mammary gland, and ovary.
 

16
 
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
 

17
 
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.
 

18
 
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
 

19
 
procedures for analyses of the incidence of tumors (Fisher exact
 

tests, Cochran-Armitage tests, etc.) were followed.
 

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

incidence of tumors. Curves of the proportions surviving without an
 

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.
 

20
 
The lower and upper limits of the confidence interval of the
 

relative risk have been included in the tables of statistical analyses. The interpretation of the limits is that in approximately 95
 

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

of the 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.
 

21
 
III. CHRONIC TESTING RESULTS: RATS
 

A. Body Weights and Clinical Observations
 

During this bioassay, slight depression of mean group body weight
 

relative to controls was apparent for high dose male rats but not for
 

low dose male rats. Among female rats, a slight depression of mean
 

group body weight was observed for the low dose group after week 46
 

and for the high dose group after week 22 (Figure 2).
 

Palpable subcutaneous masses were the only frequent clinical
 

observation. They were observed in ten high dose control females,
 

two high dose control males, three high dose females and two low dose
 

males. One high dose male had a firm nodule on the ear. Sporadic,
 

isolated observations included: One high dose male with an eye infection, one high dose control female with eye discoloration and another
 

with exopthalmia, one low dose control male with an encrusted lesion
 

on the dorsal surface, a brown vaginal exudate in one high dose female, and alopecia in one high dose control female. No other clinical
 

abnormalities were recorded for male or female rats.
 

B.	 Survival
 

The estimated probabilities of survival for male and female rats
 

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

For male rats the Cox tests did not indicate positive associations between increased dosage and accelerated mortality. From each
 

of the treated groups and from the untreated controls, five rats were
 

sacrificed in week 78. In week 100, 64 percent (32/50) of the high
 

22
 
750 750 

- 600 

-450 

— 300 

-150 

TIME ON TEST (WEEKS) 

120 

750­ -750 

600 — -600 

(- 450­

LU 

5 

g 300­

m 

-450 

-300 

150­ -150 

FEMALE RATS 

15 30 45 

~ 

60 

I 

75 

TIME ON TEST (WEEKS) 

90 

FIGURE 2 

GROWTH CURVES FOR HYDRAZOBENZENE CHRONIC STUDY RATS 

I 

105 120 

23 
PROBABILIT

Y OF SURVIVAL 

p

p b KJ 

I 

CO 

i 

8 

I 30 s

§ O ­ 30 C5 rxs > C N 33 O m 

m m N m A CO 

m 

O 30

 

O 

z 

o 00 

c 

D 

t*' 

3D 

m r 

a-

3D 

1 1 1 0 O 0 

' 

PROBABILITY OF S

O 

n 

o 

z 

8- H 

— Ul " 

m 

O 

2 

m 

§ 

m 

m 

|~ 

s­

33 

1 1 

O O 

' 
dose, 78 percent (39/50) of the low dose, 73 percent (36/49) of the
 

high dose controls and 70 percent (35/50) of the low dose controls
 

were still alive. There were adquate numbers of male rats at risk
 

from late-developing tumors.
 

For female rats the Cox test indicated that the mortality was
 

significantly (P < 0.001) greater in the high dose group than in the
 

high dose control. For both the treated and untreated female rats,
 

five animals from each group were sacrificed in week 78. There were
 

adequate numbers of female rats at risk from late-developing tumors
 

with 50 percent (25/50) of the high dose, 74 percent (37/50) of the
 

low dose, 86 percent (43/50) of the high dose control and 78 percent
 

(39/50) of the low dose control surviving in week 100.
 

C.	 Pathology
 

Histopathologic findings on neoplasms in rats are summarized in
 

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

summarized in Appendix C (Tables Cl and C2).
 

The incidence of neoplasms in most organs in the treated rats
 

was comparable to that of the control groups. However, hydrazobenzene
 

feeding increased the development of neoplasms of the liver, auditory
 

sebaceous gland, and mammary gland (females).
 

Hepatocellular carcinomas were found in 36 treated male rats as
 

early as week 72 (5/49 [10 percent] low dose and 31/49 [63 percent]
 

high dose), in none of the treated female rats, and in 1/95 (1 percent) control male rats. Neoplastic nodules found in treated males
 

25
 
and females and in control males involved a few lobules, were well
 

demarcated, and compressed the adjacent normal liver cells in some
 

areas. Cells in these nodules were large, and cytoplasm either
 

acidophilic or basophilic. Nuclei were vesicular. The hepatocellular carcinomas involved one or many lobes of the liver. The tumors
 

appeared well-differentiated. Lobular architecture was distorted
 

within the neoplasms, and the liver plates were several cells thick.
 

In some tumors, the cells were arranged in a glandular pattern, and
 

there appeared to be a slight pleomorphism in size of transformed
 

hepatocytes. The cytoplasm of a majority of cells was acidophilic;
 

in some it was vacuolated. Nuclei were large and vesicular, and
 

nucleoli prominent. Both normal and abnormal mitotic figures were
 

present. Sinusoids were distended and engorged with blood, and in
 

some there were a few hematopoietic cells. Small areas of necrosis,
 

cellular debris, and inflammatory cells were observed. Hepatocellular carcinomas metastasized to the lung in six treated male rats.
 

Neoplasms of the Zymbal's gland or ear canal occurred in 1/95
 

(1 percent) control male rats, 2/50 (4 percent) low dose males, 7/49
 

(14 percent) high dose males, 0/98 control females, 1/50 (2 percent)
 

low dose females, and 2/50 (4 percent) high dose females. The tumors
 

in eight of the treated rats were considered to be squamous-cell
 

carcinomas, one was a squamous-cell papilloma (high dose male), and
 

the remaining two were sebaceous adenocarcinomas. A squamous-cell
 

carcinoma in one low dose male rat had metastasized to the lung.
 

26
 
Adenocarcinomas of the mammary gland were found in 1/48 (2 percent) low dose control females and in nine treated females (3/50 [6
 

percent] low dose and 6/50 [12 percent] high dose). This tumor had
 

metastasized to the lung in one high dose female. The incidence of
 

fibroadenoma of the mammary gland in treated female rats was lower
 

than in female controls.
 

Neoplasms of the clitoral gland in treated females and the preputial gland in treated males as well as adrenal pheochromocytomas in
 

high dose males appeared to be slightly increased when compared with
 

control animals.
 

There was a variety of nonneoplastic lesions observed, but none
 

appeared to be compound-related.
 

Based upon this histopathologic examination evidence was provided for the carcinogenicity of hydrazobenzene in Fischer 344 rats
 

as dietary administration of the compound was associated with a dose-


related increase in the incidence of hepatic neoplasms and auditory
 

sebaceous gland neoplasms and a moderately increased incidence of
 

mammary gland adenocarcinomas.
 

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 is included for
 

every type of tumor in either sex where at least two such tumors were
 

observed in at least one of the control or hydrazobenzene-dosed groups
 

27
 
___ 

___ 

___ 

___ 

___ 

_ __ 

___ 

___ 

___ 

TABLE 3
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN MALE RATS TREATED WITH HYDRAZOBENZENE£
 

LOW DOSE HIGH DOSE LOW
 

TOPOGRAPHY: MORPHOLOGY CONTROL CONTROL DOSE
 

Skin excluding Skin of Ear: Squamous-


Cell Papillomab 1/47(0.02) 0/48(0.00) 1/50(0.02)
 

P Values0 N.S.
 

Relative Risk (Control) 0.940
 

Lower Limit 0.012
 

Upper Limit 72.331
 

Weeks to First Observed Tumor 108 90
 

ro
 

oo
 Subcutaneous Tissue: Fibroma 7/47(0.15) 3/48(0.06) 2/50(0.04)
 

P Values0 N.S.
 

Relative Risk (Control) 0.269
 

Lower Limit 0.028
 

Upper Limit 1.325
 

Weeks to First Observed Tumor 98 95 90
 

Lung: Alveolar/Bronchiolar Adenoma or
 

Alveolar/Bronchiolar Carcinoma^3 1/47(0.02) 1/48(0.02) 3/49(0.06)
 

P Values0 N.S.
 

Relative Risk (Control) 2.878
 

Lower Limit 0.241
 

Upper Limit 147.900
 

Weeks to First Observed Tumor 108 109 107
 

HIGH
 

DOSE
 

3/49(0.06)
 

N.S.
 

Infinite
 

0.590
 

Infinite
 

106
 

1/49(0.02)
 

N.S.
 

0.327
 

0.006
 

3.898
 

106
 

1/48(0.02)
 

N.S.
 

1.000
 

0.013
 

6.886
 

106
 
___ 

___ 

_ __ 

___ 

___ 

___ 

___ 

__ _ 

TOPOGRAPHY: MORPHOLOGY
 

Hematopoietic System: Leukemia or
 

Malignant Lymphomab
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Spleen: Hemangiosarcoma
 

P Values0
 

Relative Risk' (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Liver: Hepatocellular Carcinoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

LOW DOSE
 

CONTROL
 

5/47(0.11)
 

84
 

0/47(0.00)
 

0/47(0.00)
 

HIGH DOSE
 

CONTROL
 

6/48(0.13)
 

—_
 

92
 

0/48(0.00)
 

—_
 

1/48(0.02)
 

109
 

LOW
 

DOSE
 

2/50(0.04)
 

N.S.
 

0.376
 

0.037
 

2.172
 

102
 

0/49(0.00)
 

N.S.
 

_—
 

5/49(0.10)
 

P = 0.031
 

Infinite
 

1.212
 

Infinite
 

107
 

HIGH
 

DOSE
 

1/49(0.02) '
 

N.S.
 

0.163
 

0.004
 

1.274
 

106
 

3/49(0.06)
 

N.S.
 

Infinite
 

0.590
 

Infinite
 

86
 

31/49(0.63)
 

P < 0.001
 

30.367
 

5.548
 

1170.322
 

72
 
TABLE 3 (Continued) 

TOPOGRAPHY : MORPHOLOGY 

LOW DOSE 

CONTROL 

HIGH DOSE 

CONTROL 

LOW 

DOSE 

HIGH 

DOSE 

Liver: Neoplastic Nodule or 

Hepatocellular Carcinoma 5/47(0.11) 1/48(0.02) 13/49(0.27) 37/49(0.76) 

P Values0 P = 0.040 P < 0.001 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

2.494 

0.914 

8.254 

38.204 

7.290 

1395.851 

Weeks to First Observed Tumor 108 109 102 72 

u> 

o 

Adrenal: Pheochromocy toma or , 

Pheochromocytoma, Malignant 

P Values0 

7/47(0.15) ___ 8/47(0.17) ___ 7/48(0.15) 

N.S. 

16/46(0.35) 

P = 0.042 

Relative Risk (Control)

d 

Lower Limit 

Upper Limit 

0.979 

0.318 

3.019 

2.043 

0.922 

4.939 

Weeks to First Observed Tumor 108 106 74 78 

Pituitary: Adenoma NOS or

Adenoma^ 

P Values0 

 Chromophobe 

8/47(0.17) 9/38(0.24) ___ 8/46(0.17) 

N.S. 

2/29(0.07) 

N.S. 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

_ __ 1.022 

0.365 

2.856 

0.291 

0.033 

1.267 

Weeks to First Observed Tumor 108 85 78 106 
TOPOGRAPHY: MORPHOLOGY 

Thyroid: C-Cell Carcinoma 

P Values0 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

Thyroid: C-Cell Adenoma or C-Cell 

Carcinoma" 

P Values0 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

Pancreatic Islets: Islet-Cell Adenoma

Islet-Cell Carcinomab 

P Values0 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

TABLE

 or 

3 (Continued) 

LOW DOSE 

CONTROL 

2/46(0.04) 

108 

4/46(0.09) 

108 

4/45(0.09) 

85 

HIGH DOSE 

CONTROL 

1/48(0.02) 

109 

1/48(0.02) 

109 

0/46(0.00) 

LOW 

DOSE 

2/41(0.05) 

N.S. 

1.122 

0.085 

14.845 

102 

2/41(0.05) 

N.S. 

0.561 

0.053 

3.687 

102 

4/47(0.09) 

N.S. 

0.957 

0.189 

4.845 

107 

HIGH 

DOSE 

2/44(0.05) 

N.S. 

2.182 

0.118 

125.735 

82 

4/44(0.09) 

N.S. 

4.364 

0.454 

209.675 

82 

1/45(0.02) 

N.S. 

Infinite 

0.055 

Infinite 

106 
___ 

___ 

___ 

___ __ _ 

TOPOGRAPHY : MORPHOLOGY
 

Testis: Interstitial-Cell Tumor
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Zymbal's Gland: Squamous-Cell Carcinoma"
 

P Values0
 

OJ 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Ear Canal, Zymbal's Gland, and Skin of
 

the Ear: Squamous-Cell Carcinoma or
 

Squamous-Cell Papilloma^
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 3 (Continued) 

LOW DOSE HIGH DOSE
 

CONTROL CONTROL
 

44/47(0.94) 42/47(0.89)
 

N.S.
 

_ —
 

78 78
 

0/47(0.00) 0/48(0.00)
 ___ • ___
 

1/47(0.02) 0/48(0.00)
 

87
 

LOW
 

DOSE
 

46/49(0.94)
 

N.S.
 

1.003
 

0.907
 

1.109
 

74
 

1/50(0.02)
 

N.S.
 

Infinite
 

0.050
 

Infinite
 

107
 

2/50(0.04)
 

N.S.
 

1.880
 

0.101
 

108.696
 

68
 

HIGH
 

DOSE
 

43/48(0.90)
 

N.S.
 

1.002
 

0.869
 

1.156
 

78
 

5/49(0.10)
 

P = 0.030
 

Infinite
 

1.237
 

Infinite
 

86
 

7/49(0.14)
 

P = 0.007
 

Infinite
 

1.904
 

Infinite
 

78
 
Table 3 (Concluded) 

LOW DOSE HIGH DOSE LOW HIGH 

TOPOGRAPHY : MORPHOLOGY CONTROL CONTROL DOSE DOSE 

Body Cavities: Mesothelioma NOS 2/47(0.04) 2/48(0.04) 3/50(0.06) 5/49(0.10) 

P Values 

Q 

N.S. N.S. 

Relative Risk (Control) 1.440 2.449 

Lower Limit 0.173 0.416 

Upper Limit 16.632 24.226 

Weeks to First Observed Tumor 95 106 100 78 

Treated groups received time-weighted average doses of 0.008 or 0.03 percent in feed. 

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

The probability level for the Fisher exact test for the comparison of a treated group with the
 u> control group is given beneath the incidence of tumors in the treated group when P < 0.05; otherwise, not significant (N.S.) is indicated. A negative designation (N) indicates a lower incidence
 

in the treated group than in the control group.
 

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

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN FEMALE RATS TREATED WITH HYDRAZOBENZENE^
 

LOW DOSE HIGH DOSE LOW HIGH 

TOPOGRAPHY : MORPHOLOGY CONTROL CONTROL DOSE DOSE 

Hematopoietic System: Leukemia 3/48(0.06) 5/50(0.10) 0/50(0.00) 0/50(0.00) 

P Values0 P = 0.028(N) P = 0.028(N) 

Relative Risk (Control) 0.000 0.000 

Lower Limit 0.000 0.000 

Upper Limit 1.596 10.793 

Weeks to First Observed Tumor 107 104 

Liver: Neoplastic Nodule 0/47(0.00) 0/50(0.00) 0/50(0.00) 6/50(0.12) 

P Values0 N.S . P = 0.013 

Relative Risk (Control) Infinite 

Lower Limit 1.560 

Upper Limit Infinite 

Weeks to First Observed Tumor 77 

Pituitary: Adenoma NOS or Chromophobe
 

Adenoma^1 18/46(0.39) 17/40(0.43) 21/47(0.45) 13/37(0.35)
 

P Values0 N.S. N.S.
 

Relative Risk (Control) 1.142 0.827
 

Lower Limit 0.675 0.434
 

Upper Limit 1.946 1.538
 

Weeks to First Observed Tumor 94 78 78 69
 
TOPOGRAPHY: MORPHOLOGY
 

Adrenal: Pheochromocytoma or
 

Pheochromocytoma, Malignant"
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Thyroid: C-Cell Carcinoma
 

P Values0
 

Ln 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor
 

Thyroid: C-Cell Adenoma or C-Cell
 

Carcinoma"
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 4 (Continued) 

LOW DOSE 

CONTROL 

HIGH DOSE 

CONTROL 

LOW 

DOSE 

HIGH 

DOSE 

1/47(0.02) 3/49(0.06) 3/49(0.06) 

N.S. 

3/49(0.06) 

N.S. 

108 109 

2.878 

0.241 

147.907 

107 

1.000 

0.140 

7.126 

95 

3/46(0.07) 1/45(0.02) 3/50(0.06) 

N.S. 

