1978 

OFFICIAL NTP
 

FILE COPY
 

National Cancer Institute 

CARCINOGENESIS 

Technical Report Series DO NOT REMOVE FROM THIS OFFICE 

No. 91 

BIOASSAY OF 

CLONITRALID 

FOR POSSIBLE CARCINOGENICITY 

CAS No. 1420-04-8 

NCI-CG-TR-91 

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

Public Health Service 

National Institutes of Health 

BIOASSAY OF
 

CLONITRALID
 

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) 78-1341
 

REPORT ON THE BIOASSAY OF CLONITRALID
 

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 clonitralid 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 clonitralid was conducted by Hazleton
 

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

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

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

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

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

Dr. R. W. Voelker (3), Dr. W. A. Olson (3,4) and Dr. W. M. Weatherholtz (3). Chemical analysis was performed by Dr. C. L. Guyton (3,5)
 

and the analytical results were reviewed by Dr. N. Zimmerman (6); the
 

technical supervisor of animal treatment and observation was Ms. K.
 

J. Petrovics (3).
 

Histopathologic examinations were performed by Dr. D. A. Banas
 

(3) and Dr. R. H. Habermann (3) and reviewed by Dr. R. W. Voelker
 

(3) at the Hazleton Laboratories America, Inc., and the diagnoses
 

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

L. Schueler (7).
 

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

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

111
 
methods selected for the Carcinogenesis Testing Program by Dr. J. J.
 

Gart (9).
 

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

Walker (6), biochemist Mr. S. C. Drill (6), and technical editor Ms.
 

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

participating organizations.
 

The following other scientists at the National Cancer Institute
 

were responsible for evaluating the bioassay experiment, interpreting
 

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

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

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

Squire (1,10), Dr. J. M. Ward (1), and Dr. C. E. Whitmire (l).
 

1.	 Carcinogenesis Testing Program, Division of Cancer Cause and
 

Prevention, National Cancer Institute, National Institutes of
 

Health, Bethesda, Maryland.
 

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

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

Vienna, Virginia.
 

4.	 No wit w h the Center for Regulatory Services, 2347 Paddock Lane,
 

Reston, Virginia.
 

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

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

Boulevard, McLean, Virginia.
 

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

Maryland.
 

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

Rockville, Maryland.
 

9.	 Mathematical Statistics and Applied Mathematics Section, Biometry
 

Branch, Field Studies and Statistics Program, Division of Cancer
 

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

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

University, School of Medicine, Traylor Building, Baltimore,
 

Maryland.
 

v
 

SUMMARY
 

A bioassay for possible carcinogenicity of clonitralid was conducted using Osborne-Mendel rats and B6C3F1 mice. Clonitralid was
 

administered in the feed, at either of two concentrations, to groups
 

of 50 male and 50 female animals of each species. Twenty animals of
 

each sex and species were placed on test as controls. The time-


weighted average high and low dietary concentrations of clonitralid
 

were, respectively, 28,433 and 14,216 ppm for rats and 549 and 274
 

ppm for mice. After a 78-week period of compound administration,
 

there was an additional observation period of 32 to 33 weeks for rats
 

and 13 to 14 weeks for mice.
 

Adequate numbers of male rats, female rats, and female mice survived long enough to be at risk from late-developing tumors. Because
 

of inadequate survival among male mice, however, results obtained
 

from observation of the male mouse groups cannot be considered conclusive.
 

The incidences of mammary adenocarcinomas in treated female rats
 

were not significantly higher than the incidences observed in control
 

female rats. However, the incidences of this lesion in dosed female
 

rats were greater than or equal to 22 percent, while the highest incidence observed in 15 control groups at this laboratory was only 10
 

percent with a mean incidence of 2.6 percent. The occurrence in high
 

dose female rats (2/45) of carcinomas in the glandular portion of the
 

stomach with metastases to other sites was not statistically significant. This incidence, however, is much greater than the historical
 

control incidence and suggests an association between administration
 

of clonitralid and the development of these tumors.
 

No statistically significant increased tumor incidences were
 

observed among male rats or mice of either sex dosed with clonitralid.
 

Under the conditions of this bioassay, there was no convincing
 

evidence that clonitralid was carcinogenic to Osborne-Mendel rats or
 

to female B6C3F1 mice. Poor survival of male mice did not permit an
 

evaluation of carcinogenicity in these animals.
 

Vll
 

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 8
 

F. Experimental Design 9
 

G. Clinical and Histopathologic Examinations 12
 

H. Data Recording and Statistical Analyses 14
 

III. CHRONIC TESTING RESULTS: RATS 19
 

A. Body Weights and Clinical Observations 19
 

B. Survival 19
 

C. Pathology 22
 

D. Statistical Analyses of Results 24
 

IV. CHRONIC TESTING RESULTS: MICE 34
 

A. Body Weights and Clinical Observations 34
 

B. Survival 34
 

C. Pathology 37
 

D. Statistical Analyses of Results 37
 

V. DISCUSSION 42
 

VI. BIBLIOGRAPHY 44
 

APPENDIX A SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

RATS TREATED WITH CLONITRALID A-l
 

APPENDIX B SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

MICE TREATED WITH CLONITRALID B-l
 

APPENDIX C SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN RATS TREATED WITH CLONITRALID C-l
 

APPENDIX D SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN MICE TREATED WITH CLONITRALID D-l
 

IX
 
LIST OF ILLUSTRATIONS 

Figure Number Page 

1 CHEMICAL STRUCTURE OF CLONITRALID 2 

2 GROWTH CURVES FOR CLONITRALID CHRONIC 

STUDY RATS 20 

SURVIVAL COMPARISONS OF CLONITRALID 

CHRONIC STUDY RATS 21 

GROWTH CURVES FOR CLONITRALID CHRONIC 

STUDY MICE 35 

SURVIVAL COMPARISONS OF CLONITRALID 

CHRONIC STUDY MICE 36 

LIST OF TABLES 

Table Number Page 

DESIGN SUMMARY FOR OSBORNE-MENDEL RATS— 

CLONITRALID FEEDING EXPERIMENT 10 

DESIGN SUMMARY FOR B6C3F1 MICE—CLONITRALID 

FEEDING EXPERIMENT 11 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN MALE RATS TREATED WITH 

CLONITRALID • 25 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN FEMALE RATS TREATED 

WITH CLONITRALID 28 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN MALE MICE TREATED WITH 

CLONITRALID 38 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

AT SPECIFIC SITES IN FEMALE MICE TREATED 

WITH CLONITRALID 39 

Al SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

MALE RATS TREATED WITH CLONITRALID A-3 
LIST OF TABLES .(Concluded) 

Table Number Paee 

A2 SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

FEMALE RATS TREATED WITH CLONITRALID A-6 

Bl SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

MALE MICE TREATED WITH CLONITRALID B-3 

B2 SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

FEMALE MICE TREATED WITH CLONITRALID B-6 

Cl SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN MALE RATS TREATED WITH CLONITRALID C-3 

C2 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN FEMALE RATS TREATED WITH CLONITRALID C-8 

Dl SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN MALE MICE TREATED WITH CLONITRALID D-3 

D2 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN FEMALE MICE TREATED WITH CLONITRALID D-6 

xi
 

I. INTRODUCTION
 

Clonitralid (Figure l) (NCI No. C00431), a powerful molluscicide
 

and lamprey killer, was selected for bioassay by the National Cancer
 

Institute because of the large potential for human exposure resulting
 

from the direct application of the compound for control of sea lamprey
 

larvae in tributaries to the Great Lakes (Menzie, 1977) and the widespread application of clonitralid for the control of water snails
 

(Sturrock, 1974; Sturrock et al., 1974; Magendantz, 1974). The lack
 

of data on the toxic effects of chronic exposure to clonitralid was
 

an additional factor in the selection of this compound for testing.
 

