1978 

National Cancer Institute 

CARCINOGENESIS 

Technical Report Series 

No. 127 

BIOASSAY OF 

5-NITRO-o-ANISIDINE 

FOR POSSIBLE CARCINOGENICITY 

CAS No. 99-59-2 

NCI-CG-TR-127 

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

Public Health Service 

National Institutes of Health 

BIOASSAY OF
 

5-NITRO-o-ANISIDINE
 

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) 78-1382
 

REPORT ON THE BIOASSAY OF 5-NITRO-o-ANISIDINE
 

FOR POSSIBLE CARCINOGENICITY
 

CARCINOGENESIS TESTING PROGRAM
 

DIVISION OF CANCER CAUSE AND PREVENTION
 

NATIONAL CANCER INSTITUTE, NATIONAL INSTITUTES OF HEALTH
 

FOREWORD: This report presents the results of the bioassay of
 

5-nitro-o-anisidine 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 5-nitro-o-anisidine was conducted by
 

Mason Research Institute, Worcester, Massachusetts, initially under
 

direct contract to the NCI and currently under a subcontract to
 

Tracer Jitco, Inc., prime contractor for the NCI Carcinogenesis
 

Testing Program.
 

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

principal investigators for the contract were Dr. 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. D. W. Hayden
 

(3), and Dr. Yoon (3) at the Mason Research Institute, the pathology
 

narratives were written by Dr. R. L. Schueler (4), 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 (6,7) using
 

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

Gart (8).
 

ill
 
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 (6,9), 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 (1,9),
 

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

Dr. T. W. Orme (1), Dr. R. A. Squire (1,10), Dr. S. F. Stinson (1),
 

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

1.	 Carcinogenesis Testing Program, Division of Cancer Cause and
 

Prevention, National Cancer Institute, National Institutes of
 

Health, Bethesda, Maryland.
 

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

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

Boulevard, McLean, Virginia.
 

7.	 No wit w h the Solar Energy Research Institute, Cole Boulevard,
 

Golden, Colorado.
 

8.	 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.
 

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

Washington, D.C.
 

iv
 
10.	 Now with the Division of Comparative Medicine, Johns Hopkins
 

University, School of Medicine, Traylor Building, Baltimore,
 

Maryland.
 

v
 

SUMMARY
 

A bioassay of 5-nitro-o-anisidine for possible carcinogenicity
 

was conducted using Fischer 344 rats and B6C3F1 mice. 5-Nitro-oanisidine was administered in the feed, at either of two concentrations, to groups of 50 male and 50 female animals of each species.
 

The dietary concentrations used in the chronic bioassay for low and
 

high dose rats were 0.4 and 0.8 percent, respectively. Dose A and
 

B mice were fed dietary concentrations of 0.8 and 1.6 percent when
 

initially placed on test, but after week 15 the concentration fed to
 

dose B mice was reduced to 0.4 percent. After a 78-week period of
 

chemical administration, observation of rats continued for up to an
 

additional 28 weeks and observation of mice continued for up to an
 

additional 19 weeks. For each species, 50 animals of each sex were
 

placed on test as controls for the group receiving the higher concentration and 49 to 50 animals of each sex were placed on test as
 

controls for the group receiving the lower concentration.
 

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

Feeding of 5-nitro-o-anisidine to rats was associated with increased incidences of tumors of the integumentary system. Basal-cell
 

carcinomas, trichoepitheliomas, squamous-cell carcinomas and sebaceous
 

adenocarcinomas each occurred in the skin of high dose male rats at
 

statistically significant incidences. For both male and female rats,
 

carcinomas (the combined incidence of sebaceous adenocarcinomas,
 

ceruminous carcinomas and squamous-cell carcinomas) of the Zymbal's
 

gland or the skin of the ear were significant in the high dose groups.
 

In the clitoral gland of dosed female rats, the incidence of carcinomas and the incidence of adenomas were each significant.
 

Among mice, the incidence of hepatocellular carcinoma was statistically significant for dose B females when compared to their
 

appropriate controls.
 

Under the conditions of this bioassay, dietary administration of
 

5-nitro-o-anisidine was carcinogenic in Fischer 344 rats, causing tumors of the integumentary system in males and females and of the clitoral gland in females. The compound was also carcinogenic to female
 

B6C3F1 mice, causing hepatocellular carcinomas.
 

vii
 

TABLE OF CONTENTS
 

Page
 

I. INTRODUCTION 1
 

II. MATERIALS AND METHODS 5
 

A. Chemicals 5
 

B. Dietary Preparation 5
 

C. Animals 6
 

D. Animal Maintenance 6
 

E. Selection of Initial Concentrations 10
 

F. Experimental Design 11
 

G. Clinical and Histopathologic Examinations 15
 

H. Data Recording and Statistical Analyses 16
 

III. CHRONIC TESTING RESULTS: RATS 21
 

A. Body Weights and Clinical Observations 21
 

B. Survival 23
 

C. Pathology 23
 

D. Statistical Analyses of Results 27
 

IV. CHRONIC TESTING RESULTS: MICE 46
 

A. Body Weights and Clinical Observations 46
 

B. Survival 46
 

C. Pathology 49
 

D. Statistical Analyses of Results 50
 

V. DISCUSSION 57
 

VI. BIBLIOGRAPHY 59
 

APPENDIX A SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

RATS TREATED WITH 5-NITRO-o-ANISIDINE A-l
 

APPENDIX B SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

MICE TREATED WITH 5-NITRO-o-ANISIDINE B-l
 

APPENDIX C SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN RATS TREATED WITH 5-NITRO-o-


ANISIDINE C-l
 

APPENDIX D SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN MICE TREATED WITH 5-NITRO-o-


ANISIDINE D-l
 

ix
 
LIST OF ILLUSTRATIONS 

Figure Number Page 

1 CHEMICAL STRUCTURE OF 5-NITRO-o-ANISIDINE 2 

2 GROWTH CURVES

STUDY RATS 

 FOR 5-NITRO-o-ANISIDINE CHRONIC 

22 

SURVIVAL COMPARISONS OF 5-NITRO-o-ANISIDINE 

CHRONIC STUDY RATS 24 

GROWTH CURVES

STUDY MICE

 FOR 5-NITRO-o-ANISIDINE CHRONIC 

 47 

SURVIVAL COMPARISONS OF

CHRONIC STUDY MICE

 5-NITRO-o-ANISIDINE 

 48 

LIST OF TABLES 

Table Number 

DESIGN SUMMARY FOR FISCHER 344 RATS—5-NITROo-ANISIDINE FEEDING EXPERIMENT 12 

DESIGN SUMMARY FOR B6C3F1 MICE—5-NITRO-oANISIDINE FEEDING EXPERIMENT 13 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN MALE RATS TREATED WITH 

5-NITRO-o-ANISIDINE 28 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN FEMALE RATS TREATED 

WITH 5-NITRO-o-ANISIDINE 35 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN MALE MICE TREATED WITH 

5-NITRO-o-ANISIDINE 51 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN FEMALE MICE TREATED 

WITH 5-NITRO-o-ANISIDINE 53 

Al SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

MALE RATS TREATED WITH 5-NITRO-o-ANISIDINE A-3 
LIST OF TABLES (Concluded) 

Table Number Page 

A2 SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

FEMALE RATS TREATED WITH 5-NITRO-o-ANISIDINE A-8 

Bl SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

MALE MICE TREATED WITH 5-NITRO-o-ANISIDINE B-3 

B2 SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

FEMALE MICE TREATED WITH 5-NITRO-o-ANISIDINE B-7 

Cl SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN MALE RATS TREATED WITH 5-NITRO-oANISIDINE C-3 

C2 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN FEMALE RATS TREATED WITH 5-NITROo-ANISIDINE C-12 

Dl SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN MALE MICE TREATED WITH 5-NITRO-oANISIDINE D-3 

D2 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN FEMALE MICE TREATED WITH 5-NITROo-ANISIDINE D-9 

xi
 

I. INTRODUCTION
 

5-Nitro-o-anisidine (Figure 1) (NCI No. C01934), a tri-substituted
 

benzene derivative used as an intermediate in the synthesis of dyes,
 

was selected for bioassay by the National Cancer Institute along with
 

other dye intermediates in an attempt to determine which chemicals may
 

be responsible for the increased incidence of bladder cancer observed
 

among workers in the dye manufacturing industry (Wynder et al., 1963;
 

Anthony and Thomas, 1970). Aromatic nitro and amino compounds are
 

thought to contribute to the increased cancer risk in this industry
 

(Wynder et al., 1963).
 

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

*
 

name for this compound is 2-methoxy-5-nitro-benzenamine. It is also
 

known as 3-amino-4-methoxy-nitrobenzene; 2-methoxy-5-nitroaniline;
 

2-amino-4-nitroanisole; Fast Scarlet R; and C.I. (Colour Index) Azoic
 

Diazo Component 13 (C.I. No. 37130).
 

5-Nitro-o-anisidine is a chemical intermediate in the production
 

of C.I. Pigment Red 23 which is used as a colorant in a wide variety
 

of commodities including printing inks, interior latex paints, lacquers, rubber, plastics, floor coverings, paper coatings, and textiles
 

(Schlapfer, 1973; Society of Dyers and Colourists, 1971b; Society of
 

Dyers and Colourists, 1971c as cited in Urso, 1977). 5-Nitro-o-anisidine can also be used, along with certain C.I. coupling components, to
 

k
 

The CAS registry number is 99-59-2,
 
OCH3 

NO2 

FIGURE 1 

CHEMICAL STRUCTURE OF 5-NITRO-o-ANISIDINE 
produce various red, brown, yellow, and violet hues on cotton, silk,
 

acetate, and nylon (Society of Dyers and Colourists, 1971a as cited
 

in Urso, 1977).
 

Production statistics for dye intermediates bearing Colour Index
 

classifications are reported independently from those for identical
 

compounds bearing chemical names. In 1975, five U.S. companies reported production of 191 thousand pounds of C.I. Azoic Diazo Component 13, salt, the stabilized diazonium salt of 5-nitro-o-anisidine.
 

In that same year, C.I. Azoic Diazo Component 13, base, was produced
 

in commercial quantities (in excess of 1000 pounds or $1000 in value
 

annually) by only one of these companies; production statistics for
 

the base are therefore considered proprietary and are not available
 

(U.S. International Trade Commission, 1977a). Domestic production
 

of Azoic Diazo Components as a class appears to be declining, with
 

decreases of 25.7 and 19.6 percent noted from 1974 to 1975 for bases
 

and salts, respectively (U.S. International Trade Commission, 1977a).
 

5-Nitro-o-anisidine itself was not listed in Synthetic Organic
 

Chemicals, U.S. Production and Sales, 1975 (U.S. International Trade
 

Commission, 1977a), implying that it was not produced commercially in
 

that year. The compound was, however, included in the 1977 Directory
 

of Chemical Producers, U.S.A. (Stanford Research Institute, 1977) and
 

is presently manufactured on a commercial scale by two companies, one
 

of which also produces C.I. Azoic Diazo Component 13.
 
Imports of 5-nitro-o-anisidine through principal U.S. customs
 

districts amounted to 75 thousand pounds in 1975 and included 45
 

thousand pounds of C.I. Azoic Diazo Component 13, base (U.S. International Trade Commission, 1977b as cited in Urso, 1977). This quantity
 

represents a 45 percent decrease over the 1974 figure of 136 thousand
 

pounds (U.S. International Trade Commission, 1976 as cited in Urso,
 

1977).
 
II. MATERIALS AND METHODS
 

A. Chemicals
 

5-Nitro-o-anisidine was purchased from Carroll Products, Wood
 

River Junction, Rhode Island. Chemical analysis was performed by
 

Mason Research Institute, Worcester, Massachusetts. The experimentally determined melting point of 118°C was identical with that
 

reported in the literature (Weast, 1977). Infrared analysis was consistent with the structure of the compound. Thin-layer chromatography
 

did not show the presence of impurities. The evidence suggests that
 

the purchased compound was of high purity.
 

Throughout this report the term 5-nitro-o-anisidine is used to
 

represent this compound.
 

B. Dietary Preparation
 

The basal laboratory diet for both dosed and control animals consisted of Wayne Lab-Blox®(Allied Mills, Inc., Chicago, Illinois).
 

5-Nitro-o-anisidine was administered to the dosed animals as a component of the diet. Proper amounts of the chemical were removed from
 

the stock bottle under an exhaust hood. The compound was blended in
 

an aluminum bowl with an aliquot of the ground feed. Once visual homogeneity was attained, the mixture was placed into a 6 kg capacity
 

Patterson-Kelley twin shell V-blender along with the remainder of the
 

meal. The blender was sealed and operated for 20 minutes. The mixtures were placed into plastic bags and stored in the dark at 4°C.
 

The dietary preparations were used for only 1 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, mice assigned to the dose B group
 

(see p. 14), and all control animals were obtained from Charles River
 

Breeding Laboratories, Wilmington, Massachusetts. Low dose rats and
 

dose A mice were obtained from ARS/Sprague-Dawley, Madison, Wisconsin.
 

Dosed and control animals for both species were received in separate
 

shipments.
 

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 average body
 

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

D.	 Animal Maintenance
 

All animals were housed by species in rooms having a temperature
 

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

15/40 denier Dacron(R) 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, high dose rats and their controls
 

were housed in galvanized- or stainless-steel wire-mesh cages (Fenco
 

Cage Products, Boston, Massachusetts) suspended above newspapers.
 

6
 
Low dose rats and their controls were held in galvanized- or stainless-


steel wire-mesh cages during quarantine and for the first 7 months of
 

study. Newspapers under cages were replaced daily and cages and racks
 

washed weekly. For the remainder of the study, all rats were held in
 

suspended polycarbonate cages (Lab Products, Inc., Garfield, New Jersey)
 

equipped with disposable nonwoven fiber filter sheets. Clean cages
 

and bedding were provided twice weekly. Low dose rats and their controls were provided with SAN-I-CEL^ corncob bedding (Paxton Processing
 

Company, Paxton, Illinois) while in polycarbonate cages. High dose
 

(R}
 rats were provided with SAN-I-CEL for 7 months, after which Aspen
 

hardwood chip bedding (American Excelsior Company, Baltimore, Maryland)
 

was used for these animals for the remainder of the study. Stainless
 

steel cage racks were cleaned once every 2 weeks, and new disposable
 

filters were installed at that time.
 

Mice were housed by sex in polycarbonate cages. During quarantine and dosing periods, cages were fitted with perforated stainless steel lids. During the final observation period, stainless steel
 

wire bar lids were used. Both types of lids were from Lab Products,
 

Inc. Nonwoven fiber filter bonnets were used over cage lids. Dose B
 

mice and their controls were housed ten per cage for the first 11
 

months and five per cage thereafter. Dose A mice and their controls
 

were reduced to five per cage after 18 months. Clean cages, lids,
 

filters, and bedding were provided three times per week when cage
 

populations were ten and twice per week when cage populations were
 
(3


reduced to five. Ab-sorb-dri hardwood chip bedding (Wilner Wood
 

Products Company, Norway, Maine) was used for 1 month (for dose B mice
 

and their controls) and 7 months (for dose A mice and their controls).
 

(R) (R)
 SAN-I-CEL was used for the next 12 months. Bed-o-Cobs corncob
 

bedding (The Andersons Cob Division, Maumee, Ohio) was used for the
 

next 8 months, and Aspen bedding was used for the remainder of the
 

study. Reusable filter bonnets and pipe 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 supplied
 

ad libitum.
 

(R)
 Pelleted Wayne Lab-Blox was supplied to low dose rats and their
 

controls during the quarantine period and to all rats and mice during
 

the final observation period. During the 78-week dosing period, all
 

®
 animals were supplied with Wayne Lab-Blox meal containing the appropriate concentration of 5-nitro-o-anisidine. Control animals had
 

untreated meal available. Throughout the study, meal was supplied to
 

(R)
 all mice and to low dose rats and their controls in Alpine^ aluminum
 

feed cups (Curtin Matheson Scientific, Inc., Woburn, Massachusetts)
 

equipped with stainless steel baffles. High dose rats and their
 

(R)
 controls were supplied food from Alpine feed cups for the first 11
 
months of study and thereafter 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.
 

All dosed rats were housed in a room with other rats receiving
 

&
 

diets containing 3-amino-4-ethoxyacetanilide (17026-81-2); l-amino2-methylanthraquinone (82-28-0); 4-nitroanthranilic acid (619-17-0);
 

and 5-nitroacenaphthene (602-87-9). All control rats were in a room
 

with other rats receiving diets containing 3-nitro-p-acetophenetide
 

(1777-84-0); amitrole (61-82-5); and 2-methyl-l-nitroanthraquinone
 

(129-15-7).
 

Dose B mice were housed in a room with other mice receiving
 

diets containing 2,5-toluenediamine sulfate (6369-59-1); 5-nitro-otoluidine (99-55-8); hydrazobenzene (530-50-7); 1-nitronaphthalene
 

(86-57-7); 3-amino-9-ethylcarbazole hydrochloride; 6-nitrobenzimidazole (94-52-0); and 2,4-diaminoanisole sulfate (615-05-4). Dose A
 

mice and all control mice were housed in a room with other mice
 

receiving diets containing l-amino-2-methylanthraquinone (82-28-0);
 

N,N-dimethyl-p-nitrosoaniline (138-89-6); 2,5-toluenediamine sulfate
 

(6369-59-1); 2,4-dinitrotoluene (121-14-2); 1-nitronaphthalene (86­

57-7); 3-amino-9-ethylcarbazole hydrochloride; 2-aminoanthraquinone
 

* .
 CAS registry numbers are given in parentheses.
 
(117-79-3); 3-amino-4-ethoxyacetanilide (17026-81-2); 5-nitroacenaphthene (602-87-9); 2,4-diaminoanisole sulfate (615-05-4); amitrole
 

(61-82-5); 3-nitro-p-acetophenetide (1777-84-0); 4-nitroanthranilic
 

acid (619-17-0); and APC (8003-03-0).
 

E. Selection of Initial Concentrations
 

In order to establish the maximum tolerated concentration of
 

5-nitro-o-anisidine for administration to dosed animals in the
 

chronic studies, subchronic toxicity tests were conducted with both
 

rats and mice. Animals of each species were distributed among five
 

groups, each consisting of five males and five females. 5-Nitro-oanisidine was incorporated into the laboratory diet and supplied ad
 

libitum to four of the five groups of each species in concentrations
 

of 0.05, 0.1, 0.2, and 0.4 percent. The fifth group of each species
 

served as a control group, receiving only the basal laboratory diet.
 

The dosed dietary preparations were administered for 7 weeks, followed
 

by a 1-week observation period during which all animals were fed the
 

basal laboratory diet. All survivors were sacrificed at the end of
 

the observation period and gross necropsies were performed.
 

The highest concentration causing no deaths, no compound-related
 

gross abnormalities, and no mean body weight depression in excess of
 

10 percent relative to controls during the 8-week subchronic test was
 

selected as the high concentration utilized for the chronic bioassay.
 

In rats no deaths occurred at any doses tested. The single
 

gross abnormality observed, a darkened spleen, was encountered in two
 

10
 
female rats receiving 0.4 percent 5-nitro-o-anisidine. A dietary concentration of 0.2 percent produced no mean body weight depression in
 

male or female rats. A dietary concentration of 0.4 percent produced
 

mean body weight depressions of 0.7 and 10.0 percent in male and female rats, respectively. The high concentration selected for administration to rats in the chronic bioassay was 0.4 percent.
 

No deaths occurred in mice at any doses tested. A dietary concentration of 0.4 percent produced 7.9 percent mean body weight depression in male mice and no mean body weight depression in female
 

mice. The high concentration selected for administration to mice in
 

the chronic bioassay was 0.8 percent.
 

F. Experimental Design
 

The experimental design parameters for the chronic study (species,
 

sex, group size, concentrations administered, duration of treated and
 

untreated observation periods, and the time-weighted average concentrations) are summarized in Tables 1 and 2.
 

All rats were approximately 6 weeks old at the time the test was
 

initiated. The initial concentrations of 5-nitro-o-anisidine in diets
 

were 0.4 and 0.2 percent. The rat group receiving a concentration of
 

0.2 percent was sacrificed after 16 weeks and no histopathologic examinations were performed because the dose level was considered, on the
 

basis of mean body weight depression, to be too low. A new rat group,
 

receiving a dietary concentration of 0.8 percent, was started approximately 7 months after the initiation of the chronic study. Throughout
 

11
 
TABLE 1
 

DESIGN SUMMARY FOR FISCHER 344 RATS
 

5-NITRO-o-ANISIDINE FEEDING EXPERIMENT
 

INITIAL 

GROUP 

SIZE 

5-NITRO-oANISIDINE 

CONCENTRATION 

(PERCENT) 

OBSERVATION PERIOD 

TREATED UNTREATED 

(WEEKS) (WEEKS) 

MALE 

LOW DOSE CONTROL 50 0 0 108 

HIGH DOSE CONTROL 49 0 0 109 

LOW DOSE 50 0.4 

0 

78 

28 

HIGH DOSE 50 0.8 

0 

78 

24 

FEMALE 

LOW DOSE CONTROL 50 0 0 108 

HIGH DOSE CONTROL 50 0 0 109 

LOW DOSE 50 0.4 

0 

78 

28 

HIGH DOSE 50 0.8 78
 

0 28
 

12
 
TABLE 2
 

DESIGN SUMMARY FOR B6C3F1 MICE
 

5-NITRO-o-ANISIDINE FEEDING EXPERIMENT
 

5-NITRO-oINITIAL ANISIDINE OBSERVATION PERIOD TIME-WEIGHTED 

GROUP CONCENTRATION TREATED UNTREATED AVERAGE 

SIZE (PERCENT) (WEEKS) (WEEKS) CONCENTRATION" 

MALE
 

DOSE A CONTROL 50 95
 

DOSE B CONTROL 50 0 0 96 0
 

DOSE A 50 0.8 78 0.8
 

0 18
 

DOSE B 50 1.6 15 0.6
 

0.4 63
 

0 18
 

FEMALE
 

DOSE A CONTROL 50 0 0 96 0
 

DOSE B CONTROL 50 0 0 96 0
 

DOSE A 50 0.8 78 0.8
 

0 19
 

DOSE B 50 1.6 15 0.6
 

0.4 63
 

0 18
 

S (concentration X weeks received)
 T-imp— TJPI oVi f-pd a vpra CTP c•r»nr pnt-ra h n nn =
 

receiving chemical)
 

13
 

3 
this report those rats receiving the 0.8 percent concentration are
 

referred to as the high dose groups and those receiving the 0.4 percent concentration are referred to as the low dose groups. A high
 

dose control group was started approximately a week before the high
 

dose group was initiated. The dosed rats were supplied with feed
 

containing 5-nitro-o-anisidine for a total of 78 weeks, followed by
 

an observation period of up to 28 weeks.
 

All mice were approximately 6 weeks old at the time they were
 

placed on test. The initial concentrations of 5-nitro-o-anisidine
 

in diets were 0.8 and 0.4 percent. The mouse group receiving 0.4
 

percent was sacrificed after 16 weeks and no histopathologic examinations were performed because, based on the absence of weight
 

depression, the dose level was considered to be too low. A new
 

group, receiving 1.6 percent, and a control group were started
 

approximately 7 months after the initiation of the chronic study.
 