3/46(0.07) 

N.S. 

109 109 

0.920 

0.129 

6.556 

107 

2.935 

0.247 

0.618 

94 

3/46(0.07) 

109 

2/45(0.05) 

109 

5/50(0.10) 

N.S. 

1.533 

0.317 

9.398 

107 

3/46(0.07) 

N.S. 

1.467 

0.176 

6.894 

94 
TOPOGRAPHY : MORPHOLOGY 

Mammary Gland: Adeno carcinoma NOS 

P Values0 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

Mammary Gland: Fib ro adenoma 

P Values0 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

Uterus: Endometrial Stromal Polyp 

P Values0 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

TABLE 4 (Continued) 

LOW DOSE 

CONTROL 

1/48(0.02) 

109 

14/48(0.29) 

76 

15/47(0.32) 

78 

HIGH DOSE 

CONTROL 

0/50(0.00) 

19/50(0.38) 

106 

10/50(0.20) 

78 

LOW 

DOSE 

3/50(0.06) 

N.S. 

2.880 

0.241 

148.100 

92 

9/50(0.18) 

N.S. 

0.617 

0.261 

1.381 

107 

10/50(0.20) 

N.S. 

0.627 

0.282 

1.337 

78 

HIGH 

DOSE 

6/50(0.12) 

P = 0.013 

Infinite 

1.600 

Infinite 

87 

9/50(0.18) 

P = 0.022(N) 

0.474 

0.211 

0.983 

46 

5/48(0.10) 

N.S. 

0.521 

0.150 

1.540 

78 
TABLE 4 (Continued) 

LOW DOSE 

TOPOGRAPHY : M3RPHOLOGY CONTROL 

Uterus: Endometrial Stromal Sarcoma 0/47(0.00) 

P Values0 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

Uterus and Uterus Endometrium: Carcinoma 

NOS or Aden o carcinoma NOS" 2/47(0.04) 

P Values0 

UJ Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 109 

Clitoral Gland: Adenoma NOS 0/48(0.00) 

Q 

P Values 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

HIGH DOSE 

CONTROL 

1/50(0.02) 

104 

1/50(0.02) 

109 

2/50(0.04) 

104 

LOW 

DOSE 

3/50(0.06) 

N.S. 

Infinite 

0.566 

Infinite 

88 

5/50(0.10) 

N.S. 

2.350 

0.407 

23.761 

89 

0/50(0.00) 

N.S. 

HIGH 

DOSE 

1/48(0.02) 

N.S. 

1.042 

0.014 

80.093 

82 

3/48(0.06) 

N.S. 

3.125 

0.262 

160.536 

106 

6/50(0.12) 

N.S. 

3.000 

0.569 

29.250 

68 
TABLE 4 (Concluded) 

Treated groups received time-weighted average doses of 0.004 or 0.01 percent in feed. 

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

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

in the treated group than in the control group. 

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

u> 

CO 
and where such tumors were observed in at least 5 percent of the
 

group. Cochran-Armitage tests were not used in these analyses.
 

High incidences of hepatocellular carcinomas in male rats and
 

neoplastic nodules of the liver in rats of both sexes were observed.
 

For males tne Fisher exact test indicated a significantly elevated
 

incidence of hepatocellular carcinomas in the high dose (P < 0.001)
 

compared to its controls. The Fisher exact test comparing the low
 

dose male rats with their controls had a significance level of P =
 

0.031, a marginal result which was not significant under the Bonferroni criterion. For high dose female rats, the Fisher exact test
 

was significant (P = 0.013) when the high dose incidence was compared to that of the high dose control. Based upon these statistical results, the administration of hydrazobenzene was associated
 

with an elevated incidence of neoplastic nodules in female rats and
 

hepatocellular carcinomas in male rats.
 

In male rats an elevated combined incidence of squamous-cell
 

carcinomas or squamous-cell papillomas of the Zymbal's gland, the
 

ear canal, or the skin of the ear was observed. The Fisher exact
 

test comparing the incidence of these tumors in the high dose to
 

that in the high dose control was significant (P = 0.007). Based
 

upon these statistical results, administration was associated with
 

an elevated incidence of squamous-cell neoplasms in male rats.
 

The elevated incidence of adrenal pheochromocytomas in the
 

high dose male rats relative to the high dose controls yielded a
 

39
 
significance level of P = 0.042 for the Fisher exact test, a marginal
 

result which was not significant under the Bonferroni criterion.
 

In female rats a high incidence of mammary gland adenocarcinomas
 

was observed. The Fisher exact test indicated a significantly (P =
 

0.013) elevated incidence of these tumors in the high dose group relative to that in the high dose controls. The historical incidence of
 

mammary adenocarcinomas in female Fischer 344 rats observed at Mason
 

Research Institute for the NCI Carcinogenesis Testing Program was
 

8/585 (1 percent).- Based upon these statistical results the administration of hydrazobenzene was associated with an elevated incidence
 

of mammary gland adenocarcinomas in female rats.
 

Negative associations resulted from the Fisher exact test comparing the incidence of leukemia or malignant lymphoma in both low
 

and high dose female rats to the incidence in their respective controls. These results appear to be spurious.
 

In female rats negative results were obtained from the Fisher
 

exact test comparison of the high dose rats to the high dose controls
 

for the incidence of mammary gland fibroadenomas. The incidence of
 

this tumor in the high dose controls (19/50 or 38 percent) was high
 

compared to the 115/585 (20 percent) observed in the Fischer 344
 

historical untreated female control rats maintained at Mason Research
 

Institute for the NCI Carcinogenesis Testing Program.
 

In summary, the administration of hydrazobenzene was associated with increased incidences of hepatocellular carcinomas and of
 

40
 
squamous-cell neoplasms of the Zymbal's gland, ear canal, and skin of
 

ear in male rats and with increased incidences of neoplastic nodules
 

of the liver and of mammary adenocarcinomas in female rats.
 

41
 
IV. CHRONIC TESTING RESULTS: MICE
 

A.	 Body Weights and Clinical Observations
 

No distinct pattern of mean body weight depression was evident
 

for low dose mice of either sex. Depression of mean group body weight
 

relative to controls was observed, however, for male and female high
 

dose mice after week 28 (Figure 4).
 

No clinical abnormalities were recorded for treated or control
 

mice of either sex.
 

B.	 Survival
 

The estimated probabilities of survival for male and female mice
 

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

For male mice the Cox test indicated a significantly greater
 

mortality in the high dose group than in the high dose control. Five
 

male mice were sacrificed from the high dose group and from each control group in week 78. In addition, five mice from the high dose
 

male control group were sacrificed in week 49. There were adequate
 

numbers of male mice at risk from late-developing tumors with 66
 

percent (33/50) of the high dose, 88 percent (44/50) of the low dose,
 

78 percent (39/50) of the high dose control, and 86 percent (43/50)
 

of the low dose control surviving to the termination of the study.
 

For female mice the Cox test indicated a significantly greater
 

mortality in the high dose group than in the high dose control. As
 

with the male mice, five high dose females and five females from
 

42
 
50- -50 

40 — -40 

V) 

cc 

30— -30 

>­

Q 

 20­ -20 

O 

CO LOW CONTROL 

< 

LU HIGH CONTROL 

10— -10 

LOW DOSE 

MALE MICE HIGH DOSE 

~i 1 r -| 1 1 1 r 15 30 45 60 75 90 105 120 

TIME ON TEST (WEEKS) 

50­ -50 

40 — -40 

in 

< 

-30 

> 20­ Q 

O LOW CONTROL 

-20 

CO 

HIGH CONTROL 

10— LOW DOSE -10 

HIGH DOSE 

FEMALE MICE 

15 30 45 60 75 90 105 120 

TIME ON TEST (WEEKS) 

FIGURE 4 

GROWTH CURVES FOR HYDRAZOBENZENE CHRONIC STUDY MICE 

43 
PROBABILITY OF SURVIVAL PROBABILITY OF 

c3 0 p o o —' C3 O O 

c5 Kj -^ en bo b CD kj * 

1 1 1 1 . i I 1 ! , i 1 i 1 I 

1 1 1

J ! 5i 1 

w _. ! 1 1

;

 i

 

J 0 C 01 — CJ11 

33 

i

i > . j... •

5 £ * r '" I r _ r

 

0 i 0 < 

_ fil l < I I : 1

S

 = 

fi I i-> -> _ O •*• o ° g 0 f3 1- o °

 m o ' > 

z § g s i 

O S 5 :H TM "1 Z -)

0 CJ m H 3 0 g- D o8 31 0 3) , 

•o ? O r- r 

33 

8 J^ Jfc ^ 

M d d g g

- U1 

s o s m 

I O . j O 

•n m 

Z < -n Z O — _i H 33 C) S rn en > C £/) 0) _ c/) g — N 33 H O H ° 

O m — 00 W 5 S 

m Z m ­ m m i 

m 

N /* ^ m c/) t/2 

Z ~ ^ i •sj 

•*J __ m 3- rfi i Ull 

O IT 

_r«: 1 

33 r 

Z 

J Hf '

 

O 

j

i jJ

,'

 ' (O

Q 

<q g-J O 

8- 

-n r =-/ | 

0 m ^ ;/ I < 2 ­ 2 5* S r~ > o m 1— m o m ag

- S 01 2 O 

m o 

m 

M 

0 1

 1 ' i ' i * i ' 1 1 1 1 1 1 

O0 

c3 O O 

= k> '** CD 00 b cD fsj '** 

= 0 O O o 
each of the control groups were sacrificed in week 78 with an additional five from the high dose control being sacrificed in week 49.
 

Survival of the female mice was relatively good with 52 percent
 

(26/50) of the high dose, 79 percent (37/47) of the low dose, 76 percent (38/50) of the high dose control, and 72 percent (36/50) of the
 

low dose control animals living until the termination of the study.
 

Thus, there were adequate numbers of female mice at risk from late-


developing tumors.
 

C. Pathology
 

Histopathologic findings on neoplasms in mice are summarized in
 

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

summarized in Appendix D (Tables Dl and D2).
 

There was an increased incidence of hepatocellular carcinomas
 

in female mice (20/43 [47 percent] high dose, 4/39 [10 percent] low
 

dose, and 3/97 [3 percent] controls). Hepatic neoplasms in these
 

mice involved a part or a whole lobe and compressed the surrounding
 

normal hepatic parenchyma.
 

The cytoplasm of the neoplastic hepatocytes varied in tinctorial
 

quality. It was usually basophilic or acidophilic and was occasionally vacuolated. Nuclei were large and vesicular, and mitotic figures were infrequently observed. In some cells, there were nuclear
 

inclusions. Sinusoids and blood vessels in the tumors were dilated.
 

Hepatocellular carcinomas metastasized to the lung in one low dose
 

male, two control males, and one control female.
 

45
 
These mice had a variety of other neoplastic and nonneoplastic
 

lesions, but none appeared to be compound-related.
 

Based upon this histopathologic examination hydrazobenzene
 

is carcinogenic to female B6C3F1 mice as there was a dose-related
 

increase in the incidence of hepatocellular carcinomas. The compound does not appear to be carcinogenic to male 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 is included
 

for every type of tumor in either sex where at least two such tumors
 

were observed in at least one of the control or hydrazobenzene-dosed
 

groups and where such tumors were observed in at least 5 percent of
 

the group. Cochran-Armitage tests were not used in these analyses
 

since the low dose groups and their controls we^e started at different times from the high dose groups and their controls.
 

Female mice in the treated groups showed increased incidences
 

of hepatocellular carcinoma. The Fisher exact test comparing the
 

incidence in the high dose group with that in the high dose control
 

was significant (P < 0.001). The Fisher exact test comparing the
 

high dose to the high dose control for the increased incidence of
 

hepatocellular adenoma or hepatocellular carcinoma was also significant (P < 0.001).
 

46
 
TABLE 5 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT 

SPECIFIC SITES IN MALE MICE TREATED. WITH HYDRAZOBENZENE' 

LOW DOSE HIGH DOSE LOW HIGH 

TOPOGRAPHY : MORPHOLOGY CONTROL CONTROL DOSE DOSE 

Lung: Alveolar/Bronchiolar Carcinoma 5/50(0.10) 5/49(0.10) 1/47(0.02) 0/46(0.00) 

P Values0 N.S . P = 0.033(N) 

Relative Risk (Control) 0.213 0.000 

Lower Limit 0.005 0.000 

Upper Limit 1.804 0.843 

Weeks to First Observed Tumor 95 96 95 

Lung: Alveolar/Bronchiolar Adenoma or 

Alveolar/Bronchiolar Carcinoma^ 5/50(0.10) 10/49(0.20) 2/47(0.04) 3/46(0.07) 

P Values0 N.S. P = 0.046(N) 

Relative Risk (Control) 0.426 0.320 

Lower Limit 0.042 0.060 

Upper Limit 2.454 1.149 

Weeks to First Observed Tumor 95 96 95 95 

Hematopoietic System: Malignant 

Lymphoma 5/50(0.10) 5/49(0.10) 4/49(0.08) 1/48(0.02) 

P Values0 N.S. N.S. 

Relative Risk (Control) 0.816 0.204 

Lower Limit 0.171 0.004 

Upper Limit 3.567 1.732 

Weeks to First Observed Tumor 74 96 95 95 
TABLE 5 (Continued) 

LOW DOSE 

TOPOGRAPHY : MORPHOLOGY CONTROL 

Hematopoietic System: Malignant Lymphoma 

or Leukemia" 5/50(0.10) 

P Values0 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 74 

Liver: Hepatocellular Carcinoma 12/50(0.24) 

P Values0 

oo Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 94 

Adrenal: Pheochromocytoma 0/49(0.00) 

P Values0 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

HIGH DOSE 

CONTROL 

5/49(0.10) 

96 

6/48(0.13) 

78 

1/44(0.02) 

96 

LOW 

DOSE 

4/49(0.08) 

N.S. 

0.816 

0.171 

3.567 

95 

11/47(0.23) 

N.S. 

0.975 

0.433 

2.169 

87 

1/45(0.02) 

N.S. 

Infinite 

0.058 

Infinite 

95 

HIGH 

DOSE 

3/48(0.06) 

N.S. 

0.613 

0.100 

2.965 

85 

8/46(0.17) 

N.S. 

1.391 

0.460 

4.492 

78 

3/42(0.07) 

N.S. 

3.143 

0.265 

0.893 

92 
TABLE 5 (Concluded) 

a

Treated groups received time-weighted average doses of 0.008 or 0.04 percent in feed. 

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

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 < 0.05; other- i.
 

wise, not significant (N.S.) is indicated. A negative designation (N) indicates a lower incidence
 

in the treated group than in the control group.
 

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

TABLE 6
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN FEMALE MICE TREATED WITH HYDRAZOBENZENE'
 

TOPOGRAPHY : MORPHOLOGY 

Hematopoietic System: Malignant Lymphoma 

P Values0 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

Hematopoietic System: Malignant Lymphoma 

or Leukemia 

P Values0 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

Lung: Alveolar/Bronchiolar Adenoma or 

Alveolar /Bronchiolar Carcinoma 

P Values0 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

LOW DOSE 

CONTROL 

5/48(0.10) 

96 

7/48(0.15) 

83 

2/46(0.04) 

, 

96 

HIGH DOSE LOW 

CONTROL DOSE 

2/50(0.04) 10/40(0.28) 

, N.S. 

2.400 

0.818 

8.189 

96 76 

2/50(0.04) 10/40(0.28) 

N.S. 

1.714 

0.649 

4.795 

96 76 

3/50(0.06) 3/38(0.08) 

N.S. 

1.816 

0.219 

20.760 

78 95 

HIGH 

DOSE 

5/44(0.11) 

N.S. 

2.841 

0.492 

8.601 

67 

6/44(0.14) 

N.S. 

3.341 

0.635 

32.410 

67 

2/40(0.05) 

N.S. 

0.833 

0.072 

6.902 

95 
TABLE 6 (Concluded)
 

TOPOGRAPHY : MOKPHOLO GY 

LOW DOSE 

CONTROL 

HIGH DOSE 

CONTROL 

LOW 

DOSE 

HIGH 

DOSE 

Liver: Hepatocellular Carcinoma 

P Values0 

2/47(0.04) 1/50(0.02) 4/39(0.10) 

N.S. 

20/43(0.47) 

P < 0.001 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

2.410 

0.366 

25.430 

23.260 

4.027 

922.300 

Weeks to First Observed Tumor 94 96 95 78 

Liver: Hepatocellular Adenoma or 

Hepatocellular Carcinoma^ 

P Values0 

2/47(0.04) 1/50(0.02) 

—­

4/39(0.10) 

N.S. 