The Chemical Abstracts Service (CAS) Ninth Collective Index
 

(1977) name for this compound is 5-chloro-N-(2-chloro-4-nitrophenyl)­

&
 

2-hydroxybenzamide compound with 2-aminoethanol (1:1). It is also
 

called 2',5-dichloro-4'-nitrosalicylanilide compound with 2-aminoethanol (1:1); clonitralide; Bayer 25648; Bayer 73; and SR 73.
 

Clonitralid is used alone or in a mixture with 3-trifluoromethyl4-nitrophenol sodium salt by the U.S. Fish and Wildlife Service in the
 

Great Lakes sea lamprey (Petromyzon marinus) control program (Menzie,
 

1977). Sea lampreys, which become trapped in the lakes after gaining
 

access in large numbers through canals with locks, have attacked and
 

drastically reduced the large, and commercially important lake trout
 

populations, at one point reducing the annual catch from 15 million
 

*
 

The CAS registry number is 1420-04-8.
 
OolM V \ 

FIGURE 1 

CHEMICAL STRUCTURE OF CLONITRALID 
to 0.5 million pounds (Storer et al., 1972). Clonitralid is introduced in Great Lakes tributaries, in amounts which vary with the rate
 

of flow of the streams, to eradicate the burrowing lamprey larvae
 

(Menzie, 1977).
 

Clonitralid is used extensively for the control of water snails,
 

especially in tropical areas (Farm Chemicals Handbook, 1976; Sturrock,
 

1974; Sturrock et al., 1974; Magendantz, 1974). These snails are
 

intermediate hosts for blood and liver flukes, which produce schistosomiasis and fascioliasis, respectively, thereby constituting a significant health hazard (Farm Chemicals Handbook, 1976; Morris, 1976).
 

The control of water snails by clonitralid is achieved, not only by
 

direct introduction into the aqueous environment, but also by application of the compound to vegetation and soils (Sturrock, 1974; Sturrock
 

et al., 1974). Clonitralid is extremely toxic to water snails, giving
 

a total kill of mature Australorbis glabratus after 24 hours of exposure to concentrations as low as 0.1 ppm (Fox et al., 1966), and lowering populations of Biomphalaria choanomphala by 90 to 100 percent,
 

under favorable conditions, when applied to lake and stream bottoms
 

at a rate of 3.75 kg/ha (Magendantz, 1974).
 

Clonitralid has also been examined, with favorable results, for
 

use as a fish toxicant in streams or impounded water (Farm Chemicals
 

Handbook, 1976).
 

Specific production figures for clonitralid are not available.
 

Commercial production (in excess of 1000 pounds or $1000 in value
 
annually) was last reported in 1975 by one U.S. company (Stanford Research Institute, 1976).
 

The potential for exposure to clonitralid is greatest for pest
 

control workers, especially those who apply the compound as a spray.
 

Workers in production facilities may also be exposed. The major
 

route of population exposure is presumably dermal contact with or
 

ingestion of treated water or ingestion of contaminated fish.
 

Clonitralid levels in both cases are probably quite low because the
 

high toxicity of the compound to fish and other nontarget organisms
 

(Farm Chemicals Handbook, 1976) necessitates extremely low and well-


controlled practicable concentrations. Toxic levels of clonitralid
 

for a number of species of fish tested were well below 0.5 ppm (Farm
 

Chemicals Handbook, 1976).
 

Clonitralid residues do not: accumulate to a detectable degree in
 

bananas when applied to the soil surrounding the plants at concentrations of 0.125 to 1.250 ppm (Sturrock et al., 1974), thus precluding
 

ingestion when used in this manner. Clonitralid sprayed on vegetation, however, remained toxic to Biomphalaria glabrata for 8 weeks,
 

and residues persisted in mud for over a year, in both cases under
 

tropical conditions (Sturrock, 1974).
 

No data on the toxicity of this compound to humans are available.
 

In contrast to the efficacy of clonitralid against mollusks, fish,
 

and lampreys, experiments with rats have indicated a relatively low
 

mammalian toxicity (Farm Chemicals Handbook, 1976).
 
II. MATERIALS AND METHODS 

A. Chemicals
 

Technical-grade clonitralid (21 ,5-dichloro-4'-nitrosalicylanilide
 

ethanolamine salt) was purchased from Chemagro Corporation, Kansas
 

City, Missouri, and analyzed by Hazleton Laboratories, Inc., Vienna,
 

Virginia. The experimentally determined melting point of 192° to
 

200°C suggested the presence of impurities due to the wide spread
 

and difference from the reported literature value of 216°C. Purity,
 

determined by a colorimetric method (suggested by the manufacturer)
 

utilizing ultraviolet absorbance at 416 nm, was shown to be greater
 

than 90 percent. A similar chemical analysis was performed twelve
 

months later. A change in the experimentally determined melting
 

point (186° to 190°C) did suggest some change, although it cannot
 

be ascertained whether this was attributable to alterations in the
 

impurities, the major compound, or both. Colorimetry results of the
 

second analysis were similar to those of the first.
 

Throughout this report the term clonitralid is used to represent
 

this technical-grade material.
 

B. Dietary Preparation
 

The basal laboratory diet for treated and control animals con­

®
 sisted of Wayne Lab-Blox meal (Allied Mills, Inc., Chicago, Illinos)
 

®
 plus 2 percent Duke's corn oil (S. F. Sauer Company, Richmond, Virginia) by weight. Fresh mixtures of clonitralid in corn oil were
 

prepared each week and stored in the dark. The clonitralid mixtures
 
were incorporated into the appropriate amount of laboratory diet in a
 

twin-shell blender fitted with an accelerator bar.
 

C. Animals
 

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

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

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

carcinogenesis bioassays and has proved satisfactory in this capacity.
 

Rats and mice of both sexes were obtained through contracts of
 

the Division of Cancer Treatment, National Cancer Institute. The
 

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

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

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

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

dosed and control groups.
 

D. Animal Maintenance
 

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


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

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

conditioning system in the laboratory provided filtered air at a rate
 

of 12 to 15 complete changes of room air per hour. Fluorescent lighting was provided on a 12-hour-daily cycle.
 