The dietary concentration administered to this group was lowered to
 

0.4 percent after 15 weeks of chemical administration. Throughout
 

this report those mice initially receiving a concentration of 1.6
 

percent are referred to as the dose B groups and those receiving a
 

concentration of 0.8 percent are referred to as the dose A groups.
 

The terms dose A and dose B groups are used instead of the more
 

common terms, high and low dose groups, because the groups that
 

initially received a higher dietary concentration of the compound
 

ultimately received a lower time-weighted average concentration, due
 

14
 
to dose changes. The dosed mice were supplied with feed containing
 

5-nitro-o-anisidine for a total of 78 weeks, followed by an observation period of up to 19 weeks.
 

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.
 

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.
 

15
 
Slides were prepared from the following tissues: skin, subcutaneous tissue, lungs and bronchi, trachea, bone marrow, spleen, lymph
 

nodes, thymus, heart, muscle, salivary gland, liver, gallbladder
 

(mice), pancreas, esophagus, stomach, small intestine, large intestine, kidney, urinary bladder, pituitary, adrenal, thyroid, parathyroid, testis, prostate, brain, Zymbal's gland, uterus, mammary gland,
 

and ovary.
 

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
 

16
 
that bear on the possibility of carcinogenicity are discussed in the
 

statistical narrative sections.
 

Probabilities of survival were estimated by the product-limit
 

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

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

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

missing; animals dying from natural causes were 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.
 

17
 
The purpose of the statistical analyses of tumor incidence is to
 

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

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

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

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

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

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

significance level of 0.05 may be made. The Bonferroni inequality
 

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

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 (Armitage,
 

1971, pp. 362-365) was not used.
 

A time-adjusted analysis was applied when numerous early deaths
 

resulted from causes that were not associated with the formation
 

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

tumor was observed were excluded by basing the statistical tests
 

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

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

early tumor was found, comparisons were based exclusively on animals
 

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

was found. Once this reduced set of data was obtained, the standard
 

procedures for analyses of the incidence of tumors were followed.
 

18
 
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
 

c
 

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

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

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

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

larger proportion in the treated group than in the control.
 

The lower and upper limits of the confidence interval of the
 

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

19
 
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.
 

20
 
III. CHRONIC TESTING RESULTS: RATS
 

A. Body Weights and Clinical Observations
 

Mean body weight depression was observed in all dosed groups
 

when compared to their control groups. The mean body weight difference between high dose groups and their control groups was greater
 

than that between low dose groups and their controls (Figure 2).
 

Fluctuations in the growth curve may be due to mortality; as the size
 

of the group diminishes, the mean body weight may be subject to wide
 

variations.
 

Twelve high dose males and two high dose females had subcutaneous and/or cutaneous growths. Similar subcutaneous and/or cutaneous
 

growths were observed in two low dose males and three low dose
 

females. Two high dose control male rats developed subcutaneous
 

and/or cutaneous growths, as did 10 high dose female rats. Similar
 

growths were recorded in four low dose control male rats and six low
 

dose control female rats. A crusted lesion was observed on the dorsal surface of one high dose male, and one high dose male displayed
 

bleeding from the ear canal. White discoloration of the eyes was
 

recorded in the two high dose females, one high dose control female,
 

and three low dose control females. Exopthalmia was observed in one
 

high dose control female and alopecia was present in one female of
 

the same group. No other clinical abnormalities were observed.
 

21
 
750- -750 

600 — -600 

h- 450­ -450 X 

LU 

g 300­ — 300 

CO 

LOW DOSE CONTROL 

150­ HIGH DOSE CONTROL -150 

LOW DOSE 

MALE RATS 

HIGH DOSE 

I I 

15 30 45 60 90 105 120 

TIME ON TEST (WEEKS) 

750­ •750 

LOW DOSE CONTROL 

HIGH DOSE CONTROL 

600 — -600 

LOW DOSE 

HIGH DOSE 

(- 450 — X -450 

C3 

LU 

§ -300 

00 

150­ -150 

FEMALE RATS 

I T I \ \ 

15 30 45 60 75 90 105 120 

TIME ON TEST (WEEKS) 

FIGURE 2 

GROWTH CURVES FOR 5-NITRO-o-ANISIDINE CHRONIC STUDY RATS 

22 
B. Survival
 

The estimated probabilities of survival for male and female
 

rats in the control and 5-nitro-o-anisidine-dosed groups are shown in
 

Figure 3. For both males and females the Cox tests indicated that
 

both the high dose and the low dose group had significantly greater
 

mortality than their respective controls.
 

For males the survival of both dosed groups was good for at
 

least 65 weeks, followed by sharp increases in mortality with 17
 

high dose rats dying in weeks 71 and 72. Five high dose controls
 

were sacrificed in week 78 and five low dose controls in week 80.
 

Adequate numbers of males were at risk from late-developing tumors as
 

at least 39 males from each of the groups survived on test at least
 

70 weeks.
 

For females five high dose controls were sacrificed in week 78
 

and five low dose controls in week 80. Adequate numbers of females
 

were at risk from late-developing tumors with 58 percent (29/50) of
 

the high dose, 74 percent (37/50) of the low dose, 86 percent (43/50)
 

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

control surviving on test at least 85 weeks.
 

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).
 

23
 
PROBABILITY OF SURVIVAL PROBABILITY OF SUR

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Neoplasms attributed to the administration of 5-nitro-o-anisidine
 

were found in the integument and associated glands. Some dosed rats
 

had tumors at more than one body site.
 

Basal-cell carcinoma of the skin occurred in 1/48 (2 percent)
 

low dose control, 7/50 (14 percent) low dose, and 30/48 (63 percent)
 

high dose males, and in 1/50 (2 percent) high dose control and 1/46
 

(2 percent) high dose female rats. This tumor was defined as a
 

tumor of undifferentiated cells of the basal layer of the epidermis
 

growing down into the dermis, sometimes in a polarized arrangement
 

or forming a lacey pattern. Tumor cells were small with oval, deep-


staining nuclei and scanty indistinct cytoplasm (Zackheim, 1973).
 

Sebaceous adenocarcinoma occurred in 5/50 (10 percent) low dose
 

and 21/48 (44 percent) high dose male and in 1/49 (2 percent) low
 

dose and 2/46 (4 percent) high dose female rats while none occurred
 

in the control animals. This tumor arose either in the skin or in
 

the ear canal (Zymbal's gland). It consisted of a variable mixture
 

of basal cells proliferating from the wall of a sebaceous gland, of
 

fully differentiated sebaceous cells with abundant foamy cytoplasm,
 

and of intermediate cells having a small amount of foamy cytoplasm.
 

Tumors arising in the ear canal usually exhibited superficial squamous differentiation (Pliss, 1973).
 

Trichoepitheliomas occurred in 20/50 (40 percent) low dose and
 

9/48 (19 percent) high dose male rats, while none occurred in the control groups. This tumor also arose from basal cells of the epidermis
 

25
 
and contained cystic structures composed of whorled squamous cells
 

surrounded by undifferentiated basal cells and exhibiting no zone of
 

transition (Zackheim, 1973). In some instances the contents of the
 

cyst resembled those of a normal hair follicle.
 

Squamous-cel1 carcinoma occurred in 1/48 low dose control males,
 

3/50 low dose and 12/48 high dose males, and in 2/49 low dose and
 

3/46 high dose females. The tumor was characterized by invasion of
 

adjacent tissue by whorls and strands of neoplastic cells, some of
 

which exhibited keratinization and some of which might be small and
 

poorly differentiated (Zackheim, 1973). Mitoses were often abundant.
 

Mammary adenocarcinoma was observed in 0/99 control, 11/49 low
 

dose and 5/46 high dose female rats. This was defined as a tumor
 

arising from glandular epithelium of the mammary gland and having a
 

more or less well-marked acinar pattern. Nuclei were round with
 

prominent nucleoli. Cytoplasm was usually acidophilic and occasionally contained large secretion vacuoles. Secretion was often found
 

in the lumens of acini.
 

The incidences of adenoma NOS, carcinoma NOS, squamous-cell carcinoma, or cystadenoma NOS of the preputial gland (2/96 [2 percent]
 

controls, 3/50 [6 percent] low dose, 6/48 [13 percent] high dose) in
 

male rats and of adenoma NOS, carcinoma NOS, squamous-cell papilloma,
 

squamous-cell carcinoma, adenocarcinoma NOS, or papillary adenoma of
 

the clitoral gland (4/99 [4 percent] controls, 13/49 [27 percent]
 

low dose, 14/46 [30 percent] high dose) in female rats was elevated.
 

26
 
The morphology of these tumors was similar in both sexes. The characteristic feature of these tumors was the presence of at least a few
 

large cells having abundant cytoplasm packed with coarse, brightly
 

acidophilic granules. Some of these tumors exhibited enough superficial squamous differentiation to be classified as squamous-cell
 

neoplasms. Other tumors with minimal squamous change were classified
 

as adenomas if they consisted of masses of large, often granulated
 

cells with large, rounded vesicular nuclei surrounding a central
 

cavity and having a well-defined intact outer border. If the outer
 

border of the tumor was irregular and consistent with invasion of
 

surrounding tissue, the tumors were classified as adenocarcinotnas or
 

carcinomas. The boundary lines between the varieties of these tumors
 

were not distinct so that there was a continuous spectrum from adenoma
 

to carcinoma.
 

Nonneoplastic cutaneous lesions were observed among male rats in
 

both dosed groups and included cystic and hyperplastic lesions.
 

Based upon this histopathologic evaluation, 5-nitro-o-anisidine
 

was carcinogenic to Fischer 344 rats. Benign and malignant neoplasms
 

of the skin and its glands were induced by oral administration of the
 

compound.
 

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 malignant tumor in either sex where at least two such
 

27
 
TABLE 3
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN MALE RATS TREATED WITH 5-NITRO-o-ANISIDINEE
 

TOPOGRAPHY : MORPHOLOGY
 

Skin: Basal-Cell Carcinoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

to
 

00 Skin: Trichoepithelioma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Skin: Papilloma NOS or Squamous-


Cell Papillomab
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

LOW DOSE HIGH DOSE LOW HIGH 

CONTROL CONTROL DOSE DOSE 

1/48(0.02) 0/48(0.00) 7/50(0.14) 30/48(0.63) 

P = 0.034 P < 0.001 ___ ___ 

6.720 Infinite 

0.914 10.044 

296.013 Infinite 

107 74 48 

0/48(0.00) 0/48(0.00) 20/50(0.40) 9/48(0.19) 

P < 0.001 P < 0.001 __._ Infinite Infinite 

6.208 2.633 

Infinite Infinite 

76 68 

1/48(0.02) 0/48(0.00) 2/50(0.04) 4/48(0.08) 

N.S. N.S. ___ 

1.920 Infinite 

0.103 0.928 

110.993 Infinite 

108 81 71 
TOPOGRAPHY :MORPHOLOGY
 

Skin: (Excluding skin of ear):
 

Squamous-Cell Carcinoma"
 

P Values
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Skin: (Excluding skin of ear) :
 

Sebaceous Adenocarcinoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Skin: (Excluding skin of ear):
 

Sebaceous Adenoma or Sebaceous
 

Adenocarcinoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 3 (CONTINUED) 

LOW DOSE HIGH DOSE


CONTROL CONTROL


0/48(0.00) 0/48(0.00)


0/48(0.00) 0/48(0.00)


0/48(0.00) 0/48(0.00)


 LOW
 

 DOSE
 

 3/50(0.06)
 

N.S.
 

Infinite
 

0.578
 

Infinite
 

60
 

 5/50(0.10)
 

P = 0.031
 

Infinite
 

1.212
 

Infinite
 

66
 

 14/50(0.28)
 

P < 0.001
 

Infinite
 

4.192
 

Infinite
 

60
 

HIGH
 

DOSE
 

11/48(0.23)
 

P < 0.001
 

Infinite
 

3.325
 

Infinite
 

66
 

18/48(0.38)
 

P < 0.001
 

Infinite
 

5.770
 

Infinite
 

47
 

23/48(0.48)
 

P < 0.001
 

Infinite
 

7.533
 

Infinite
 

47
 
TABLE 3 (CONTINUED)
 

TOPOGRAPHY: MORPHOLOGY

LOW DOSE 

 CONTROL 

HIGH DOSE

CONTROL

 LOW 

 DOSE 

Skin or Adnexa (Excluding skin of 

ear): Squamous-Cell Carcinoma, 

Basal-Cell Carcinoma, Trichoepithelioma, Sebaceous Adenocarcinoma, or 

Sweat-Gland Carcinomab 1/48(0.02) 0/48(0.00) 30/50(0.60) 

P Values0 P < 0.001 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

28.800 

5.228 

999.999 

Weeks to First Observed Tumor 107 60 

Skin and Subcutaneous Tissue: 

Fibromab 2/48(0.04) 3/48(0.06) 3/50(0.06) 

P Values0 N.S. 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

1.440 

0.173 

16.632 

Weeks to First Observed Tumor 99 95 79 

Lung: Alveolar/Bronchiolar Adenoma or 

Alveolar/Bronchiolar Carcinomab

P Values0 

 0/48(0.00) 1/48(0.02) 4/49(0.08) 

N.S. 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

Infinite 

0.909 

Infinite 

Weeks to First Observed Tumor 109 98 

HIGH
 

DOSE
 

40/48(0.83)
 

P < 0.001
 

Infinite
 

13.896
 

Infinite
 

1/48(0.02)
 

N.S.
 

0.333
 

0.006
 

3.976
 

101
 

1/48(0.02)
 

N.S.
 

1.000
 

0.013
 

76.887
 

47 

100 
TABLE 3 (CONTINUED)
 

TOPOGRAPHY :MORPHOLOGY
 

Hematopoietic System: Leukemia or
 

Malignant Lymphoma"
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Liver: Neoplastic Nodule
 

c
 

P Values
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

b
 

Pituitary: Adenoma NOS
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

LOW DOSE
 

CONTROL
 

6/48(0.13)
 

98
 

1/48(0.02)
 

99
 

1/41(0.02)
 

108
 

HIGH DOSE
 

CONTROL
 

6/48(0.13)
 

92
 

0/48(0.00)
 

_—
 

9/38(0.24)
 

_ —
 

85
 

LOW
 

DOSE
 

0/50(0.00)
 

P = 0.012(N)
 

0.000
 

0.000
 

0.600
 

3/48(0.06)
 

N.S.
 

3.000
 

0.252
 

154.112
 

96
 

8/44(0.18)
 

P = 0.019
 

7.455
 

1.070
 

321.866
 

66
 

HIGH
 

DOSE
 

0/48(0.00)
 

P = 0.013(N)
 

0.000
 

0.000
 

0.624
 

1/48(0.02)
 

N.S.
 

Infinite
 

0.054
 

Infinite
 

75
 

5/39(0.13)
 

N.S.
 

0.541
 

0.157
 

1.624
 

72
 
___ 

___ 

___ 

TABLE 3 (CONTINUED)
 

TOPOGRAPHY: MORPHOLOGY
 

Adrenal: Cortical Adenoma or Cortical
 

Carcinoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Adrenal: Pheochromocytoma or
 

Pheochromocytoma, Malignant"
 

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
 

LOW
 

DOSE
 

1/48(0.02)
 

N.S.
 

0.979
 

0.013
 

75.277
 

98
 

6/48(0.13)
 

N.S.
 

0.588
 

0.191
 

1.634
 

83
 

4/47(0.09)
 

N.S.
 

Infinite
 

0.775
 

Infinite
 

100
 

HIGH
 

DOSE
 

5/48(0.10)
 

P = 0.030
 

Infinite
 

1.237
 

Infinite
 

70
 

14/48(0.29)
 

N.S.
 

1.714
 

0.745
 

4.262
 

71
 

2/47(0.04)
 

N.S.
 

2.043
 

0.110
 

117.920
 

72
 

LOW DOSE
 

CONTROL
 

1/47(0.02)
 

106
 

10/47(0.21)
 _._
 

99
 

0/39(0.00)
 

N.S.
 

HIGH DOSE
 

CONTROL
 

0/47(0.00)
 

—_
 

8/47(0.17)
 

—__
 

106
 

1/48(0.02)
 

N.S.
 

109
 
_ __ 

TABLE 3 (CONTINUED) 

TOPOGRAPHY iMORPHOLOGY
 

Pancreatic Islets: Islet-Cell Adenoma
 

or Islet-Cell Carcinoma
 

P Values
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Preputial Gland: Adenoma NOS or
 

Carcinoma NOSb
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Testis: Interstitial-Cell Tumor
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

LOW DOSE
 

CONTROL
 

3/45(0.07)
 

N.S.
 

85
 

2/48(0.04)
 

103
 

45/47(0.96)
 

80
 

HIGH DOSE
 

CONTROL
 

0/46(0.00)
 

N.S.
 

0/48(0.00)
 

42/47(0.89)
 

78
 

LOW
 

DOSE
 

4/44(0.09)
 

N.S.
 

1.364
 

0.245
 

8.822
 

98
 

2/50(0.04)
 

N.S.
 

0.960
 

0.072
 

12.789
 

96
 

37/47(0.79)
 

P = 0.014(N)
 

0.822
 

0.754
 

0.980
 

61
 

HIGH
 

DOSE
 

0/46(0.00)
 

N.S.
 

5/48(0.10)
 

P = 0.028
 

Infinite
 

1.263
 

Infinite
 

70
 

16/48(0.33)
 

P < O.OOl(N)
 

0.373
 

0.281
 

0.542
 

72
 
TABLE 3 (CONCLUDED)
 

TOPOGRAPHY :MORPHOLOGY 

LOW DOSE 

CONTROL 

HIGH DOSE 

CONTROL 

LOW 

DOSE 

HIGH 

DOSE 

Zymbal's Gland or Skin of Ear: Sebaceous Ad eno carcinoma, Ceruminous 

Carcinoma, or Squamous-Cell 

Carcinoma" 1/48(0.02) 0/48(0.00) 2/50(0.04) 10/48(0.21) 

P Values0 N.S. P = 0.001 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

1.920 

0.103 

110.993 

Infinite 

2.979 

Infinite 

Weeks to First Observed Tumor 99 79 63 

Body Cavities: Mesothelioma NOS or 

Mesothelioma, Malignantb 1/48(0.02) 2/48(0.04) 5/50(0.10) 2/48(0.04) 

P Values0 _— N.S. N.S. 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

_— —— 4.800 

0.566 

222.171 

1.000 

0.075 

13.306 

Weeks to First Observed Tumor 108 106 71 72 

Treated groups received doses of 0.4 or 0.8 percent in feed.
 

Number of tunor-bearing animals/number of animals examined at site (proportion).
 •^
 

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

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 on the relative risk of the treated group to the control group.
 
___ 

TABLE 4
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN FEMALE RATS TREATED WITH 5-NITRO-o-ANISIDINE*
 

LOW DOSE
 

TOPOGRAPHY rMORPHOLOGY CONTROL
 

Skin (Excluding skin of ear) : Squamous-


Cell Carcinoma, Basal-Cell Carcinoma,
 

or Sebaceous Adenocarcinoma^3 0/49(0.00)
 

P Values
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 CO
 

Ui
 

Lung: Alveolar/Bronchiolar Adenoma
 

or Alveolar/Bronchiolar Carcinomab 0/49(0.00)
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pituitary: Adenoma NOSb

 18/44(0.41)
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor 89
 

HIGH DOSE
 

CONTROL
 

1/50(0.02)
 

109
 

1/50(0.02)
 

109
 

17/40(0.43)
 

78
 

LOW
 

DOSE
 

2/49(0.04)
 

N.S.
 

Infinite
 

0.296
 

Infinite
 

59
 

5/49(0.10)
 

P = 0.028
 

Infinite
 

1.262
 

Infinite
 

92
 

10/46(0.22)
 

P = 0.041(N)
 

0.531
 

0.250
 

1.071
 

84
 

HIGH
 

DOSE
 

5/46(0.11)
 

N.S.
 

5.435
 

0.640
 

250.926
 

75
 

1/43(0.02)
 

N.S.
 

1.163
 

0.015
 

89.179
 

87
 

5/41(0.12)
 

P = 0.002(N)
 

0.287
 

0.093
 

0.720
 

84
 
TOPOGRAPHY : MORPHOLOGY
 

Adrenal: Cortical Adenoma or
 

Cortical Carcinoma"
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Adrenal: Pheochromocytoma
 

u> 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
 

TABLE 4 (CONTINUED) 

LOW DOSE 

CONTROL 

HIGH DOSE 

CONTROL 

LOW 

DOSE 

HIGH 

DOSE 

1/49(0.02) 1/49(0.02) 2/49(0.04) 

N.S. 

3/44(0.07) 

N.S. 

104 109 

2.000 

0.108 

115.581 

102 

3.341 

0.280 

171.218 

86 

2/49(0.04) 3/49(0.06) 3/49(0.06) 

N.S. 

6/44(0.14) 

N.S. 

108 109 

1.500 

0.180 

17.316 

84 

2.227 

0.508 

13.017 

86 

3/40(0.08) 2/45(0.04) 3/49(0.06) 

N.S. 

1/42(0.02) 

N.S. 

108 109 

0.816 

0.116 

5.802 

92 

0.536 

0.009 

9.894 

106 
TOPOGRAPHY : MORPHOLOGY
 

Clitoral Gland: Carcinoma NOS
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Clitoral Gland: Carcinoma NOS or
 

Squamous-Cell Carcinoma*5
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Clitoral Gland: Adenoma NOS or
 

Papillary Adenoma or Carcinoma NOS
 

or Squamous-Cell Carcinoma*3
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 4 (CONTINUED) 

LOW DOSE HIGH DOSE


CONTROL CONTROL


0/49(0.00) 0/50(0.00)


0/49(0.00) 0/50(0.00)


1/49(0.02) 2/50(0.04)


108 104


 LOW
 

 DOSE
 

 0/49(0.00)
 

N.S.
 

 1/49(0.02)
 

N.S.
 

Infinite
 

0.054
 

Infinite
 

60
 

 12/49(0.24)
 

P = 0.001
 

12.000
 

1.894
 

499.415
 

 77
 

HIGH
 

DOSE
 

7/46(0.15)
 

P = 0.004
 

Infinite
 

2.112
 

Infinite
 

67
 

9/46(0.20)
 

P = 0.001
 

Infinite
 

2.860
 

Infinite
 

67
 

14/46(0.30)
 

P < 0.001
 

7.609
 

1.887
 

65.553
 

57
 
___ 

___ 

___

TOPOGRAPHY: MORPHOLOGY
 

Uterus: Endometrial Stromal Polyp
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 4 (CONTINUED)
 

LOW DOSE HIGH DOSE
 

CONTROL CONTROL
 

12/49(0.24) 10/50(0.20)
 

80


Uterus and Uterus/Endometrium: Carcinomain-Situ NOS, Undif ferentiated Carcinoma, Adenocarcinoma NOS, Adenoca
 

in Adenomatous Polyp, or Papillary
 

Adenocarcinoma'3
 

c
 

P Values
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Cerebrum and Brain: Astrocytoma, Glioma
 

NOS, Oligodendroglioma, or Medulloblastomak
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

2/49(0.04)


—_


——


108


1/49(0.02)


92
 

 78
 

 1/50(0.02)
 

—_
 

_—
 

 109
 

 0/50(0.00)
 —_
 

LOW
 

DOSE
 

14/49(0.29)
 

N.S.
 