22/43(0.51) 

P < 0.001 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

2.410 

0.366 

25.434 

25.581 

4.493 

1006.419 

Weeks to First Observed Tumor 94 96 95 78 

Treated groups received time-weighted average doses of 0.004 or 0.04 percent in feed. 

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

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

in the treated group than in the control group.
 

The 95% confidence interval of the relative risk of the treated group to the control group.
 
Based upon these statistical results the administration of hydrazobenzene was associated with an elevated incidence of hepatocelcellular carcinoma in female mice.
 

For male mice the Fisher exact test comparing the high dose animals to the high dose control for alveolar/bronchiolar carcinoma was
 

not significant under the Bonferroni criterion. Similarly, when incidences of both alveolar/bronchiolar adenoma and alveolar/bronchiolar
 

carcinoma were combined the results of the Fisher exact test were not
 

significant under the Bonferroni criterion.
 

52
 
V. DISCUSSION
 

Adequate numbers of animals in all groups of rats and mice in
 

this study survived sufficiently long to be at risk from late-


developing tumors.
 

The methods used for chemical analyses were not quantitative,
 

and, therefore, did not indicate the purity of the compound. No
 

qualitative analyses for identification of impurities were performed,
 

The liver was a target organ for carcinogenicity in the rat.
 

The incidence of hepatocellular carcinomas was significantly increased in dosed male rats and the incidence of neoplastic nodules
 

of the liver was significantly increased in dosed female rats. The
 

combined incidence of squamous-cell carcinomas and squamous-cell
 

papillomas of the Zymbal's gland, ear canal or skin of the ear was
 

statistically significant in male rats. The incidence of mammary
 

gland adenocarcinoma was significantly increased for female rats.
 

The liver was the target organ for carcinogenicity in female
 

mice. The incidence of hepatocellular carcinoma in the high dose
 

female mice was significant. No significantly increased incidences
 

of liver tumors were, however, observed among dosed male mice when
 

compared to controls.
 

The evidence for carcinogenicity of hydrazobenzene in this bioassay is supported by positive results for carcinogenicity reported
 

in other studies. Hydrazobenzene was found to be carcinogenic in
 

both rats and mice following exposure via subcutaneous injection
 

53
 
(Pliss, 1974). Tumors of the uterus, mammary gland, concha cimbae
 

(Zymbal's gland), and liver were noted in rats while subcutaneous
 

sarcomas and hepatic tumors were noted in mice (Pliss, 1974). Concurrent subcutaneous administration of benzidine sulfate and hydrazobenzene to rats resulted in an increased incidence of bladder cancer
 

and a decreased latent period when compared to the tumor incidences
 

and latent periods noted following administration of the individual
 

compounds (Kurlyandskii et al., 1976).
 

Under the conditions of this bioassay, hydrazobenzene was carcinogenic to Fischer 344 rats of both sexes, causing increased incidences of hepatocellular carcinoma and Zymbal's gland squamous-cell
 

neoplasms in male rats, neoplastic nodules of the liver in female
 

rats, and mammary adenocarcinomas in female rats. Hydrazobenzene was
 

also carcinogenic to female B6C3F1 mice, causing an increased incidence of hepatocellular carcinomas. The compound was not carcinogenic
 

to male B6C3F1 mice.
 

54
 
VI. BIBLIOGRAPHY
 

Anthony, H.M., and G.M. Thomas, "Tumors of the Urinary Bladder: An
 

Analysis of the Occupations of 1,030 Patients in Leeds, England."
 

Journal of the National Cancer Institute 45:879-895, 1970.
 

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

J. Wiley & Sons, New York, 1971.
 

Bannister, D.W., "Dyes and Dye Intermediates." Kirk-Othmer Encyclopedia of Chemical Technology, Second edition, Volume 7, p. 491,
 

1967.
 

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.
 

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.
 

International Agency for Research on Cancer, "Aromatic Amines." IARC
 

Monographs on the Evaluation of Carcinogenic Risk of Chemicals
 

to Man, Volume 1, pp. 69-91, 1972.
 

International Agency for Research on Cancer, "Hydrazine and its
 

Derivatives." IARC Monographs on the Evaluation of Carcinogenic
 

Risk of Chemicals to Man, Volume 4, p. 81, 1974.
 

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

Observations." Journal of the.American Statistical Association
 

53^:457-481, 1958.
 

Kurlyandskii, B.A., A.G. Medvedowskii, V.M. Voronin, and F.D. Moshbits,
 

"Experimental Study on the Combined Effect of Some Diphenylamino
 

Derivatives with Regard to the Prevention of Occupational Urinary
 

Bladder Growths." Gig. Tr. Prof. Zobal 5:34-38, 1976.
 

55
 
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.
 

Pliss, G.B., "Carcinogenic Properties by Hydrazobenzene." VOPR. ONKOL.
 

_20_:53-57, 1974.
 

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
 

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

Stanford Research Institute, 1977 Directory of Chemical Producers,
 

U.S.A. Menlo Park, California, 1977.
 

Sutton, W.L., "Heterocyclic and Miscellaneous Nitrogen Compounds."
 

Industrial Hygiene and Toxicology: Volume II. Toxicology,
 

Second edition. F.A. Patty, editor. Interscience, New York,
 

pp. 2171-2234, 1967.
 

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

6^:679-682, 1975.
 

Weast, R.C., editor, The Handbook of Chemistry and Physics, 52nd
 

edition. The Chemical Rubber Company, Cleveland, Ohio, 1972.
 

Wenner, W., "Malonic Acid and Derivatives." Kirk-Othmer Encyclopedia
 

of Chemical Technology, Second edition, Volume 12, p. 857, 1967.
 

Wynder, E.L., J. Onderdonk, and N. Mantel, "An Epidemiological
 

Investigation of Cancer of the Bladder." Cancer 16:1388-1407,
 

1963.
 

56
 
APPENDIX A
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS
 

IN RATS TREATED WITH HYDRAZOBENZENE
 

TABLE A1

 

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

LOW DOSE CONTROL (UNTR ) 

01-0055 

HIGH DOSE CONTRO L IJNTR ) 

01-0118 

L OU CCS E 

01-0050 

HIG H tf E 

01-0092 

F. 

«NIMAL S 

SNIMSL S 

SNIMAL S 

JNIMAL S 

INITIALL Y I K STUD Y 

MISSIN G 

NECROPSIE D 

EXAMINE D HISTOPATHCLCGICML Y 

50 

2 

1(7 

** U7 

M9 

i»8 

US 

50 

50 

«9 

50 

1(9 

119 

INTEGUMENTARY SYSTEM 

SQUAMOU S CEI L PHPILLCM * 

BASAL-CEL L CARCINOM A 

LEIOKYCSAECOd A 

(2K) 

(2*) 

;50) 

1 (2!!) 1 (8*) 

*SUECU T TISSUE 

UNDTFFEPENTIATED CAPCINOMA 

BASAL-CELL CARCINOMA 

SARCOMA, NOS 

FIBR01A 

FIBPOS»RCOM A 

MYOEPITHELIOMA 

(47) 

1 (2*) 

(«8) (50) 

2 ;»») 

(U9) 

1 I2K) 

BESPIPATORY SYSTEM 

•LUNG 

SQUHfOUS CELL CARCINOMA 

SQUAMODS CELL CAPCTNOMA, NETASTA 

HEPATCCEILULAR CARCINOMA, KITAST 

ALVEOLAR/BPONCHIOLfR ADENOMA 

ALVEOLAF/BRONCHIOLAB CARCINOMA 

_PHEOCHBOMOCYTOMA, METASTATIC 

SAPCOMA, NOS, METASTATIC 

1 (2X) 2 ;«*) 

HEMATOPOIETIC SYSTEM 

•MULTIPLE ORGANS 

MALIGNANT LYMPHCMA, NOS 

LEUKSMIA.NOS 

UNDIFFERENTIATED LE'IKENIA 

1 (2%) 1 (2X) 

;50) 

I NUMBE R O F ANIMAL S WIT H TISSUE EXAMINE D

* NUMBER OF ANIMAL S NECROPSIED 

"EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

 MICROSCOPICALL Y 

A-3 
• SPLEEN


MUCINOUS ADENOCAPCTNCMA, METASTA


HEMANGIOSAPCOMA
 

MSLIG. LYMPHOPA, LYHFHOCYTIC TYPE


MALIG.LYNPHOMA, HTSTIOCYTIC TYFF
 

*MEDIASTINAL I. NODE


MUCINOaS ADENOCARCTNOMA , METASTA


*CARDIOVASCULAR SYSTE


ONDIFFERENTIfTED LEUKEMIA
 

• LIVER


KUPFFER-CELL SAPCOMA
 

*THY»US


THYMOM A
 

CIRCULATORY SYSTEM
 

• HEART
 SQUAMOUS CELL CAFCINOMA, ME"ASTA
 

DIGESTIVE SYSTEM
 

*ORAL CAVITY


FIBPCSAECCMA


• SALIVARY GIAND


ADENOCARCINORA, NOS


SARCOMA, NOS


•LIVES


NEOPLASTIC NODUIF
 HEPATOCELLOLAR CARCINOMA


• PANCREAS


MUCINOUS ADSNOCAPCINCMA, METASTA
 ACINAS-CELL ADENOMA
 

• STOMACH
 SOUAMOUS CELL PAPIILOMA


TABLE A1 (CONTINUED)
 

LOW DOSE
 CONTFOL(DNTP)


01-0055


 (47)


 1 (2X)
 

1 (2H)
 

 (4?)


 1 (2*)
 

(47)


 (47)


 (42)


 [47)


 J47)


 1 (2%}
 

(46)


 (47)


 5 (11%)
 

(45)


 1 (2%)
 

(47)


 HIGH DOSE
 CONTROL (UNTR)
 

 C1-0118
 

 (48)
 

 (44)
 

 (48)
 

 (48)
 

 (23)
 

 '48)
 

 ;48)
 

 (47)
 

1 (2X)
 

1 \2%)
 

 (48)
 

1 (2%)
 

 (46)
 

 (48)
 

. .. .
 

f	 NUMBER OE ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBE" OF ANIMALS NECFOPSIED
 

A-4
 

LOW DOS! HIGH CCSF
 

01-0050 01-0092
 

(49)	 (48)
 

3 (6X)
 

1 I2X)
 

(48) (27)
 

(50)	 (49)
 

1 (2%)
 

(49) (49)
 

(31) (20)
 

1 (2*)
 

;50) ;49)
 

(46) (46)
 

(49)	 (49)
 8 (16%) 6 (12*)
 

5 !10«) 31 (63)!)
 

(47)	 (45)
 

1 (2«)
 

(49) (47)
 
TABLE Al (CONTINUED) 

LOW DOSE CONTROL (UNTB) 

01-0055 

HIGH DOSE 

CONTROL (UNTE) 

01-0118 

LOW DOSE
 

01-0050
 

HIGH CCSE
 

01-0092
 

MUCINOOS SDENOCAECINCMA, METAS1A 1 (2K) 

ISBALL INTESTINE 

LEIOMYOSARCOHA 

(47) (46) (48)
 (39)
 

• 3»)
 

tILEUB 

SAECOBA, NOS 

(47) (46) 

1 (2X) 

(48)
 (39)
 

•COLON 

BUCINQUS ADENOCABCINOBA 

(46) 

1 (2*) 

(46) (47)
 (33)
 

UFINAEY SYETEB
 

tKIDNEY 

LIPCBA 1 (2%)
 

:»8,
 ;49)
 ;48)
 

INDOCEINE SYSTEM
 

tPITUITAEY (38)
 

ADENOMA, NOS 8 (17*) 9 (24*)
 CHBOMOPHOBF ADENOMA
 

(46)
 

6 (13*)
 

(29)
 

2 (7K)
 

*ADEENAI (47) (47)


7 PHEOCHBOBOCYTOMA 7 (15*) (15*)
 PHEOCHBOBOCYTOMA, MALIGNANT 1 ;2*)
 

(48)
 

7 (1?*)
 

(46)
 

16 IBS*)
 

ITHYPOIC (46) (48)
 

FOLLICUIAR-CELL ADENCBA
 

C-CELL ADENOMA 2 ;4*)
 C-CFLL CARCINOMA 2 ;4*>
 

(41)
 

2 ;5*)
 

(44)
 

1 (2*)
 

2 (5*)
 2
 

(24) (28)
 

ADENOMA, NOS 1 (4*)
 

(19)
 (22)
 

IPAMCREATIC ISLETS (45) (46)
 

ISLET-CELL ADENCM A
 

ISLET-CELL CARCINOMA
 

(47)
 

4 (9«)
 

(45)
 

1 (2K)
 

FEPBODOC1IVE SYSTEM
 

*PBEPUTTA L GLf.N D ;48) 

C_A£CJ KCMi^Jifii 

;49)
 

* NUMBEE OF ANIMALS HITH TISSUE EXAMINED BICPOSCOPICAI1Y
 

* NUMEEE OF ANIBAL S NECECESIED
 

A-5
 
TABLE A1 (CONTINUED) 

ADENCBA , NOS

*SEMINA L VESICL F

KOCINOU S ADENOCSPCINOBA ,

tTESTIS

INTERSTITIAI-CELL TUMOR

 METAST A

LOW DOSE 

CONTRO L (ONTR ) 

01-0055 

1 (2t) 

 (47) 

1 (2%) 

 (47) 

 44 (94*) 

HIGH DOSE CCNTBCL (UNTR )

01-0118

(48)

(47) 42 (89%)

 LOU DOSE 

 01-005 

1 (2% ) 

 (50) 

 (49) 46 (94*) 

HIG H DOSE 

01-0092 

2 («*) 

(49 ) 

(48;

43 |90)t) 

NFHVOU S SYSTE M 

•BRAI N

SAHCCflA , NCS , HETASTATI C 

GLTOMA , NO S 

OLIGODENDROGLIOM A 

 |45) |48)

1 (2*)

 149) 

1 ;2*) 

116) 

1 (2*) 

SPECIA L SENS E ORGAN S 

•EA R CANA L

SQDAMCU S CEL L PAPILLCM A 

SQUAMOU S CELL CARCINOM A

*ZYHEAI' S GLAN D

SQUAMOU S CELL CARCINOM A 

 [47) 

1

 (47) 

 (2*) 

|48)

(48)

 ;50) 

 (50)

1 ;2*| 

;49)

1

(49 ) 

5

 (2X) 

 110*) 

BUSCULOSKELETA L SYSTE M 

*STEPNriM

MUCINOU S ADENOCAPCINCMA , METAST A

 ;47) 

1 (2)1) 

;48> ;50) :«, 

EOC Y CHVITIF. S 

*EOD Y CAVITIE S

HESOTHET.IOKA .

MESOTHELIUMA ,

 NO S 

 MALIGNAN T

 (47) 

2 (U*) 

(48)

2 (4*) 

 (50) _ (49) 

5 |10*) 

4 LL O^H^ F SYSTEM S 

SITE UNKNOW N 

» NUMBE R O F BNIHAL 1 :

* NUMBE R O F ANIMAL S

 WIT H TISSU E

 NEC°CES!E D 

 EXAMINE D MICROSCOPICALL Y 

1 

A-6 
TABLE Al (CONCLUDED)
 

LOW DOSE HIGH DOSE
 CONTROL (UNTB) CONTROL (DNT R) LOW DOSE HIGH DCS!
 

01-0055 01-0118 01-0050 01-0092
 

;»!I«IJL DISPOSITION SUMMAF Y
 

ANIMUS INITIALLY IN STUDY. 50 »9 50 50
 

NATURA L DEATHS 2 6 7 15
 

MORIBUND SATFIFICE 8 8 6 6
 

SCHEDULED SACRIFICE 5 5 5 5
 

ACCIDENTALLY KIIIED
 TERMINAL SACPIFT

CE 33 30 32 2U
 ANIMA L MISSING 2
 

a. INCLUDES ADTOLYZED ANTMALS
 

TCMOP SUMMAR Y
 

TOTAL ANIMALS KITH PRIMAR Y IUHOES* 17 1)4 «8 M7
 

TOTAI PRIMAR Y TUTORS 96 80 99 137
 

TOTAL ANIMALS WITH BENIGN TUMOFS 17 13 16 16
 

TOTAL EENIGN TUMORS 76 62 72 77
 

TOTAL »tHNJ>.LS WITH MALIGNANT TDMOBS 11 17 10 ri
 TOTAL HAIIGNANT TMMCES 15 18 16 1(9
 

TOTAL ANIMAL S WITH SECONDARY TUMOBS* 1 1 2 7
 

TOTAI SECONDARY Tljurps 6 1 2 8
 

TOTAL ANIMALS WITH TUMCRS UNCEBTAINBEMIGN OR MAIIGNA.NT 5 10 10
 

TOTAL lINCiETATN TJ1KCFS 5 11 11
 

TOTAL ANIMAL S WITH TUMCRS ONCEETAINPRIMARY OR METASTA"IC
 TOTAL UNCERTAIN TOSCSS
 

* PBIMAFY TOHOBS: AIL TOMORS EXCEPT SECONDARY TUMORS
 

* SECONDAFY TUMOBS: MSTASTATIC TDMOPS OR TOMORS INVASIVE INTO AN ADJACENT ORGAN
 

A-7
 
TABLE A2

 

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

LOW DOSE HIGH DOSE 

CONTROL (UNTP ) CONTROL (UNTR ) LO W CCS E HIG H CCE E 

02-0055 02-0118 02-0052 02-0091 

ANIMAL S INITIAII Y I K STTD Y 50 50 50 50 ANIMAL S MISSIN G 2 

ANIMAL S NECFOPSIE E K 8 50 50 50 fNIMAL S EXAMINE D HIS T OPATHOIOGICALL Y * 50 50 50 

INTEGUMENTAR Y SYSTE M 

*SKIN :«8) ;50) ;50) ;50> BASAL-CELL CAPCINO>"A

 1 (2*) 

*SUBCO T TISSU E (K8 ) (50) (50) (50) FIESOM A 1 (2*)

 FIBP.OSA9COM A

 

FESPISATOS Y SYSTEM . 