The rats were individually housed in suspended galvanized-steel
 

wire-mesh cages with perforated floors, while mice were housed by sex
 

in groups of ten in solid-bottom, polypropylene cages equipped with
 

filter tops. Sanitized cages with fresh bedding (Sanichips , Pine-


wood Sawdust Company, Moonachie, New Jersey) were provided once each
 

week for mice. Rats received sanitized cages with no bedding with
 

the same frequency. Food hoppers were changed and heat-sterilized
 

once a week for the first 10 weeks and once a month thereafter, while
 

fresh heat-sterilized glass water bottles and sipper tubes were provided three times a week. Food (Wayne Lab-Blox meal) and water were
 

available ad libitum.
 

The clonitralid-dosed and control rats were housed in the same
 

*
 

room with other rats receiving diets containing dioxathion (78-34-2);
 

mexacarbate (315-18-4); pentachloronitrobenzene (82-68-8); amitrole
 

(61-82-5); nitrofen (1836-75-5); endosulfan (115-29-7); and trifluralin (1582-09-8).
 

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

housed in the same room as other mice receiving diets containing trifluralin (1582-09-8); chlorobenzilate (510-15-6); dioxathion (78-34­

2); sulfallate (95-06-7); p,p'-DDT (50-29-3); methoxychlor (72-43-5);
 

p,p'-DDE (72-55-9); p,p'-TDE (72-54-8); dicofol (115-32-2); pentachloronitrobenzene (82-68-8); nitrofen (1836-75-5); endosulfan (115­

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

(53-96-3); and safrole (94-59-7).
 

X

CAS registry numbers are given in parentheses.
 
E. Selection of Initial Concentration
 

In order to establish the maximum tolerated concentrations of
 

clonitralid for administration to treated animals in the chronic
 

studies, Subchronic toxicity tests were conducted with both rats and
 

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

each consisting of five males and five females. Clonitralid was
 

premixed with a small amount of corn oil. This mixture was then
 

incorporated into the laboratory diet and fed _ad_ libitum to five of
 

the six rat groups in concentrations of 2150, 4640, 10,000, 21,500,
 

and 46,400 ppm. Five of the six mouse groups were given concentrations of 464, 1000, 2150, 4640, and 10,000 ppm. The sixth group of
 

each species served as a control group, receiving only the mixture of
 

corn oil and laboratory meal. The dosed dietary preparations were
 

administered for a period of 6 weeks, followed by a 2-week observation
 

period during which all animals were fed the basal laboratory diet.
 

A concentration inducing no mortality and resulting in a depression in mean group body weight of approximately 20 percent relative
 

to controls was selected as the initial high concentration for the
 

chronic study. When weight gain criteria were not applicable, mortality data alone were utilized.
 

At both 10,000 and 21,500 ppm, mean body weight depression was
 

17 percent in male rats. In female rats, mean body weight depression
 

was 14 percent at 10,000 ppm, and 21 percent at 21,500 ppm. The
 

initial high concentrations selected for male and female rats in the
 
chronic bioassay were 17,500 and 19,000 ppm, respectively. The high
 

concentration actually used for rats of both sexes in the chronic
 

bioassay was 18,250 ppm.
 

Mean body weight depression was first evident at a concentration
 

of 464 ppm for both male and female mice. At that concentration mean
 

body weight depression was 27 percent for the male mice and 14 percent
 

for the female mice. At 1000 ppm, the mean body weight depression for
 

male mice was 34 percent, and for female mice was 55 percent. The
 

initial high concentration selected for both male and female mice in
 

the chronic bioassay was 380 ppm.
 

F

' Experimental Design
 

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

treated and untreated observation periods, and the time-weighted
 

average concentrations) are summarized in Tables 1 and 2.
 

The treated and control rats were all approximately 6 weeks old
 

at the time they were placed on test and all shared the same median
 

date of birth. The concentrations of clonitralid initially utilized
 

for male and female rats were 18,250 and 9125 ppm. Throughout this
 

report those rats initially receiving the former concentration are
 

referred to as the high dose groups, while those initially receiving
 

the latter concentration are referred to as the low dose groups. In
 

week 8, the high and low concentrations administered to the male and
 

female rats were increased to 25,000 and 12,500 ppm, respectively.
 
TABLE 1
 

DESIGN SUMMARY FOR OSBORNE-MENDEL RATS
 

CLONITRALID FEEDING EXPERIMENT
 

INITIAL OBSERVATION PERIOD TIME-WEIGHTED 

GROUP CLONITRALID TREATED UNTREATED AVERAGE 

SIZE CONCENTRATION3 (WEEKS) (WEEKS) CONCENTRATION 

MALE
 

CONTROL 20 0	 110 0
 

LOW DOSE 50 9,125 7 14,216
 

12,500 8
 

15,000 63
 

0	 32
 

HIGH DOSE 50	 18,250 7 28,433
 

25,000 8
 

30,000 63
 

0	 33
 

FEMALE
 

CONTROL 20	 110
 

LOW DOSE 50 9,125 7 14,216
 

12,500 8
 

15,000 63
 

0	 33
 

HIGH DOSE 50	 18,250 7 28,433
 

25,000 8
 

30,000 63
 

0	 33
 

Concentrations given in	 parts per million.
 

b . .	 . £(concentration X weeks received)
 Time-weighted average concentration = -rrr : :—: ; : r\
 Z(weeks receiving chemical;
 

10
 
TABLE 2
 

DESIGN SUMMARY FOR B6C3F1 MICE
 

CLONITRALID FEEDING EXPERIMENT
 

INITIAL 

GROUP CLONITRALID 

SIZE CONCENTRATION3 

MALE 

CONTROL 20 0 

LOW DOSE 50	 190
 

225
 

300
 

0
 

HIGH DOSE 50	 380
 

450
 

600
 

0
 

FEMALE
 

CONTROL 20	 0
 

LOW DOSE 50	 190
 

225
 

300
 

0
 

HIGH DOSE 50	 380
 

450
 

600
 

0
 

a
 

Concentrations given in parts per
 

OBSERVATION PERIOD TIME-WEIGHTED
 

TREATED UNTREATED AVERAGE ,
 

(WEEKS) (WEEKS) CONCENTRATION
 

91	 0
 

8 274
 

15
 

55
 

13
 

8 549
 

15
 

55
 

13
 

91	 0
 

8 274
 

15
 

55
 

14
 

8 549
 

15
 

55
 

14
 

million.
 

S (concentration X weeks received)
 

(weeks receiving chemical)
 

11
 
In week 16, the dosages were again increased, to 30,000 ppm for the
 

high dose male and female rats, and to 15,000 ppm for the low dose
 

male and female rats. These concentrations were maintained for the
 

remainder of the period of compound administration. After the period
 

of 78 weeks, observation of all rats used in the chronic bioassay
 

continued for an additional period of up to 33 weeks.
 

The treated and control mice were all approximately 6 weeks old
 

at the time the experiment began and all shared the same median date
 

of birth. The concentrations administered to male and female mice for
 

the first 8 weeks were 380 and 190 ppm. Throughout this report those
 

mice initially receiving the former concentration are referred to as
 

the high dose groups, while those mice initially receiving the latter
 

concentration are referred to as the low dose groups. In week 9,
 

the high and low concentrations were increased to 450 and 225 ppm,
 

respectively. In week 24, dosages were again increased, this time
 

to 600 and 300 ppm for the high and low dose groups, respectively.
 