1.167
 

0.560
 

2.469
 

61
 

3/49(0.06)
 

N.S.
 

1.500
 

0.180
 

17.316
 

106
 

2/49(0.04)
 

N.S.
 

2.000
 

0.108
 

115.581
 

58
 

HIGH
 

DOSE
 

6/44(0.14)
 

N.S.
 

0.682
 

0.221
 

1.890
 

5/44(0.11)
 

N.S.
 

5.682
 

0.670
 

262.045
 

100
 

3/44(0.07)
 

N.S.
 

Infinite
 

0.684
 

Infinite
 

76 

68 
TOPOGRAPHY : MORPHOLOGY
 

Zymbal's Gland or Skin of Ear:
 

Sebaceous Adenocarcinoma^
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Zymbal's Gland or Skin of Ear:
 

Sebaceous Adenocarcinoma or
 

Squamous-Cell Carcinoma
 

P Values0
 

Relative Rd.sk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tuir.or
 

Mammary Gland: Adeno carcinoma NOS
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 4 (CONTINUED)
 

LOW DOSE HIGH DOSE


CONTROL CONTROL


0/49(0.00) 0/50(0.00)


0/49(0.00) 0/50(0.00)


0/49(0.00) 0/50(0.00)


 LOW
 

 DOSE
 

 1/49(0.02)
 

N.S.
 

Infinite
 

0.054
 

Infinite
 

-tj
 I 1
 

 3/49(0.06)
 

N.S.
 

Infinite
 

0.602
 

Infinite
 

60
 

 10/49(0.20)
 

P =* 0.001
 

Infinite
 

2.976
 

Infinite
 

84
 

HIGH
 

DOSE
 

4/46(0.09)
 

P = 0.049
 

Infinite
 

1.008
 

Infinite
 

75
 

7/46(0.15)
 

P = 0.004
 

Infinite
 

2.112
 

Infinite
 

32
 

4/46(0.09)
 

P = 0.049
 

Infinite
 

1.008
 

Infinite
 

81
 
TABLE 4 (CONTINUED) 

LOW DOSE HIGH DOSE LOW HIGH 

TOPOGRAPHY : MORPHOLOGY CONTROL CONTROL DOSE DOSE 

Mammary Gland: Adenocarcinoma NOS, 

Papillary Adenocarcinoma or 

Adenoma NOSb 2/49(0.04) 0/50(0.00) 11/49(0.22) 4/46(0.09) ___ ___ 

P Values0 P = 0.007 P = 0.049 ___ 

Relative Risk (Control) 5.500 Infinite 

Lower Limit 1.288 1.008 

Upper Limit 48.873 Infinite 

Weeks to First Observed Tumor 108 84 81 

Mammary Gland: Fibroadenoma 16/49(0.33) 19/50(0.38) 4/49(0.08) 0/46(0.00) 

P Values0 P < O.OOl(N) P < O.OOl(N) ___ ___ Relative Risk (Control) 0.250 0.000 

Lower Limit 0.066 0.000 

Upper Limit 0.708 0.178 

Weeks to First Observed Tumor 80 106 84 

Hematopoietic System: Leukemia or 

Malignant Lymphoma" 7/49(0.14) 5/50(0.10) 0/49(0.00) 4/46(0.09) 

P Values0 P = 0.006(N) N.S. 

Relative Risk (Control) 0.000 0.870 

Lower Limit 0.000 0.183 

Upper Limit 0.515 3.788 

Weeks to First Observed Tumor 106 104 84 
TABLE 4 (CONCLUDED) 

a
 

Treated groups received doses of 0.4 or 0.8 percent in feed.
 

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

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

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 on the relative risk of the treated group to the control group.
 
tumors were observed in at least one of the control or 5-nitro-oanisidine-dosed groups and where such tumors were observed in at
 

least 5 percent of the group. Caution must be used in interpreting
 

the low dose results since the low dose groups were from a different
 

supplier than the low dose control groups.
 

For both male and female rats numerous carcinomas of the Zymbal's
 

gland or the skin of the ear were observed in the high dose groups.
 

For males, the Fisher exact comparisons of the high dose group to the
 

high dose control indicated that the combined incidence of sebaceous
 

adenocarcinomas, ceruminous carcinomas, and squamous-cell carcinomas
 

was significantly (P = 0.001) greater in the high dose group. Similarly, for females the combined incidence of sebaceous adenocarcinomas
 

and squamous-cell carcinomas were significantly (P = 0.004) greater in
 

the high dose group than in the high dose control. Based upon these
 

results the administration of 5-nitro-o-anisidine was associated with
 

the increased incidence of carcinomas of the skin of the ear or of the
 

Zymbal's gland in both male and female rats.
 

In male rats large numbers of skin tumors were observed in the
 

dosed groups, with basal-cell carcinomas observed as early as week
 

48 and sebaceous adenocarcinomas observed as early as week 47. For
 

the incidence of basal-cell carcinomas, the incidence of trichoepitheliomas, the incidence of squamous-cell carcinomas, and the incidence
 

of sebaceous adenocarcinomas, each had a significantly (P <_ 0.001)
 

greater incidence in the high dose than in the high dose control.
 

42
 
When the combined incidences of all carcinomas of the skin or adnexa
 

(excluding skin of the ear) were considered, both the high dose and
 

the low dose comparisons were significant (P < 0.001) with 83 percent
 

(40/48) of the high dose, 60 percent (30/50) of the low dose, none of
 

the 48 high dose control, and 2 percent (1/48) of the low dose control males with one or more of these tumors. Based on these results
 

the administration of 5-nitro-o-anisidine was associated with an increased incidence of carcinomas of the skin in male rats.
 

In female rats numerous clitoral gland neoplasms were observed.
 

The Fisher exact test indicated a significantly (P = 0.004) higher
 

incidence of carcinomas NOS in the high dose than in the high dose
 

control. When the combined incidence of carcinomas NOS or adenomas
 

NOS or papillary adenomas or squamous-cell carcinomas was considered,
 

both the low dose and the high dose Fisher exact test were significant (P <_ 0.001). Based upon these results, the administration of
 

5-nitro-o-anisidine was associated with an increased incidence of
 

neoplasms of the clitoral gland in female rats.
 

For females the Fisher exact test indicated a significantly
 

(P = 0.001) higher incidence of mammary adenocarcinomas NOS in the
 

low dose than in the low dose control group. The high dose comparison had a probability level of P = 0.049, a marginal result. It
 

must be reiterated, however, that the low dose rats were obtained
 

from a different supplier than the low dose control, the high dose,
 

and the high dose control group.
 

43
 
For male rats the incidence of pituitary adenomas NOS was significantly (P = 0.019) higher in the low dose than in the low dose
 

control. The high dose comparison was not significant, however.
 

In historical control data compiled by this laboratory for the NCI
 

Carcinogenesis Testing Program, 35/334 (10 percent) of the untreated
 

male Fischer 344 rats had either an adenoma NOS or a chromophobe
 

adenoma, compared to 1/41 (2 percent) of the low dose control and
 

8/44 (18 percent) of the low dose group in this bioassay.
 

For male rats the high dose to high dose control comparison had
 

marginal positive results both for adrenal cortical neoplasms and for
 

preputial neoplasms. Similarly for low dose females marginal test
 

results were noted for lung neoplasms.
 

For females the Fisher exact test comparison of the low dose to
 

the low dose control group indicated a significant negative association between dose and the incidence of malignant lymphomas or
 

leukemia. The incidence in the high dose group, however, was not
 

significantly different from that in the high dose control.
 

Both for leukemia or malignant lymphomas and for interstitial-


cell tumors of the testes in the males and both for pituitary adenomas NOS and for mammary fibroadenomas in the females, the possibility
 

of a negative association between chemical administration and incidence was observed. It must be noted, however, that a relatively
 

high mortality from tumors was observed in the dosed rats.
 

44
 
Thus, based upon these statistical results the administration
 

of 5-nitro-o-anisidine was associated with the increased incidence of
 

carcinomas of the skin and of the Zymbal's gland in male rats and of
 

carcinomas of the Zymbal's gland and of clitoral neoplasms in female
 

rats. There also seemed to be some possibility of an association
 

between compound administration and the incidence of mammary adenocarcinomas in female rats.
 

45
 
IV. CHRONIC TESTING RESULTS: MICE
 

A. Body Weights and Clinical Observations
 

Moderate mean body weight depression was evident in all treated
 

groups, except dose A male mice, when compared to control groups
 

(Figure 4). Fluctuations in the growth curve may be due to mortality;
 

as the size of the group diminishes, the mean body weight may be subject to wide variations.
 

No clinical abnormalities were noted in mice of any group.
 

B. Survival
 

The estimated probabilities of survival for male and female mice
 

in the control and 5-nitro-o-anisidine-dosed groups are shown in Figure 5. For males there were no statistically significant differences
 

between the mortality of the dosed and control groups. For females
 

the Cox tests indicated significantly greater mortality in each of
 

the dosed groups than in their respective control group.
 

For males five dose B control mice were sacrificed in week 49;
 

in addition, five mice from each group but the dose B group were
 

sacrificed in week 78. Adequate numbers of males were at risk
 

from late-developing tumors as 88 percent (44/50) of the dose B
 

group, 84 percent (42/50) of the dose A group, 80 percent (40/50)
 

of the dose B control, and 86 percent (43/50) of the dose A control
 

survived on test until the end of the study.
 

For females five dose B control mice were sacrificed in week 49;
 

five mice from the dose A control, five from the dose B control, and
 

46
 
MEAN BODY WEIGHT (GRAMS)
 MEAN BODY WEIGHT 

o o 

J_ _L 

30 

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3D 

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01 

o 

z 

30 m m c/)

 

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30 

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o 

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D 

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m 

8 
PROBABILITY OF SURVIVAL PROBABILITY OF SURV

cD O O o o

 

c3 NJ -^ en co 0

1 1 1 1 1 1 1 1 1 - , 1 1 1 1 l 

-n r S 

(n m I >

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30

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5 S 

c/J m m

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:

! 

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D

 D

O O o o o D O O O m _, § c/i w en o o o W W w 8 m m rn m o— m m m m CJl ro > ro > CO > CO > o o 0 0 o 8 o z z z z H H H H 33 3) 33 31 0 0 O O ir 

r~ 

fO — 0 1 | 1 | 1 | I | l 1 I 1 ' 1 

cD O O O O ^ o p p 

c3 Kj ^ O) 00 0 
six from the dose A group were sacrificed in week 78. Of the 19
 

dose A females that died in weeks 43 through 45, 17 had nephrosis
 

of the kidney and 1 was autolyzed. Adequate numbers of females were
 

at risk from late-developing tumors as 66 percent (33/50) of the dose
 

B group, 32 percent (16/50) of the dose A group, 76 percent (38/50)
 

of the dose B control and 72 percent (36/50) of the dose A control
 

survived on test until the end of the study.
 

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).
 

The incidence of tumors of the liver in male and female mice is
 

summarized below:
 

MALES 

Dose A

Control

 Dose B 

 Control Dose A Dose B 

Number of animals with tissues 

examined histopathologically (50) (48) (48) (47) 

Liver 

Hepatocellular Carcinoma 12 

Hemangioma
 

Hemangiosarcoma
 

Nodular Hyperplasia
 

FEMALES
 

Number of animals with tissues
 

1

0

2 

6 

0

1

1 

25 

0

1 

2 

3

0 

2

1 

examined histopathologically (47) (50) (41) (43) 

Liver 

Hepatocellular Carcinoma 

Nodular Hyperplasia 

2 

0 

1 

0 

0 

0 

49
 
An increased incidence of hepatocellular carcinoma as compared
 

to controls was noted among dose B female and dose A male mice.
 

Hepatocellular carcinoma was defined as a neoplasm within the liver
 

parenchyma composed of hepatocytes arranged in an irregular fashion
 

so that all traces of normal liver architecture were obscured. Cells
 

varied in size but tended to be large with abundant, usually eosinophilic cytoplasm. Nuclei varied in size and shape both within a
 

given tumor and between various tumors. Nuclear morphology ranged
 

from normal to nuclei with hyperchromatism, abnormal chromatin
 

patterns, and atypical mitoses. Occasionally, fat vacuoles were
 

found in the cytoplasm.
 

With the exception of renal and hepatic lesions, the nonneoplastic lesions which occurred in both control and dosed mice were the
 

usual types observed in aging B6C3F1 mice. There were elevated incidences of toxic nephrosis in dosed females and of hepatocyte hyperplasia and degeneration in dosed males.
 

Based upon this histopathologic evaluation, there appeared to
 

be an association between liver carcinomas in dosed male and female
 

B6C3F1 mice and the dietary administration of 5-nitro-o-anisidine.
 

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 malignant tumor in either sex where at least two such
 

tumors were observed in at least one of the control or 5-nitro-oanisidine-dosed groups and where such tumors were observed in at
 

50
 
___ 

TABLE 5
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN MALE MICE TREATED WITH 5-NITRO-o-ANISIDINE£
 

TOPOGRAPHY : MORPHOLOGY
 

Lung: Alveolar/Bronchiolar Carcinoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Lung: Alveolar/Bronchiolar Carcinoma
 

or Alveolar/Bronchiolar Adenoma"
 

P Values0
 

d
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Hematopoietic System: Leukemia or
 

Malignant Lymphoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

DOSE A
 

CONTROL
 

5/50(0.10)
 

_—
 

95
 

5/50(0.10)
 

95
 

5/50(0.10)
 

74
 

DOSE B
 

CONTROL
 

5/49(0.10)
 

96
 

10/49(0.20)
 

_—
 

96
 

5/49(0.10)
 

96
 

DOSE A
 

1/48(0.02)
 

N.S.
 

0.208
 

0.005
 

1.768
 

96
 

5/48(0.10)
 

N.S.
 

1.042
 

0.255
 

4.243
 

96
 

5/48(0.10)
 

N.S.
 

1.042
 

0.255
 

4.243
 

96
 

DOSE B
 

1/47(0.02)
 

N.S.
 

0.209
 

0.005
 

1.767
 

96
 

2/47(0.04)
 

P = 0.017(N)
 

0.209
 

0.023
 

0.912
 

96
 

6/49(0.12)
 

N.S.
 

1.200
 

0.327
 

4.654
 

89
 
TABLE 5 (CONCLUDED)
 

TOPOGRAPHY: MORPHOLOGY 

DOSE A 

CONTROL 

DOSE B 

CONTROL DOSE \ DOSE B 

Liver: Hepatocellular Carcinoma 12/50(0.24) 6/48(0.13) 25/48(0.52) 3/47(0.06) 

P Values0 P = 0.004 N.S. 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

___ 

2.170 

1.202 

4.081 

0.511 

0.087 

2.239 

Weeks to First Observed Tumor 95 78 78 96 

Liver: Hepatocellular Carcinoma or 

Hepatocellular Adenoma" 

P Values0 

12/50(0.24) 8/48(0.17) ___ 25/48(0.52) 

P = 0.004 

3/47(0.06) ___ 

Relative Risk (Control) 

Lower Limit 

Upper Limit 

___ 2.170 

1.202 

4.081 

0.383 

0.069 

1.485 

Weeks to First Observed Tumor 95 78 78 96 

Treated groups received time-weighted average doses of 0.8 or 0.6 percent in feed.
 

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

£

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

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 on 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 5-NITRO-o-ANISIDINE*
 

TOPOGRAPHY rMORPHOLOGY
 

Lung: Alveolar/Bronchiolar Carcinoma
 

or Alveolar/Bronchiolar Adenoma"
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Hematopoietic System: Leukemia or
 

Malignant Lymphoma"
 

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
 

DOSE A
 

CONTROL
 

2/46(0.04)
 

96
 

7/48(0.15)
 

83
 

2/47(0.04)
 

94
 

DOSE B
 

CONTROL
 

3/50(0.06)
 

78
 

2/50(0.04)
 

96
 

1/50(0.02)
 

96
 

DOSE A
 

0/42(0.00)
 

N.S.
 

0.000
 

0.000
 

3.685
 

5/43(0.12)
 

N.S.
 

0.797
 

0.214
 

2.692
 

78
 

0/41(0.00)
 

N.S.
 

0.000
 

0.000
 

3.854
 

DOSE B
 

2/44(0.05)
 

N.S.
 

0.758
 

0.066
 

6.300
 

96
 

10/45(0.22)
 

P = 0.008
 

5.556
 

1.270
 

49.760
 

80
 

8/43(0.19)
 

P = 0.008
 

9.302
 

1.325
 

401.243
 

80
 
TABLE 6 (CONCLUDED)
 

DOSE A DOSE B 

TOPOGRAPHY :MORPHOLOGY CONTROL CONTROL DOSE A DOSE B 

Pituitary: Adenoma NOS or Chromophobe 

Adenoma" 5/43(0.12) 3/42(0.07) 1/31(0.03) 4/33(0.12) 

P Values0 

_ __._ N.S. N.S. ___ Relative Risk (Control) 

Lower Limit —— 0.277 0.006 

1.697 

0.307 

Upper Limit 2.300 10.800 

Weeks to First Observed Tumor 95 96 97 96 

Treated groups received time-weighted average doses of 0.8 or 0.6 percent in feed.
 

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

£

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

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 on the relative risk of the treated group to the control group.
 
least 5 percent of the group. Caution must be used in interpreting
 

results involving the dose A groups, since these dosed mice were
 

obtained from a different supplier than the dose A control groups.
 

Numerous incidences of hepatocellular carcinomas were observed
 

in both male and female mice. For males the Fisher exact test indicated a significantly (P = 0.004) higher incidence of hepatocellular
 

carcinomas in the dose A group than in the dose A control. It must
 

be noted, however, that the dosed mice were from a different supplier
 

than the control mice. When the incidence in the dose B group was
 

compared with the incidence in the dose B controls, it was not significant. For female mice the comparison of dose B to dose B control
 

was significant (P = 0.008). In historical control data collected by
 

this laboratory for the NCI Carcinogenesis Testing Program, 51/350
 

(15 percent) of the male and 13/350 (4 percent) of the female untreated B6C3F1 mice had a hepatocellular carcinoma. Based upon these
 

results, the administration of 5-nitro-o-anisidine was associated
 

with the increased incidence of hepatocellular carcinomas in female
 

mice.
 

For female mice the Fisher exact test indicated a significantly
 

(P = 0.008) higher incidence of leukemia or malignant lymphomas in
 

the dose B group than in the dose B control group. It must be noted,
 

however, that the incidence rate of 10/45 (22 percent) observed in
 

the dose B group did not greatly differ from the 7/48 (15 percent)
 

55
 
observed in the dose A control group, which came from the same supplier and was kept in the same room.
 

For males the Fisher exact test indicated a significantly (P =
 

0.017) lower incidence of alveolar/bronchiolar neoplasms in the dose
 

B group than in the dose B control.
 

56
 
V. DISCUSSION
 

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

Among rats, feeding of 5-nitro-o-anisidine was associated
 

primarily with increased incidences of tumors of the skin and its
 

glands. Incidences of the following skin tumors: basal-cell
 

carcinomas, trichoepitheliomas, squamous-cell carcinomas, and sebaceous adenocarcinomas—were each significant in high dose male rats.
 

For both male and female rats, carcinomas (the combined incidences of
 

sebaceous adenocarcinomas, ceruminous carcinomas, and squamous-cell
 

carcinomas) of the Zymbal's gland or the skin of the ear were significant in the high dose groups. The incidences of tumors of
 

the preputial and clitoral glands were elevated in dosed rats. The
 

incidences of carcinomas NOS and the incidences of adenomas (the
 

combined incidences of adenomas NOS and papillary adenomas) were each
 

significant in the clitoral gland of dosed female rats. The incidence
 

of mammary adenocarcinomas was significant in low dose female rats,
 

and the incidence of pituitary adenomas was significant in low dose
 

male rats, when each was compared to its respective control group.
 

It should be noted that low dose rats were received from a different
 

supplier than the low dose controls. It is, therefore, not certain
 

that all differences in tumor incidences between low dose rats and
 

their controls are attributable to administration of the compound.
 

57
 
The incidences of hepatocellular carcinomas were statistically
 

significant in dose A male mice and in dose B female mice when compared to their respective controls. It should be noted that the
 

dose A mice were received from a different supplier than the dose A
 

control mice. This reduces the weight of the evidence for attributing
 

the increase in hepatocellular carcinomas in dose A male mice to the
 

administration of the test compound. Liver neoplasms in dose B female
 

mice were considered to be due to the administration of 5-nitro-oanisidine. The number of female mice with either leukemia or malignant
 

lymphomas was significantly increased in the dose B group, when compared to the dose B controls. However, this incidence was comparable
 

to that observed in the dose A control group and, therefore, not considered attributable to the compound.
 

Under the conditions of this bioassay, dietary administration
 

of 5-nitro-o-anisidine was carcinogenic in Fischer 344 rats, causing
 

Zymbal's gland carcinomas in both sexes, integumentary carcinomas in
 

males, and clitoral gland neoplasms in females. The compound was
 

also carcinogenic in female B6C3F1 mice, causing hepatocellular carcinomas.
 

58
 
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.
 

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, B.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.
 

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

Observations." Journal of the American Statistical Association
 

5^:457-481, 1958.
 

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., "Tumours of the Auditory Sebaceous Glands." Pathology
 

of Tumours in Laboratory Animals, V.S. Turusov, editor. International Agency for Research on Cancer, Lyon, France, 1973.
 

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

Kaufman, "Respiratory Tract Carcinogenesis in Hamsters Induced
 

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

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

59
 
Schlapfer, L.A., "Naphthal Red and Brown." Pigment Handbook, Volume
 

I. T.C. Patton, editor. Wiley-Interscience, New York, pp. 473­

481, 1973.
 

Society of Dyers and Colourists. Colour Index, 3rd edition, Volume 1.
 

Yorkshire, England, p. 1570, 1971a.
 

Society of Dyers and Colourists. Colour Index, 3rd edition, Volume 3.
 

Yorkshire, England, p. 3303, 1971b.
 

Society of Dyers and Colourists. Colour Index, 3rd edition, Volume 4.
 

Yorkshire, England, p. 4036, 4750, 4706, 1971c.
 

Stanford Research Institute, 1977 Directory of Chemical Producers,
 

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

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

62^:679-682, 1975.
 

Urso, S., Research Analyst, Chemical-Environmental Program, Chemical
 

Industries Center, Stanford Research Institute, Menlo Park,
 

California. Personal communication, 1977.
 

U.S.	 International Trade Commission, Imports of Benzenoid Chemicals
 

and Products, 1974. USITC Publication 762, U.S. Government
 

Printing Office, Washington, D.C., 1976.
 

U.S.	 International Trade Commission, Imports of Benzenoid Chemicals
 

and Products, 1975. USITC Publication 806, U.S. Government
 

Printing Office, Washington, D.C., 1977b.
 

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

Production and Sales, 1975. USITC Publication 804, U.S. Government Printing Office, Washington, D.C., 1977a.
 

Weast, R.C., editor, CRC Handbook of Chemistry and Physics, 58th
 

edition. CRC Press, Cleveland, Ohio, 1977.
 