• LUN G ;K7 ) ;50) ;sO) ;50) SOUAMOUS CEII CAPCINCM A 1 (2%) 

SQUAMOU S CEI L CARCINOMA , METAST A 

ADENOCASCINOMA , NO S , BETASTATIC 

ALV^CLAR/BPONCHIOLP P ACENOM A 1 (2*) 2 ;K«) 1 !2K) 

LEIOMYOSAKCOMA , METASTATI C 1 J2» ) 

HEMATOPOIETI C SYSTE M 

• MULTIPL E ORGAN S ;<t8) ;50) ;50) ;50)

 MALIGNAN T IYBPHCMA , KCS

 

UNDIFFEBENTTATE E LEUKEMI A 1 (2*)

 MYELOMONOCYTI C LEUKEMI A 3 ;6K) 3 ;6X)

 

•SPLEEN (U7 ) (K8) (50) (50) PHEOCHROMOCYTOMA , METASTATI C 1 (2%) UNDIFFEPENTIATE D LEUKEMI A 

• LYMP H NOE E ,uo, (K7 ) (UK ) (31) 

C-CKLL CABCINOMA , METASTATIC 1 (3*) 

ITHYMU S (39) (3K) (38) (2K) 

I NUBBF P O F ANIMAL S WIT H TISSUE EXAMINE D MICROSCOPICALL Y 

* NUMBE R OF ANIMAL S NECROPSIE D 

"EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

A-8 
TABEL A2 (CONTINUED) 

LOW DOSE CONTROL (DNTR )

02-0055

 HIGH DOSE CONTROL (UNTR )

 02-0118

 LOW DCS!

 02-0052

 HIGH CCE E 

 02-0091 

CIRCULATOR Y SYSTEM 

NONE 

DIGESTIVE SYSTEM 

*LIVER

NF.OPUSTIC NODDLE

PHEOCHHOMOCYTCMA, METASTATIC

• STOMACH

SQOANOU S CELL PAPILLCK A

IILEU M

LEIOMYOSAflCOM A

 ;47)

 (46)

 (46)

 ;50)

 (48)

 (48)

 1 (2)8) 

 ;50)

 (50)

 1 (2X) 

 (50)

 ;50) 

6 (12*) 

1 ;2K) 

 (44) 

 (40) 

URINARY

NONE 

 SYSTEM 

ENDOCRINE SYSTEM 

•PITUITARY 

CAOCISOMA.NOS 

ADENOMA, NOS 

CHFOMOFHOBE ADENOMA 

•BDRENIU 

CORTICAL ADENOMf. 

CORTICAL CARCINOMA 

PREOCHROMOCYTOMA 

PHEOCHROfOCYTOMA, MALIGNANT 

•ADRENA L MEDULL A 

GANGLICNEU"0(1 A 

•THYROI E 

FOLLICULAR-CEL L CARCINOM A 

C-CELL ADENOM A 

C-C1!LL CA3CINOM A 

(U6) 

2 (tt*) 

16 139%) 

(47) 

3 (6*) 

1 (2%) 

[47) 

(46) 

3 

(10) 

(49) 

1 

(49) 

(45) 

1 

(2%) 

(2*) 

(2%) 

(47) 

11 ;23«) 

10 ;2U) 

(49) 

1 (2%) 

3 ;6*) 

(49) 

(50) 

(37) 

13 [35*) 

(49) 

(49) 

(46) 

*PANCRE!TIC ISLETS (46) (18) (49) (46) 

« NUMBE R OP ANIMAL S WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBE R OF ANIMAL S NECROPSIED 

A-9 
REPRODUCTIV E SYSTE M 

*MAMMAP Y GLAN D 

ADENOMA , NO S 

ADENOCARCINOMA , NO S 

PAPILLAR Y CYSTADENOMA , NO S 

LEIOMYOSfRCOM A 

FIBRCADENCM A 

*CLITOPBL GLAN D 

CARCINOMA , NOS 

SQUAMOtlS CFLL PAPILLCKA 

ADENOMA , NO S 

*VAGIN A 

SARCOMA , NO S 

IOTEFO S 

CARCINOMS,NO S 

ADENOMA , NO S 

ADENCCAPCINOMA , NO S 

LEIOMYOSAHCOM A 

ENDOMETPIA L STF01AL POLYP 

ENDOMETBIA L STROMA L SARCOM A 

• DTERUS/ENCOMETPIO M 

CARCINOMA , NOS 

ADENOCAPCINOMA , NO S 

HOVAP Y 

GPANULO?A-C"LL TOHCR 

GPANULOSA-CEL L CARCINOM A 

CFRVOU S SYSTE M 

«EPAT N 

GLIOMA , NO S 

ASTROCYTOM A 

CLIGODENDROGIIOM A 

SPECIAL SENFE PPGANS
 

*ZYMEHL'S GLAND
 

* NUfPE P O F ANIMAL S HTT H TISSUE

* NUMBE R O F ANI»AL S NECFOFSIE C 

A-IO 

TABLE A2 (CONTINUED) 

LOW DOSE HIGH DOSE CONTROL (UNTR ) CONTFOL (ONTR ) 

02-0055 02-0118 

(18) (50) 

1 ;2X) 

1 (2 %) 

14 (29%) 19 (38*) 

(48) (50) 

1 (2X) 

2 ;«%) 

(48) (50) 

1 ;2«) 

(47) (50) 

2 (4*) 1 (2X) 

15 (32*) 10 ;20«) 

1 !2X) 

(47) (50) 

(46 ) (49) 

1 (2X) 

1 (2% ) 

;47) ;50> 

1 (2% ) 

 EXAMINE D MICROSCOPICALL Y 

LOW DOSE 

02-0052 

(50) 

3 ;e%) 

9 ;i8)i) 

(50) 

(50) 

(50) 

1 (2X) 

2 ;4%) 1 ;2%) 10 ;20*> 

3 ;e%) 

(50) 

1 (2X) 1 ;2%) 

(50) 

1 (2%) 

;50) 

2 (4% ) 

;50)
 

JiiL
 

HIGH tCSE 

02-0091 

(50) 

2 

6 

1 

9 

(50) 

1 

6 

(50) 

(48) 

1 

2 

5 

1 

(48) 

1 

(45) 

1 

:i9>

1 

1 

!50) 

(4*) 

(12*) 

;2*) 

(18X ) 

(2% ) 

(12X) 

;2K) 

(4*) 

;10») 

!2t) 

(2% ) 

(2*) 

(2*) 

;2*) 
TABLE A2 (CONTINUED) 

LOft DOSE HIGH DOSE 

CONTP01 (UNTR ) CONTRO L (ONTR ) LOW DCSE HIGH DOSE 

02-0055 02-0118 02-OC5 2 02-0091 

SEBACEOUS »DENCCA°CIKCWA 2 (UK)
 

HUSC010SKEIETHL SYSTEM
 

V'ONE
 

ECDY CAVITIES
 

NONE
 

AIL OTHER SYSTEMS
 

SITE UNKNOWN
 SQUAMOUS CELL CAPCTVOHA
 

(NIMSL DISPOSITION
 

ANIMALS INITIALLY IN STUD? 50 50 50 50.
 

NATURA L DEATH* 3 2 10
 BORTBUUD SACRIFICE 6 6 14
 

SCHECaiED EACRiriCE 5 5 5
 

ACCIDENTALLY KILLED
 

TERMINAL SACRIFICE 3U 37 21
 

ANIMA L MISSIS'S 2
 

j INCLODES > UTQLIZEP ANI H Aj,S
 

t NUMBER OF ANIMALS MITH TISSUE EXAMINED MICROSCOPICALLY
 

* NDMEE" OF ANIMAL S SECPCESIED
 

A-ll 
TABLE A2 (CONCLUDED)
 

LOW DOSE HIGH DOSE CONTFO L JDNTP ) i CCNTRC L ;UNTE ;| LOH D03E HIG H DOSE 

C2-CC5 5 02-0118 02-OOS 2 02-0091 

Tone s SUMMAR Y 

TOTA L ANIMAL S WIT H PRIMAR Y T UMCPS * «2 38 39 36

 

TOTAL PPIMAP Y TUMOPS •70 73 69 66 

TOTA L ANIMAL S KITH 3FNIG N TUMOR S 33 35 3« 30 

TOTA L EENIG N TUMOR S 53 59 119 39 

TOTAL ANIMAL S WIT H MALIGNAN T TUMOP S 1U 12 18 17

 

TOTAL PALxGNAN T T'JNOS S )1 13 19 20

 

TOTAL JNIMAL? WITH SECONDABY TtJMCPS* 1
 

TOTAL SECONDARY T'IMOSS 1
 

TOTAL ANIMAL S HTTH TDMOBS UNCERTAINBENIGN OR MALIGNAN T 1 1
 

TOTAL UNCERTAIN TUMORS 1 1
 

TOTAL ANIMALS WITH TUMORS ONCERTUNPPTMAP Y OP METASTATIC
 

TOTAL UNCERTAIN TUMCRS
 

* PRIMAR Y TUMOPS: ALL TUMORS EXCEPT SECONDARY TUMORS
 

» SECONDARY TUMORS: METASTATIC TUMORS OP TUMORS INVASIVE IN^O AN ADJACENT OEGAH
 

A-12
 
APPENDIX B
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS
 

IN MICE TREATED WITH HYDRAZOBENZENE
 

TABLE Bl 

. SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE TREATED WITH HYDRAZOBENZENE 

LOW DOSE HIGH DOSE
 CONTROL ttJNTR) CONTROL [UNIS) LOW DCSE HIGH COSE
 

05-0070 05-0118 05-0050 05-0093
 

ANIMALS INITIALLY IK STUDY 50 50 50 50
 ANIMALS MISSING 1 1 2
 

ANIMALS NECROPSIED 50 19 19 18
 

ANIMALS EXAMINED HI STOFATHCLOGICALLY ** 50 19 17 16
 

INTEGUMENTARY SYSTEM
 

*SUBCUT TISSUE (50) (19) (19) (18)
 

SARCOMA, SOS 1 ;2X)
 

BESPIRATCRY SYSTEM
 

*LUNG	 ;so) ;i9) ;i7)
 NEOPLASM, NCS, METASTATIC 1 (2%)
 HEPATOCEILULAB CARCINOMA, METAST 1 (2%) 1 !2X) 1 ;2»)
 

ALVEOLAB/BRONCHIOLAR ADENOM A	 3 ;7X>
 

ALVEOLAR/BSONCHIOLAR CARCINOMA 5 ;10*) 5 ;10%) 1 ;2t)
 

HIMATOPOIETIC SYSTEM
 

'MULTIPLE ORGAN?
 ,,
 MALIGNAN T LYMOHCMA, NCS 2 (If) 

MALIG.LYMPHOMA, HISTIOCYTIC TYPE 1 (2%) 3 (6%) 1 ;2*) 

UNDTFFERENTIATED LEUKEMIA 

LYMFHOCYTIC LEUKEMIA !21f) 

•SPLEEN	 (50) (19) (17) (11)
 HEMAN3IOSARCOMA 1 (2%)
 

MALIG.LYMPHOMA, RISTIOCYTTC TYPE
 

•LYMPH	 NOEE (15) (12) (38)
 

MALIG.LYMPHOM H , HTSTICCYTIC TYPE 2 (1%) 1 (2%) 1 |2»)
 

»MESENTERIC L. NODE (15) (12) (11) (38)
 

MAL T G.LYMEHOMA , HISTICCYTIC TYPE 1 (2*)
 

•PEYERS	 PATCH (19) (19) (17) (12)
 

MALIG.LYMPHOMA , HISTIOCYTTC TYPE 1 (2K)
 

CIRCULATORY SYSTEM
 

NO.N_E
 

* NUMBE R OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALLY
 

* NUMBE R O F ANIMAL S NECRCPSIE D 

**EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

B-3 
TABLE Bl (CONTINUED)
 

LOW DOSE HIGH DOSE 

CONTROL (ONIR) CONTROL (ONTP) 

05-0070 05-0118 

DIGESTIVE SYSTE M 

• LIVE? (50) (18) 

HEPATOCEILOLA R ADENOM A 2 (IS) 

HEPATOCELLDLAR CARCINOM A 12 ;21«) 6 ;13«) 

HEMANGIOM A 1 ;2X) 

HEMANGIOSARCONA , UNC PRI M CR MET 1 ;2X) 

•JEJDNU M (19) ((19) 

ADENOCARCINOHA , NO S 

U F T NAR Y SYSTE M 

• KIDNE Y ;i»9) ;U9) 

TUBUHR-CEL L AEENOCARC I NOM A 

ENDOCRIN E SYSTEM 

• ADRENA L ;19) JKU ) 

CORTICA L ADENOM A 

PHEOCHROMOCYTOM A 1 ;2X) 

•THYPOIt (10) (15) 

ADENOCARCINONA , NOS 1 (3X) 

FOLLICtJLAR-CEL L ADENOM A 

• PANCREATI C IS1ETS (16) (17) 

ISLET-CFLL ADENOM A 1 (2*) 

FEFRODOCTIV! SYSTE M 

• PBEPOTTA L GLAN D J50) t19) 

ADENCMA , NO S 

•T^STIS (50) (18) 

INTERSTITIf L-CEL L TUMC R 

NE8VOO S SYSTE M 

NON E 

• NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBF" OF ANIMAL S NEC5CPSIED
 

LOW DOSE 

05-0050 

(17) 

11 [23%) 

(17) 

-.17) 

1 (2X ) 

;15)

1 (2X ) 1 [2X) 

(15) 

(17) 

1 (2%) 

•19) 

1 (2*) 

(17) 

—— --' ­ 1IGH tCEE 

05-0093 

(16) 

1 (2S) 8 (17*) 

(12) 

1 (2K ) 

(16) 

(12) 

3 ;7») 

(36) 

1 |3«) 

(11) 

;i8) 

(16) 

1 (2») 

B-4
 
TABLE Bl (CONTINUED) 

LOW DOSE HIGH DOSE CONTPOL (UNTH ) CONTRO L (UNT E ) LOU DOSE HIG H CCS E 

05-0070 05-0118 05-0050 05-0093 

SEFCIAL SENSE ORGAN S 

'HARD^IAN GLAN D (50) (U9) (U9) (48) 

SDEN01S, NOS 1 (2*) 

MUSCULOSKELE'TAL SYSTEM 

NONE 

EOtY CAVITIFS 

NONE 

SI1 OTHEP SYSTEMS 

*Ht'lTIPL E CSGAN S (50) (49) (a? ) («8) 

HEUEOFrEROSSBCOPl l 1 (2«) 

!NIH> L CISECSITIC N S01PB5 Y 

ANIMAL S 1NIIISLL Y IN STUD Y ^0 50 50 50 

NHTM5A L DEATB3 I 2 3 7 

BORIEUN D SACFIFTCE 2 3 

SCHiniLF D SBCPI7IC' 5 10 5 

ACCIDENTALL Y KILLE D 

TEPMINA I SACFI^ICE a3 39 ou 33 •MINU L 11SSING 1 1 2 

I NUMBE R OF ANIMAL S HTT H TISSUE FX1MINE D HI CPOSCOPIC *I L 1 

* NUM9E H C F ANIMAL S NECFOFSIE C 

B-5 
TABLE Bl (CONCLUDED) 

LOW DOSE HIGH DOSE CONTRO L tUNTR ) CONTRO L ;UNTR ) LO W DOSE HIG H DOSE 

05-0070 05-0118 05-0050 

TUMOR SUMMAR Y
 

TOTAL ANIMALS WITH PRIMARY TUMCPS* 23 22 18 17
 

TOTAL PRIMARY TUMOPS 27 26 22 22
 

TOTAL ANIMALS WITH BENIGN TUMOSS 3 8 5
 7
 

TOTAL E1TNIGN TUMORS 3 8 5 9
 

TOTAL ANIMALS WITH "ALIGNANT TUMORS 22 15 15 13
 

TOTAL MALIGNANT T'lMCPS 20 17 17 13
 

TOTAL ANIMAL S WITH SECCNDAB 1 Y THMOES * 1 1 

TOTAL SECONDAR Y TTINORS 1 1 2 

TOTAL ANIMALS WITH TUTORS DNCERTAINE5NIGN OP MALIGNANT
 

TOTfL UNCERTPIN T'lMORS
 

TOT.'L ANIMALS WITH ^OMORS UNCERTAINPBIMARY CH METASTATTC 1
 

1PTAL UNCERTAIN TflMORS 1
 

* PRIMARY 1IIMOES: «LL TUMORS EXCEPT SECONDARY TUMORS
 

I SECONDLY TUMORS: J1ETASTATIC TUMOBS OR TUMORS INVASIVE INTO AN ADJACENT ORGAN
 

B-6 
TABLE B2

 

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

LOW DOSE
 CONT50L(U!ITB)
 

»NIMALS INITIALLY IN STUDY


BNIMALS NECBOPSIED


•NIMAIS EXAMINED HISTOFA THOLOGICALLY **


TNTEGOMEN'MPY SYSTEM
 

•SKIN


SARCOWA, NO?
 