These concentrations were maintained for the remainder of the 78-week
 

period. Observation of all mice continued for up to 14 weeks after
 

the period of compound administration.
 

G. Clinical and Histopathologic Examinations
 

Animals were weighed immediately prior to initiation of the
 

experiment. Body weights, food consumption, and data concerning
 

appearance, behavior, signs of toxic effects, and incidence, size,
 

and location of tissue masses were recorded at weekly intervals for
 

12
 
the first 10 weeks and at monthly intervals thereafter. From the
 

first day, all animals were inspected daily for mortality. The presence of tissue masses was determined by observation and palpation of
 

each animal.
 

During the course of this bioassay several pathology protocols
 

were in effect, each for different periods of time. The minimum protocol required that, if possible, certain tissues were to be taken
 

and examined histopathologically from all control animals, from any
 

animal in which a tumor was observed during gross examination, and
 

from at least 10 grossly normal males and 20 grossly normal females
 

from each treated group. In addition, any tissues showing gross abnormalities were to be taken and examined histopathologically. Under
 

later protocols, some tissues were taken from additional dosed animals,. The number of animals in each group from which a tissue was
 

examined is indicated in Appendices A through D.
 

A necropsy was performed on each animal regardless of whether it
 

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

bioassay. The animals were euthanized by exsanguination under sodium
 

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

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

Tissues were preserved in 10 percent buffered formalin, embedded
 

in paraffin, sectioned, and stained with hematoxylin and eosin prior
 

13
 
to microscopic examination. An occasional section was subjected to
 

special staining techniques for more definitive diagnosis.
 

Slides were prepared from the following tissues from selected
 

animals: skin, subcutaneous tissue, lungs and bronchi, trachea, bone
 

marrow, spleen, lymph nodes, thymus, heart, salivary gland, liver,
 

gallbladder (mice), pancreas, esophagus, stomach, small intestine,
 

large intestine, kidney, urinary bladder, pituitary, adrenal, thyroid,
 

parathyroid, testis, prostate, brain, tunica vaginalis, 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,
 

14
 
1969). Data tables were generated for verification of data transcription and for statistical review.
 

These data were analyzed using the statistical techniques described in this section. Those analyses of the experimental results
 

that bear on the possibility of carcinogenicity are discussed in the
 

statistical narrative sections.
 

Probabilities of survival were estimated by the product-limit
 

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

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

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

missing; animals dying from natural causes were 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 Tarone1s (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
 

15
 
tumors), or when lesions could have appeared at multiple sites (e.g.,
 

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

The purpose of the statistical analyses of tumor incidence is to
 

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

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

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

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

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

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

significance level of 0.05 may be made. The Bonferroni inequality
 

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

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

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

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

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

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

when appropriate. Under the assumption of a linear trend, this test
 

determined if the slope of the dose-response curve is different from
 

zero at the one-tailed 0.05 level of significance. Unless otherwise
 

noted, the direction of the significant trend was a positive dose relationship. This method also provides a two-tailed test of departure
 

from linear trend.
 

16
 
A time-adjusted analysis was applied when numerous early deaths
 

resulted from causes that were not associated with the formation of
 

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

tumor was observed were excluded by basing the statistical tests on
 

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

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

tumor was found, comparisons were based exclusively on animals that
 

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

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

Cochran-Armitage tests, etc.) were followed.
 

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

incidence of tumors. Curves of the proportions surviving without an
 

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
 

17
 
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
 

t c t
 

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

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

c
 

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

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

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

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

larger proportion in the treated group than in the control.
 

The lower and upper limits of the confidence interval of the
 

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

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

of the risk in a treated group of animals to that in a control group
 

would be within the interval calculated from the experiment. When
 

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

can be inferred that a statistically significant result (a P < 0.025
 

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

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

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

the lower limit indicates the absence of a significant result while
 

the upper limit indicates that there is a theoretical possibility
 

of the induction of tumors by the test chemical which could not be
 

detected under the conditions of this test.
 

18
 
III. CHRONIC TESTING RESULTS: RATS
 

A. Body Weights and Clinical Observations
 

In treated females there was distinct, compound-related mean
 

body weight depression when compared to controls. This was not,
 

however, true for males (Figure 2).
 

For the duration of compound administration, appearance and behavior were generally comparable for all groups with the exception
 

of slightly higher incidences of hunched appearance, abdominal urine
 

stains, and/or rough fur in the treated rats when compared to the
 

untreated controls. From cessation of compound administration to
 

termination of the study, clinical signs were noted at comparable
 

rates in control and treated animals. These signs included hunched
 

appearance; abdominal urine stains; rough fur; body sores; cloudy,
 

squinted, or reddened eyes with discharge or crust; and alopecia.
 

As the animals aged, the incidence of these signs gradually increased
 

in all groups. Respiratory signs characterized by labored respiration, wheezing and/or nasal discharge were observed at a low to
 

moderate incidence during the study. During the last three months
 

of the study, palpable subcutaneous nodules were observed at a
 

greater frequency in the high dose male and female rats than in the
 

other groups.
 

B. Survival
 

The estimated probabilities of survival for male and female rats
 

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

19
 
750 750 

— 600 

— 450 

-300 

(-150 

15 120 

TIME ON TEST (WEEKS) 

750 750 

- 600 

-450 

-300 

-150 

r 

15 45 60 75 90 

TIME ON TEST (WEEKS) 

FIGURE 2 

GROWTH CURVES FOR CLONITRALID CHRONIC STUDY RATS 

20 
PROBABILITY OF SURVIVAL PROBABILITY OF SURVI

p 

b 

V) 

c 

3) 

oo_ O o 

O 

-o 

> 

33 

s 

r-o -i 

O c m Z3J to 

Hm ^ 

5" ^ j> m 

m 

O 

X 

3] 

O 

O 

CO 

O 

2 

o — 

<J1 

3 

1 1 O O 
For male rats the Tarone test for positive association between
 

increased dosage and accelerated mortality was not significant. There
 

were adequate numbers of male rats at risk from late-developing tumors
 

with 92 percent (46/50) of the high dose, 80 percent (40/50) of the
 

low dose and 85 percent (17/20) of the controls living at least 90
 

weeks.
 

For female rats the Tarone test did not show a significant association between increased dosage and accelerated mortality. With 64
 

percent (32/50) of the high dose, 76 percent (38/50) of the low dose,
 

and 70 percent (14/20) of the control animals alive until the termination of the experiment, there were adequate numbers at risk from late-


developing tumors.
 

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).
 

Adenocarcinomas of the mammary gland occurred in 1/20 (5 percent)
 

control males, 3/49 (6 percent) high dose males, 1/20 (5 percent)
 

control females, 12/50 (24 percent) low dose females, and 11/50 (22
 

percent) high dose females. Microscopically, the adenocarcinomas of
 

the mammary gland consisted of irregularly formed acini lined with
 

anaplastic epithelial cells. A piling up of cells around the acini
 

accompanied by papillary projections into the lumen was often observed.
 