Wynder, E.L., J. Onderdonk, and N. Mantel, "An Epidemiological Investigation of Cancer of the Bladder." Cancer 16:1388-1407, 1963.
 

Zackheim, H.S., "Tumours of the Skin." Pathology of Tumours in Laboratory Animals, V.S. Turusov, editor. International Agency
 

for Research on Cancer, Lyon, France, 1973.
 

60
 
Review of the Bioassay of 5-Nitro-o-Anisidine* for Carcinogenicity
 

by the Data Evaluation/Risk Assessment Subgroup
 

of the Clearinghouse on Environmental Carcinogens
 

June 29, 1978
 

The Clearinghouse on Environmental Carcinogens was
 

established in May, 1976, in compliance with DREW Committee
 

Regulations and the Provisions of the Federal Advisory
 

Committee Act. The purpose of the Clearinghouse is to
 

advise the Director of the National Cancer Institute (NCI)
 

on its bioassay program to identify and to evaluate chemical
 

carcinogens in the environment to which humans may be exposed.
 The members of the Clearinghouse have been drawn from
 

academia, industry, organized labor, public interest groups,
 State health officials, and quasi-public health and research
 

organizations. Members have been selected on the basis of
 

their experience in carcinogenesis or related fields and,
 collectively, provide expertise in chemistry, biochemistry,
 biostatistics, toxicology, pathology, and epidemiology.
 Representatives of various Governmental agencies participate
 

as ad hoc members. The Data Evaluation/Risk Assessment
 

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

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

on the bioassay of 5-Nitro-o-Anisidine for carcinogenicity.
 

The reviewer agreed with the conclusion that 5-Nitro-o-
 Anisidine was carcinogenic in both mice and rats, he pointed
 

out a number of experimental deficiencies. Despite the
 

flaws, he said the compound should be considered carcinogenic
 

under the conditions of test. A motion was made by the
 

reviewer that the report on the bioassay of 5-Nitro-o-
 Anisidine be accepted as written. The motion was accepted
 

without objection.
 

Clearinghouse Members present:
 

Arnold L. Brown (Chairman), Mayo Clinic
 

Paul	 Nettesheim, National Institute of Environmental
 

Health Sciences
 

Verne Ray, Pfizer Medical Research Laboratory
 

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

Louise Strong, University of Texas Health Sciences Center
 

*	 Subsequent to this review, changes may have been made
 

In the bioassay report either as a result of the review
 

or other reasons. Thus, certain comments and criticisms
 

reflected in the review may no longer be appropriate.
 

61
 

<tU.S. GOVERNMEN T PRINTIN G OFFICE : 1978-260-899/318 1
 

F
 

APPENDIX A
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS
 

IN RATS TREATED WITH 5-NITRO-o-ANISIDINE
 

TABLE A1

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS TREATED WITH 5-NITRO-o-ANISIDINE 

LOW DOSE HIGH DOSE —, —, 

CONTRO L (UNTR )

01-0070 CONTRO L (U NT R)  01-0118 LOil BOSE 

01-0069 HIGH DOSE 

01-0121 

ANI.IAL S ANIMAL S

ANIMAL S

ANIMAL S

 INITIALL Y I N STUD Y MISSING

 NECROPSISD

 EXAMINE D HISTO PATHOLOGIC A LL Y

 50 1 

 48

 ** 4 8

 <BSO 

 U8 

 48 

50 

5J 

0 3 

50 

48 

48 

INTEGUMENTAR Y SYSTE M 

•SKIN (48) (4B) (50) (td) PAPILLOMA , NO S 1 (2*) 3 (6*) 

SQUAMO'J S CFLL PAPILLOM A 1 (21) 1 (2*) 2 (4*) 

SQUANOU S CFLL CARCINOM A 1 (2*) 3 (6*) 12 (25H) BASAL-CEL L CABCINOH A 1 (21) 7 (1UX) 30 (631) TSICHOSPITHELIOM A 20 (40%) 9 (19*) 

SW?:AT GLAN D CARCINOM A 1 (21) SEBACEOUS ADENOm 9 (18*) 8 (17*) SEBACEOU S ADENOCARCINOM A 5 (10*) 21 (44*)< ­

KEPHTOACANTHOM A 2 (<•« ) 2 (41) 

FIBROM A 3 (6!f) 1 (21) "IEBOSARCOM A 2 (4X ) 

*SU3C'JT TISSUE (US) (43) (50) <U8 ) 

SAP.COMA , NOS 1 (2%) 

FIBPOM A 2 (4X ) 3 (6H) 

FIEBOSARCOM A 1 (2JS) 

PESPTRATCP Y SYSTE M 

»L''N G (48 ) (43 ) (49 ) (48) 

CAFCINOMA , NOS, METASTAII C 1 (25 ) 

SSUAMOU S CELL CAFCINOMA , .IETASTA 1 (2H) ALV-CLAR/BFONCHIOLA H ADENOM A 3 (6*) ALVSCLA3/BSONCHIOLA 8 CASCINOM A 1 (2%) 1 <2X ) 1 (2X ) SEEACFOUS ADFNOCASCINO^IA , METAST 1 (2%) PHEOCH50M5CYTOHA , METASTATIC 1 (2X) 1 (21) 

OSTECStRCOrA , CETASTATI C 1 (2J) 

•M'JLTIPI'' ORGANS	 (48) (48) (50) (48)
 

* '1U13E3 OF ASIMJLS WITH TISSUE EXA1IN3D 1IC8 J5COPICALLY
 

* Nn«3"5 OF ANIMAL S NECROPSI2D
 

<- MULTIPLE OCCUaHENCi OF MORPHCL03Y IN THE SAMS ORGAN TISSUES IS COUNTED ONCE ONLY
 

"EXCLUDES PARTIALLY AUTOLYZED ANIMALS
 

i SO ANI1SLS WrSE INITIALLY IN TH^ STUDY, BUT ONS ANI1AL WAS FOUND TO 3E A FEMALE IN A MALE GROUP.
 

A-3
 
LETKI'KIA.NO S 

"YELCIONOCYTI C L5JKEMI A 

*SPLEE!I 

OSTECSAPCGKA , I1ETASTAIIC 

CIPCOLAICR Y SYSTE M 

BONE 

D n rSTIVF SYSTE M 

»SALIVAP Y GLAN E 

ADINOCARCINOMA , 10S 

SARCCfA , NO S 

tLivr " NFCELASTI C NODUL 3 

1'EPATOCELLULA R CARCINOM A 

»J£JJNU H 

A DFNCCAPCIN01A , NOS 

»H.*'JB 

SARCOMA , NO S 

*COInNI C SL'BIUCTS A 

"IBRO'A 

UHINHR Y SYSTE M 

"KIDNE Y T USriA=-CPL L ADENOCARCINOI A 

*KID1EY/PFLVI S 

TFAfSITIONAL-CIL L CARCIHOH A 

KUSINAH Y ELADDi P 

""UNSITIONHL-ri'L L PAPILL01 A 

ENDOrPIN I SYSTFK 

• PITUIIAP Y 

iDiJisaij. ^05 

» N-J13EP 0 ? ANIMSL S KIT H TISSU E 

* MM5EP OF ANIIAL S NSCROPSIE 3 

TABLE A1 (CONTINUED) 

CONTRO L (UNTR ) CONTROL (UNTR ) 

11-0071 01-0118 

1 (2*) 1 (2*) 

5 (11S) 4 (8*) 

(48) (48) 

1 (21) 

(to ) (47 )

 

1 (2X) 

1 (2*) 

(18) (48 )

 

1 (2J) 2 (4%) 1 (2%)

 

(45) (46) 

(45 ) (46 ) 

1 (2%) 

(41) (46 ) 

(18) (48 ) 

(18) (48) 

(46) (13) 

1 (2J) 

(tl) (38) 

._l.li^J 2 __12SSJ 

1ICPOSCOPICALL Y 

L O U DOSE 

01-0069 

(47) 

(15) 

(48 ) 

3 (6*) 

(46) 

1 (2S) 

(16) 

(45 ) 

(48) 

1 (2«) 

(48) 

(48) 

1 (2*) 

(44 ) 

8 (18X1 

HIGH DOSE 

01-0121 

(18) 

(16) 

(18) 

1 (2J) 

2 (IX) 

(17) 

(17) 

(39) 

1 <3X) 

(48) 

1 <2X ) 

(18) 

1 (2X) 

(18) 

(39) 

5 i33Xl 

A-4 
TABLE AI (CONTINUED) 

_ „__,. „- ....... 

LOW DOSE HIGH DOSE CONTROL (UNTH ) CONTROL (UNTH)

31-3070 01-0118

«A3"MAL (1*7, (47)

CCRTICA L ADENOM A 1 (2? ) 

CORTICAL CARCINOM A 

PHF.OCHHOMOCYTOM A 10 (21i5 ) 7 (15X ) PHEOCHROMOCYTOMA , HALIGSAN T 1 (2J5) G A NGLIOTEUPOM A ' (2« ) 

• THYROI D (39 ) (43 )

ADENOCARCINOMA , NO S 

FOLUCULAP-CFL L ADENOM A 

FOLLICULAR-CEL L CARCINOM A 

C-C!ILL ADFNOM A 

C-CELL CAHCINOM A 1 (2%) PAPILLAR Y CYSTADE«OCARCINOMA,NO S 

*EARATHYPOI D (23) (28) ADFNOMA , NO S 1 (4%) 

tPA^C'EATI C ISLET S (45 ) (46 )

ISLET-CELL ADENOM A 3 (-n>) ISLET-CEL L CARCINOM A 

R^PPODGCTTV E SYSTE M 

*«AK«AR Y GLAN D (48) (48 ) PAPILLAR Y ADENCCARCINOM 4 1 (2'.) 

"IBP.CADEHCM A 1 (2X ) 

• PREPUTIA L GLAN D (1*8) (48)

CASCI10MA,NO S 2 C*^) 

SQUANOU S CELL CAPCINOM A 

ADENCMA , NOS 

CYS7ADENOMA , NO S 

•TESTI S (47 ) (47 )

INTERSTITIAL-CEL L TQflO R 4 5 (96%) 42 (89*)

NERVOU S SYSTE M 

»CERESR'J1 (1*7) (48)

USTROCYTOM A 

(147) 

* NUMBE R OF ANIKAL S ilITH IISSU? EXA.1INSD MICROSCOPICALLY 

* N'JIBE R OF ANIMAL S N3CROPSIE D 

 LOU COSE 

 01-0069 

 (48) 

1 (2%) 

6 (131) 

 (47 ) 

1 (2X) 

1 (2X) 

2 (4% ) 

2 (4X ) 

1 (2%) 

 (25) 

 (41*) 

3 (7X) 

1 (2*) 

 (50 ) 

 (50) 

1 <2X ) 

2 (4%) 

 (47 ) 

 37 (79X) 

 (48 ) 

HIGH DOSE 

01-0121 

(48) 

4 (8%) 

1 (2*1 

13 (27% ) 

1 (2%) 

(47) 

2 (4X ) 

1 (2% ) 

1 (2% ) 

(19) 

(46) 

(48 ) 

(U 9 , 

2 (4%) 

3 (6 S) 

1 (2%) 

(48) 

16 (33%) 

(48) 

1 (2%) 

(48) 

' A-5 
TABLE Al (CONTINUED) 

CONTHOL (UNTS)

01-0070 CONTROL (UNTR) 01-0118 LOH COSE 01-0069

 HISH DOSE 

 01-0121 

•C^RFORA L CORTE X

AST30CYTOMA

 (U7 ) (U3 ) (18)

 1 (2%} 

 (»8) 

SPFCIA L SENS E CFGAN S 

*Z!"-:SAL'S 3LAN D

SCUAtinn S CH1L CARCINOI A

^"BAC^OU S ADENOCAECINOM A

c^auniious CARCINOM A

 (K8 ) (48) (50)

 1

 1

 (2% ) 

(2%)

 (1(8) 

2

 1

 (418) 

 (2<) 

KUSC'JLOSK rLETAL SYSTE;1 

N01" 

BODY CAVITIES 

*BOPY CAVITI"S

A,

U,

 NOS

 .".ALIGNANT

 (U8)

 1 (2*)

 («8)

 2 (tX)

 (50)

 <t

 1

 (8*)

 (2*)

 (18) 

1

 1

 (2*) 

 (2%) 

A L L OTH^ ? SYSTEM S 

N0'lr 

ANIMA L DISPOSITION SU1HABY 

ANI1AL5 INITIALLY IN STUDY 

NAT'IFAL DEATHS 

10RTEUND SACRIFICE 

SCHFCUL^D SACRIFICE 

ACCIDENTALLY KILLED 

UNIKAL MISSING ANIMA L DELETED (WBONG SEX) 

51 

5 

5 

5 

3U 

1 

50 

6 

8 

5 

30 

1 

50 

9 

38 

50 

5 

45 

* N"BEr" OF ANIMALS dlTH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBE R OF ANIMAL S NECROPSIED 

A-6
 
TABLE A! (CONCLUDED)
 

--—­ LOW DOSE HIGH DOSE CONTRO L (UNTR ) CONTROL (UNTE ) L OW COSE HIGH DOSE 

J1-3371 01-0118 01-0069 01-0121 

TUMOR SUMMAR Y 

TOTAL ANIMAL S WIT H PRIMAR Y TUCORS* U5 44 47 48 TOTA L PRIMAR Y TU10RS 31 80 137 156 

TOTAL ANIMAL S WITH BENIG N TU»CRS 45 43 44 34 

TOTA L BENIG N TUMOR S 66 62 I C O 69 

TOTAL ANIMAL S WITH 1ALIGNHN I

 TU10RS 10 17 28 45 TOTA L MALIGNAN T TUMOR S 13 13 30 85 

TOTAL ANIMALS WITH SECONDARY TUMOSS* 2 1 1
 

TOTAL SECONDARY mUMO'i3 3 1 1
 

TO"AL ANIMALS MI^H TJMORS UNC'EHIN­ B-NIGN OR MA1I3NANT 2 7
 

TOTAL UNCERTAIN "UMORS 2 7
 

TOTAL ANI1ALS WITH TUMOES UNCIRTAINPFIMAKY 08 METASTATIC
 

'OTAL UNC^PTAIN "U10RS
 

* PRIMAR Y TUMORS: ALL TUMOrtS EXCEPT SECONDARY TU10RS
 

* SDCONDAFY TUMORb: M£TASTATIC TUMORS OR TUMORS INVASIVE INTO AN ADJACENT ORGAN
 

A-7
 
TABLE A2

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE RATS TREATED WITH 5-NITRO-o ANISIDINE 

LOW DOSE HIGH DOSE 

CJNTEOL(UNTS )

02-1070

 CONTRO L (UM8)

 02-0118

 LOW DOSE

 02-0069

 HIGH DOSE 

 02-0121 

ANI-HLS INITIALL Y IN STUD Y 50 50 50 50 

mmLS MISSIN G 1 

ANIMM 3 NECKOPSIt D 49 50 49 46 

AM1ALS "X^IINF D HISTOPATHOLOGICALL Y ** 4 9 50 49 46 

IN^GUMENTAEY SYSTEM
 

*S<TN (U9) (51) (49) (46)
 

SQU^MO'JS CELL CA3CIN01A 2 (4X) 3 (7%)
 

BASAL-CFLL CARCINOMA 1 (2X) 1 (2X)
 S'B^CnUS ADENOCARCINOMA 1 (2X) 2 (4X)
 SASCPMA, NOS 1 (2X)
 

"131QV\ 1 (2%)
 

'TOHOSA'iCOI'A 1 (2X)
 

'SJ3CJT TISSUE (49) (50) (49) (46)
 FIBPC"U 1 (2X)
 FI3ROSARCOMA 1 <2X)
 

SYSTE1
 

*LUM 3 (49) (50) (49) (13) 

SCJAMOJ S CFLL CARCINOMA , METAST A 1 (2*) 

ALVEOHF/BRONCHIOLA R ADENOM A 1 <2X ) 4 (8S) 

ALVTCLAF/aPONCHIOLAii CASCINCMA 1 <2X) 1 <2X) BHABD01YOSAi:COMA, BETfSHTIC 1 (2«) 

OSI-OIA 1 (2X) 

HEMATOFOI^TIC SYSTEM
 

*l'JLTIPLr OP»AdS (49) (50) (49) (46)
 

•IALIGN\NT LYMPHCMA, NOS 2 (4*) 1 (2X)
 

UNCIFF^PENTIATED LEUKE1IA 1 (2X) 2 (4X)
 

fYELCIONOCYTIC LEUKEMIA 5 (10X) 3 (6X)
 

(U9) (4B) (48) (43)
 

H'lAN^IOSArCOWA 1 (2X)
 

t NUKBE T 0" ANIMAL b WIT H TISSUE 2XA1INSD MICROSCOPICALLY 

* NUMBE R OF ANIMAL S S-CROPSIED 

**EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

A-8
 
TABLE A2 (CONTINUED) 

LOW DOSE HIGH DOSE CONTfiO L (UNTS )

02-0073

 CONTRO L (UNTR ) 

 U2-0118 

LOrf TOSE 

32-0069 

'JNCI^RFNTtATr D LEJK--I A 1 (2S) 

CI=CULATOP Y SYSTF 1 

« H ­: A a r (49) (53) (4"} "I3=OSARCOI« A 

DIGESTIV E SYSTT I 

*SALIVAS Y GLAN H (•49 ) (50 ) (45 ) CARCINOMA , NOS 

* L I V E =t 

NSOPLA5TI C NODUL E 

(149)

2 (4i) 

 (53) (49) 

w^PATOCILLULA " CARCIN01 A 1 (2J) 

IHL'.W (49 ) (M) (49 ) 

LTICI Y D^AFCur A 1 (2% ) 

UPI'IAR Y SYSTE M 

«K:PN: Y (I. 3) (50) (49 ) 

CA^CINOCA.NO S 1 (2« ) TPANSiriONAL-Cil L CAFCINOM A 

tUfllA^ Y 3HDCFS (49) (« 6) (4b ) 

^iUAHO'J S CrL L CARCINOM A 

-aANSITICNAL-CFL L PAPILLO-1A 

EVDOCOIN E SYSTF.1 

^PI-UITA^ Y 

• DEM01H , NOS 

(1414 )

H (U1X )

 (4J) 

 17 (431 ) 

(-16) 

10 (22*) 

HAD1ENA L (149) (49 ) (49 ) COBTICA L APCNOK A 1 (2%) 2 (U« ) rORTICU L CAPCINOM A 

PHrocHSnNOCY^OI A 

1 (2? ) 

2 («1) J (6S) 3 (6S) 

*A3PE"A L 1"DOLLA (49 ) (49 ) (49 ) 

* NU1BE R 0' ANI1AL S JITH TISSUE EXAMINE D MICROSCOPICALL Y 

* M^IBF S O F ANIMAL S NECROPSIE D 

HI3H DOSE 

32-1121 

(13) 

1 (24 ) 

(143 ) 

1 (24) 

(414) 

1 (251) 

1 (2%) 

(4 j) 

(15) 

1 (21) 

(12) 

1 (2? ) 

1 (21) 

(141) 

5 (121) 

(11) 

3 (71) 

6 (141) 

(11) 

A-9 
TABLE A2 (CONTINUED) 

LOW DOSE HIGH DOSE 

CONTtiOL (UNT3) CONTROL (UNTR) 

02-1370 02-0118 

•i K?:TC (4") (45)

 

"OLIIC'ILAS-CSLL CAPCINOH A 1 (U) 1 (2*)

 

C-C-LL ADENOM A 2 (5? ) 1 (2*)

 

C-CILL CASCTNOKA 1 (3«) 1 (2*)

 

PAPILLAR Y C/STADriOilA, NOS 

«;A1CB"*T'r i* --s (47) (48)

 

ISLET-JTLL ADFN01A 1 (2%) 2 (4X)

 

Rr PnOCJCIIVr SYSTE M 

* WMA'Y ,LAN D (49 ) (50)

 

£PINCM!\, NC5 2 (4?)

 

AD 3NOC ^ "'CINO^A , N05 

^PILU'Y ADiNCrASCINP.M P

I dR° A")r >.0fl 3 16 (33-4 ) 19 (38*) 

*C^ITJ-»AL TLAND (49) (50)

 

r»'CI\'TU, NOS 

SiLiA-'OI'S CrLL 3APILLOBA 1 (2X1

 

S^JfOJ; CELL C«BCINC1A 

Ar^uclA , KC3 1 (2X ) 2 <4X)

 

AD~NCC",:;CINnm, NOS 

PJiTLH^Y ADENOM A 

*VAT I 4A (49) (50)

 

-••AtiaHP-LFLi T'JMOt, B'NISN 

mr^Rus (49) (50)

 

CA^:INOMA-TN-S!TJ , NOS 

'J\3:iTF-'i;r>JTIA"rD CARCINOMA 

Ar'NccARnt'OMA, ios 1 (2X) 

>D""TTA IN ADrNOMATOUS POLYf 

'APILUBY ; C"NOCAPCINO(1A 

SASTCH , NOS L'ICIYT'APCOfIA 1 (2*.) 

JNDOI;T~UAL ST^OWA L POLYP 12 (24*) 10 (20*) 

"NCOM'TilAL STfOMAL SAKCOMA 1 (2*) 

*n-"»15/EMD01'-I'?IUW (49 ) (50)

 

AP'MCC^RCINOIA, MOS 2 (4*)

 

(47) (49)

 

*

O"-"IN.,L.S&-C-U ujflM

 

» J'I*IF"R T ,'Mi'ALS WITH TISSUE EXAMINED 1ICROSCOPICALLY
 

* NJ»L<- I JF ANIMAL S NECROPSIED
 

, 

LOU DOSE 

02-0069 

HIGH DOSE 

02-0121 

(49) 

3 (6X) 

(42) 

1 <2X) 

1 (2X) 

1 <2X) 

(44) (43) 

(49) (46) 

10 (20*) 

1 (2*) 

4 (8X) 

4 (9X) 

1 (2X) 

(49) (46) 

7 (15X) 

1 (2X) 

9 (18X) 

1 <2X) 

4 <8X) 

2 (4X) 

5 (11X) 

(49) (46) 

1 (2X) 

(49) 

2 (4X) 

1 (2X) 

1 (2X) 

2 <4X) 

14 (29X) 

1 <2X) 

(44) 

1 (2X) 

1 (2X) 

2 <5X) 

1 (2*) 

6 (14*) 

(49) (44) 

(48) (45) 

A-IO
 
r

 

TABLE A2 (CONTINUED) 

ROL(UNTS )

02-0070

 CONTRO L (UNTR )

 02-0118

 LO H COSE

 02-0069

 HIG H DOSE 

 02-012 1 

NEPV1US SYSTEM 

*C2REBRU M

ASTRCCYTCM A

*3PAI N

GLICBA , NOS

OLIGODSNDRCGLIOMA

.1EDU1L53LASTOH A

 (49 )

 (1(9)

 1 (2*) 

 (5'">)

 (50)

 (t« )

 (49 )

 1 (2% )

 1 (2S) 

( K1*) 

2 (5% ) 

 (14) 

1 (2*) 

SPECIA L SENS E ORGAN S 

*ZYMBAL' S GLAN D

SCDAMOU S CELL CARCINOMA

SEBACEOU S ADENOCARCINOM A

 («9 ) (50 ) (U9 )

 2 (t«)

 (1*6) 

2

 U

 (UJ) 

 (9*) 

ML'SC'JLOSK'IL'ilTAL SYSTE M 

*MOSCL 3 HIP/THIGH

RHABDCflYOSARCOM A

 (19) (50 ) («9 ) («6) 

1 (2% ) 

BOD Y C^VITI^ S 

tPEHITONKaM

lESOTH^LIOWA, NOS

 (1(9)

 1 (2?) 