*SMECUT TISSUE


TINDTBFIBENTIATED CARCINOMA
 

RESPIR'TORY SYSTEM
 

ILUNG


HEPATOCEILULAB CUPCINOMA, METf.ST
 ALVEOLAP/BPONCHIOIJP .ADENCMA
 

^LVEOLAE/BRONCHIOLSS CARCINOMA


OSTEOSSECOMA, META?TATIC


HEMATOPOIBTTC SYSTEM
 

• MULTIPLE ORGANS


MALIGNANT LYMPHCIH, NCS


MALIG.T.YMPHOMA, HISTIOCYTIC TYPE
 

ONDTfERENIIATE t LIDKEMI A 

LYMPHOCYTI C LEUKEMI A 

E?YTH"OCYTIC LEUKEMI A 

tSPLEEN 

HFM».NGIOSAPCOMA
 

MALIGNANT LYMPHOBA, NOS
 

MfLTG.LYHPHOMA, HISTIOCYTIC TYPE
 

*(1ESENTERIC L. NODE
 

MfLTG.LYMPHOMA, HISTIOCYTIC TYPE
 

06-0070
 

 50
 

 18
 

 17
 

 (18)
 

 (18)
 

 ;116)
 

1 (251)
 

2 ;«%)
 

1 ;2X)
 

 ;H8)
 

2 (1*)
 

1 (2%) 

(17)
 

1 (2*)
 

(36)
 

1 (3%)
 

« NUMBE E OF ftNIMALS WIT H TISSUE EXAMINE D MICEOSCOPICALL Y 

* ND«EF P OF SNIMAL S NECPOPSIE D 

**EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

B-7 

CONTBOL (UNTR)


06-0118


50


50


50


(50)


(50)


;soj


2 (1*)


;50)


2 (14%)


(19)
 

(11)
 

(50)
 

 LOU DOSE HIGH tCSE
 

 06-0052 06-0093
 

 «7 50
 

 10 HI
 

 39 13
 

 (10)	 (11) _
 

 (KO) (11)
 

1 [3%)
 

 ;3«)
 

•> (5*)	 2 (5*)
 

 ;«0)
 

2 (5%)	 1 (2%)
 

2 I5K)
 

(39 ) (11) 

(33 ) 

1 (?*)
 

(39)	 (13)
 

l__L21i_
 
TABLE B2 (CONTINUED) 

LOW DOSE HIGH DOSE CONTRO L (UNTR ) COHTFOL (ONTB ) LOU CCSE HIGH CCS E 

06-0070 06-0118 06-0052 06-0093 

tPEYERS PATCH (45) <«8) (39) (37) 

M1LTGNAN T LYMPHOMA , NO S 1 (2X ) 

MHIG . LYMPHOMA , HISTIOCYTIC TYPE 2 ;5X) 

CIRCULATOR Y SYSTE M 

NON E 

DIGESTIV E SYSTE M 

•LIVEP (47) (50) (39) (U3 ) 

N£OPI»5M , NO S 1 (2K) 

HE?ATOrElLULA R ADENOM A 2 I5X ) 

HEPATOCEIIUIA E CARCINOM A 2 (U%) 1 (2X ) 4 ;10*) 20 ;l|71!) 

URINAR Y SYSTE M 

NON E 

fNCOCPIN E SYSTE M 

(1?) (42) (3« ) (9) 

ADENOMA , NO S 5 (12% ) 1 (2X) 3 (9%) 

CHPOMOPHOB E ADENOM A 2 :5%) 

MDPENJ L (47) (48) (39) (40) 

COP"ICPL ADENOM A 1 (2%) 

PHEOCHROMOCYTOM A 1 ;3X) 

REPRODUCTIV E SYSTE M 

fUTEPU S ;43> ;«7) ;34) ;33) LETOMYON A 1 (2%) 

ENDOMETSIA L STFONU I POLY P 1 (3X ) 

tCFPVI X TJTEP I (U3 ) (47) (34) (33) 

SARCOMA , NOS 1 (3X ) 

*nVARY/OVIDOC T (43) (47) (34 ) (33) 

f NIJMBF P OF ANIKAI S «TTH TISSUE EXAMINE D MICBOSCOPICALLY 

* NUME' R O F AJIMAL S NECBOPSIE D 

B-8 
TABLE B2 (CONTINUED) 

LOW DOSE CONTROL (UNT" )

06-0070

 HIGH DOSE CONTROL (UHTR )

 06-0118 LOU DOS! OS-OOS2

 HIGH tOSI 

 06-0093 

HOVAPY

GPANULOSA-CEI1 TUMCR

 («5) (1»8) (37) (31) 

1 (3*) 

NERVOU S SYSTEM 

NONE 

SPECIAL SENSE ORGANS 

*H&RDEI!IAN GLAND

ADENOMA, NOS

PAPTLLA3Y UDENOMS

 (18)

 1 ;2*) 

 (50)

 1 ;2%) 

 (1(0) («") 

BUSCULOSKELETAL SYSTE1 

NONF 

BODY CAVITIES 

NONE 

A ll OTH?P SYETfCS 

OMENTUd 

HFHSNGinSAECOHA 1 

JNIMA L DTSFCS1TION SrjMPAE Y 

ANIMAL S :NITIAIL Y m STUD Y

NATIIPA L DEATH S

MOBIBUN D SACRIFICE

SCHEEUIf D SACRIFIC E

ACCTD?NTALI Y KIII1?D

TEBHINA I SSCRIFICE ANTKA L KISSTN G 

 so 6

 3

 5

 36

 50

 2

 10

 38

 i»7

 8

 2 

 37

 so 1« 

3 

5 

2 

 26 

a INCLUDES AUTOIYZED 'NIMAlj 

I NUMBER OF ANIMALS WITH TISSUE EXAMINED HICROSCOPICAILY 

* NUMBER OF ANIMUS NECPCFSIED 

B-9
 
TABLE B2 (CONCLUDED)
 

LOW DOSE HIGH DOSE
 CONTROL (UNTR) CCNTRCL (UNTE) LOW DOSE HIGH DOSE
 

06-0070 06-0118 06-0052 06-0093
 

TUflCF SUMMAF Y
 

TOTAL ANIMALS WITH PRIMARY TUMCRS* 18 10 19 29
 

TOTAL ESIMAPY TUMOPS 22 11 23 3K
 

7 7 U
 

TOTAL EE NIGN TUMORS 9 7 7 «
 

"O^AL ANIMALS WITH MALIGNANT TUMORS 12 H 15 25
 

TOTA1 MALIGNANT TUMOBS 13 U 16 28
 

TOTAL ANIMALS WITH S'CCNDARY TUMCPS» 2
 

TOTAL SECONDARY TUMOES 2
 

TOTAL ANIMALS WITH TUMOSS ONCERTAINBENIGN OR MALTGNAU T
 

TOTAL UNCERTAIN TUMORS
 

TOTAL ANIMAL? WITH TUMORS UNCERTAINPRIMARY OP METASTATIC
 TOT>.L UNCERTAIN TUMORS
 

•"OT.'L ANIMAL S WITH BFNIGN TUMCSS 8
 

* PRIMARY TUMORS: AIL TUMORS EXCEPT SECONDARY TUMORS
 

« SECONDARY TUMORS: M8TASTATIC TUMORS OR TUMOSS INVASIVE INTO t.V SDJACENT ORGAN
 

B-IO
 
APPENDIX C
 

SUMMARY OF THE INCIDENCE OF NONNEOPLA3TIC
 

LESIONS IN RATS TREATED WITH HYDRAZOBENZENE
 

TABLECl

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS 

TREATED WITH HYDRAZOBENZENE 

LOW DOSE HIGH DOSE CONTROL (UNTR ) CONTROL (UNT R ) LOU DOSE HIG H ECS F 

01-0055 01-0118 01-0050 01-0092 

JNIMAL S INITIALL Y IN STUD Y 50 19 50 50 

ANIMAL S HISSIN G 2 

ANIMAL S NECROESIF D U7 U8 50 19 

JNIMAL S EXAMINE D HISTOPATHOIOGICSLLY* * 17 18 U9 U 9 

INTEGUMENTAR Y SYSTE M 

*SKIN ;i7) ;i»8) ;50) ;i9) EPIDERMA L INCLUSIO N CYS T 1 (2%) 1 (2*) 1 (2*) INFLAMMATION , NECROTIZIN G 1 (2U) 

INFLAMMATION , ACUT E FOCA L 1 ;2*) 

HYPERKEPAinsi S 1 \2%) ACANTHOSI S 1 ;2%) 1 (2%) 

• SUBCU T TISSU E (17) (U8) (50) (U9 ) FIBROMATOSI S 1 ;2K ) METAPLASIA , OSSFCU S 1 (2)6) 

RESPIRATOR Y SYSTE M 

*TP»CHE A ;i!6) |lt8) ;i8) ;i6) INFLAMMATION , NCS 2 (1%) 1 (8*) 1 (2» ) 

• LUNG/ERONCHU S (U7 ) (1(8) (19) (18) ERONCHIFCTASI S 1 (2*) 1 (2%) INFLAMMATION , NOS 7 ;15%) 1 [2%] 8 (17*) 

INFLAMMATION , FOCA L 1 [2%) 

INFLAMMATION , NECHOTTZIN G 2 (<!*) 

ABSCESS , NOS 1 (2% ) 

• LUNG/BRONCHIOL E (U7 ) (KB ) (19) (18)

 INFLAMMATION , NC S 1 (2% )

 

• LUN G (U7 ) (US ) (19) (US ) 

CONGESTION , NO S 1 (2t) HEMORRHAG E 1 ;2») INFLAMMATION , NO S 1 ;2H ) INFLAMMATION , FOCA L i ;2%) ^2 ;2_1%J ±6 ;.3 .3.11 

O F ANIMAL S WIT H TISSU E EXAMINE D MIfEOSCIFIC»LI Y 

* NUMBE R O F ANIMAL S N3CFOFSIE C 

**EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

C-3 
TABLE Cl (CONTINUED) 

LOW DOSE 

CONTROL (UNTR ) 

01-0055 

HIGH DOSE CONTROL (UNTR )

01-0118

 LOU DCSE 

 01-0050 

HIGH CCSE 

01-0192 

INFLAMMATION , BECRCTIZIN G 

ABSCESS, NOS 

PNEUMONIA , CHRONIC MURIN E 

FIBROSIS, FOCAL 

HYPEBPIASIA , EPITHELIA L 

KYPERPLASIA , ALVFOLA R EPITHELIU M 

1 (2X ) 

1 (2% ) 

1 (2X) 3

1

 ;6X) ;2«) 

2 («*) 1 J2X ) 3 :e») 

1 !2X) 

1 f2K ) 

4 !8X) 

HIMATOPOIETIC SYSTEM 

IBONE MARRO H 

MYELCFIBROSIS 

MEGAKARYOCYTOSIS 

;it6) 

ISPLFF.N

HEMORRHAGE 

INFLUMMATION, NOS 

FIBFOSIS 

NECROSIS, FOCAL 

INFABCT HEMORRHAGIC

HEMOSIDEROSIS 

HYPERPLAEIA, HEMATOPOIETIC 

HYPERPLASIA, ERYTHROID 

HYPERPLASIA, RETICULUM CELL 

 (47) 

1 (2*) 

(48)

1 (2)1) 

1 '2%) 9 ;19X) 10 ;21%)

 (49) 

3 J6%) 10 ;2ox> 23 I47X) 

(48)

1 

2 

1 

2 

2 

8 

8 

1 

(2X) 

!"*) ;2*i

;4K) 

[4X) 

|17!() 

(17X) 

!2*) 

ILYMPH NOCE

HEMORRHAGE 

INFLAMMATION, NOS 

HYPERPLASIA, NOS 

RETTCULOCYTOSIS 

LYMPHOCYTOSIS 

PLASMACYTOSTS 

KYPEPPLASIA, PLASMA CELL 

HYPEBPLASIA, LYMPHOID 

 (42) (44)

1 (2X) 

1 »2X) 

3 ;7!6)

 (48) 

1 (2%) 2 ;4«) 

1 ;2X) 

2 -4X) 

1 ;2«) 5 ;10X) 

(27) 

1 

1 

1 

(4X) 

;4*) 

[4*) 

• PANCREATIC L.NODE

INFLAMMATION, ACOT E/CHRCNIC

 (42) 

1 (2*) 

(44) (48) (27) 

•ILEOCOLIC LYBEH NODE

LYMPHSCENOPATHY

 (42) 

1 (2*) 

(44) (48) (27) 

CISCULATORY SYSTEM 

•MYOCARDIUM

JNFL.AHJJATICN, NOS 

 ;47) ;u8) ;49)

5 (1pt) I47) 

> NUMBE" OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICAI1Y 

* NUMBER OF ANIMAL S NECPCFSIED 

C-4
 
TABLE Cl (CONTINUED)
 

« LOW DOSE CONTROL (ONTR) 

HIGH DOSE CONTRO L (UNTR ) 

01-0055 01-0118 

INFLAMMATION ,

FIBROSIS 

 INTERSTITIAL 23

12

 (48*) 

 ;25*) 

DEGENERATION , NOS 10 ;21«) 

*AOKTA (47) (48) 

MINERALIZATIO N 

*PULMONAR Y ABTER Y (47) (48) 

MINERALIZATION 1 ;2*| 

DIGESTIVE SYSTEM
 

•LIVES	 |48) 

FIBROSIS SFETA L LIVE R 2 (4%) 

DEGENERATION , HYALIN E 

NECROSIS , FOCAL 1 '2*( 

NECROSIS, COAGULATIV E 

METAMORPHOSI S FATT Y 3 [6*) 

CYTOPLASHIC VACUOLIZATIO N 

FOCAL CEILULA R CHANG E 12 [26*) 

HYPEFPIASIA , FOCAL 15 

HYPERPHSIA , DIFFUSE 

• NGIECTASI S 1 (2%) 

HEMATOPOIESI S 

• LIVEB/CEN'RIIOEtlLA R («7) (48) 

NECROSIS , NO S 1 (2*) 

*EILE DUCT (47) (48) ^ 

INFLAMMATION , NO S 

HYPEBPLSSIA , NO S 

• PANCREA S (45) (46) 

INFLAMMATION, NOS 17 (37*) 

INFLAMMATION, ACOTE/CHRONTC 

•PANCREATIC	 DUCT [46)
 

HYPERPLASIA, NOS
 

•PANCREATIC	 ACINUS (45) (46)
 

JTPOPHY, NOS 3 (7*)
 ATROPHY, FOCAL
 

HYPERTROPHY, FOCAL
 

I NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMF.ER OF ANIMALS NECFOPSTED
 

C-5
 

L OW DOSE 

01-0050 

39 (80*) 

21 I43X) 

(50)

1 ;2*) 

(50) 

'.49)
 

4
 

3
 

1
 

4
 

20
 

1
 

(49)
 

(50)
 

1
 

25
 

(47)
 

15
 

(47)
 

(47)
 

1 (2*)
 

HIG H DCSE 

01-0092 

14 (1C*) 

23 (49») 

(49) 

[49) 

;49)
 

2 [141!)
 