Some of the tumors were very cellular and were composed of tightly
 

22
 
packed epithelial cells in sheets or nests forming small acini. Some
 

acini contained eosinophilic amorphous material. The neoplastic cells
 

invaded and replaced the surrounding tissues, and lung metastasis was
 

evident in one low dose and two high dose female rats.
 

Mammary fibroadenomas occurred in 1/49 (2 percent) high dose
 

males, 4/20 (20 percent) control females, 13/50 (26 percent) low dose
 

females, and 11/50 (22 percent) high dose females. The fibroadenomas
 

were composed of nests and islands of mammary gland acini separated
 

by broad bands of collagenous and fibrous connective tissue. The
 

fibroadenomas were well-circumscribed from the surrounding tissues.
 

Gastric carcinomas of the glandular region of the stomach occurred in 2/45 (4 percent) high dose females. Microscopically, one
 

of the tumors had irregular glandular acini of various sizes. The
 

epithelial cells were supported by bands of fibrous connective tissue.
 

In some areas, only scattered clumps of anaplastic epithelial cells
 

were seen, typical of a scirrhous carcinoma. The neoplastic cells
 

invaded and replaced the tunica muscularis and serosa of the stomach
 

and metastasized to the pancreas, liver, urinary bladder, and esophagus. The other gastric carcinoma was characterized by nests and
 

cords of poorly differentiated, large, plump cells with a scanty
 

fibrous connective tissue stroma. Many bizarre mitotic figures were
 

present. This tumor metastasized to the liver and lung. Nonneoplastic proliferative lesions (hyperkeratosis and acanthosis) were
 

23
 
observed in the stomachs of 7/41 (17 percent) low dose and 7/44 (16
 

percent) high dose female rats.
 

A greater incidence of thyroid neoplasms was observed in the
 

treated rats than in the control groups; however, the overall incidence
 

was not large. Both follicular-cell and C-cell tumors were observed.
 

There was no evidence of increased numbers of preneoplastic proliferative thyroid lesions, and thyroid neoplasms have been observed with
 

greater frequency in historical control groups. Therefore, these
 

neoplasms are not considered to be compound-related. Pancreatic
 

islet-cell tumors and endometrial stromal polyps were observed among
 

treated rats, but not in associated control groups. These relatively
 

common tumors were considered spontaneous.
 

Other neoplastic lesions occurred with similar frequency in the
 

control and treated groups and were considered to have occurred naturally.
 

The inflammatory, degenerative, and hyperplastic lesions that
 

occurred were similar in kind and number to those that occur spontaneously in aged Osborne-Mendel rats.
 

Based upon the results of this histopathologic examination clonitralid was carcinogenic in female rats, causing an increased incidence of adenocarcinomas of the mammary gland and possibly carcinomas
 

of the glandular portion of the stomach.
 

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
 

24
 
TABLE 3
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN MALE RATS TREATED WITH CLONITRALID*
 

TOPOGRAPHY : MORPHOLOGY
 

Subcutaneous Tissue: Fibroma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Circulatory System: Hemangiosarcoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pituitary: Chromophobe Adenoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

CONTROL
 

0/20(0.00)
 

P = 0.039
 

2/20(0.10)
 

N.S.
 

100
 

5/19(0.26)
 

P = 0.034(N)
 

97
 

LOW
 

DOSE
 

0/49(0.00)
 

N.S.
 

5/49(0.10)
 

N.S.
 

1.020
 

0.188
 

10.204
 

97
 

7/40(0.18)
 

N.S.
 

0.665
 

0.217
 

2.371
 

96
 

HIGH
 

DOSE
 

4/49(0.08)
 

N.S.
 

Infinite
 

0.394
 

Infinite
 

93
 

3/49(0.06)
 

N.S.
 

0.612
 

0.078
 

6.996
 

71
 

2/33(0.06)
 

N.S.
 

0.230
 

0.025
 

1.267
 

111
 
TOPOGRAPHY : MORPHOLOGY
 

Thyroid: Follicular-Cell Adenoma
 

or Follicular-Cell Carcinoma^3
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pancreatic Islets: Islet-Cell
 

Adenoma or Islet-Cell Carcinoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Mammary Gland: Adenocarcinoma NOS
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

CONTROL
 

0/20(0.00)
 

N.S.
 

0/18(0.00)
 

N.S.
 

1/20(0.05)
 

N.S.
 

91
 

LOW
 

DOSE
 

4/42(0.10)
 

N.S.
 

Infinite
 

0.460
 

Infinite
 

60
 

3/41(0.07)
 

N.S.
 

Infinite
 

0.277
 

Infinite
 

94
 

0/49(0.00)
 

N.S.
 

0.000
 

0.000
 

7.624
 

HIGH
 

DOSE
 

3/35(0.09)
 

N.S.
 

Infinite
 

0.358
 

Infinite
 

96
 

2/32(0.06)
 

N.S.
 

Infinite
 

0.174
 

Infinite
 

96
 

3/49(0.06)
 

N.S.
 

1.224
 

0.108
 

62.958
 

88
 
TABLE 3 (CONCLUDED)
 

o
 

Treated groups received time-weighted average doses of 14,215 or 28,433 ppm in feed.
 

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

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

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

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

control group is given beneath the incidence of tumors in the treated group when P < 0.05;
 

otherwise, not significant (N.S.) is indicated. For both Cochran-Armitage and Fisher exact
 

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

in the control group.
 

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

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN FEMALE RATS TREATED WITH CLONITRALID£
 

HIGH
 

DOSE
 

0/50(0.00)
 

N.S.
 

0.000
 

0.000
 

7.475
 

2/45(0.04)
 

N.S.
 

0.889
 

0.050
 

51.294
 

111
 

10/44(0.23)
 

N.S.
 

0.909
 

0.337
 

3.031
 

88
 

TOPOGRAPHY: MORPHOLOGY
 

Subcutaneous Tissue: Fibroma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Circulatory System: Hemangiosarcoma
 OO
 

P Values0
 

Relative Risk (Control)d
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pituitary: Chromophobe Adenoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

CONTROL
 

1/20(0.05)
 

N.S.
 

96
 

1/20(0.05)
 

N.S.
 

110
 

5/20(0.25)
 

N.S.
 

93
 

LOW
 

DOSE
 

3/50(0.06)
 

N.S.
 

1.200
 

0.106
 

61.724
 

97
 

3/42(0.07)
 

N.S. •
 

1.429
 

0.129
 

74.895
 

110
 

7/42(0.17)
 

N.S.
 

0.667
 

0.215
 

2.392
 

101
 
TOPOGRAPHY: MORPHOLOGY
 

Thyroid: Follicular-Cell Adenoma or
 

Follicular-Cell Carcinoma^
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Thyroid: C-Cell Adenoma or C-Cell
 

Carcinoma^5
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Mammary Gland: Adenocarcinoma NOS
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 4 (CONTINUED)
 

CONTROL
 

1/20(0.05)
 

N.S.
 

90
 

0/20(0.00)
 

P = 0.040
 

1/20(0.05)
 

N.S.
 

110
 

LOW
 

DOSE
 

4/41(0.10)
 

N.S.
 

1.951
 

0.214
 

93.623
 

110
 

1/41(0.02)
 

N.S.
 