 (50) (49) («6) ­

ALL OTHER SYSTEMS 

SI'S UNKNOWN 

SQUAMOJS CFLL CARCINOMA 1 

ANIPUL DISPOSITION SUMMARY 

ANIMALS INITIALLY IN STUDY

NATURAL DEATHS

.10P.IEUND SAC2IFICS

SCHECULED SACRIFICE

ACCIDENTALLY KILLED 

TERMINAL SACRIFICE

ANIMA L KISSING

 50

 3

 7

 5

 35

 50

 5

 3

 5 

 37

 50

 13

 26

 11

 50 

 15 

 23 

 11 

1 

* NUHEER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBE R OF ANIMAL S NECROPSIED 

A-l l
 
TABLE A2 (CONCLUDED)
 

HIGH DOSE CONTROL (UNTR ) CONTROL (UNTR) L OH COSE HIG H DOSE 

02-0070 02-0118 02-0069 02-0121 

rjioa SUMIHR Y 

TOTAL ANIMAL S WITH PRIMAR Y TUMORS* 45 38 45 «1

 

TOTA L PRIMAR Y TUMOR S 73 73 8« 78

 

TO^H L ANIMAL S KIT H BENIG N TUMOR S 37 35 27 21 

TOTAL BENIG N TUMORS 5« 59 55 29 

TO-AL FINAL S WIT H MALIGNAN T TUMOR S 13 12 29 36

 

TOTAL MALIGNAN T TUMORS 16 13 29 US

 

"OTAL ANI1ALS KITH SECONDARY TUMORS* 1
 

TT"A1 35CONDARY TUMORS 1
 

'OTAL ANIMALS BITH TUBOFS UNCFRTAINB^NIGN 03 MALIGNANT 3 1
 

TOTAL UNCEPTAIN T'JMORS 3 1
 

TOTAL ANIMAL S WITH TUMORS UNCESTAINFDI1A! 1Y )R MTTASTATIC
 

"OTAL 'JNCERTAI!" TUMORS
 

* FFT-UFY ru-HORS: ALL TUMOE3 EXCEPT SECONDARY TUMORS
 

« SECONDARY TUflORS: M5TASTATIC TUMORS OR TUMORS INVASIVE INTO AN ADJACENT ORGAN
 

A-12
 
T
 

APPENDIX B
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS
 

IN MICE TREATED WITH 5-NITRO-o-ANISIDINE
 
r

 
TABLE Bl

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE TREATED WITH 5 NITRO-o-ANISIDINE 

DOSE A DOSE B 

CONTROL (US T t) CONTROL (UNTF ) 

05-0070 05-0116 

ANIMAL S INITIALL Y IN STUDY 50 50 

ANIMALS MISSING 1 

ANIMAL S NECSOPSIED 50 19 

ANIMAL S FXAMINE D rilSTCPATriOLOGIC A LL Y ** 50 19 

INTEGUMENTAR Y SYSTEM 

HONE 

RESPIRATOR Y SYSTE M 

ILUN G (50) (19) 

HhPAIOCELLULA R CABCINOBA , METAE T 1 (2%) 1 <2X ) 

ALVEOLAB/3RONCHIOLA R ADENOM A 5 (10%) 

ALVEOLAR/BRONCHIOLA K CARCINOM A 5 (10?) 5 (1C)S) 

HEMANSIObAFCOMA , 1ETAST ATIC 

HEHATOPOIZTIC SYbTEC 

*MULTIPLF ORGANS 

MALI3NAN T LYMPriOMA , NO S 

HALI&.I YMPHOKA , HISTIOCYTIC T1PE 

(DO) 

2 (15) 

1 (2S) 

(19) 

3 (61) 

MASI-CILL "HMOS 

GRANOLOCYTIC LECKEMIA 

tSPLEEN (50) (19) 

HEMANGIOSARCOM A 

HALIS.LYMt'HONA. HISTIOCYTIC TYPE 

1 <2S) 

1 (2%) 

«LYMPH NODS 

MALIG.LYHPHOBA , HISTIOCYTIC TYPE 

(15) 

2 (IX) 

(12) 

1 (2X ) 

iHESENTEBIC L. NODI (15) (U2) 

MALIG.LYMPaOMA , HISTIOCYTIC TYPE 

tPEYEES PATCH (19) (19) 

BALIG1.ANT LYMPHOMA , NOS 

tEUODENU M (19) (49) 

fliiJafiiNT LYHPHOBA . fiOS__ _ 

t NUMBE R OF ANIMAL S KITH TISSUE EXAMINE D MICSOSCOtICALLY 

* SOBBER. Of ANIHALS NECBOPSIED 

"EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

DOSE A

05-0071

50

18

48

(46)

5 (10X ) 

4 (8X)

1 <2X )

1 (2X) 

(48)

1 (2X)

(45)

1 (2X) 

(10)

(10)

3 (8X)

(18)

(48)

1 (2X) 

 DOSE B 

 05-0102 

 50 

 49 

 18 

 (17) 

1 (2X) 

1 (2X) 

 (19) 

1 (2X) 

1 (2X) 

1 (2X) 

1 (2X) 

 (17) 

 (10) 

 (40) 

2 (5X) 

 (45) 

1 <2X) 

 (45) 

B-3 
TABLE Bl (CONTINUED) 

DOSE A

CONTROL (UnTK )

05-0070

 DOSE B 

 CONTROL (UNTu )

 05-011fi

 DOSE A 

 05-0071 

CIRC ' -A n- v ,\ 3TJr 

NON J 

*LIVi(

i rpATOL-'LLuLA ? Au£SOM A

i^pATnL-^LlJU t CA"CIMO(H

,-IAI.aiOr A

n^'A N .ijbUCOK A

-MUSoIOSA'COriA , UN C PtilM Of MET

 (50)

 12 (24« )

 1 (2X ) 

 (M8 )

 2 (4X ) 

o (13%}

 1 (2S) 

 (48) 

 25 (52X) 

1 (2X) 

»Sir»A n

l i b \1PI S _ rLl. tAPILLOM A

 (i)9) («8) (H8) 

1 (2it) 

URIbAS Y r-vSTr " 

NON 5 

^NDOr^IN L fYST1"1 

tPIIJITAF Y

>D-N3»A , UOS 

»ADF:NA L cOIiTICAL ADENOM A 

FHEOCMROHCCYTOMA

 (1)6)

 (u9j

 (tO)

 (n«)

 1 (2*) 

 (Ml) 

 («u) 

'THYROID

Al)ENOCARCI!!C«A, NOS

"jLLICULAk-CtLL CAPCINOMA 

 (110)

 1 (3X) 

 (15) (H7) 

«PANC'itA''IC ISLETS

ISLc.T-Cc.LL \D£hOHA

 (116)

 1 (2S) 

 (17) (U7) 

EEPRCDUCTIVi SfSTS.1 

• TrsTIS

INTLr STITIAL-CELL TUMOS

 (50) (<49) (H8) 

1 (2X) 

NEFV'JUS bYSPtl 

DOSE B 

05-0102 

3 <6X) 

2 (4X) 

(U6) 

1 <2X) 

(38) 

3 (8X) 

C<6) 

1 (2X) 

(«7) 

1 (2X) 

(43) 

(16) 

1 (2X) 

* NUKBt P OF ANI1ALS WITH IISSOE EXAKIHE D MICROSCOPICALLY 

* NUMBr S CF AMIKAL S NBCPOPSITD 

B-4 
TABLE Bl (CONTINUED) 

DOSE A DOSE B 

CONTROL (U Nil-) CONTROL ( (JNTIi) ' DOSE A DOSE B 

05-0070 05-0118 05-0071 05-0102 

SPECIA L 5_hS T 0 D TAN S 

•hAFOctlA r CLAN D (50) (4J) (49) (1*9) 

ADLNCMA , NOS 1 (2*) 

PAPILLA" ? ADJNOr A 1 (2*) 

MUSCULOSK^LZ'AL SYSTjJK 

NO» "! 

BODY CAVI'lES 

NO'.T 

ALL OTH H3 SYSTEfS 

»rULriPLr OFGAN S (50) (49) (US) («9) 

\")UPOFIBhr3AFCO« A 1 (2X) 

ANIMA L DISIOSITION SU1MA3Y 

ANI.IALS INITIALLY IN STUDY 30 50 50 50 

N-IT'JFSL DEATH* 2 3 6 

"JfilP'JMD SACPIFICE 

SCriSJIILED iAC5IFICE 5 10 5 

ACLlTflTALLY KILLFD 

"Jr/IKA L SULFIFICI 43 39 42 U« AM«AL 

t NUMEL f OF AMhALS WITH TISSUE 'XA«INtD (SICSOSLOPZCALL Y 

* KUHBT S OF ANI'iAL S NECPOPSIilD 

B-5 
r

 

TABLE Bl (CONCLUDED)
 

DOSE A DOSE B 

CONTROL (UNIfi ) CONTROL (U N T.i) DOSE A DOSE B 

05-0070 05-0118 05-0071 05-0102 

TUBO ? SU1MAE I 

TOTAL ANIMAL S WIT H PSIMAR V TUhOPS * 2 3 22 32 17 

TOTA L PtUMAR X TU10FS 27 26 40 21 

TOTAL ANIMAL S WIT H EEKIG N TIIMOB S 3 8 7 7 

TOTA L BENIG N TUMOR S 3 8 7 7 

TOTAL ANIMAL S WITH MALIGNAH T TUMOR S 72 15 29 1 1 

TOTA L KA1IJNAN T TUMOR S 24 17 33 13 

TOTA L A1TMAL S MTT H SECONDAR Y TUMOHS * 1 1 b

 

TOTA L .SECOMDAE I TUMOR S 1 1 6 

TOTAL ANIMAL S WITH TUMORS UNChRTAIN-


BtNIGN JK MALIGNAN T
 

TOTAL UNCERTAIN TUMORS
 

TOTAL ANIMALS WITH TUMORS UNCEBTAINFn.iAfiy OR K-;TASTATI C 1 

TOTA L USCE.-iTHI N TUMC3 S 1 

* PnllAi- ? 7U1O?3: AL L TUMOk S SXCEP 1 SE^CItL'A& Y TUM03 S 

* StCQNDAP Y TUMOfS : METASTATI C TUMOiiS OF . TUMOE S INVASIV 3 INT O A N ADJACEN T ORGA N 

B-6 
TABLE B2

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICE TREATED WITH 5-NITRO-o-ANISIDINE 

DOSE A 

CONTROL (UN IK) 

06-0070 

DOSE B 

CONTROL (DNTF)

06-0118

 DOSE A

 06-0071

 DOSE B 

 06-0102 

ANIKAL J

ANIKAL S

ANIMAL S

ANIMAL S

 INITIALL Y I K STUD Y 

 MISSIN G 

 NECPKSIE D 

 EXAMINE D HISTOPA THOLOGIC A LL Y 

50 

Ui) 

** 47 

50

50

50

 50

1 

 43

 43

 50 

 45 

 44 

INTEGUMENTAR Y SYSTE M 

*SKI'J 

17

 13 HO E A B CO HA 

(48 ) (50) (43)

1 (2X )

 (45) 

1 (2X) 

&ESFIRA:0= Y SYSTE M 

*LIJNT 

tIJOPLASM , Nd S 

HEEMCCELLULA 8 CARCINOMA , .l^TAST 

ALV"OLAa/ERONCHIGLA R ADENOM A 

lUVEOLAR/BRONCriMIA R CARCINOM A 

ODT.:OS Apco;iA, METHSTATI C 

(46) 

1

2

1

 (2%} 

 (4*) 

 (2*) 

(50)

2

1

 (4%)

 (2% )

 (42) (44) 

1 (2X) 

1 <2X ) 

1 (2X) 

1 (2X) 

HEMATOPOIFTIC 5Y3TEK 

*«ULTIPL E ORGAN S 

fiALIGNAU T LYMPHOMA , NO S 

MAL I j.LYKPhO M A, HISTIOCYTIC TYP E 

LY"?HOCYri C LEUKEMI A 

iPYfrlrOCYII C LEUKEMI A 

GRAJJLOCYTI C LEUKEMI A 

(48 ) 

2 (45) 

1 (2*) 

1 (2%) 

(50)

2 (4X)

 (43)

 2 (5t)

1 <2X )

 (45) 

3 (7X) 

5 (11X) 

1 (2X) 

*SFL2F N 

HIIANGIOSARCOM A 

1ALIGNAN T LYMPHCflA , NO S 

(47) 

1 (2%) 

1 (2%) 

(49 ) (38)

1 <3S) 

1 (3*) 

 (43) 

tMESiNITFIC L. NOD E 

MALIG.LY1PH0.1A , HISTIOCYIIC TYP E 

(36) 

1 (3*) 

(44 ) (27) (30) 

tPEY^r a PATCH (45) (48) (33) (41) 

t NLR3Ec , OE ANI1AL S KITH TISSUE EXAMINE D

* NU13E S O F ANI1AL S NECFOPSIE D 

**EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

 MI CHOSCOPICALLY 

B-7 
TABLE B2 (CONTINUED) 

DOSE A

CONTROL (U (IT 6)

06-0070

 DOSE B 

 CONTROL (ONT8 )

 06-0118 DOSE A  06-0071 DOSE B 

06-0102 

"ALIG. LY HPHOMA ,

*KIDNE Y

PALI'J.LYKPHOMA ,

 HISTIOCYIIC

 HISTIOCYTI T

 TYP S 

 TYP E

 (45) (50 ) (42) 

1 (2*) 

1 (2X) 

(43) 

CIPCULATOF Y SYSTE M 

my. 

DIGESTIV E SYSTE M 

*LIV=: P hEFAIOCTLLULA ? CAPCINOM A

*GALi.BLADDS 3

PAPILL01A , NO S 

• STOMAC H

SjjAICU S v-FLL PfFILLOM A

 (47 ) 2 (4% )

 (4« )

 (45)

 (50 ) 1 (2% ) 

 (50)

 (49)

 (41 ) 

 (43) 

 (30) 

1 (3X) 

(43) 

d (19X) 

(45) 

1 <2X) 

(40) 

UPINAS Y SYSTE M 

NOUS 

ESDOCBISIE SYSTE M 

• PITai'AP Y

ADENOMA , NOS

CHROMOPHOB H ADENOM A

 (43)

 5 (12*)

 (4^ )

 1 (2X)

 2 (5X) 

 (31) 

1 (3X) 

(33) 

4 (12X) 

*ADfNA L

COPTICA L ADENOM A

 (47 )

 1 (2%) 

 (43) (34) (40) 

• THYhOI D

FOLLICULAR-CLL L ADENOM A 

 (41 ) (44 ) (34 ) (43) 

1 (2») 

EEPPODUCTIV E SYSTEM 

IliTERII S

ADENOCARCINCf1* x NOS

 (43) (47)

 _ 

 (32) (36) 

1 (3X1 

t NUMBE P OF ANIMALS KITH TISSUi) ^XAMINED MICROSCOPICALLY 

* NUMBES CF ANIMAL S NECFOFSISD 

B-8
 
TABLE B2 (CONTINUED) 

DOSE A DOSE B
 

CONTROL (UNTR) CONTROL (UNIB) DOSE A


06-0070 06-0118 06-0071

LEIOMYO A 1 (2*) 

«OVARY/OVIDUCT (13) (47) (32)


PAPILLARY ADENOMA 1 (21) 1 (2%)
 

fOVAB Y (U5) (KB) (30)

GHANULOSA-CELL TUMOR

TOBULAP ADENOMA


NERVOUS SYSTZI
 

NONE
 

SPECIAL SENS F ORGAN S 

*HARDEBIA N Gi.AND (UK) (SO) (U3)

ADENOMA , NOS 1 (2X) 

PAPILLAPY ADENOM A 1 (2X) 

HUSCOLOSKELETAL SYSTEM 

NONE 

BODY CAVITIES
 

NONE
 

ALL OTHEP SYSTEMS
 

CHENTDH
 

BERANGIOHA
 

HEHAHGIOSABCOHA
 

( VOHBdB OF ANL1ALS RITH TISSUE EXACINED BICEOSCOPICALLY
 

** iOMBEK OF ANIMALS NICF.OPSIED
 

 DOSE B
 

 06-0102 

 (36)
 

 (37) 

1 (3X) 

2 (5X)
 

 (45) 
TABLE B2 (CONCLUDED) 

DOSE A DOSE B 

CONTROL (UNTE) CONTROL (UNTE) DOSE A DOSE B 06-0070 06-0118 06-0071 06-0102 

ANIMA L DISPOSITION SUMMA5 Y 

ANIMAL S INITIALL Y IN STUJY 50 50 50 50 

NATURA L DEATHS b 2 18 15 

KOFIfaUSD SACRIFICE 3 10 1 

SCHEDULE SACRIFICE 5 10 5 

AC-CIDTNTALL Y KILLE D 1 

TERMINA L SACRIFICE 36 38 16 33 

ANIfA L MISSING 1 

A INCLUD- S AUTOLYZI D AIII.1ALS 

TUMO R SUMMAR Y 

TOTAL ANIMAL S KITH EFIMA3 Y T'JHOhS* 18 10 9 22 

TuTAL PRIMAP Y TU10ES 22 11 9 35 

TOTAL ANIMAL S KITH BENISN TUMOh S 9 ^ 2 9 

TOTAL S'NISN TUKOES 9 7 2 11 

TOTAL ANIMAL S WITH MALIGNAN T TUKCSS 12 » 7 16 

10TAL MALISNAN T TU«OPS 13 H 7 22 

TOTAL ANIMAL S WITH SECONDARY TUMORS* 2 1 

TOTAL SFCOHOARY TUMORS 2 1 

TOTAL AKIMALS WITH TUHOhS IIHC2RTAINBEHIG N CF MALIGNAN T 2 

TOTAL UNCERTAI N TUM08S 2 

TOTAL ANIMALS WITH TUMORS UHCEPTAINEFI1AFY Ot RET* STATIC

 

TOTAl UNCERTAIN TUMORS 

«> PBIHAfc* TUKORS: ALL TUMOES EXCEPT SECONDARY TUBORS 

t SFCONDAEY TUHORS: M5TASTATIC TUROSS OR TUMORS INVASIVE INTO AN ADJACENT OHGAH 

frflO 
T
 

APPENDIX C
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN RATS TREATED WITH 5-NITRO-o-ANISIDINE
 

TABLE Cl

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS 

TREATED WITH 5-NITRO-o-ANISIDINE

 

LOW DOSE 

CONTROL(UNTR) 

ni-0070 

"HIGH"DOS£ CONTROL (UNTR)

01-0118

 LOW COSE 

 01-0069 

HIGH DOSE 

01-0121 

ANIMALS INITIALLY IN STUDY 

ANIMAL S MISSING 

ANIMALS HECROPSIED 

ANIMALS EXAMIKED HISTOPATHOLOGICHLLY 

50 

1 

48 

** 48 

S50

48

48

 50 

 50 49 

50 

48 

48 

INTEGUMENTARY SYSTEM
 

*SKIN (48) (48) (50) (48)
 

CYST, NOS 1 (2%)
 

EPIDERMAL INCLUSION CYST 3 (6%) 2 (4%)
 

SEBACEOUS CYST 3 (6*)
 ABSCESS, NOS 1 (236)
 INFLAMMATION ACTIVE CHRONIC 1 (2*)
 

ATYPIA, NOS 1 (21)
 HYPERPLASIA, NOS 1 (2*) 1 (2%)
 

•SUBCUT TISSUE (48) (48) (50) (48)
 ABSCESS, NOS 1 (2%)
 METAPLASIA, OSSEOUS 1 <2X)
 

RESPIRATORY SYSTEM
 

KTRACHF.A (45) (48) (47) (48)
 INFLAMMATION, NOS 2 (4*)
 INFLAMMATION, ACUTF/CHRONIC 13 (40S) 3 (6X)
 INFLAM4TION, CHRONIC 1 (2%)
 

KLUNG/EFONCHUS (43) (49) (49) (48)
 BRONCHIECTASIS 3 (6*) 1 (2*) 1 (2*) 2 (4 t)
 

INFLAMMATION, NOS 7 (15*)
 

*L'JNG/BFONCHIOLE (48) (48) (49) (48)
 

HYPEEPLASIA, FOCAL 1 (2*)
 

• LUNG (48) (48) (49) (48)
 

CONGESTION, NOS 1 (2?) 1 (2%)
 FDEMA, NOS 1 (2%)
 

HEMOBHHAGF 1 (2*)
 

^
 aa
 t NUMBE R O F ANIMAL S WIT H TISSOF EXAMINE D MICROSCOPICALL Y 

* NU1BEP OF ANIMAL S NECROPSIED **EXCLUDES

 PARTIALLY AUTOLYZED ANIMALS 

3 50 4NINALS WER E INITIALLY IN 'HE STUDY, 3UT ONE ANIMA L WAS FOUND TO BE A FEMALE IN A MALE GROUP. 