10 (20*)
 

8 !16*)
 

2 [4*)
 

4 (8*)
 

(49)
 

(49)
 

21
 

(45)
 

^ (16*)
 

(45)
 

1 (2*)
 

(45)
 

1 ;2*)
 
HYPEBPIASIA, FOCAL
 

•ESOPHAGUS
 

DYSP1ASIA, SOS
 

•STOMACH
 

INFLAMMATION, NOS
 

INFLAMMATION, FOCAL
 

HYPEPPLASIB, BASAL TELL
 

HYPEPKERATOSIS
 

ACANTHOSIS
 

PAPAKERATOSIS
 

•GASTRIC	 MUCOSA
 

HYPERPLASIA, FOCAL
 

•PEYERS	 PATCH
 

HYPEBPLASIA, NOS
 

»ILEUM
 

INFLAMMATION , NOS
 

KCOLON
 

NEMATOCIASIS
 

PARASITISM
 

U5INAEY SYSTEM
 

•KIDNEY
 

CYST, NOS
 

GLOMESULONEPHRT TIS, NCS
 

INFLAMMATION , CHEONIC
 

FTBEOSI?, DIFEOSE
 

• IIPTNAPY ELADDER
 

HYPERPLASIA, EPITHELIAL
 

HYPEPPLASIA, DIFFUSE
 

ENDOCRINE SYSTEM
 

•PITUITARY
 

HYPEPPLASIA, NOS
 

^YPEPP], A SIA^ FOCAL
 

S NOMEE S OF ANIMAL S WIT H TISSU E

* NUME5 R O F ANIMAL S NECPOPSIE D 

TABLECl (CONTINUED) 

LOW DOSE	 HIGH DOSE
 CONTROL (ONTP) CONTROL (UNTR)
 

01-0055 01-0118
 

1	 (2*)
 

(46)	 (45)
 

1 (2%)
 

(47)	 (48)
 

1 (2*)
 

1	 ;2*i
 2	 ;4X)
 

2 ;4%)
 

(47)	 (48)
 

(47)	 (46)
 

12 (26%)
 

(47)	 (46)
 

2 (4*)
 

(46)	 (46)
 

3 ;7«)
 

J47) ;48)
 

1 (2*)
 

47 (98%)
 

39 ;83K)
 

6 -13S)
 

(47)	 (43)
 

1 (2%) 1 (2%)
 

;i7) ;38)


1	 0*)
 3 J6%) 2	 '5%j__
 

 FXAPINE D MICROSCOPICALL Y 

C-6 

LOW CCSI
 

01-CC50
 

2 (1%)
 

[48)
 

(49)
 

4 (8% )


1 ;2K)
 

1 |2S)
 

4 ;8*)
 

(49)
 

(48)
 

8 (17J)
 

(48)
 

(47)
 

1 (2*)
 

[49)
 

46 (98K)
 

(46)
 

3 (7%)
 

1 (2%)
 

[46)
 

2 r41i)
 

HIGH CCSE
 

01-0092
 

(39)
 

(47)
 

1 (2*)
 

4 ;9K)
 

10 (21*)
 

17 ;36%)
 

1 !2%)
 

(47)
 

4 (9K)
 

(39)
 

7 (18*)
 

(39)
 

(33)
 

3 ;9*)
 

;48)
 

44 |92»)
 

8	 [17*)
 

(40)
 

1 (3t)
 

;29)
 

1 (3K)
 

2_ J7K)
 
TABLE Cl (CONTINUED) 

LOW DOSE CONTRO L (UNTR ) HIGH DOSE CONTROL (UNTR ) LOU CCS! HIG H CCS E 

01-0055 01-0118 01-OC^ O 01-0092 

•ADRENA L 

HYPF.RPLBSIA ,

HYPBBPLASIA ,

 NO S 

 FOCA L 

(»•»)

1 ;2*) 

(I"1) (IS) (16) 

1 (2K ) 

•ADRFNP L CORTF X (17) (47) (18) (46) 

HYPEBPLASIA , FOCAL 1 (2% ) 

»AD"ENA L MEDUIL A 

HYP^RPIASIP , NODULA B 

HYPEBPLAEIA , FOCAL 

(47)

3 (6*)

 (47) 

1

4

 (2X ) 

 ;9«) 

(18) 

7 (15X) 

(46)

2 (4X ) 

2 !U*) 

KTHYROT C 

FOLLICULB R CYST , NO S 

HYP^BPLASIS , C-CELL 

(4B)

i ;9X)

 (48) 

3 ;6X) 

(11) 

1 ;2t) 

(44) 

2 (51) 

»PARATHY"CI D (24 ) (28) (19) (22 ) 

HYPERPLB.SIA ,

HYPEBPIB.SIB ,

 NO S 

 FOCAL 

1 (IX) 1 (IX ) 2 19*) 

• PANCREATI C ISLF.TS 

HYPFRPLASIB , NO S 

(15) (46)

1 (2*) 

(47) 

3 (6X ) 

(15) 

1 (2X ) 

FEPRODUC1IV E SYSTE M 

*MAMMAB Y GLPN D 

GPLACTCCEL E 

HYPERPLASIB , NO S 

;i7)

1

1

 (2X)

 ;2X)

 ;48) ' 2

 4

 (4*) 

-.8%) 

;50) 

1

11

 (3% ) 

 ;22t) 

!19) 

1 tSX) 

*PBEPUTIA L GLAN D (47) (48) (50) (19) 

INFLAMMATION , ACUT E 

HYPEEPLJSIA , NO S 

1 I2X) 

1 ;2t) 

*PaOSTATT 

INFLAMI15TION , NOS 

(46) (44) 

17 (39X) 

(48) 

25 (52X ) 

(13) 12 (28X ) INFLAMMATION, ACUTE 

INFLAMMATION, BTUTE FOCSL 

INFLAMMATION, ACUTE/CHRONIC 

HYPF.RPLASIB, FOCAL 2 ;4X) 1 ;2*) METAPLASIA, SQOAMOUS 

*TSSTIS (U7) (47) (19) (48) 

HINEBSIIZATION 1 (2X) 

» NUMBE R O F BNIMAL S HIT H TISSUE EXAMINE D MICBOSCQPICALL Y 

* N1JMBEB OF ANIMAL S NECPOPSIE D 

C-7 
TABLE Cl (CONCLUDED) 

LOW DOSE CONTROL (UNTF)

01-0055

 HIGH DOSE CONTROL (UNTB)

 01-0118

 LOW DOSE

 01-0050

 HIGH CCSE 

 01-0092 

ATHOFHY , NOS

HYPEHPHSIA , INTERSTITIA L CEI L

 f,

 3

 (13*)

 ;6JS)

 (I

 7

 (8«)

 ;1!t*)

 6

 3

 (13K ) 

 ;6«) 

ITESTIS/TUEULE

MINERillTZATION

DEGENEBHTIflN, NOS

 (U7)

 1 (2%)

 (U7) (1(9)

 1 (2*)

 3 [6%) 

 (18) 

2 (««) 

NFPVODS SYSTEM 

IBHMN

HEHORRHSGE

NECPOSIS, NOS

 ;«5) ;l)8) ;U9)

 1 (2K) 

1 ;2)f) 

 I«6) 

SEHCIAL SENSE ORGANS 

•EAR CANA1

METAPLASIA, SQUAMOWS

 ;H7) ;t8) ;t>0)

 1 (2K) 

 ;«9) 

MOSCItLOSKELFTAL SYSTE1 

NONE 

BODY CAVITIES 

NONE 

A LL OTHEH SYSTEMS 

OBENTU B 

NECROSIS , FAT 2 1 

SPECIAL BOBPHOLOGY SUMf".PY 

flNIBAL HISSING/N O NFCROPS Y

AOTO/NECBOPSY/N O HISTO

AUTOIYSTS/NC NECPOPSY

» NUMBE R O F ANIBAL S WIT H TISSOE

* NOMBE S O F ANIBAL S NECBOPSIE D 

 EXAMINE D

 2 

1 1

 KICSOSCOPIC A LL Y 

1 

1 
TABLE C2

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS 

TREATED WITH HYDRAZOBENZENE 

— LOW DOSE HIGH DOSE CONTRO L (UNTR ) CONTRO L (UNTR ) 

02-0055 02-0118 

9NIMM S INITIAII Y I K STUD Y 50 50 

ANIMAL S MISSIN G 2 

BNIMAI S NECROPSIE D u e 50 

JNIMAL S F.XAHINF D HISTOP«.THClOnlCA LLY <'* 47 50 

INTEGUMENTARY SYSTEM
 

•SKI!!	 ;ue) 

INFLAMMATION , NC S 1 (2% ) 

*SUBCUT TISSUE (48) (50)
 

MINFRSLIZATION 1 (2%)
 

ABSCESS, NOS 1 ;2%)
 NECROSIS, NOS
 

RESPTBMCBY SYSTEM
 

ITRACHEA ;U7) ;i»9)
 TNFLA'HTICN, NCS
 

ILUNG/FRONCHUS (47) (50)
 

INFLAMMATION, NOS 3 (6%)
 

INFLAMMATION , FOC\L
 

ILUNG	 (U7) (50)
 

INFLAMMATION, FCCiL
 

INFLAMMATION, INTERSTITIAL 6 ;12)S)
 

PNEUMONIA, CHRONIC MURINE
 

HYPERPLASIA, EPITHELIAL 1 (2X)
 HYPERPLASIA, ALVEOLAR EPITHFLIUM
 

HFMA^OPOTETIC SYSTEM
 

*BONE MARRO U |15) •46)
 

OSTEOSCLEBOSIS 1 (2t)
 

KYELOSCIEROSIS
 

tSPLEEN (U7) (K8)
 

_INFL8MMAIIO!)4 NOS
 

« NOMBE R OF SNIMRL S WIT H TISSUE EXUBINE D MICROSCOPICALL Y 

* NUMBE R OF ANIMAL S NECSOPSIED 

**EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

C-9 

LOH ECS E HIG H I:TSE 02-0052 02-0091 

50 50 

50 50

 

50 50

 

ISO ) ;50) 

(50) (50)
 

1 ;2K)
 

•50)	 ;47)
 

5 (10t)
 

(50)	 (50)
 

2 (1*) 2 (UK)
 

1 ;2%)
 

(50) (5C)
 

2 (««)
 29 ;58^) 7 (14*)
 

1 ;2*)
 

5 ;iox) 1 ;2»)
 

;49) ;«6)
 

1 (2J)
 

(50)	 (50)
 

6 J12X)
 
TABLE C2 (CONTINUED) 

- _ _
 HIGH DOSE
 CONTROL (UNTR) CONTROL (UNTR)
 

02-0055 02-0118
 

HEMOSIEEPOSIS 12 (25*)
 

EBYTHROPHAGOCYTOSTS
 

HYPEBPLASIA, HFMATOPOIETIC 25 ;52»)
 

HYPEBPLASIA, EEYTHSOID 19 ;uo*)
 HEMATTPOIESIS 1 [?.%)
 

ISPTFNIC CAPSULE (47) (48)
 

HF.MORBHAGIC CYST 1 (2X)
 

ILYMPH NODE (40) (47)
 

HEMORRHAGE
 

INFLAMMATION, NOS
 

HYPERPLAEIA, NOS
 

RETIC'ILOCYTOSIS
 

LYMPHOCYtOSIS
 

PLASMACYTOSIS 1 ;2%)
 HYPEFPLAEIA, BETTCOLUK CELL
 

HYPEBPLASIA, LYMPHOID	 4 ;9X)
 

CIBC'JLATORY SYSTEM
 

IMYOCARDIUM ;47) ;so)
 INFLAMfATICN, NOS 1 (2X)
 

INFLAMMATION, TNTSRSTITI JL 23 ;«6t)
 

FIBROSIE 15 ;30*)
 

DEGENEPATION, NOS 7 ;15%)
 

IfNDOCARCITIM (47) (50)
 

INFLAMMATION, NOS 1 (2X)
 

DIGESTIVE SYSTEM
 

ISALIVABY GLAND ;u6) ;so)
 HYPFRPLASIA, INTEAEUCTAL
 

•LIVER	 (47) (50)
 

DEGENERATION, NOS 1 (2%)
 

NECROSIS, FOCAL 2 ;uxj
 NECROSIS, COASULATIVE
 

METAMORPHOSIS FATTY 2 («*) 6 ;12X)
 CYTOPLA.SMIC VACUOLTZATION
 

FOCAL CELLULAR CHANGE 25 (53X)
 

HYPEBPIA JIA. FOCAL 38 .r76?ll_
 

» NOMBEF OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NOMBER OF ANIMALS NECEOESIID
 

C-IO
 

LOW DCSH
 

02-CC52
 

12 |2MX)
 

1 [2*)
 38 ;76X)
 

tO (30*)
 

(50)
 

(44)
 

t ;9X)
 1 ;2X)
 2 ;5%)
 

3 ;7X)
 

1 ;2X)
 12 ;27K)
 

;so)
 

t1 [82%)
 

11 |22X)
 

(50)
 

;50)
 

1 (2X)
 

(50)
 

6 '12X)
 

1 [2X)
 

1 ;2X)
 

1 ;2%)
 

31_[i2XJ
 

HIGH DCSI
 

02-0091
 

9 He*)
 

21 (42»)
 

20 (UOX)
 

(50)
 

(31)
 

1 (3X)


1 ;3X)
 

1 (3X)
 

u [13X)
 

tt(1 3f)
 

2 ;6X)
 

;50)
 

2 ftlU)
 

17 (34t)
 

(50)
 

148)
 

(50)
 

2
 »(J^»
 

2 'ttT )
 10 (201)
 

2j>_a5231
 
. : .
 

FYPEBPIASIA, DIFFUSE
 

HYPESFIASIA, EBYTHBOIE
 

HEHATOPOIESIS
 

*EILF COCT
 

INFLAMHATION, NOS
 

HYPEBPLASIA, NOS
 

HYPEfPLAEIA, FOCAL
 

•PBNCREBS
 

INFLAMMATION, NOS
 

•PANCREATIC	 DUCT
 

HYPFRPIASIA, NOS
 

•PANCREBTIC ACINUS
 

ATROPHY, NOS
 

HYPERIROPHY, FOCAL
 

HYPEIPIASIA, FCCAL
 

•STOMACH
 

INFLAMMATION, NOS
 

INFLAMMATION, FOCAI
 

HYPEBDLASIA, BASAL CELL
 

HYPEBKEBATOSIS
 

ACAN1HOSIS
 

•PEYEBS	 PBTCH
 

"YPESPLASIA, NOS
 

•COLON
 

PARASITISM
 

OBINAB Y SYSTEM
 

•KIDNEY
 

HYDRnNEFHROSIS
 

GLOMERULCNEPHRITTS, NOS
 

INFLAMMATION, INTERSTITIAL
 

INFLAMMATION, CHRONIC
 

FIBPOSIS, FOCAL
 

FTBPOSIS, DIFFUSE
 

POSTMOBTEM CHANGE
 

• tlPINAPY BLADDER
 

_ , HYPEBPIASI.LI EPT-^HELIAI
 

* NUMBE " OF ANIM»L S WITH TISSUE

* NUMEF " O F ANIMAL S NECP.OPSIE D 

TABLE C2 (CONTINUED) 

LOW DOSE HIGH DOSE
 CONTROL (UNTR) COFTFOL (DNTB)
 

02-0055 02-0118
 

1 '2%)
 

2 ;«*)
 

(«8) (50)
 

1 '2%)
 

2 ;4%) 32 J64*)
 

1 ;2%)
 

(46)	 (48)
 

6 (13%)
 

(46) (48)
 

(46)	 (48)
 

8 (17%)
 

(46)	 (48)
 

1 (2%)
 

2	 ;«*)
 

(46)	 (48)
 

15 (31%)
 

(45)	 (46)
 

2 (4%)
 

;47) ;50)
 

43 (86%)
 

29	 (62%)
 

1 ;2%)
 i ;2%)
 

(47) (i»6)
 

 EXAMINE D BICPOS^OPICHLLY 

C-ll 

LCH DOSE
 

02-0052
 

1 (2%)
 

5 :10%)
 

(50)
 

1 J2%)
 

27 ("ii.%)
 

(US)
 14 129*)
 

(49)
 

1 (2%)
 

(49)
 

1 (2%)
 

1 ;2%)
 

(50)
 

1 |2%)
 

3 ;6%)
 

6 ;12%)
 

(50)
 

8 (16%)
 

(49)
 

5 (10%)
 

.50)
 

46 (92%)
 

3 ;6%)
 

1 ;2%)
 

(46)
 

. .-2,.J4%1
 

HIGH DOSE
 

02-0091
 

3 (61)
 

3 ;s*)
 

(50)
 

24 [48%)
 