Infinite
 

0.027
 

Infinite
 

111
 

12/50(0.24)
 

N.S.
 

4.800
 

0.803
 

200.027
 

69
 

HIGH
 

DOSE
 

3/44(0.07)
 

N.S.
 

1.364
 

0.120
 

69.919
 

111
 

5/44(0.11)
 

N.S.
 

Infinite
 

0.597
 

Infinite
 

103
 

11/50(0.22)
 

N.S.
 

4.400
 

0.722
 

184.751
 

86
 
TOPOGRAPHY : MORPHOLOGY
 

Mammary Gland: Fibroadenoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Mammary Gland: Fibroadenoma or
 

Adenocarcinoma NOS
 

P Values0
 

OJ 

o Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Uterus: Endometrial Stromal Polyp
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 4 (CONTINUED) 

CONTROL
 

4/20(0.20)
 

N.S.
 

90
 

4/20(0.20)
 

N.S.
 

90
 

0/20(0.00)
 

P = 0.036
 

LOW
 

DOSE
 

13/50(0.26)
 

N.S.
 

1.300
 

0.474
 

4.977
 

107
 

21/50(0.42)
 

N.S.
 

2.100
 

0.846
 

7.538
 

69
 

2/41(0.05)
 

N.S.
 

Infinite
 

0.150
 

Infinite
 

96
 

HIGH
 

DOSE
 

11/50(0.22)
 

N.S.
 

1.100
 

0.384
 

4.321
 

104
 

20/50(0.40)
 

N.S.
 

2.000
 

0.799
 

7.225
 

86
 

6/44(0.14)
 

N.S.
 

Infinite
 

0.758
 

Infinite
 

106
 
TABLE 4 (CONCLUDED)
 

«a
 

Treated groups received time-weighted average doses of 14,216 or 28,433 ppm in feed.
 

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

£

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

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

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

control group is given beneath the incidence of tumors in the treated group when P < 0.05;
 

otherwise, not significant (N.S.) is indicated. For both Cochran-Armitage and Fisher exact
 

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

in the control group.
 

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

u>
 
every type of malignant tumor in either sex where at least two such
 

tumors were observed in at least one of the control or clonitraliddosed groups and where such tumors were observed in at least 5 percent of the group.
 

For male rats the Cochran-Armitage test indicated a significant
 

(P = 0.039) positive association between dosage and the incidence of
 

fibromas of the subcutaneous tissue and a significant (P = 0.034)
 

negative association between dosage and the incidence of pituitary
 

chromophobe adenomas. In both cases, however, the Fisher exact tests
 

were not significant.
 

Similarly, for female rats the Cochran-Armitage test indicated
 

significant associations both between dose and the combined incidence
 

of C-cell adenomas and C-cell carcinomas of the thyroid and between
 

dose and the incidence of uterine endometrial stomal polyps. Once
 

again, the Fisher exact tests were not significant.
 

Mammary adenocarcinomas NOS were seen in unusually high numbers
 

in dosed rats of both sexes. In historical data collected by this
 

laboratory for the NCI Carcinogenesis Testing Program 5/389 (1 percent) of the untreated male and 11/388 (3 percent) of the untreated
 

female Osborne-Mendel rats had an adenocarcinoma NOS of the mammary
 

gland. These tumors were observed in 24 percent (12/50) of the low
 

dose and 22 percent (11/50) of the high dose females, but never in
 

excess of 10 percent (2/20) in the 15 historical female control
 

32
 
groups. A life-table analysis performed for this tumor was not significant.
 

In females carcinomas of the stomach were observed in 2/45 (4
 

percent) of the high dose rats. These rare tumors were observed in
 

none of the 350 historical control rats.
 

To provide additional insight into the possible carcinogenicity
 

of this compound, 95 percent confidence intervals on the relative
 

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

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

Tables 3 and 4, the value one is included; this indicates the absence
 

of statistically significant results. It should also be noted that
 

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

indicating the theoretical possibility of tumor induction in rats by
 

clonitralid that could not be established under the conditions of this
 

test.,
 

33
 
IV. CHRONIC TESTING RESULTS: MICE
 

A. Body Weights and Clinical Observations
 

Only very slight depression in mean group body weight was observed in treated mice when compared to controls (Figure 4).
 

Throughout the study, appearance and behavior were generally
 

comparable for treated and control mice except that body sores, alopecia and a hunched appearance were more frequently observed in the
 

males. A rapidly declining survival rate was noted in treated and
 

control males beginning in week 46 and continuing to termination of
 

the bioassay. The low survival and clinical signs were apparently
 

attributable to fighting in the group-housed male mice since in the
 

females survival and a comparatively lower incidence of clinical
 

signs were observed throughout the study. Other clinical signs commonly observed in laboratory mice were noted at a comparable rate in
 

control and treated groups. These signs included external genital
 

irritation, squinted or reddened eyes, rough or stained fur, and
 

palpable nodules.
 

B.	 Survival
 

The estimated probabilities of survival for male and female mice
 

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

For male mice the Tarone test for positive association between
 

increased dosage and accelerated mortality was not significant. Five
 

control mice drowned in week 63. There were not adequate numbers of
 

male mice at risk from late-developing tumors, with only 32 percent
 

34
 
50- -50 

40 — -40 

CO 

£30- -30 

X 

o 

£20- -20 

o 

CO 

CONTROL 

10- LOW DOSE — 10 

MALE MICE HIGH DOSE 

I I 1 I T \ 

15 30 45 60 75 90 105 120 

TIME ON TEST (WEEKS) 

50­ -50 

co 

40 — — 40 

£30- -30 

> 20­ Q 

O 

CO 

10— 

FEMALE MICE 

15 30 45 60 

TIME ON TEST (WEEKS) 

90 

_

..

_

I 

 CONTROL 

 LOW DOSE 

 HIGH DOSE 

105 

-20 

—10 

120 

FIGURE 4 

GROWTH CURVES FOR CLONITRALID CHRONIC STUDY MICE 

35 
PROBABILITY OF SURV

PROBABILITY OF SURVIVAL 

0) 

3D 

> 8­ I— 

n 

o 

30 

8 

o 

z 

m 

3 8­

o 

o 

DC 

30 

O 

O 

I 

o 

o 

m 

TO 

I 

o 

m 

I 

O 
(16/50) of the high dose, 42 percent (21/50) of the low dose, and 20
 

percent (4/20) of the vehicle controls alive at week 70.
 

For female mice the Tarone test did not show a significant association between increased dosage and accelerated mortality. With 90
 

percent (45/50) of the high dose, 92 percent (46/50) of the low dose,
 

and 90 percent (18/20) of the control mice alive until the termination of the experiment, adequate numbers of female mice were at risk
 

from late-developing tumors.
 

C. Pathology
 

Histopathologic findings on neoplasms in mice are summarized in
 

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

summarized in Appendix D (Tables Dl and D2).
 

A low incidence of neoplasms was observed in both control and
 

treated mice. These neoplasms were of the usual number and type
 

observed in mice of this age and strain.
 

Other degenerative, proliferative, and inflammatory lesions observed were also of the usual number and kind observed in aged B6C3F1
 

mice and were essentially comparable in incidence between control and
 

treated mice.
 