03 
TABLECl (CONTINUED) 

LOW DOSE HIGH DOSE 

CONTROL (DNTH) CONTROL (UNTH)

01-0070 01-0118

INFLAMMATION , INTERSTITIA L 1 (8X) 

INFLAMMATION , NECROTIZIN G 1 (2X) 

ABSCESS, NOS 1 (2%) 

PNEUMONIA , CHRONI C MU8IN E 1 (2*) INFLAMMATION , CHRONIC 

GRANULOMA, NOS 1 (2X) 

GPANULOBA , FOREIG N BODY 

PERIVASCULITI S 

HYP"RPLASIA , EPITHELIA L (211) HYPEBPLASIA , FOCAL 1 (2*) 

HYPERPLASIA , ALVEOLA R EPITHELIU M 1 (2%) 

HEMATOfOIETIC SYSTEM
 

#3UNE MAFROW (47)
 THROMBOSIS, NOS
 

NSCROSIS, FOCAL
 

"YELCFI3FOSIS 1 (2X)
 HYPE3FLASIA, HEMATOEOIETIC 1 (2*)
 MYPSRPLASIA, JKANULOCYTIC 1 (2*)
 HYPEPPLASIA, MEGAKARYOCYTIC 1 (2X)
 MYELCPOI5SIS
 

• SPLEEN- (i*8) (US)
 

FIBROSIS 1 (2X)
 

FIBROSIS, FOCAL
 

HEICSIDSROSIS 1 (2X)
 

ATROPHY, NOS
 

LYMPHOID DEPLETION
 

HYP?SPLASIA, NOS
 

HYPEHPLASIA, HEMATOEOIETIC 9 (19X)
 

HYPERPLASIA, ERYTHROID 10 (21*)
 

HYPERPLASIA, HETICULBfl CELL
 

HEMATOPOIESIS
 

"RYTHROPOIESIS 1 (2%)
 

• LYMPH NODE (t2)
 HEMORRHAGE 1 (2X)
 

ATROPHY, NOS
 

PLASMACYTOSIS 1 (2X)
 

HYPERPLASIA, LYMPHOID 3 (7X)
 

tMANDIBULAR L. NODE <K2)
 

NUMBER OF ANIMALS KITH TISSUE EXAMINED MICROSCOPICALLY
 

NUMBER OF ANIMALS NECROPSIED
 

C-4
 

 LOU DOSE 

 01-0069 

1 <2X)
 

1 <2X)
 

(17)
 

2 (IX)
 

1 (2X)
 

1 <2X)
 

U (9X)
 

(38)
 

(38)
 

HIGH DOSE 

01-0121 

1 (2X)
 

1 <2X)
 

2 (UX)
 

(16)
 

1 <2X)
 

1 (2X)
 

1 (2X)
 

3 (7X)
 

2
 

(18)
 

6 (13X)
 

2 (IX)
 

1 (2X)
 

16 (33X)
 

1 (3X)
 3 <8X)
 

(10)
 
TABLE Cl (CONTINUED)
 

HIGH DOSE
 CONTROL (UNTR) (:ONTROL (ONTB) L3rf DOSE
 

01-0070 01-0118 01-0069
 

ATROFHY, NOS
 

HYPEBPLASIA, NOS 1 (2%)
 

*MEDIASTINAL L.NODE (42) (44) (38)
 ATROEHY, NOS
 

»PANCREATIC L.NODE (42) (44) (38)
 HYPERPLASIA, NOS 1 (31)
 

tNESENTERIC L. NODE (U2) (44) (38)
 ATROEHY, NOS
 

tTHYMIC CORTEX (24) (23) (34)
 HEMORRHAGE
 

CIBCULATCRY SYSTEM
 

*H3ART (48) (48) (49)
 FIBBOSIS, FOCAL 11 (23X)
 FISROSIS, DIFFDSE 1 (21)
 PERIARTERITIS
 

*H*ART/ATRIUM (48) (48) (49)
 

THROMBOSIS, NOS
 

1MYOCARDIUM (48) (48) (49)
 THROMBOSIS, NOS
 

INFLAMMATION, INTERSTITIAL 2 (HI) 23 (48*)
 INFLAMMATION, ACUTE/CHRONIC 3 (6*)
 FIBROSIS 12 (25*)
 FIBROSIS, FOCAL 2 (4%)
 DEGENERATION, NOS 1 (2%) 2 (4*)
 

IENDOCARDIUM (48) (48) (49)
 INFLAMMATION, ACUT VCHRONIC 1(21)
 

*CARDIAC VALVE (48) (48) (49)
 INFLAMMATION, ACDTE/CHRONIC 1 (2*)
 

•COPONAPY ARTERY (48) (48) (50)
 P3RIVASCULITIS 1 (2X)
 

•PULMONARY ARTERY (48) (48) (50)
 

t NUMBER OF ANIMALS KITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

HIGH DOSE
 

01-0121
 

1 (3*)
 

(40)
 

2 (5%)
 

(40)
 

(40)
 

1 (3*)
 

(32)
 

1 131}
 

(48)
 

1 (2*)
 

(43)
 

1(21}
 

(48)
 

1 (2*)
 

15 (31*)
 

(48)
 

(48)
 

(48)
 

(48)
 

C-5
 
TABLE Cl (CONTINUED) 

LOW DOSE HIGH DOSE 

CONTROL(UNTH ) 

01-0070 

CONTROL (UNTR) 

01-011 8 

THROMBOSIS , SOS 

.1ZDIAL CALCIFICATIO N 

"J1ESSMTERIC ART3R Y 

PERIARTERITI S 

(18) (18) 

ARTERIOSCLEROSIS , NOS 

DIG-STIVE SYSTEM
 

*SALIVARY GLAND (U7)
 ATROPHY, NCS
 

HYPEPPLASIA, FOCAL
 

•LIVEP	 (18)
 INFLAMMATION, FOCAL
 

INFLAMMATION, NSCROTIZING 1 (2*)
 FIBHOSIS SEPTAL LIVFR 2 (lit)
 

DEGENERATION, NOS
 NECROSIS, NOS
 

NECROSIS, FOCAL 9 (17*) 2 (IX)
 METAMORPHOSIS FATTY U (8*)
 HYP-JRPLASIA. NOS
 HYPZRPLASIA, FOCAL 8 (17*) 15 (31*)
 

^NGIECTASIS 2 (*«) 1 (2*)
 FFYTHFOPOIFSIS 1 (2J)
 

•LIVER/CENTRILOBULAR	 (118) (18)
 DEGENERATION. NOS
 

DEGENr?ATICN, FOSINOPHILIC 2 (»*)
 •iECRCSIS, NOS 1 (2*)
 

1ETAM01PHOSIS FATTY
 

tLIVER/KUPFFER CELL (48) (18)
 PIGMENTATION, NOS
 HYPERPIGHENTATION
 

*LIVSR/HEPATOCYTES (18)
 DEGENERATION, BALLOONING
 NECROSIS, NOS
 

•BILE DUCT (19)
 INFLAMMATION, NOS 3 (6*)
 HYPSRPLASIA, NOS 6 (13X) U3 (90X)
 

•PANCREAS	 (U5) (16)
 

17_il2SL..
 

# NUMBE R OF ANIMAL S WITH TISSUE EXAMINED MICROSCOPICALLY
 * NMM3EP OF ANIMALS NECROPSIED
 

L OW DOSE 

01-0069 

1 <2X)
 

(50)
 

1 <2X)
 

1 (2X)
 

(15)
 

<2X)
 

(18)
 

1 (2*)
 

1 (2X)
 

1 (2X)
 

6 (13X)
 

1 <2X)
 

(18)
 

2 (H)
 

1 (2*|
 

(18)
 

(18)
 

(50)
 

(11)
 

HIGH DOSE 

01-0121 

<2X)
 

(18)
 

(D6)
 

1 (2X)
 

(18)
 

1 (2X)
 

1 (2X)
 

2 <«*)
 

1 (2X)
 

1 <2X)
 

(18)
 

14 (8X)
 

U (8X)
 

(18)
 

1 (2X)
 

1 (2*)
 

(18)
 

1«	 (29*)
 

1 (2*)
 

(18)
 

<2X)
 

(16)
 

C-6
 
TABLE Cl (CONTINUED) 

-(UNTR) 

01-0070 01-0118 

INFLAMMATION, ACUTE/CHRONIC 6 (13X) 

PERIARTERITIS 

CYTOLOGIC DEGENERATION 

ATROPHY, FOCAL 1 (2*) HYPSRPLASIA, FOCAL 

•PANCREATIC ACINUS C*5) 

METAMORPHOSIS FATTY 

ATROPHY, NOS 

HYPERTROPHY, NOS 

HYPSHPLASIA, FOCAL 1 (2*) 

•FSOPHAGUS (45) 

DYSPLASIA, NOS 1 (2*) 

•STOMACH (48) 

EPIDERMAL INCLUSION CYST (2X) INFLAMHA-ION, NOS 1 (2%) ULCER, NOS 

HYPEHPLASIA, EPITHELIAL 

HYPERPLASIA, FOCAL 

HYPERPLASIA, BASAL CELL <2X) 

HYPERK^RATOSIS 2 <4X> 

ACAVTHOSIS 2 <«»> 

»GASTRIC MUCOSA (48) (48) 

SCLEROSIS 

CALCIFICATION, NOS 

•S1ALL INTESTINE (45) (16) ATROPHY, NOS 

HYPERPLASIA, EPITHELIAL 

tPEYERS PATCH 

HYPEEPLASIA, NOS 

(46) 

12 (26X ) HYPEFPLASIA, RETICULOM CELL 1 (2*) 

•DUODENAL MOCOSA (45) (46) PIGMENTATION, NOS 

•ILEUM (46) 

INFLAMMATION, NOS 2 (4«) 

HYPERPLASIA, LYHPHOID 1 (2*) 

•COLON (U6 ) 

a_i2U.
 

NU1BER OF ANIMALS WITH TISSUE EXA.1IN3D MICROSCOPICALLY
 

NUMBE3 OF ANIMALS NECROPSIED
 

LOU DOSE
 

01-0069
 

1 (2%) 

1 (2X)
 

1 (2X)
 

2 <5X)
 

(46)
 

(46)
 

1 <2X>
 

1 (2X)
 

(46)
 

1 (2X)
 

(46)
 

1 <2X)
 

1 (2X)
 

(46)
 

(46)
 

(46)
 

(45)
 

HIGH DOSE
 

01-0121
 

1 (2%)
 

(46)
 

4 (9*)
 

(44)
 

(48)
 

1 <2X)
 

1 (2*)
 

1 <2X)
 

(48)
 

10 (21X)
 

(47) 

(47) 

(47)
 

1 (2X)
 

(47)
 

(39) 

C-7
 
r
 

TABLE Cl (CONTINUED)
 

cWmPf (UNTR)
 

11-0070
 

(44)
 

(18)
 

3 (6*)
 

1 (2*)
 

41	 (85X)
 

(48)
 

1 (2S)
 

(46)
 

1 (2*)
 

(11)
 

3 (7*)
 

(17)
 

1 (2%)
 

3 (6X)
 

(17)
 

1 (2*)
 

(47)
 -__~»__«.»— •-___— »_M~<
 

HIGH DOSE
 CONTROL (UNTH)
 

01-0118
 

3 (7*)
 

(46)
 

(18)
 

47 (98*)
 

6 (13*)
 

(48)
 

(43)
 

1 (2*)
 

(38)
 

1 <3X>
 2 (5»)
 

(47)
 

(47)
 

(47)
 

.1—Uii
 

LOU COSE
 

01-0069
 

1 (2»)
 

(45)
 

(48)
 

9 (19*)
 

2 (4*)
 

34 (71«)
 

(48)
 

1 (2*)
 

(48)
 

(44)
 

3 <7«)
 

(48)
 

1 (2«)
 

(48)
 

1 (2*)
 

1 (2*)
 

(48)
 

2 IIS)
 

HIGH DOSE
 

01-0121
 

2 <5X)
 

(39)
 

1 (3*)
 

(48)
 

1 (2»)
 

47 (98*)
 

(48)
 

(48)
 

(39)
 

(48)
 

(48)
 

1 (2X)
 

1 (2*)
 

2 (4X)
 

(48)
 

PARASITISM
 

PIGMENTATION, NOS
 

HYP'SPPLASIA, EPITHELIAL
 

HCOLONIC .lucous NEMBR
 PIGMENTATION, NOS
 

URINAR Y SYSTEM
 

•KIDNEY
 

•3LOMERULONEPHRITIS, NOS
 

INFLAMMATION, INTERSTITIAL
 

INFLAMMATION, ACUT1/CHROSIC
 

FI880SIS, DIFFUSE
 

NEPHROPATHY
 

NEPHBOSIS, NOS
 

• KIDNEY/PELVIS
 

MINEKALIZATION
 

HYPERPLASIA, EPITHELIAL
 

((URINARY BLADDER
 

HYPERPLASIA, EPITHELIAL
 

METAPLASIA, SQUAMOUS
 

ENDOCRINE SYSTEM
 

IPITUITARY
 

HYPERPLASIA, NOS
 

HYPEBPLASIA, FOCAL
 

IADRENAL
 

THROMBOSIS, NOS
 

METAMORPHOSIS FATTY
 

ANGIECTASIS
 

•ADRENAL	 CORTEX
 

THROMBOSIS, NOS
 

HENORRHAGT
 

NODULE
 

HYPEHPLASIA, FOCAL
 

#ADPENAL 1EDULLA
 

. _ HYEIiELASIA^MM!ILA.B_
 

» NUMBE R OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIHALS NECROPSIED
 

C-8
 
, ——_—. 

HYPERPLASIA , NO S 

HYPERPLASIA , FOCAL 

*THYSOI D 

COLLCID CYST 

HYPERPIGMENTATIO N 

HYPERPLASIA , C-CELL 

HYPESPLASIA , FOLLICOLAH-CEL1 

OTHYROID FOLLICLE
 

PIGMENTATION, NOS
 

HYPERPIGMENTATION
 

»PARATHY3QID
 

HYPERPLASIA, NOS
 

HYPERPLASIA, DIFF3S3
 

•PANCREATIC	 ISLETS
 

HYPEEPLASIA, NOS
 

HYPERPLASIA, FOCAL
 

REPRODUCTIVE SYSTEM
 

*MAflMARY GLAND
 

5ALACT3CSLE
 

HYPERPLASIA, NOS
 

»P.93P'JTIAL GLAND
 

MYEIA, NOS
 

IPROSTAT1:
 

INFLAMMATION, NOS
 

INFLASMATICN, FOCAL
 

INFLAMMATION, ACUTE
 

INFLAMMATION, ACUTE FOCAL 

INFLAMMATION, ACUTS/CHHONIC 

INFLAMMATION, CHRONIC 

INFLAMMATION, CHRONIC FOCAL 

DEGENERATION, NOS 

ATROPHY, NOS 

*SE«INAL VESICLE
 

ATROFHY, NOS
 

*TESTI3
 

S£A 1,1341101
 

* NUMBE R 0 " ANIMAL S rflTH TISSUE

* NUMBE 3 OF ANIMALS NECROPSIED 

TABLE Cl (CONTINUED) 

LOW DOSE HIGH DOSE CONTROL (UNTR) CONTROL (UNTH) 

01-0070 01-0118 

4 (9X) 

(39) (48) 

1 (3X ) 3 (6X) 

(39) (48)
 

(23)	 (28)
 

1 <4X)
 

(45) (46)
 

1 (2X)
 

1 (2*)
 

(48) (48) 

1 (2*) 2 (4*) 

3 (6*) 4 (8%) 

(48) (48) 

(43 ) (44 ) 

17 (39%) 

1 (2*) 

6 (14*) 

8 (19X) 

2 (5X) 

(48) (48) 

2 (4*) 

(47) (47) 

1.I2SI. 

 EXAMINE D MICROSCOPICALL Y 

LOM COSE 

01-0069 

(47) 

11 (23X) 

(47)
 

(25)
 

(44)
 

(50) 

(50) 

(47) 

1 (2*) 

8 (17*) 

1 (2X) 

(2*) 

(2X) 

1 (2*) 

2 (4X) 

(50) 

8 (16*) 

(47 ) 

HIGH DOSE 

01-0121 

1 (2X) 4 (8*) 

(47) 

1 (2X) 

1 (2X) 

1 (21) 

(47)
 

39	 (83*)
 

1 <2X)
 

(19)
 

1 (5*)
 

(46)
 

(18) 

6 (13*) 

C»8) 

1 (2X ) 

(45) 

3 (7*) 

3 (7*) 

(48) 

8 (17*) 

(48) 

C-9 
r
 

TABLE Cl (CONTINUED)
 

LOW DOSE CONTRO L (U»ITR )

01-0070

 HIGH DOSE CONTROL (UNTR ) 

 01-0118 

DEGEhElATION , NOS 

ATROPHY , NO S 

HYPSRPLASH , UTSaSTITIAL CFLL 

39

1

 (83*) 

 (2X)

6 (13X) 

3 (6X) 

*T3STIS/T'JBULE 

DEGE-iFHATION , NO S 

C*7) (47) 

NERVOU S SYSTE M 

IBHAI N (47) (48) 

MALACIA 

SP'CIAL SENSE OnGAN S 

, D Y r (48) (48) 

rlE^OSRHAG E 

P U S 

CATARAC T 

• SYE/COBMS A (48) (48) 

INFHJ1.1ATION, S'JPPURATIVE 

*SYF/RETIV A (48) (48) 

D'3EXSaA?ION , NO S 

(1USCOIOSKCLETAI. SYSTE1 

SON1; 

BODY CAVITIE S 

*A3CCMI!HL CAVITY (43) (48) 

NECROSIS , FA T 

• PLFUR A (48) (48) 

FIBROSIS , FOCAL 

'MES^NTES Y (43) (48) 

HYPEEPLJSIA , FOCA L 

ALL OTH2B SYSTEMS
 

ADIPOSE TISSU2
 

IHIiiJt lilIJ2JSx_A<:fll£/£ ^BaHIi 

* N'JflSEh OF ANIHAL S WIT H TISSUE EXA1INE D WICiiOSCOPICALLY 

* H'J«3^R 0" ANIMAL S NECHOPSIED 

C-IO 

L OM DOSE 

01-0069 

5 (1115) 

(47) 

8 (17X) 

(48) 

(50) 

1 <2X ) 

3 (6X) 

(50) 

(50) 

2 (IX) 

(50) 

1 (2X) 

(50) 

(50) 

HIGH DOSE 

01-0121 

2 (UK) 

1 (2X) 

(48) 

34 (71X) 

(US) 

1 (2X) 

(48) 

1 (2X) 

(48) 

1 (2X) 

(48) 

(48) 

1 (2X) 

(48) 

1 <2X) 

(48) 

1 <2X ) 
TABLE Cl (CONCLUDED) 

LOW DOSE HIGH DOSE 

CONTEOL (UHTR) CONTROL (UNTS) LOh COSE HIGH DOSS 

01-0118 11-0069 01-012 1 

NECOQSIS, F'T 

SPHCUL SUIHAHI 

ANI1AL CISSING/XO NECROPSY 

HUIO/N3CROESY/HISIO PEFF 

AL'TOLY3IS/NO NECKOPSV 

* NUMBE R 0 " ANIMAL S WIT H TISSU E EXAMIN E MICROSCOPICALL Y 

« N'llBS P 0" ANIMAL S NECROPSIE D 

C-ll 
TABLE C2

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS 

TREATED WITH 5-NITRO-o-ANISIDINE 

LOW DOSE HIGH DOSE —————— CONTROL(ONTR ) CONTROL (UNTR ) LOrf DOSE HIGH DOSE 

02-0070 02-0118 02-0069 02-0121 

AN-KAL S INITIALL Y I N STUS Y 5 0 50 50 50 ANTIAL S 1I3SING 1 A N T 1AL S NFTROPSI^ D <*9 50 49 46 ANI1ALS CXMIHFD HISTOPATHOL03ICALL; ** U9 50 «9 1*6 

IN" THI S N^AS Y SYSTE M 

*"IN (19) (50) (19) (16) 

;

c

ic -RIA L INCLU C I O N CYST 1 (2!!) 

S^B A C"JU S CYST 1 (2X) IN*l A1MA-ICN , NCS 1 (2X) 

*?UBCU T TISSU E (1*9) (50) (19) (1*6)

 MIN E RALIZ^TJO N 1 (2X)

 A»S C ^53, NOS 1 (2X)

 

Bi:c;?rEATOPY SYSTEM
 

•NASAL	 CAVITY (50) (19) (16) , ACUT r (2X) 

(19) (19) 

:N*LAMATICN. ACUT-/CHPONIC 15 (31S) 1 (2V

 

\, SQUAMOUS
 

(50) (19)

 

ar :NCHIECTAS:S 2 (4*)

 INFLAP11ATION, ^S 3 <6X)

 IMFLAB1A?ICfc, ACUTE/CHRONIC 1 (2%)

 

(19) (50) (19) 

INFLMMATION, FOCAL 1 (2X) 

IN^L AK^IA^ION, N^CROTIZING 1 (2X) 

1YP5FPLASIA, LYKPHOID 1 <2X) 

*L JN T	 (15) (50) (19) (13) CC11GT 3IION, CnFONIC PASSIV E < 2V 

(2*) 

2-JiSi 

* MU135 E 0' ANI1»L S KIT H T ISSUE EXAMINE D MICROSCOPICALL Y 

* lUIP ^ < 0" ANIMAL S NECROPSIE D 

**EXCLUDES PARIIALLY AUTOLYZED ANIMALS 

C-12 
T

 

TABLE C2 (CONTINUED) 

LOW DOSE HIGH DOSE 

CONTPOL(UNTR ) CONTRO L (UNT8 ) 

02-0070 02-011 3 

INFLAMMATION , INTERSTITIA L 2 (<4<S ) 6 (12%) 

PNEUMONIA , ASPIRATIOK 

3RCNCH3PNEUKONIA , ACUTE 

INFLAMMATION , ACUT E NECROTIZIN 3 

HYPEPPLASIA , EPITHELIA L 1 (2*) 

HYPESPLASIA , ALVEOLA R EPITHELIU M 1 (2/4 ) 

HEMATOEOIDTIC SYSTEMtBONF NARfOfc (U6)
 HYPCP1ASIA, NOS
 

OST20SCLEEOSIS 1 (2%) 1 (2X)
 

HYPHRBLASIA, HEMATOPOIETIC
 

HYPERPLASIA, RETICULUM CELL
 

HYELOP3IESIS
 

»SPLES»I	 (1*8)
 

INFLAPltlATION, NECROTIZING
 

INFLAM1ATION PSOLIFEP ATIVT
 

FIBFOSIS
 

-lEMCSIDIROSIS 12 (25X)
 LYNPHOID DEPLETION
 

HYPERPLASIA, NOS
 

HYPEBPLASIA, HFM ATOPOIETIC 1 <2X ) 25 (52X)
 HYPERPLASIA, ETYTHROID 19 (40X)
 HYP^RPLASIA, 5FTICULOB CELL 1 (2«)
 MEflATOPOIESIS
 

1YILOPOIESIS
 

•SPLENIC	 CAPSULE (148)
 

H5MOSPHAGIC CYST 1 (2X)
 

*SPLENIC ^ED P"LP (=18)
 

ATROPHY, NCS
 

VOD E (42) (147)
 

LYMPHOID DEPLETION
 

PLAS1ACYTOSIS 1 (2X)
 HYPEFPLASIA, RETICULUN CELL
 

HYPIRPHSIA, LYBPHOID 14 (9*)
 

ILB1BAR LY1PH NODE C*2) (147)
 

____ H £
 

« NJM3E R OF ANIMAL S KITH TISSUE EXAMINED MICROSCOPICALLY
 

* N'J"BE^ 0" ANI'ALS NECROP5IED
 

L O U COSE 

02-0069 

2 <«*>
 1 (2*)
 

1 (2*)
 

(147)
 

1 (2X)
 

1 (2%)
 

(ttS)
 

2 (H%)
 

1 (2X)
 

1 (2%)
 

7 (15%)
 

12 (25«)
 

1 (2*)
 

(148)
 

(43)
 

HISH DOSE 

02-0121 

1 (2%)
 

(1414)
 

2 (51)
 3 (7%)
 

2 (5%)
 

(143)
 

1 (2S)
 1 (2X)
 

1 (2%)
 

1J (30X)
 

(43)
 

1 (2X)
 

C*3)
 

1 (2*)
 

1 (2%)
 

(143)
 

C-13
 
r

 

TABLE C2 (CONTINUED) 

LOW DOSE HIGH DOSE 

CONTROL(UNTR ) CONTROL (UNTR) LOrf EOSE 

02-007 3 02-0118 02-0069 

e-Ij^NToTC L. NOE2 (42) (47) (43) 

I\ 1 JH>"HTION, NCS 

C'TJ^AI ­ Y SYS"r « 

»! "A"" (49 ) (50) (48) 

FINIS'S, FOUL 1 (25) 

FI30STS, TIFFOS r 1 (2? ) 