1	 ;2%)
 

(46)
 

6 (13%)
 

(46)
 

(46)
 

2 (4%)
 

(U4)
 

2 |5*)
 

2 ;5*)
 

5 (11*)
 

(40)
 

8 (20*)
 

(37)
 

4 (11%)
 

.50)
 

1 (2%)
 

37 [74%)
 

(38)
 
TABLE C2 (CONTINUED) 

HIGH DOSE CONTROL (ONTR) CONTROL (ONTR ) 

02-0055 02-0118 

ENDOCRIN E SYSTE M 

tPITOITAR Y (46) (40)

 

PEPIVASCULITI S 1 (3%)

 

HYPERPLASIS , FOCAL i ;2X) 3 !8X)

 

• ADRENA L (47) (49) 

METAMORPHOSI S FATTY 1 (2X )

 

LIPOIDOSI S 

• ADRENA L COPTEX (47) (49)

 

CYST, NOS 1 (2% )

 

DEGENERATION , NOS 3 ;6K)

 

METAMORPHOSI S FATT Y 1 ;2X)

 

HYPERPHSIA, NODULA R 2 ;4X)

 

HYPESPLASIA , FOCAL 3 ;6K)

 

• ADRENA L MEDULL A (47) (49)

 

THROMBOSIS, NOS 1 (2X)

 

HYPERPLSSIA , NODULA R 3 ISX)

 

HYPESPlftSIA , FOCUL 1 ;2X) 3 I6X)

 

•THYPOTC (46) (45) 

CYSTIC FOLLICLES 1 (2*) 

HYP'RPLASIA , PAPILLAR Y 

HYPEBPIASIP , C-CFLL 4 (9%) 1 (2X ) 

BEPRODUCTIV E SYSTE M 

*MA«1MABY GLAS D '48) ISO)

 

DILAT > TIOK/DUCTS ' 3 (6X)

 

GAL»CTOCEL F 7 ;15X) 16 ;32%)

 

HYPSBPLASIA , NO S 4 ;8K) 8 ;16%)

 

*M»f!MA? Y CUCT (48) (50) 

FTB80SI5 2 ;4*> 

*CLITO"IS (48) (50)

 

NECROSIS , NO S 

•CLTTOPJ L GLAN D (48) (50)

 

HYP'IFPLASIA, NOS 

• V8GIN A (48) (50)

 

INFL. A.M.M.^TJON^ J10? "

 

» NUMBER OF ANIMAL S WITH TTSSHE EXAMINED MICBOSCOPICALLY
 

* NUKPEF OF ANIM»I S NSCBCESIJC
 

C-12
 

LOW ECS! 

02-0052 

(47) 

2 [4%) 

(49) 

(49) 

(49) 

3 ;6X) 

(50) 

2 ;4X) 

;50> 

18 [36X) 

20 ;40X) 

(50) 

(50) 

(50) 

(50) 

HIG H CCS! 

02-0091 

(37) 

(49) 

1 !2X) 

(49) 

1 ;2X) 

(49) 

3 [6X) 

(46) 

1 (2X) 1 ;2t) 

[50) 

9 (18X) 

8 J16X) 

(50) 

(50)

1 ;2») 

(50)

1 J2X) 

(50) 

_ 1 ^J21 
TABLE C2 (CONTINUED)
 

LOW DOSE HIGH DOSE CONTPO L (UNTR ) CONTRO L (UfiTP ) LCVi DCSE HIG H DOSE 

02-0055 02-0118 02-0052 02-0091 

• UTERU S •U7 ) ;50) ;50) IIS) 

HYEBCMETS A 2 (U*) ABSCESS , NO S 3 (€*) KYP1BPLASIA , ADENOMATOU S 1 '2*) 2 (1*) HYPEBPL.ASIA , STPIJUL 1 ;2%) METAPLASIA , SQUAMTU S

 1 ;2«) 

1 UTEPHS/ IN COME T RU'M («7) (50) (50) (18) 

INFLAMMATION , NO S 22 (i»U% ) 28 (56*) 16 |33*) 

INFLAMMATION , SUPPU">ATIV E 2 ;u*) INFLAMMATION , ACUT E 9 (19*) ABSCESS , NOS 2 ;u*) INFLAMMATION , CHRONIC 1 (2*) HYPEPPLASIA , NO S 6 ;12*) 9 118*) 9 (19*) 

HYPEBPLASIA , FOCAL 1 (21) HYPEBPLASIA , CYSTIC 3 (6*) u ;a*) 1 '8*1 HYPEBPLASIA , ADENOMATOD S 1 '2 *) HYPERPLASIA, STS^MAL 1 ;2*i 

HOVARY/OVICUC T (117) (50) (50) (US ) INFLAMMATION , NO S 10 (20%) 7 |1a*) 1 (2*) INFLAMMATION , SUPPURATIV E 2 ;t%) 2 ;<t*) INFLAMMATION , ACUTE 3 ;6K) INFLAMMATIO N ACTIVE CHRONI C 1 ;2K) INFLAMMATION , CHRONIC 1 ;2K) 

»OVAR Y (U6 ) (15) (50) (15) CYST, NOS 8 (16X) 10 (20*) INFLAMMATION , NOS 1 !2*) INFLAMMATION , CHRONI C 1 (2%) 

DEGENERATION , CYSTIC 1 ;2*> 

tOVARY/FOLLICL ? (U6 ) (19) (50) (15)

 

HYPEBPIASIA , NOS 1 (2*)

 

KERVOU S SYSTFB 

NON E 

SPECIAL SENSE ORGANS
 

(50) (50) (50)
 CATABACT
 

* NUMBEP OF ANIHALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* N01EEB OF ANIMAL S NECBOFSIED
 

C-13
 
TABLE C2 (CONCLUDED)
 

LOW DOSE HIGH DOSE CONTRO L (ONTR ) CONTRO L (DHI P )

02-0055 02-0118

'EYE/RETINA It8) ISO)

ATROPHY , NO S 1 (2X )

 

*HARDERIA N GLAR E («8) (50)

HYPERPIASIA , NO S 

BOSCULOSKELETA L SYSTE M 

*BONE ;«8 ) ISO)

CSTECSCLEROST S 1 (2*)

 

EOD Y CAVITIE S 

*PLEt!P A («8) (50)

INFLAMMATION , CHRONI C 1 ;2t)

 

*IPICBRCIU « (U8 ) (50)

INFLAMMATION , CHRONI C 1 \2%) 

A IL OTHER SYSTEM S 

OMENTU M 

NECPOSIS , FA T 1

 

SPECIA L MORPHOLOG Y SHMMAR Y 

ANIMA L fISSTNT/N O NECROPS Y 2

 

AIJTO/NECROPSY/HIST O PEP F 

AUTO/NECPOPSY/NO HISTO 1

 

* NUMBE R OF ANIMAL S WITH TISSUE FXAFISED MICFOSTOPICALLY
 

* NU1EEE OF ANIMSIS NFC5CPSIF.D
 

C-14
 

 LCS DCSE HIG H DOSE 

 02-0052 02-0091 

 150) 150) 

 (50) (50) 

 ISO) ISO) 

 (50) [50) 

 (50 ) [50) 

2 
APPENDIX D
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN MICE TREATED WITH HYDRAZOBENZENE
 

TABLED!

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE MICE 

TREATED WITH HYDRAZOBENZENE 

fNIBAL S INITIAIL Y T N STUD Y 

ANIMAL S HISSING 

PNIMAL S NECFOPSIE D 

JNIMAL S EXAMINE D HISTOPATHOIOGICALL Y 

INTEGUMENTARY SYSTEM
 

*SKIN
 

INFLAMMATION, NCS
 

ULCER, NOS
 INFLAMMATION, FOCAL
 

INFLAMMATION, NECSOTIZING
 ABSCESS, NOS
 

ACANTHOSTS
 

*SUECUT TISSUE
 

ABSCESS, NOS
 

NECRCSIS, NOS
 

NECROSIS, FAT
 

METAPLASIA, OSSEOUS
 

RESPIRATORY SYSTEM
 

ILUNG/BSONCHUS


INFLAMMATION, FCCM
 

*LUNG/B"ONCHIOLE


INFLAMMATION, NOS


INFLAMMATION, FOCAL
 

PERIVISCULITIS


U'JNG


HEMORRHAG E

INFLAMMATION , NO S • INFLAMMATION , INTFPSTITIA L 

HYPERPH.SIA , ALVEOLA R EPITHELIU M

HEMATOPOIEITC SYSTEM
 

•SPLEEN
 

c8NTDR°OL(UNTH) 

05-0070 

50 

50 

** 50 

;50)
 

2 ;«
 

(50)
 

(2X)
 

 ;50)
 

 (50)
 

1 (2t)
 

1 ;2K)
 

 (50)
 

2 (1*) 

2 ;1%) 

HIGH DOSE 

CONTROL (ONTRi I

05-0118 

50 

1 

19 

19 

1 (2%) 

1 (2%) 

(19)
 

1 (2%) 

(19) 

(19) 

10 (20%) 

« NOMBE P 0 ? ANTMAL S WIT H TISS'JE EXAMINE D MICPOSCOPICAIL Y 

* NUMBE R O F JNIMAL S NEC>=CFSIF D 

**EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

D-3
 

 LOW DOSE HIGH CCSF. 

05-0050 05-0093 

50 50 

1 2

 

19 18 

17 16

 

1 (2*)
 1 (2%)
 

(19)	 (18)
 

1 (2%)


2 ;ixi
 

1 [2*1
 

(17) (16) 

(17) (16) 

1 

3 ;e 5 

111)
 
TABLE Dl (CONTINUED) 

________—__—	 ______—_
 

LOW DOSE HIGH DOSE
 CONTROL (DNTR) COMTHOL (UNTP) LOW DOSE HIGH DOSE
 

05-0070 05-0118 05-0050 05-0093
 

PETICaiCCYTOSIS 1 (2%)
 HYPFPPLASIA, HEM.1TOPOIETIC 5 (10X) 6 (13*) I) (9K)
 

HYPEBPLASIA, ESYTHROID 3 ;6*i
 HYPEHPLASIA, LYMPHOID 1 (2%) 1 ;2X) 3 (7*)
 

•SPLENIC FOILICIES (50) (H9) (17) (11)
 HYPERPLBSIA, NOS 2 («*)
 

•HEMOLYBPH	 NODES (50) (U9) (17) (11)
 INFLAMMATION, NOS 2 (1*)
 

•LYMPH	 NODE (15) («2) (11) (38)
 HEMORRHAGE 1 (9*) 1 13*)
 INFLAMMATION, NOS 10 ;2»«) 15 ;3<t*) 5 11 3X)
 HYPEFPLASIA, NOS	 1 ;2*) 1 -2*) 1 13*)
 RETICULOCYTOSTS	 2 ;5*) 5 [11*1 1 ;3*)
 LYMPHOCYTOSIS 5 (11*)
 HYPEFPLA5IA, HEMA'"OPOIETIC 1 !2X)
 HYPEFPLASIA, LYMPHOID 3 !7X) 5 (11*) 3 •8X)
 

•MESFNTERIC	 L. NODE (15) (12) (11) (38)
 

HYPTRPL5SIA, RETICULUM CEII 1 (2X)
 

CIPCOtATOPY SYSTEM
 

•HEART	 ;i9) ;i9) ;«7)
 111)
 

MINERALIZATION 1 (2%1
 

*HEART/VEN~PICIE (19) (19) (17) (11)
 MELANIN 2 (5*)
 

• AOPTIC VMVE (19) (19) (17) (11)
 INFLAMMATION, AC"' E/CH8CNIC 1 (2*)
 

*ARTEP.Y (50) (i»9) (19) (18)
 

INFLAMMATION, NOS 2 (1*)
 

DIGESTIVE SYSTEM
 

•SALIVA3Y TLAND	 ;i9> ;i8) ;is) :i5)
 PESTVASCULITTS	 1 (2X)
 

•LIVER	 (50) («8) (17) (16)
 _I£EC.£0. si.s__ FO c. ji 1 (2*1 9 (19*) -- .-__JJ_5J, 10 J2211
 

• NUMEE ? O F ANTM»L S WIT H TISSUE EXAMINE D MICPOSCOPICALL Y 

* NUMEF P OF ANIMAL S HECPOFSIED 

D-4 
TABLE Dl (CONTINUED) 

LOW DOSE CONTPOI (UNTR ) 

HIGH DOSE CONTBOL(UNTP ) 

05-0070 05-0118 

NECFOSIS , COMULATIV E 

METAMORPHOSI S PATT Y 

HEPATOCYTOMEGAL Y 

DEPLETIO N 

HYPEBPLASIA ,

HYPEB°LASTI C

 NODULAR 

 NODDL E :<>*) 1 [2%) 

HYPEBPLASIA ,

HYPFPPLASIA ,

 FOCAL 

D T FFtJS E 

HEMATOPOIESI S 

•LIVER/CENTBILOBULAR (50) 

NECROSIS, NOS 1 (2%) 

»LTVFB/KUPFFtF CEL1 (50) (U8) 

HYPERPLASIP, NOS 1 (2%) 

«P«NCRF. AS (U7 ) 

INFLAMMATION, NOS 

INFtA»MATION, FOCAL 1 

1 (2%) 

•STOMACH (1)9) (1*8) 

INFLAMMATION , NOS 

INFLAMMATION, FOCAL 2 

INFLAMMATION, NECHOTIZING 

HYPEBPLAEIt, FOCAL 

HYPERKF^AIOSIS 

AONTHOSIS 

IGASTPIC MUCOSA (1*9) (H8) 

INFLAMMATION , FCCA L 1 (2*) 

tPEYEFS fATCH (1*9) 

HYPFRPIASIA, NOS 1 (2*) 

*ILIUM <U9, (U9) 

INFLAMMATION, NECROTIZING 

•COLON (U6) (1*3) 

GBANOLOMA, NOS P«R». SITISM 1 (2%) 3 ;7%) 

DFTNAP Y SYSTE M 

•KIDNEY 

* NUMBER OF ANlrlALS HITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBFB OF ANIMAIS NECBCESIED
 

D-5 

L O W ECS E 

05-OC50 

1 (2*) 

2 ;«%) 

(U7 ) 

(1*7) 

1 (2%) 

(U7 ) 

5 (11%) 

3 ;6 

(U7 ) 

'T ,6<)
 

(U7)
 

(U5)
 

J_L251 

HIG H trsi 

05-0093 

(U6 ) 

(UK ) 

(7*) 

3 (75!) 

(41*) 

(U2)
 

fi (1M^)
 

(t2)
 

1 (2*)
 

(39)
 

1 [3K)
 

(1*6) 
TABLE Dl (CONTINUED) 

GIOMIPULCNEFHBITIS , NOS 

INFLAMMATION , INTERSTITIA L 

INFLAMMATION , CHPONI C 

•URINAR Y EL'DDE E 

TNFLAMM«TIOW , NO S 

HYPERPLASIf , EPITHELIA L 

LOW DOSE CONTRO L CTNTR.)

05-0070

3 (6*)

(47)

 HIGH DOSE CONTRO L (UNTR )

 05-0118

2 (*%)

 16 (33%}

 (48)

 LO» DOSE 

 05-0050 

1 («*) 

 12 (26%} 

1 ;2X) 

 (47 ) 

HIG H DOSE 

05-OC93 

1 

1 

(45) 

2 

1 

(2*) 

:2X) 

(4*) 

ENDOCRIN E SYSTEM 

•ADRENA L 

HYPEPPIASI A , NOS 

•ADPENAL/CAPSUL E 

HYF'BPIASIA , NOS 

•ADRENA L CORTE X 

HYPFBTBOPHY , FOCAL 

*PA«!CRFJTIC ISLETS 

HYPEBPIASH , NO S 

;49)

(49 )

(49)

(46)

 ;44)

3 (7*) 

 (44)

3 (7%)

 (44)

 (I'M

 ;45) 

 (45) 

2 <4X ) 

 (45 ) 

 (47) 

1 (2% ) 

:«2) 

(42 )

1 

(42) 

1 

(44) 

,2T, 

(2*) 

BEPRODUCTTV E SYSTEM 

*PBEPOTIA L GLAN D 

ABSCESS, NOS 

N"CPOSIE, NOS 

tPPOSTATE 

INFLAMMATION , NOS 

*SEMIN?.L VESICLE 

MTNFRHLTZATIO N 

NECROSIS , NOS 

•TESTIS/TUEULE 

MINERALIZATIO N 

•FPTCTDYNI S 

INFLAMMATION , NO ? 

;so)

(49 )

(50)

(50)

(50)

 ;49)

1 (2K )

 (49)

 (49)

 (48)

 (49)

 ;49) 

2 (4X) 

 (41) 

 (49) 

1 (2% ) 

 (47 ) 

 (49) 

;48) 

(41) 

1 

(48) 

(16) 

(48) 

(2*) 

NERVOU S

NON E

 SYSTEM 

 _.. . . . _ . 