Based upon the results of this histopathologic examination,
 

clonitralid was neither toxic nor carcinogenic to B6C3F1 mice.
 

D. Statistical Analyses of Results
 

The results of the statistical analyses of tumor incidence in
 

mice are summarized in Tables 5 and 6. The analysis is included for
 

37
 
TABLE 5
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN MALE MICE TREATED WITH CLONITRALID*
 

LOW HIGH
 

TOPOGRAPHY: MORPHOLOGY CONTROL DOSE DOSE
 

Subcutaneous Tissue: Fibrosarcoma 0/16(0.00) 2/43(0.05) 3/43(0.07)
 

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

Relative Risk (Control) Infinite Infinite
 

Lower Limit 0.116 0.237
 

Upper Limit Infinite Infinite
 

Weeks to First Observed Tumor 62 68
 

u>
 00 Treated groups received time-weighted average doses of 274 or 549 ppm in feed,
 

D

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

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

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

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

control group is given beneath the incidence of tumors in the treated group when P < 0.05;
 

otherwise, not significant (N.S.) is indicated. For both Cochran-Armitage and Fisher exact
 

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

in the control group.
 

The 95% confidence interval on the relative risk of the treated group to the control group.
 
TABLE 6
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN FEMALE MICE TREATED WITH CLONITRALID2
 

TOPOGRAPHY: MORPHOLOGY
 

Lung: Alveolar /Bronchiolar Adenoma
 

P Values0
 

Relative Risk (Control)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Hematopoietic System: Malignant Lymphoma
 

P Values0
 

Relative Risk (Control)d
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

CONTROL
 

2/20(0.10)
 

N..S.
 

91
 

4/20(0.20)
 

P = 0.014(N)
 

85
 

LOW
 

DOSE
 

0/49(0.00)
 

N.S.
 

0.000
 

0.000
 

1.372
 

4/49(0.08)
 

N.S.
 

0.408
 

0.086
 

2.022
 

89
 

HIGH
 

DOSE
 

1/47(0.02)
 

N.S.
 

0.213
 

0.004
 

3.909
 

92
 

1/47(0.02)
 

P = 0.025(N)
 

0.106
 

0.002
 

1.003
 

92
 

Treated groups received time-weighted average doses of 274 or 549 ppm in feed.
 

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

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

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

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

control group is given beneath the incidence of tumors in the treated group when P < 0.05;
 

otherwise, not significant (N.S.) is indicated. For both Cochran-Armitage and Fisher exact
 

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

in the control group.
 

The 95% confidence interval on the relative risk of the treated group to the control group.
 
every type of malignant tumor in either sex where at least two such
 

tumors were observed in at least one of the control or clonitraliddosed groups and where such tumors were observed in at least 5 percent of the group.
 

In female mice the Cochran-Armitage test indicated a significant
 

(P = 0.014) negative association between dosage and the incidence of
 

malignant lymphomas. The Fisher exact test comparing the incidence
 

of these tumors in the high dose treated group with that in the
 

control mice had a probability level of P = 0.025, a marginal result
 

which was not significant under the Bonferroni criterion.
 

None of the statistical tests for any site in mice of either sex
 

indicated a significant positive association between the administration of clonitralid and tumor incidence. Thus, at the dose levels
 

used in this experiment there was no convincing evidence that clonitralid was a carcinogen in mice. It must be noted, however, that
 

poor survival precluded meaningful statistical analyses of the incidence of late-developing tumors in male mice.
 

To provide additional insight into the possible carcinogenicity
 

of this compound, 95 percent confidence intervals on the relative
 

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

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

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

of statistically significant results. It should also be noted that
 

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

40
 
indicating the theoretical possibility of tumor induction in mice by
 

clonitralid that could not be established under the conditions of
 

this test.
 

41
 
V. DISCUSSION
 

Adequate numbers of male rats, female rats, and female mice survived long enough to be at risk from late-developing tumors. Because
 

of inadequate survival among male mice, however, results obtained from
 

observation of the male mouse groups cannot be considered conclusive.
 

Although some neoplasms were observed at higher incidences in
 

dosed rats than control rats, none of these increased incidences were
 

statistically significant. The incidence of subcutaneous fibromas in
 

male rats, the combined incidence of C-cell adenomas and C-cell carcinomas of the thyroid in female rats, and the incidence of endometrial
 

stromal polyps in female rats each were found to have a significant
 

positive association with clonitralid dosage. The Fisher exact test,
 

however, did not indicate that the incidence in a dosed group was significantly greater than the incidence in the respective control group
 

in any of these cases.
 

The incidences of mammary adenocarcinomas in dosed female rats,
 

although not significantly higher than the incidences observed in
 

control female rats, were greater than or equal to 22 percent, while
 

the highest spontaneous incidence of this lesion observed in 15 control groups at this laboratory was only 10 percent with a mean incidence of 2.6 percent. The observation of mammary adenocarcinomas
 

in the dosed males (3/49) supports the findings in the females. The
 

occurrence in high dose female rats (2/45) of carcinomas of the glandular portion of the stomach with metastases to other sites was not
 

42
 
statistically significant. This incidence, however, is much greater
 

than the historical incidence (i.e., 0/350) observed in control female
 

rats maintained at Hazleton Laboratories during the NCI Carcinogenesis
 

Testing Program and suggests an association between administration of
 

clonitralid and the development of these tumors.
 

No significantly increased tumor incidences were observed among
 

mice treated with clonitralid.
 

Under the conditions of this bioassay, there was no convincing
 

evidence that clonitralid was carcinogenic to Osborne-Mendel rats or
 

to female B6C3F1 mice. Poor survival of male mice did not permit an
 

evaluation of carcinogenicity in these animals.
 

43
 
VI. BIBLIOGRAPHY
 

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.
 

Farm Chemicals Handbook, Meister Publishing Company, Willoughby,
 

Ohio, 1976.
 

Fox, I., G.A. Rivera, L.S. Ritchie, and L.P. Frick, "Effects of
 

Bayluscide Against Australorbis Glabratus of Different Ages,
 

Strains, and Cultural Conditions." Bulletin of the World
 

Health Organization 34(2):305-306, 1966.
 

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
 

_53:457-481, 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.
 

Magendantz, M., "Molluscicide Trials, and Recommendations for the
 

Reduction of Schistosomiasis Transmission Around an Urban Area
 

of Lake Victoria." Pflanzenschutz-Nachr. (American edition)
 

r7_(l):46-55, 1974; Chemical Abstracts 83 189274v.
 

Menzie, C., U.S. Fish and Wildlife Service, Washington, D.C. Personal communications, August 1977.
 

44
 
Miller, R.G., Simultaneous Statistical Inference. McGraw-Hill Book
 

Co., New York, 1966.
 

Morris, W., editor, The American Heritage Dictionary of the English
 

Language. Houghton Mifflin Company, Boston, 1976.
 

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

in Male Rats Given Subcutaneous Carbon Tetrachloride." Journal
 

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

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

Kaufman, "Respiratory Tract Carcinogenesis in Hamsters Induced
 

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

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

Stanford Research Institute, 1976 Directory of Chemical Producers,
 

U.S.A., Menlo Park, California, 1976.
 