*H" \P"/A":(lUn (49) (50) (48) 

Ti--jr>lE )SIS, NOS 1 (2X) 

»ilY ICAR^ I J1 (49 ) (50) (48) 

TN'FLAFIITION , (O S 1 (2X) 

TVFUHInTIPN , IMP P S T ITIA L 2 (4J) 23 (46X) 

INFI.AMATION , ACUT/CHiONIC 1 (2% ) 

'TDrCSIS 15 (30X) 

"IP^OSIS, FOCA L 2 (4? ) 

DEu"N"RA"ION , NO S 2 (4%) 

CALCIFICATION , "JOS 1 (2X) 

^ ^ N^^C^ I iDI0 1 (H9 ) (50) (48) T NFLA11A"ICN / NOS 1 (2X ) 

•VFHII'TION, ACUTE/CHRJHIC 

»CARr T A f J/11.VF

TNFL^H^A^TON , AC JTE/CHRONIC

 (U9 )

 1 (2% ) 

 (50) (48) 

*"'Ji. v IGNAF f AR T E R Y (49 ) (50) (49 ) 

lINtSALTZA'IO N 9 (181) 

«H-D}fT r V3I N (49 ) (50) (49) 

I NFLA11AIION , NO S 

•SJPF8IJ F «1^S"NT"FIC (49) (50) (49) 

IK,»H"1A'"-ON f SOS 

DI^-Sl'V1 7 3YSTE f 

» L* '""^ (4? ) (5J) (49) 

hZJoI^A^ <2 ' 

» MJ1E ' 0­ HtlllALi JITH ""ISSUE "XAIIN^ D 1IC 

* NJ'S" ' OF AilIIAi- S NiCROPSIE D 

HIGH DOSE 

02-0121 

(43) 

1 (2*) 

(«3) 

(43) 

1 (2*) 

(43) 

7 (16% ) 

(43) 

1 (2X) 

(43) 

(46) 

(46) 

1 (2X) 

(46) 

1 (2X) 

(44) 

. _ J_U1L__ 

C-14 
TABLE C2 (CONTINUED) 

LOW DOSE CONTRO L (UNTR ) 

02-0070 

HIGH DOSE 

CONTRO L (UNTH ) 

02-0118 

LOtf DOSE 

02-0069 

HIGH DOSE 

02-0121 

H'MCHPHAGIC C 

INFLAMMATION , 

INFLAMMATION , CIRRHOSIS , NO 

D- 13tSraA'"IO«lI 

DrGEN*"UTIOM, JincHOSIS , NOS 

YST 

NTCPOTIZIN3 

ACUT E NECSOTIZINS 3 

CYSTIC 

FOSINOPHILIC 

N-CRCSTS , FOCA L 

NECROSIS , CO A 

I;TAIORPHOSI S 

oULATIV E 

FATT Y 

SASOFHILI C JYT O CHANG E 

HYFTSTROPriY , HYP3PPLASIA , 

ANJIICTASIS 

HYP"FPLASIA , 

'I'MATOFOirSIS 

FOCAL FOCAL 

ERYTHROI D 

2

3

U

29

1

 {U%) 

 (61 ) 

 (8*) 

(59%) 

 (2*) 

2

6

38

1

2

 <UX ) 

 (12«) 

 (76*) 

 (2*) 

 («*) 

1 (2*) 

1 (2*) 

1 (2*) 

1 (2%) 

2 (it*) 

5 (10X) 

1 <2X ) 

3 (6*) 

1 

1 

1 

2 

4 

1 

2 

2 

(2%) 

(2*) 

(2*) 

(5*) 

(9*)

(2*) 

(5*) (5*) 

«H~PHTIC CAPSULE 

'I330SIS, FOCAL 

(19) (50) (19) (ii)

1 (2S) 

ON, NOS NECROSIS, NOS 

ITTAMOaPHOSIS FATTY 

(19) (50 ) (19) 

3 (6*) 

3 (6*) u (8X) 

(ii) 

2 (5J) 

IN'PLAKIATION ,

IM'Hr. 1ATION ,

 NO S 

 ACUTE/CHRONI C 

(19) (50 ) (19) 

3 (6%) 

(11) 

1

1

 (2*) 

 (2X) 

»LIVFVdr?A?OCYTE S 

NECSOSIS, FOCAL 

(19) (50 ) (19) 

1 <2X) 

(11) 

*BTLr CIJC? 

INFLAM1ATICN, NOS 

INFLA"1ATION, ACUTE/CHRONIC 

dYPEFPLASIA, NOS HYP^PPLASIA, FOCAL 

(19) 

5

1

 (10* ) 

(2%) 

(50) 

1

32

 (2S) 

(6U%) 

(19) 

1 (2%) 

(16) 

1 (2X) 

•PANCREAS THBCM30SIS, NOS 

INFLAddATION , NOS INFLAMMATION, ACUTE/CHRONIC 

ATROPHY, NOS HYPSPPLASIA, FOCAL 

1

1

 (9S) 

 (2X ) 

(13) 

6 (13* ) 

(11) 

1 (2*) 

(13) 

1 (2S) 

*PH»ICE SATIC ACINUS (17) (13) (11) (13) 

* N'llE^R O F ANIMAL S * NJME' B n" ANIMAL S WIT H TISSUE

 NECROPSIED 

 EXAMINE D MICROSCOPICALL Y 

C-15 
•STOMACH
 

INFLA11HION, NOS
 

ILCER, NOS
 

ULCER, FOCAL
 

HYPERPLASIA, EPITHELIAL
 

ACANThDSIS
 

• GASTRIC 1'JCOSA
 

SCLEHOSIS
 

»INT"STIN*L VILLUS
 

PIG1ENTA"'ION, NOS
 

•FEYEHS	 PATCH
 

HYPF5PLASIA, NOS
 

•COLON
 

NEIA^ODIASIS
 

"ARASI-TSM
 

•COLONIC	 1UCOUS BEMBR
 

PIGMENTATION, NOS
 

URI1ARY SYSTEM
 

•KIDNEY
 

1INE6»i,IZATION
 

CYST, NOS
 

GLCIER'JLONEPHRITIS, NOS
 

INFLAM1ATION, INTERSTITIAL
 

FIBROSIS, DIFFUSE
 

NEPHROSIS, NOS
 

HYPERPLASIA, EPITHELIAL
 

•KIDNEY/MEDULLA
 

HYPERPLASIA, EPITHELIAL
 

HKIDNEY/GLOMEFULUS
 

INFLAMIA^ION, MEMBRANOUS
 

tKIDNEY/TJBULE
 

PIGflrNTATION, NOS
 

tPZNAL TUBULAR BASEME
 

CALCIFICATION, N0£
 

TABLE C2 (CONTINUED)
 

CONTROL (UNTR)
 

02-0070
 

(19)
 

1 (2X)
 

(19)
 

(19)
 

(19)
 

(11)
 

2 (5X)
 

(41)
 

(19)
 

1 (2X)
 

31 (D9S)
 

(19)
 

(19)
 

(19)
 

(19)
 

HIGH DOSE
 CONTROL (UNTS)
 

02-0118
 

(la)
 

1 (2X)
 

2 (IX)
 

(18)
 

(18)
 

(18)
 

15 (31X)
 

(16)
 

2 (IX)
 

(16)
 

(50)
 

13 (86X)
 

1 (2X>
 

(50)
 

(50)
 

(50)
 

(50)
 

-_
 

10« COSE
 

02-0069
 

(19)
 

2 (4 %)
 

1 (2X)
 

(19)
 

(19)
 

(19)
 

(17)
 

(17)
 

(19)
 

1 (2X)
 

1 (2X)
 

2 (IX)
 

29 (59X)
 

(19)
 

1 (2X)
 

(19)
 

(19)
 

(19)
 

, 1 U*)
 

HIGH DOSE
 02-0121
 

(13)
 

(13)
 

2 (5X)
 

(13)
 

1 (2X)
 

(13)
 

1 (2X)
 

(39)
 

(39)
 

1 <3X)
 

(15)
 

1 (2X)
 

1 (9X)
 

21 (53X)
 

2 (IX)
 

(15)
 

(15)
 

1 <2X)
 

(15)
 

2 (IX)
 

(15)
 

» NUHB3P OF ANIRAL S WITH TISSUE "XAMINED MICROSCOPICALLY
 

* NUMBER OF ANIFALS NECROPSIED
 

C-16
 
TABLE C2 (CONTINUED) 

— LOW DOSE HIGH DOSE
 ——————-—
 CONTROL (UNTR) CONTROL (UNTR) LOW COSE
 

02-0070 02-0118 02-0069
 

#KIDNEY/EELVIS (19) (50) (19)
 HEMATOPOTESIS 1 (2X)
 

ENDOCRI1" SYSTEM
 

•PITUITARY	 (11) (10) (16)
 ABSCFSS, NOS 1 (2*)
 PERIVASCULITIS 1 (3«)
 HTYPIA, NOS
 

HYPS3PLASIA, NOS 1 (2%)
 HYPERPLASIA, FOCAL I (5%) 3 (8X)
 

•ADRENAL	 (19) (19) (19)
 HETAM03PHOSIS FATTY 3 (6?) 1 (21)
 HEMATOPOIESIS 1 (2X)
 

•ADRENAL	 CORTEX (19) (19) (i»9)
 

THROMBOSIS, NOS 1 (2*)
 METAMOHFHOSIS FATTY 3 (6*)
 HYPSRPLASIA, FOCAL 1 (2*) 3 (6%)
 

•ADRENAL MFDCLLA (19) (19) (19)
 P2BIVASCUHR CUFFING 1 (2*)
 HYPERPHSIH, NODULA3 3 (6X)
 HYPPRPLASIA, NOS 1 (2X)
 HYPE3PLASIA, FOCAL 1 (2X) 3 (6X)
 

•THYROID	 (10) (15) (19)
 CYST, NOS 1 (2*)
 CYSTIC FOLLICLES 1 <2X)
 D7GIN'3ATION, NOS 1 (2*)
 HYPFPPLASIA, C-CELL 1 (2X)
 HYPESPLASIA, FOLLICUL AR-CELI 1 (2X)
 

tTHYBOir FOLLICLE (10) (15) (19)
 PIGMENTATION, NOS
 

•PANCREATIC	 ISLETS ("7) (18) (11)
 

HVPEEPLASH, NOS
 

REPRODUCTIVE SYSTEM
 

*HAMMAR Y 3LAND	 (19) (50) (19)
 

- ~H_L«?T,OCELE . . _, t _, ,„ „,„.,.. 2_-USSl_~ .16.,^12H J t£Sl
 

* MUH3E R O F ANIMAL S IflT H TISSUE EXAMINE D MICROSCOPICALL Y 

* SIIKB2H OF ANIMAL S NECROPSIED 

HIGH DOSE
 

02-0121
 

(15)
 

(11)
 

1 (2%)
 

1 (2*)
 

(11)
 

(11)
 

5 (11X)
 

(11)
 

1 (2X)
 

6 (11J)
 

(12)
 

3 (7*)
 

(12)
 

5 (12X)
 

(13)
 

1 (2«)
 

(16)
 

C-17 
TABLE C2 (CONTINUED)
 

LOW DOSE HIGH DOSE 

CONTRO L (UNTR ) CONTRO L (UNTR ) 

02-0070 02-0118 

I t FLAIIATlrj , SUPPURAT E 

I>|PLA" 1 A T :ON , ACUT E 1 (21} FI3F35I 3 

HY? r! iPlA3IA , J

 YP F ^ F LA SI A

 NJ 3 

 ^OCAL 

23

2

 (47« ) 

 (4J) 8 (16*) 

•CLITIRH L 'SLAN D (49 ) (50) 

^BSC""Sj , NO S 

*VA3IN * TNr

L Av l'HTI'"'; , ACUTE/CHHONI C 

(19)

1 (24 ) 

(51) 

»''T"iU S 

f- >Y:i : ;i vn T PA 

(19) 

6 (12X ) 

(50) 

IN^L'^IJTION , SH I PUBA7IV E a y 0 u-Tq ^ 

H^SCES:, , NO S 

INtaairA-IC"!, ACUTF/CHRONI C 

NECROSIS , NOS 

HY?rf?LASIA , ADENC1A~OU S 1 (2*) 

• CLRVI X rjfM 

INFH1-<ATIO'I, AC'JTE/CHRONI C 

(49 ) 

2 («) 

(50) 

!' YFE?F-5SI A , NOS 

rtY° rS^LAHA ,

\CAN-h5SI S 

 3A5A L CrLL 1

1

 (2*) 

 (2*) 

*.j-rp., v :v,noK-TSIUH 

IH-LACIATICN , f'C S 

(49 ) (50) 

22 (44*) INFUK'UTIUN , SLtPJPATIV " 

IN^LAIlATIOli , JC'JTE 2 3 (47< ) A a S C r S i , N'OS TN?HI"HTION P.OLIF:FATIV : 'V?r 5PLASIA (

Hmr^FlASI f ,

 N-OL 

 CYSTIC 

5

5

 (1 T% ) 

( 11S) 

6 (12*) 

J Y?"ErHSIA , J YtTP?LA3IA ,

 AD r NLMATOU S 

 STPOMA L 

1 (2*) 

'O/AFY/'VIDUC " 

I"FLA, " 1A7ICN , NO S T N ? I t iiATio t , ^IIFPUPAT I v: 

(49 ) (50 ) m

2

 (20*) 

 (4*) I'. t :L»>"UT!OH( ACUT E 1 (21) 

> 0 ''-yI T i ,OS ^1UL.-.. 

(49 ) 

o" ;NIM;LS IJITH TISSU: DXAIINDD HICROSCOPICALLY
 C.- ANII1AL ? UTCROPSIE D 

—• • 

L OW COSE 

02-0069 

1 (2X) 2 (41) 

3 fbX ) 

(19) 

(49) 

(49) 

3 (bX) 

1 <2X > 

3 <6X) 

4 (8X) 

2 (4*) 

(49) 

(49 ) 

1 (2X) 

1 (2X) 

3 (6X) 

1 (2X) 

9 (18*) 

1 (2X) 

(49 ) 

1 (2*) 

(48) 

HIGH DOSE 

02-1121 

1 (2*) 

1 (2*) 

(46) 

1 (2X) 

(46) 

(44) 

1 (2*) 

1 (2*) 

(44) 

1 (2*) 

(44) 

5 (11*) 

1 (2*) 

1 (21) 

2 (5X) 

(44) 

(45) 

C-18 
TABLE C2 (CONTINUED) 

LOW DOSE 

CONTROL(UNTR ) 

02-OJ70 

HIGH DOSE 

CONTROL (UNTR ) 

02-0118 

L OS DOSE 

02-0069 

HIGH DOSE 

02-0121 

HTMCRRHAGE 

IK"LAMNATION, ACUTE/CHRONIC CORPUS HFNOHRHA'JICUM 

1 (2*) 

1 (2S) 

NE9VOBS SYSTEM 

• B8MN/HENIN3ES INFLAMMATION, ACUTE 

INFLAMMATION, ACUTE SUPPURA1IVE 

(U9 ) (50) (19) 

(2X) 

(2X) 

(tl) 

H33AIN 

HEMORRHAGE 

PESIVHSCULITIS 

(50) (19) 

<2X) 

(11) 

<2X ) 

tNEDULLA OBLONSATA HEMOPEHAGE (50) (19) 

1 (2X) 

(11) 

SPECIAL SENSE ORGANS 

INFLAMMATION, HSMORRHAGIC SYNSCHIA, NOS SYNECHIA, POSTERIOR CATAFA:T 

(19) 

1

1

 (2%) 

 (2X) 

(50) 

1 (2X) 

(19) 

9 (18X) 

(16) 

1 (2X) 

•EYS/CORN^A INFLAMMATION, NOS 

INFLAMMMION, SUPPUHATIVE 

INFLAMMATION, CHRONIC 

(19) 

1 (2X) 

(50) (19)

1 (2X) 

 (16) 

1 <2X) 

*EY;?/ciFTINA 

DH'IENEHATION, NOS 

ATROPHY, NOS 

DYSPLASIA, »OS 

(19) 

1 (2X) 

(5") 

1 (2X) 

(19)

8

1

 (16X) 

 (2X) 

 («6) 

•HAROEHIAN GLAND 

HYPEKPLASIA, NOS (19) (50) 1 (2X) 

(19) (16) 

*^AR CANAL 

ABSCESS, NOS (19) (50) (19) (16) 

1 (2X) 

tllJSC'JLOSKELETA L SYSTEM 

*V2RTEER A (19) (50) (19) (46) 

__l_Uii_ 

* N'JIEFR

* NUMBH H

 OP

 OF

 ANIMAL S

 ANIMAL S

 WITH TISSUE

 NECHOPSIE D 

 EXA.1INED MICROSCOPICALLY 

C-19 
TABLE C2 (CONCLUDED) 

$&?§§& (UNT8 ) 'tWrtfbT(UNTR)

02-0070 02-0118

(«9 ) (50)

OSTEOPOROSI S 1 (2*) 

*»EDCMTNA L M'ISCL2 (U9 ) (50)

INFLAM1ATION , ACUTE FOCAL

BOD Y CAVITT- S 

*«:DIASTIIU M (<(9) (50) P^RIARTSRITIS 1 (2X)
 

*EPT "EIUM (19) (50)


_AMdATTO> . PROLIFERATIVE


ALL O^HEH SYSTEMS
 

ADIPOSE TISSUE
 

INFLAC1ATION, ACUTE/CHRONIC 3
 

INFLAMMATION, CHRONIC 2
 

INFLAMMATION, GRANOLOHATODS


OflEMTO " 

1INERALIZATIO N 1 

NECrCSIS , FAT 1 

SOECIJL POSPHOLOGY SOMHARY
 

NO LESION EEPORTED


ANIPA L MISSING/NO NECROPSY


AUTCLYSIS/NO NECROPSY 1


» NJMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 * NUMBER 0" ANIMALS NECROPSIED
 

 LOU E03E HIGH DOSE 

 02-0069 02-0121 

 (U9 > («6) 

 (U9) (46) 

1 (2*) 

 (K9 ) (U6> 

 (U9) (U6)
 

1 (2%)
 

1
 

2
 

1
 

1 3
 

C-20 
APPENDIX D
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN MICE TREATED WITH 5-NITRO-o-ANISIDINE
 
f 
TABLE Dl

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE MICE 

TREATED WITH 5-NITRO-o-ANISIDINE 

DOSE A DOSE B 

CONTROL (UNI 'R) CONTROL lUNTR ) 

05-0070 05-01 16 

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

ANIMAL S NECHOPSIED 50 49 

ANIMAL S EXAMINE D HISTOPATHOLOGICALLY ** 50 49 

INTEGUMENTAR Y SYSTEM 

*SKIN (50) (49 ) 

INFLAMMATION . NO S 1 (2%) INFLAMMATION , FOCAL 3 (6%) 

INFLAMMATION , NECEOTIZIN G 1 (2%) ABSCESS, NOS 2 (U% ) 

HYPFRPLASIA, NODULAR
 

•SUBCUT TISSUE (50) (49)


NECSOSIS, FAT 1 (2%)
 

RESPIRATORY SYSTEM
 

*LUSu/BP,ONCHUS (50) (49)


BHONCHIECTASIS
 

INFLAMMATION, FOCAL 1 (2S)
 hYP£PPLASIA, EPITH3LIAL
 

*LUNG/dPONCHIOLE (50) (49)
 INFLAMMATION, NOS 1 (2%)
 INFLAMMATION, FOCAL 1 (2%)
 INFLAMaATION, ACUTE/CHRONIC
 

P3RIVASCULITIS 1 (2%)
 

tLUNG (50) (49)


HEMORRHAGE 2 (US)
 

INFLAMMATION, INTERSTITIAL 10 (20%)
 

HYPEEPLASIA, ALVEOLAR EPITHELIUM 2 (4t)
 

HEMATOPOIETIC SYSTEM 

• SPLEEN (50 ) (149) 

H. _NOS_

* NUMBE R OF ANIMAL S WITH TISSUE EXAMINE D KIC80SCOPIC*LLY 

* NUM3E P CF ANIMAL S NECBOPSIED 

**EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

DOSE A 

05-0071 

50 

48 

48 

DOSE B 

05-0102 

50 

49 

48 

(U8 ) («9) 

1 (2*) 

 (48) (49) 

 (48)

1 (2*) 

1 (2*) 

 (48)

 (48)

 ­

 (47) 

 (47) 

2 (4X) 

 (47) 

(1*5) (47) 

D-3 
r

 

= ETICULOCYTOSI S 

riYs>:B?LAs:A, H:MATOPOIETIC 

H/PEFPLAEIA . LYPPHOI D 

«SPL:NI C FOLLICLES 

lYPJPPLASIA , NO S 

« L Y IPh NODI.

 

INFLAMMATION , NO S 

hYP-FPLASIA , NOS -^T ZUJLGCYTOSIS

 

'Yr-F7i,ASH, LYHPHOI D 

« N - ^-N-ici c L. NODs 

INFLAI'HTIOM , NOS 

AMi.IECTASI S 

(lYPL'FPLASiA , RETICULU M CS'LL 

l^P^PPLASH , LYMPHOI D 

CIPC 'ILATOrY 3YSTE1 

*[•_, An!'

 

^IN^^AuIiA^IO N 

P *? if.- :~~ 'TIS "•- T'M5CJLI'IS 

<MY n;Ai JiJI

 

i Eai^r :<^TroN , NCS 

« AO f^r c v^vt 

:>l?!A M "ATirN , ACUTE/CHSONIC 

°«* 

i-:=i * *sc'jnTis 

* LT ;:­

I IFLA Jr1A~" CN, F"CAL 

M ID JL: c ~ ,~:- E <A'io'i, NO S

 

»<"C"OSIS , N'l b 

\~CP O : 'j_ "OCAL 

# NUIi' P 0 ? > »Ii v \ L S HU H IIS5U5 * M'i"J' v r ANT1AL S MCFOPSljJP 

TABLE Dl (CONTINUED) 

DOSE A DOSE B 

CONTROl(UNTB) CONTROL (C NTH) 

05-0070 05-0118 

1 (21.) 