I NUMBEB OF ANIMALS WITH TISSUE EXAMINED MICPOSCOPICALLY 

* N'IMEEP OF ANIMALS NECROPSIED 

D-6 
SPECIAL SEKSE OSG&NS
 

NONE
 

BUSC'JLOSKEIETAL SYSTEM
 

NONE
 

EODY CAVITIES
 

NONE
 

BLL OTHER SYSTTUS
 

SDTPOSE TISSUE
 

INFLftMIATION, ACUTE


OMENTUM
 

NFCECSIS , ?AT

SFFCISL "ORPHOLOG Y SUMHJS Y 

NO LFSTO N PEFOPTID

S N T H » L "1SSTNG/NO NECPOES Y

.W'O/NECPOPSY/NO HISTO

« NUMBE R O F "NIMJL S WTT H TISSUE * NUMBE R O F ANIMAI S NICPOPSIE D 

TABLE Dl (CONCLUDED) 

LOW DOSE HIGH DOSE CONTPOL(ONTR ) CONTROL(UNTP ) LOU DCSE HIGH EOSE 

05-0070 05-0118 05-0050 05-0093 

1
 

1 

 12 5 3 9 

1 1 2 ' 

2 2 

 EXAKINE D HTCEOSCOPICAI1T 

D-7 
TABLE D2

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE 

TREATED WITH HYDRAZOBENZENE 

T PNIMAL S INITIAII Y K STtlDY

/NIMALS NECROPSIED

SNIMAL S EXAMINE D HI STOPATHCLOGICALL Y

INTEGUMENTAR Y SYSTEM 

*SOBCUT TISSUE

ABSCESS , NO S 

RESPIRATOR Y SYSTE M 

•LUNG/BRONCHUS

INFLAMMATION , FOCAI 

• LONG/BRONCHIOL E

INFLAMMATION , NOS

HYPEPPLASIA , NOS 

*L'JNG

INFLAMMATION , INTERSTITIAL

HYPERPLASIA , ALVEOtA R EPITHELIU M 

HEMATOPOI3TIC SYSTEM 

KBON E MARRO W

MYEICFIEPOSI S

• SPLEEN HEMOSIDEROSI S 

HYPEBPIASIA , NO S 

HY°SRPLA5IJ, HFMATOPOIETIC 

HYPERPLASIA , ERYTHROID 

HYPERPLASIA , LYMPHOT D

• SPLFNTC FOILICIES

HYPERPI'SIA , NOS

• HEMOLYMP H NODE S

• H'JNEE1? O F ANTMAL S WIT H TISSUE

* NUMBE R TF ANIMAL S NFCROPSIE D 

LOW DOSE CONTROL (tJNTR) 

06-0070 

 5 0 

 48 

 ** 47 

 (48) 

 ;46) 

 (46 ) 

1 (2%) 

 (46 ) 

1 (2%) 

 ;46) 

1 (2%) 

 (4-j) 

1 (2%) 

 (47) 

3 (6*) 

 (47) 

HIGH DOSE CONTROL (UNTR) 

06-0118 

50 

50 

50 

(50) 

•50) 

1 (2*) 

(50) 

(50) 

14 (28%) 

:49, 

(49) 

(49) 

(49) 

 EXAMINE D MICROSCOPICALL Y 

**EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

D-8 

LOU CCSE HIGH DCS! 

06-0052 06-0093 

47 50 40

 44 39

 43 

(40 ) (44) 

;38) (40) 

(38) (40) 

(38) |«0) 

5 |13%) 3 (8*) 

2 ,'5t) 

:38) ;40) 

(39) (41) 

1 (3*) 

13 ;33«) 

(39) (41) 

(39 ) 141) 
TABLE D2 (CONTINUED)
 

LOW DOSE HIGH DOSE CONTROL (IINTS) CONTROL (UNTS )

06-0070 06-0118

HYPFRPIASIA , NO S 1 (2%) 

•LYMP H NOE F (36) (tU)

HEMORRHAG E 

INFLAMMATION , NO S 

HYPERPLASIA , NO S 

PETICULOCYTOSI S 

1

1

 ;3X) ;3%> 9 ;20X) 3 ;7«)

1 ;2«)

LYMPHOCYTOSI S 

HYPERPLASIA ,

HYPEPPL8SIA ,

HYPEFPLASIA ,

 HEMA^OPOIEII C 

 PLASM A CELL 

 LYMPHOI D 

1 ;3%) 

1

n

 ;2«)

 ;9%> 

*MEDIASTTNA L L.NOD E (36) (UU )

PLASMACYTOSI S 

«AEDO«TN«. L LYMP H NOD S (36) (UU )

PLASMACYTOSI S 1 (3*) 

CISCDLAT05Y SYSTEM
 

•HEART	 ;u«) ;50)
 MINERALIZATION
 

•HFART/VENTRICLE	 («1> (50)
 MF1BNIN
 

•MYOCARDTUM	 (U«) (50)
 INFUMMATION, FOCAL
 

FIBPOSIS, FOCAL
 

DIGESTIVE SYSTEM
 

»SAL T VAR Y GLAND
 

PEBIVA SCULITIS 3 (7*)
 

PFPIVASCaiAR CUFFING 1 (2*) 3 (6%)
 

•LIVER	 (50)
 MINFRHLIZATION
 

INFLAMMATION, ACUTE FOCAL 1 ;2<6)
 

INFLAMMATION, ACUTE/CHRONIC
 

NECROSIS, FOCAL	 2 :<*«)
 NECEOSTS, COAGOLATIVE
 

» NUMBES OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

+ NUMBER "F ANIBALS NHCRCFSI?D
 

D-9
 

 LPH ECSE 

 C6-0052 

 (31) 

5 ;15*) 1 ;3% > t J12K ) 

u ;12^) 

2 J6K) 

 (311) 

1 (3*) 

 (3«) 

;39)


(39)


(39)


;38)
 

(39)
 

11	 [28%)
 

2_J531_.
 

HIG H CfS I 

06-0093 

(33) 

1 (31)

2 ;6%) 

1 '3X)

5 IlEt)

5 ;i?t) 

1 '3*1 

1 ;3»> 

(33) 

(33) 

 ;u2)
 

 (t2)
 

 (K2)
 

(38)
 

O3)
 

1 (2f)
 
TABLE D2 (CONTINUED)
 

LOW DOSE HIGH DOSE
 CONTROL (tINTH) CCNTRCL (UNTB)
 

06-0070 06-0118
 

HYPERFIASIA, FCCAL
 

HEMATOPOIESIS
 

*EILB DUC* (18) (50)
 

INFLAMMATION, ACUTE/CHRONIC i J8*)
 HYP*:RPLASIA, NOS
 

•PANCREAS	 (13) (18)
 INFLAMMATION, NOS 1 (2*) 2 (1*)
 

INFLAMMATION, INTERSTITIAL 1 J2X)
 

PERTARTERITTS 1 (2*)
 

•PANCREATIC ACINUS (13) (18)
 

ATROPHY, NOS 1 (2*1
 

•STOMACH	 (15) (19)
 

INFLAMMATION, NOS 1 (2X)
 

INFLAMMATION, FOCAL 1 ;2X)
 ULCER, FOCAL 1 *2 %)
 

HYPERKERATOSIS
 

ACANTHOSIS 2 ;i*j
 

•PEYERS	 PATCH (15) (18)
 

HYPERPLASIA, NOS 1 (2*) 7 (15*)
 

UFINARY SYSTEM
 

•KIDNEY	 ;i5) ;50)
 MINERALIZATION
 

GLOMERULONEPHRITTS, NOS 3 (7*) it (8*)
 

INFLAMMATION, NOS
 

GLOMEROLONEPHPITIS, FOCAL 2 (IX) 1 (2%)
 INFLAMMATION, INTEBSTITIHL 1 (2*) 12 (21*)
 

GLOMERULONEPHRITIS, MEMBRANOUS 2 ;ixi
 PYELONGPHKITIS, ACUTE/CHRONIC 1 ;2K)
 GLOMERULONEPHRITTS, CHRONIC 1 (2X)
 

•KICNFY/GLOBERULDS	 (15) (50)
 

CYST, NOS
 

•KIDNEY/TUBULE	 f U 5) (50)
 

MINERALIZATION 1 (2X)
 

DEGENERATION, CYSTIC
 

•URINARY BLADDER	 I U 5) (18)
 

I NUMBER OF ANIMALS WITH TISSOE EXAMINED MICROSCOPICALLY
 

* NUMBFR OF ANIMALS NECROPSIED
 

LOW DOSE
 

06-0052
 

1 (3X)
 

3 :8X)
 

(10)
 

(38)
 

3 (8*)
 

(38)
 

(37)
 

2 (5X)
 

t	 ;1 1*)
 i	 ;ii*)
 

(39)
 

1 13*)
 

;39)
 

M (10*)
 

9 (23*)
 

(39)
 

(39)
 

1 (3X)
 

(37)
 

HIGH DOSE
 

06-0093
 

2 (5*)
 

(11)
 

1	 ;2*)
 

(39)
 

(39)
 

(39)
 

3 (8X)
 

2 ;5*)
 

(37)
 

2 15*)
 

(12)
 

1 (2*)
 

1 (2*)
 

1 (2%)
 

5	 (12*)
 

112)
 

1 (2*)
 

(12)
 

1 !2X)
 

(11)
 

D-IO
 
TABLE D2 (CONTINUED) 

CONTROL(UNTR ) ,(UNTS) 

06-0070 06-0118 

PERTSBTIRITIS 

HYPEEPLASIA, EPITHELIAL 

ENDOCRINE SYSTEM
 

•ADRENAL/CAPSULE
 

NOD') IE
 

HYPEEPLASIA, NOS
 

•ADRENAL COPIES (47) C»8)
 NODULE 1 (2)1)
 HYPESPLASIA, NOS
 

tTHYROIE (11) (It 11)
 FOLLTCULAR CYST, NOS
 

INFLAMMATION, FOCAL 1
 HYPERPHSIA, PAPILLARY 2
 HYPEEPLASIA, ADENOMATOUS
 

HYPERPLASIA, FOLLICULAR-CELL
 

ITHYPOIE FOLLICLE (H1) C*t)


NECROSIS, FOCAL
 

BEPHODDCTTVJ SYSTEM
 

*MAM!HPY GLAND
 

HYP?5PISSIA, NOS 1 (2*)
 

(13)
 

HYDSOfETPA 3 (7*) 13 (28X)
 

CYST, NOS
 

ABSCESS, NOS 2 (5*)
 

METAPLASIA, SQOAHOUS
 

« UTERUS/ ?NCOMFTPIUB (13) (U7)
 

INFLAMMATION, NOS 2 (5%) 8 (17*)
 

INFLAMMATION, SUPPO"ATIVE 2 I5«)
 

INFLAMMATION, ACU'E 6
 

INFLAMMATION, ACUTE FOCAL
 

INFLAMMATION, ACUTF/CHPCNIC 3
 HYPEBPL f SIA, NOS 1 I2U) 

HYPFBPLASIA , CYS m IC 20 ____ ,£ETA £ii SI4j._12U4I10 U « 

NUMBEP OF JNIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

NHMPFR OF ANIMALS NFCROPSIEC
 

D-l
 

LOW DCSI
 

06-0052
 

1 (3*)
 

2 :•>*)
 

(39)
 

2 (5%)
 

(31)
 

 (3<t)
 

(30)
 

1 ;3
 3 ;9
 

2 (6%)
 2 :<s*)
 

12 135%)
 

8 ;2
 

HIGH CCSE
 

06-0093
 

t»0)
 

1 (3«)
 

f [15%)
 

(33)
 

1 (3*)
 

(33)
 

1 (3K)
 

(33)
 

(33)
 
TABLE D2 (CONTINUED)
 

LOW DOSE HIGH DOSE CONTROL (UNTR) CONTROL (ONTR) LOW DCSE HIGH tCEB 

06-0070 06-0118 06-OC52 06-0093 

•OVARY/OVIDUCT (13) (17) (31) (33)

 

INFLAMMITION , NOS 1 (9X) 1 <3X)

 

INFLAMMATION, SUPPUPATIVE 1 >9X)

 

ABSCESS, NOS 1 (2%) 1 !2«)

 

HYPEBPLASIA, FOCAL ;3*>

 1 

IOVARY (45) (48) (37) (31) 

CYST, NOS 10 (21X) 5 (11X) 1 (3%) 

INFLAMMATION, NOS 4 :s«) 6 116*) INFLAMMATION , SUPPDRATIV E 6 ;13X) 4 (11*) 

ABSCESS, NOS 1 !3X)

 

INFLAMMATION , CHRONIC 1 (2t)

 

ABSCESS, CHRONIC 1 :2<)

 

PERIARTERITI S 1 (2%) 1 ',2%)

DEGENERATION, CYSTIC 3 :ex)

 

KEPVOK S SYSTEM 

• ERAIN/MENINGES ;46) ;39)

 

INFLAMPATIOH, ACUTE/CHSCNIC 1 (2*) INFLAMMATION, CfiRONIC FCCAI 1 (2K)

 

tBRATN (16) (18) (39 ) (11) 

NFCEOSIS, NOS 1 (2)1) 

SfECIAL SENSE ORGANS
 

NONF
 

MUSCULOSKELF.TAL SYSTEM
 

NONE
 

BODY CAVITIES
 

NONE
 

All OTHEP SYfTEMS
 

•MULTIPLE ORGANS ;48)	 ;«0) [11)
 

» NUMBEE OF ANIMALS WITH TISSUE F.XAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECFCESIED
 

D-12
 
TABLE D2 (CONCLUDED) 

CO)NTPOS1 (UNTP ) pLUNTF ) LO W DOSE 

06-0070 06-0118 06-0052 

SPECIA L MOBPHQIOG Y SUHCAP Y 

NO LESIO N REPORTE D 3 2 

AUTO/NECBOPSY/HISTC EEE F 1 

AUTO/NECRGPSY/N C HISTO 1 1 1 

AOTOLYSIS/N O NECPOPS Y 2 7 6 

I NIJHEE P O F ANIMAL S WIT H TISSTE EXS.BINE D «IC POSCOPIC ALL Y 

* NUMEE P O F ANIHAL ? NSCSOPSTE D 

D-I3 

Review of the Bioassayof Hydrazobenzene* for Carcinogenicity
 

by the Data Evaluation/Risk Assessment Subgroup
 

of the Clearinghouse on Environmental Carcinogens
 

The Clearinghouse on Environmental Carcinogens was
 

established in May, 1976, 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 responsibility 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 Hydrazobenzene
 

for carcinogenicity.
 

The primary reviewer said that Hydrazobenzene was
 

carcinogenic, under the conditions of test, in both sexes
 

of treated rats and in treated female mice. He opined that
 

the experimental design was adequate. In his critique he
 

noted the following: the test chemical was obtained from
 

a small laboratory and thus may not represent the chemical
 

usually found in commerce; the stability of the compound
 

in the diet mixture was not determined; the results of the
 

subchronic study were not useful in establishing the dose
 

levels for the chronic phase; and the high and low doses
 

were too far apart and varied during the course of the
 

chronic phase. Despite these shortcomings, the primary
 

reviewer considered the study to be valid, given the
 

unquestionable carcinogenic response induced in the treated
 

animals. He said that Hydrazobenzene could pose a
 

carcinogenic risk to humans.
 

The secondary reviewer concluded that, although
 

Hydrazobenzene was quite toxic, it nevertheless was carcinogenic under the conditions of test. He noted that,
 in addition to the other tumors reported in the summary,
 

57
 
there was an increased incidence of adrenal tumors in
 

several of the treated animal groups. He agreed that
 

Hydrazobenzene may pose a carcinogenic risk to humans,
 even at low dose levels.
 

The primary reviewer reiterated his concern over the
 

lack of stability data on Hydrazobenzene. He said that the
 

compound could readily oxidize to azobenzene, a known
 

hepatocarcinogen. A Subgroup member noted that, in any case,
 Hydrazobenzene probably is metabolized to azobenzene.
 

The primary reviewer moved that the report on the
 

bioassay of Hydrazobenzene be a.ccepted with the qualifications
 

expressed in his critique. The motion was seconded and
 

approved unanimously.
 

Members present were:
 

Michael Shimkin (Acting Chairman), University of California
 

at San- Diego
 

Joseph Highland, Environmental Defense Fund
 

George Roush, Jr., Monsanto Company
 

Louise Strong, University of Texas Health Sciences Center
 

John Weisburger, American Health Foundation
 

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.
 

58
 

DHEW Publication No. (NIH) 78-1342