5 (10X) 

1 (2*) 1 (2X) 

(50) <<••>> 2 (4<) 

(45) (12) 

10 (24X ) 

1 (27,) 

2 (5X) 

3 (7X) 

(45) (42) 

1 (24 ) 

(49 ) (49) 

1 (2% ) 

(-19) (49) 

(-19) (19) 

1 (2S) 

(<") (48) 

1 (2%) 

(50) (48) 

i_i2Sl _ ,_ 2_lJiSl 

 :XAMINF O hlCKOSl-Ot'ICALLY 

DOSE A 

05-0071 

3 <7X ) 

(45) 

(40) 

(40) 

1 (3X) 

1 (3X) 

(48) 

(48) 

1 <2X ) 

(48) 

(47) 

(49 ) 

14 (3X) 

1 (2X) 

1_IZS1 

DOSE B 

05-0102 

(47) 

(40) 

(40) 

1 (3X) 

(47) 

1 (2X) 

3 (6X) 

(47) 

(47) 

(46) 

(47) 

1 <2X) 

1 (2X) 

1 (2X) 

_ 1 J2X1__ 

D-4 
1 

TABLE DI (CONTINUED) 

DOSE A DOSE B 

CONTROL (UNTn ) CONTROL (UNThJ 

05-0070 05-0118 

N3CHOSIS, COAGULATIV E 

METAMORPHOSI S FATTY 2 (l*-"5)
 

HSPATOCYTOM3GAL Y 2 (U*)
 

DEPLETIO N 1 (2%)
 

HYPEPThOPHY , SOS 

HYPERTROPHY , DIFFUSE 

HYPEPPLASIA , NODULA R 2 (UX)
 

HYFERPLASTI C NOCU1E <2X)
 

HYPERPLASIA , NO S 

HYPFEPLASIA , FOCAL 1 (2SS)
 

HYPC.KPLASIA , DIFFUSE 1 (2*)
 

ANGIECIASI S 

tLIVER/CCSTHILOBULA R (50) C*8)
 

DEGENERATION , NO S 

NECSOSIS. NOS 1 (2%)
 

*LIVSVPERI PORTA L (50) ("8)
 

INFLAMMATION , SOS 

INFLAMMATION , ACUTF/CHRONIC 

*LIVER/KUPFFE R CEL L (50)
 

HfPEKPLASIA , NOS 1 (2X)
 

*I.IV EH/H E PA TOCYTES (50)
 

DEGENERATION , NO S 

HYPERTROPHY , NO S 

HYPERPLASIA , NO S 

HYP^RPLASIA , DIFFUSE 

• PANCREA S (46)
 

INFLAMMATION , NO S 1 (2X)
 

INFLAMMATION , FOCAL 1 (2X)
 

#PANCREATI C ACINU S (U6)
 

HYPERTROPHY , FOCAL 

•STOMACH (K9) (1*8)
 

INFLAMMATION , FOCAL 2 (U%)
 

INFLAMMATION , NFCROTIZIN G 1 (2X)
 

HYPSRPLASIA , FOCAL 1 (2X)
 

HYPERKEPATOSI S 1 (2X)
 

ACANTHOSI S 1 (2X)
 

•GASTPIC MUCOSA (18)
 

INFL.AJ}MAT.ICN,_FOCAL 

t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBEF OF ANIMALS NECROPSIED
 

DOSE A 

05-0071 

1 (2X)
 

2 (4X)
 

1 (2X)
 

19 (40X)
 

1 (2X)
 

(48)
 

(48)
 

(48)
 

20 (42X)
 

(47)
 

(47)
 

1 (2X)
 

(48)
 

DOSE B 

05-0102 

1 (2X)
 

3 (6X)
 

1 (2X)
 

21 (45X)
 

19 (40X)
 

(47)
 

(47)
 

3 <6X)
 

1 (2X)
 

(47)
 

(47)
 

22 (47X)
 

2 (4X)
 

3 (fcX)
 

12 (26X)
 

(43)
 

(43)
 

(46)
 

(46)
 

D-5
 
IH"LA''1ATICN, ACUTVCHRONIC 

*GA=;r 0 IC SEF03A

Pa. lAtTIP I IIS 

• EEYE.-C5 PATC H

HYPERPLASIA , >)OS

• COLO N

3RANaLO"A , NOS

PARASITIS M 

URINAR Y SYSTTI 

• KIDNE Y Gi-OirSULONEPHRIIIS , NO S 

INFLAMMATION , INtEPSTIIIAL piiBiAPTEciT^ s GLO"iF=ULOSCLEROSIS , NOS 

"KIDNEY/TUBUL E

DtGEN i RATION , NOS 

C V.CIFICATIO!., FOCAL 

PIjMLNTATION . NO S 

*UPINAR Y BLADDE R

INf LAPMATION , SUPFUHATIV E 

P^SIARTERITI S 

HVpEhrLASIA , EPITHELIAL

ENTOCKINi . SYSTE M 

ItAoFSMA L

H Y t EnPLASIA , SO S 

»ADh t'NAi./CAPSUL E

fYPIKPLASIA , NOS 

• ADRENA L COPIE X

HYc-EPPLASTI C NODUL E 

*ThY=iOI D

FOLLICULAE CYST^ NOS 

t NblbP P OF ANIP1AL S WI T H TISSUE

* NUfBE P CF ANIMAL S NLCROPSlL D 

TABLE Dl (CONTINUED) 

DOSE A DOSE B CONTROL (UNTR ) CONTROL (ONIR )

05-0070 05-0118 

 (49 ) (48) 

 ("49) (49) 1 (2)4) 7 (14X) 

 («6 ) (43) 

1 (2") 

3 (7X) 

 (49 ) (49) 

2 (4X) 3 (6S) 16 (33X) 

 (49) (49) 

 (47 ) (48 ) 

1 (2%) 4 (8*) 

 (t9) (44 ) 

3 (7X) 

 (U9 ) (44) 

3 (7X) 

 (49 ) (44) 

 (40 ) (45) 

 EXAMINi D MICROSCOPICALL Y 

DOSE A 

05-0071 

(48) 

(48) 

(47) 

(48) 

5 (10X) 

(48) 

1 (2X) 

(48) 

(44) 

(44) 

(44) 

1 <2X) 

(47) 

2.J2S1 

f 

DOSE B 

05-0102 

1 (2X) 

(46) 

1 <2X) 

(45) 

(40) 

(48) 

4 (8X) 

1 (2X) 

1 (2X) 

(48) 

1 (2X) 

1 (2X) 

(47) 

1 (2X) 

1 (2X) 

(46) 

(46) 

(46) 

(47) 

._. . .. 

0-6 
TABLE Dl (CONTINUED) 

DOSE A 

CONTROL (UNIR ) 

05-007 0 

DOSE B 

CONTROL (DNTH) 

05-0118 

DOSE A 

05-0071 

DOSE B 

05-0102 

LYMPHOCYTI C INFLAMMATOE Y INFILT E 

DKGL'NEPATION , NO S 

PIGMENTATION , NO S 

HYPF.SPI3KFNIATIO N 

HYPERPLASIA , NODULA F 

HY2FEPLASH , EPITHELIA L 

HYPERPLASIA , FOCA L 

KYPEFPLASIA , IAPILLAH Y 

KYPEP.PLASIA , AD-NOBATOU S 

HYPEBPLASIA , "OLLICULAE-CiL L 

ANGIECTASI S 

11

5

1

1

1

3

1

 (87X) 

 (11X) 

 (2X) 

 (2X) 

 (2X) 

 (6*) 

 (2*) 

3

4U

2

1

6

 <6X) 

 (9HX) 

(*%) 

 (2X) 

 (13«) 

PEPSODUCTIVE SYSTEM 

«PEEPUTIAL GLAND 

ABSCESS, NOS 

HYPEkPLASIA, NOS 

(50) 

1 (2X) 

(19) 

1 (2X) 

«PPOSTAT" 

INFLASMATICN, ACUTE/CHEOKIC 

PERIAFTFHITIS 

(19) (19) (18) (11) 

1 <2X ) 

1 (2X) 

*SEHINAL VJSICLE 

INFLAMMATION, ACUTE 

D-SGENE3ATIOH, NOS 

DE3ENE3ATION, CYSTIC 

HYP^hPLASIA, CYSTIC 

(50 ) (18) 

1

1

 (2X) 

 (2X) 

(19) 

1

1

 (2X) 

 (2X) 

*TESriS/TDBULS 

DEGENERATION, NOS 

CALCIFICATION, NOS 

(50) (13) (18) 

2 (1%) 

(16) 

1

1

 (2X) 

 (2X) 

•EFIDIDYKIS 

INFLAKMATION , NOS 

(50 ) (19) 

1 <2« ) 

(18) (19) 

NERVOU S SiSTE" 

*EEAIS/1ENINGE 5 

ISFLAftfATICN , 

(50 ) (19) (17) (16) 

1 (2X) 

SPECIA L SENS E ORGAN S 

KON i 

* NU«3i ?

* NUK3E ?

 O F

 O F

 ANIMAL S

 ANIKAL S

 WIT H TISSU E

 SECfiOPSIE D 

 EXAMINE D MICROSCOPICALL Y 

D-7 
TABLE Dl (CONCLUDED) 

DOSE A

CONTROL (DNTH)

05-0070

 DOSE B 

 CONTROL (UNTR)

 05-0116

 DOSE A

 05-0071

 DOSE B 

 05-0102 

MUSCULOSKELETA L

NO S3 

 SYSTEM 

BODY CAVITIE S 

NOU S 

ALL OTHER SYST3HS 

ADIPOSE TISSUE 

INFLAMMATION ,

OMSNT'J M 

NECPOSIS, FAT

 ACUTE 1 

1 

SPECIAL MOHPHOLOGY SUMMARY 

NO LESION REPORTED 1 <i

*NIPAL MISSING/NO NECROPSY

NECROPSY PSPF/NO HISTO PERFORMED

MJTOLYSIS/NO NECROPSY

• NUMBE R OF ANIMAL S WITH TISSUE EXAMINE D HICROSCOPICALLY 

* NUflBE1 * OF ANIMAL S NECROPSIED 

5 

1 

2

 1 

1 

0-8 
_ __ 

T 

TABLE D2 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE 

TREATED WITH 5-NITRO-o-ANISIDINE 

DOSE A DOSE B 

CONTROL (UNTK ) CONTROL (UNTE) DOSE A DOSE B 

06-0070 06-0118 06-0071 06-0102 

ANIMAL S INITIALL Y I N STUD Y 5 0 50 50 50 ANIMAL S MISSIN G 1 ANIMAL S NECROPSIE D 4 6 50 43 45 

ANIMAL S EXAMINE D H ISTOPATHOLOGICALL Y ** 47 50 43 44 

INTEGUMENTAR Y SYSTE M 

*SU3CUT TISSUE (48) (50) (43) (45)

 

A3SCSSS, NOS 1 <2X )

 

RESPIRATOR Y SYSTE M 

ILUNG/3ROSCHUS (46) (50) (42) (44)

 

INFLAMMATION , FOCAL 1 (2X)

 

tLUNG/BSONCHIOL E (46) (50) (42 ) (44) 

INFLAMMATION , NOS 1 (2%) 

INFLAMMATION , ACUTE/CHRONIC 1 (2X) HYPERPLASIA , NO S 1 (2X) 

*LUN< ; (46 ) (50) (12) (44) 

INFLAMMATION , INTERSTITIA L 1 (2S) 14 (28X) 1 (2X) 

HEMATOPOIETIC SYSTE M 

*BONE MARRO W (46) (49) (35) (40) 

MYELOFIBROSI S 1 (2X ) 15 (43X) 31 (78X) MYELOSCLEROSI S 2 (5X) 

• SPLSE N (47) (49) (38) (43) PIGMENTATION , NO S 1 (3 K ) 

ATROPHY , NOS 1 (3*) HYPERTROPHY , NO S 1 <3« ) HYPERPLASIA , NOS 9 (18X) 1 (2X) 

ANGIECTASI S 1 (3X) HYPERPLASIA , HEMATOPOIETIC 6 (12X) 

HYPEPPLASIA , LYMPHOI D 1 (2X) 2 <<**} 3 (8X) 1 (2X) 

*SPL2NIC FOLLICLES (47 ) (49) (38) (43) 

_ HYPEBP.LASIA,,. NOS _ _ __ __ __3 J6*J. 

* NUMBE K O F ANIMAL S SITH TISSUE EXAMINE D MICROSCOPICALL Y 

* NUMBE R OF ANIHAL S NECROPSIE D 

**EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

D-9 
w

 

TABLE D2 (CONTINUED) 

DOSE A DOSE E 

CONTROL (TJNTfi) CONTROL (UNTR) DOSE A DOSE B 

Ob-0070 06-0118 05-0071 06-0102 

KHEMCLYMP H NODE S 

INFLAMMATION , NO S 

(47) (49)

2 (4X) 

 (38) (43) 

HYPSPPLASIA , NOS 1 (2X) 

KLYMP F NOD E 

INFLAMMATION , NO S 

HYPtrPHSIA , NOS 

PiTICULOCYTOSI S 

(36) 

1 (3%) 

1 (3X) 

(44)

9 (20X) 

3 <7X) 

1 (2*) 

 (27) (30) 

HYP^&PLASIA , HEMATOPOIZTIC 1 (2X ) HYPSRPLASIA ,

HYPEPPLASIA ,

 PLASM A CELL 

 LYMPHOI D 

1 (3% ) 

4 (9X) 

IABDOHINA L LYBP H

PLASfACY m OSTS 

 NODE (36) 

1 (3%) 

(44) (27) (30) 

CIECU1.AT1RY SYSTE M 

•HEART (44) (50) (41) (44) 

PcRIAfTEFITI S 2 (5*) ENDOCAFDIOSI S 1 (2*) 

•MYOCARDIU M (44 ) (50) (41) (44) 

INFLAK1ATICN , FOCAL 1 (2X) FIBKOSIS, FOCAL 1 (2%) 

• ENDGCASDH,, , (44 ) (50) (41) (44) 

INFLAMMATIO N FEOLIFEKAIIV E 1 (2X) 

DIGESTIV? SYSTE M 

•SALIVAR Y GLAf D (45) (48) (31) (43) 

PFPIV^CLLITI S 3 (756) 

P^HIVASCULA H CUFFIN G 1 (2%} 3 <SX ) 

ILIVE F (47) (50) (41) (43) 

INFLAMMATION , ACJTSI FOCAL 1 (2X) INFLAMMATION , ACUTMF/CHRONIC 

NECROSIS , 'OCAL 

1

2

 <2X) 

 (4«) 7 (14X) NaCHOSIS , COAGULATIV E 1 (2X) KCTAMOKPHOSI S FATTY 1 (2X) HYPSPPLASIA , NODULA R 1 <2X) 

«LIVER/fE8IF08TA L 

nJUMOSPHflSIS FATIY 

(47) 

_. 

(50) (41) 

1.J2S1 

(43) __._ ___,_,„ j 

t NUtiSEfi OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALL Y 

* NU13E1? OF ANIMAL S NECROPSIa D 

D-IO 
TABLE D2 (CONTINUED) 

DOSE A 

CONTROL (UNTfi ) 

Oo-007 0 

DOSE B 

CONTROL (U d IH) 

06-0118 

DOSE A 

06-0071 DOSE B 

06-0102 

•LIV3E/HEPATOCYTE S 

HYPZFPLASIA , DIIFUS E 

(47 ) (50 ) (41 ) (43) 

1 (2*) 

•BILE DUCT 

INFLAH1ATICN , ACUTE/CHRONI C 

(48 ) 

U (3%) 

(50 ) (43) (45) 

*FANCtiEA S 

INFLAMMATION , NO S 

INFLAMMATION , INTERSTITIA L 

PEHIAbTEPITI S 

HYPOPLA3IA , NO S 

(43 )

1

1

1

 (2i,} 

 (25) 

 (2».) 

(43 ) 

2 (4% ) 

(32) 

1 (3*) 

(38) 

IPANCPEATI C ACINU S 

DEGENERATION , NO S 

ATFOPHK , NO S 

(43 ) 

1 (2% ) 

(4tj ) (32) 

1 (3/1) 

(38) 

tSTOIACn 

INFLAflPATICN , NO S 

INFLAK1ATION , FOCAL 

ULCEF , FOCAL 

ACANTnOSI S 

METAPLASIA , SJUANOU ? 

(45 ) 

1 (27.) 

(43 ) 

1 (2% ) 

1 (27i ) 

2 (41) 

(30) (40) 

1 (3S) 

tSMAL L IKTESTIN F 

INFLAr?ATICN , NO S 

N"Ci?OSIS , '10S 

ATEOt. JY , NOS 

(4b ) (43 ) (33) 

4 (1251) 

2 (6*) 

2 <6X ) 

(11) 

#S. IN7'.3TIKE/MUCOS A 

N3CFOSIS , NOS (15) (IB) (33) 

3 (9X ) 

(41 ) 

*PEY:ES PATC H HYPEhPLASIA , NO S (45) 

1 (2X ) 

(43 ) 

7 (15% ) 

(33) (41 ) 

URINAR Y SYSTE'1 

HYLPONEPHPOST S 

GL0.1EFULONEPHBITIS , NO S 

GLORiPULOMEPHEITIS , FOCAL 

INFLAMMATION , INTSPSTiriA L 

GL01CRULOSTHPITIS, ME13SANOU S 

(45 ) 

3 (7S) 

2 (f!) 

1 (2*) 

2 (4%) 

(50) 

4 <8K ) 

1 (2%) 

12 (24«) 

(42) 

4

ID

 (10X) 

 (36«) 

(43) 

25

35

 (58X) 

 (81*) 

* NU1BF 5

* NU.loE?

 OF

 ^F

 ANIMAL S

 ANIMAL S

 WIT H TISSUE

 NECFOPSIE D 

 IXAfllNL O 1II.RO iCOPICALL Y 

D-ll 
r

 

TABLE D2 (CONTINUED) 

1 DOSE A DOSE B 

CONTROUUNTR) CONTROL (UNTR ) DOSE A DOSE B 

06-0070 06-0118 06-0071 06-0102 

jLOiiERULONTPHRITIS , CHRONI C 1 (25) 

AU^FIOSCLHEOSIS , NO S 1 (2*1 

N'PtiiiOSIS, NO S 15 (36X) 32 (7UX) 

N'SZ'jI.GSIS, TOXIC 1 (2X) 9 (21X) 

KKI'D'JEY/GLCMEPULU S (45) (50) (42) (U3)

 N3P4EOSI3 , TOXIC 12 (29X)

 

*KIDNEY/:'JBUL E (US ) (50) (42) <U3 ) 

HIN-PALIiATIC N 1 (2%) NS?HSOSIS, NOS U (10X) N-.?H=OSIS , TOXIC 9 (21X) 1 (2X) 

*UP.I')A6Y BLADDT B (US ) (48) (33) (39)

 INFLAMMATION , CHRONIC FOCAL 1 (2X)

 

P£3H?TEhITl S 1 (2',)

 HYPCPPLASIA , EPITHELIA L 1 (2X)

 

r

NDOC8IN ^ SYSTH.1 

*ADF"iN,^/CAPS3L: (47) (48) <3U ) (UO)

 

HfPZSELASIA , NO S 5 (10X)

 

«ADPtN« L CUPT5 V (47) (48) (34) (UO)

 i, >ouL= ; 1 (2X)

 

HYPl.rn.ASIA , NOS 1 (2X)

 

*THYr.OI D (41) (4U ) (3U ) (13) 

r > :FLAi1MA"'ICN , FOCAL 

ll.r'.AMIATIO N , ACUTT/CnaONIC 1 (2X) C5G5 N r?ATIO N , NOS 12 (35X) 8 (19X) 

HiP:""?IGM5N TM ATIO N 11 (32X) HYPEPP 1 ASH, PAPILLAP Y 2 <5X) r.YP-'PLASIA , CYSTIC 1 (2X) 

.lYPZ E i'LASH, ADENOMATOU S 1 (2X) hYT-f PLA5IA , "OLLICULAK-CELL 1 (2»,) 2 (5X) 

EEP?nj'JCTIV i SYS T £! M 

*KA>:.1AFY GLAN D (48 ) (50) (U3 ) (US)

 >!Yr^?LASIA , NOS 1 (2S)

 

fuTJtjjq (43 ) (47) (32) (36) 

»YD.-,OMETE A 3 J7»l . J3 J288J. 5 M6

*!-- 4 01*1 

« N'I1j2.T r ? ANIMAL S WIT H TISSU E EXAMINE D MICROSCOPICALL Y 

* NL'MJE ? OF ANIHAL S NFC50PSITD 

D-12 
TABLE D2 (CONTINUED)
 

DOSE A 

CONTROL (U N IS) 

06-0070 

DOSE B

CONTROL (UMTS )

06-0118

 DOSE A

 06-0071

 ~ ­ — 

 DOSE B 

 06-0102 

ABSCESS, NOS 2 (5%) 

t UTERUS/ENDOMETRIU M 

INFLAMMATIO N 

INFLAMMATIO N 

INFLAMMATIO N 

INFLAMMATIO N 

INFLAMMATIO N 

HYPTRPLASIA , 

hYPEPPLASIA , 

METAPLASIA , 

, NOS 

, SUPPUSATIV- ; , ACUT 2 

, ACUTE FOCA L 

, ACUTE/CHRO'IIC 

NOS 

CYSTIC 

SQUAMOU S 

(43 ) 

^ 2 

b 

1 

3 

1 

20 

1 

(5;,) 

(5'j) 

(14*,) 

(2%) 

(7-.) 

(2',) 

(473 ) 

(27.) 

(«7 )

8 (17% ) 

9 (17% )

6 (131)

 (32)

 1 (3*) 

 17 (53X)

 (36) 

 28 (78X) 

#OVASY/OVIDUCT 

INFLAMMATION, NOS 

INFLAMMATION, SUPPUEATIV3 

ABSCESS, NOS 

(43) 

U (9S5) 

1 (25) 

(U7) 

4 (9%) 

1 (2%) 

(32) (36) 

tOVAHY 

CYST, NUS 

INFLAMIATICti, NOS 

INFLAMMATION, SUPPUPATIVF 

IHFLAMMATION, CHRONIC 

ABSCESS, CHRONIC 

PEEIAETERITIS 

DEGENERATION, CYSTIC 

ATROPHY, NOS 

HYPERPLASIA, NOS 

LUTEINIZATION 

(45) 

b (13%) 

1 (2*) 

1 (2%) 

1 (2%) 

(48) 

10 (21%) 

4 (8$) 

1 (2%) 

3 (6%) 

(30) (37) 

1 (3*) 

1 (3X) 

2 (5X) 

1 (3%) 

NEHVO'JS SYSTEM 

tEEAIN/MENINGES 

INFLAMMATION, ACU1S/CHROSIC 

INFLAMMATION, CHRONIC FOCAL 

(16) 

1 (2%) 

1 (27.) 

(34) 

SPECIAL SENSE OEGANS 

NONE 

HOSCULOSKELETAL SYSTEM 

I NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

« NUMBER OF ANIMALS NECROP3IED 

D-I3
 
TABLE D2 (CONCLUDED) 

"DOSlT 

CONTROL{UNTR)

06-0070

 CONTROL (UMk)

 06-0118

 DOSE A

 06-0071

 DOSE B 

 06-0102 

BODY CAVITIE S 

*MiSENTEPY

PmARTERITIS

 (U8) (50) (13)

 1 (2*) 

{US) 

ALL OTHEB SYSTEMS 

*HULII?LC CSGANS

PEEIVASCULITI S

 (48)

 1 (2S) 

 (50) (H3) (US) 

SPECIAI MOEPHOLOGY SUMWAE Y 

NO L^SIOli DEPORTEE

ANIMA L MISSING/NO NECBOPSY

DSCROFSY P£PF/KO HISTO PERFOEMfcC

AUTO/NECKOPSY/HISTO PERF

AUTO/KLCROFSY/NO HISTO

AUTOLYSIS/MO KECROPSY

 1 

2

 3

 1 

2

 1 

6 

1 

1 

5 

« NUMBER

* NUNB2R

 OF AMIKALS

 OF ANIMAL S

 WITH TISSUE

 NECfcOPSISD 

 EXAHINED MICROSCOPICALLY 

D-14 






DHEW Publication No. (NIH) 78-1382