Storer, T.I., R.L. Usinger, R.C. Stebbins, and J.W. Nybakken, General
 

Zoology, Fifth edition. McGraw-Hill Book Company, New York,
 

T972.
 

Sturrock, R.F., "Persistence of the Molluscicide Bayluscide (Clonitralide) Emulsifiable Concentrate on Mud Surfaces in the Tropics."
 

Annals of Tropical Medicine and Parisitology 68(4):427-434, 1974;
 

Chemical Abstracts 82 94160v.
 

Sturrock, R.F., G. Barnish, and J. Seeyave, "Field Tests on the Effects
 

of Three Molluscicidal Chemicals on Bananas." Pflanzenschutz-


Nachr. (American edition) 27_(D :56-61, 1974; Chemical Abstracts
 

83_ 189207a.
 

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

62:679-682, 1975.
 

45
 

Review of the Bioassay of Clonitralid* 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 DHEW Committee
 

Regulations and the Provisions of the Federal Advisory
 

Committee Act. The purpose of the Clearinghouse is to
 

advise the Director of the National Cancer Institute (NCI)
 

on its bioassay program to identify and to evaluate chemical
 

carcinogens in the environment to which humans may be exposed.
 

The members of the Clearinghouse have been drawn from
 

academia, industry, organized labor, public interest groups,
 

State health officials, and quasi-public health and research
 

organizations. Members have been selected on the basis of
 

their experience in carcinogenesis or related fields and,
 collectively, provide expertise in chemistry, biochemistry,
 

biostatistics, toxicology, pathology, and epidemiology.
 

Representatives of various Governmental agencies participate
 

as ad hoc members. The Data Evaluation/Risk Assessment
 

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

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

on the bioassay of Clonitralid for carcinogenicity.
 

Although the carcinogenicity of the compound was not
 

established under the conditions of test, the reviewer said
 

that the study should not be considered "definitive." He
 

pointed out the finding of mammary adenocarcinomas in both
 

the treated and control female rats, as well as in treated
 

male rats. The reviewer concluded that the evidence was
 

insufficient to declare that Clonitralid was carcinogenic
 

but that it raised "concern that the study may not have
 

been definitive." The reviewer's conclusion 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.
 

47
 

. S . GOVERNMEN T PRINTIN G OFFICE ! 1978-260-899/3194
 

APPENDIX A
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS
 

IN RATS TREATED WITH CLONITRALID
 

TABLE A1

 

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

ANTPAI S INITIALL Y IN STUD Y

ANIMAL S NECPOPSIE D

ANIMAL S FXAMISF D HISI3PATHOLOGICALLY* *

INTEGUMENTAR Y SYSTE M 

4SUBCU T TISSU E

FIBROM A

FI3ROSARCOM A

LIPOM A

HEMANGIOSARCOH A

BESPIEATOR Y SYSTE K 

#L'!NG

CORTICA L CAPCINOSA , METASTATI C

HEHA"-OPOIFTIC SYSTE M 

*MULTIPL E OR3AN S

MALIGNAN T LYMPHOMA , NO S

MALI3.LYMPHOMA , HI STIOC YT 1C TYPE

#SPLE'N


HFMAN5IOSABCOMA


CIRCOLATOPY SYS1E1
 

HCNF
 

DIGESTIVE SYSTEM
 

IPANCPFA S

HFMAN3I05ARCOM A

URINAR Y SYSTEM
 

• KIDNEY


CONTRO L (VEH ) 01-M016

 2 0

 20

 2 0

 (20 )

 (20 )

 (20 )

 1 (5%) 

(18)


 2 (11")


 (IB )

 (20)


 LOW DOSE 01-1047

 5 0

 49

 4 3

 HIG H DOSE 01-M04 8 

 5 0 

 49 

 3 6 

 (49 )

 2

 1

 1

 (4«) 

 (2%) 

 (2% ) 

 (49) 

4 (8%) 

 (43 )

 1 (2%) 

 (36) 

 (49 )

 2 («»)

 (43)

 3 (7X)

 '

 (49 ) 

1

 (35) 

3

 (2*) 

 (9%) 

 (41 )

 1 (2%) 

 (32 ) 

 (43) (35) 

t HUMBF B O F ANIMAL S WIT H TISSli ? EXAMINE D MICROSCOPICALL Y 

* HUMEE P OF ANIMAL S N'ECEOPSIEC 

"EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

A~3 
TABLE A1 (CONTINUED)
 

CONTROL(VEH ) LOK DOSE HIGH DOSE 01-MOU o 01-MOU 7 01-BOH8 

ENnrcPIN " SYSTE M 

*?ITUITA? Y (19) («0 ) (33) 

CHPOMOPHOB E ADENOM A 5 (26%) 7 (18%) 2 (6*) 

(19) (K3 ) (36) CORTICA L CARCINOM A 1 (5%) 1 (2%) 2 (6X) PHEOCHPOMOCYTOB A 1 (2%) 1 (3%) 

• THYROI D (20) (U2 ) (35) FOLLICHLAB-CEL L ADENOM A 2 (5%) 3 (9%) FOLLICULAB-CEL L CARCINOM A 2 (5%) 

C-CEI L ADENOM A 1 (2%) 3 (9*) 

#?ASCREATI C ISLETS (18) (t1) (32) ISLET-CEL L ADENOS A 3 (7%) 1 (3*) 

ISLET-CEL L CABCINOM A 1 (3S) 

REPRODUCTIV E SYSTE M 

*MAKMAP Y GLAN D (20 ) (<49 ) (U9) 

ADEKOCASCINOMA , NO S 1 (5%) 3 (6%) FIEROADENOM A 1 (2%) 

*PPE?'ITIA I GLAN D (20) (U9 ) 

ADSNOCAPCINOHA , NO S 1 (2*) 

NFPVDU P SYSTE M 

*EEAI N (20) (U2 ) (36)

 OLIGODESDROGLIOMA 1 (2X)

 

SPECIS L Si'NSS ORGAN S 

N0\'? 

SDSCi)LnsK<aETA L SYSTE M 

SOS11 

30DY CAVITIE S 

*T1JNIC A VAGIHALI S (20) (U9 ) (U9 ) 

* NUMBE R OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NHM3SR OF ANIMALS NECHOPSIED
 

A-4 
ALL OTHFR SYSTEMS
 

THORACIC CAVITY
 

LIPOSABCOHA
 

ANIMA L DISPOSITION SUMMAR Y
 

ANIMALS INITIALLY IN STUDY
 MATORA L DEATHS
 

MORIBUND SACRIFICE
 

SCHEDULED SACRIFICE
 

ACCID3NTALLY KILLED
 

TERMINA L SACRIFICE
 

ANIMA L MISSING
 

9 INCLUDES AUTOLYZED ANIMALS
 

TUHOB SUK.1A5Y
 

TABLE A1 (CONCLUDED) 

CONTRO L (VEH ) 01-HOU 6 

20
 

1 3
 

TOTAL ANIMAL S WITH PRIMAR Y TUMORS* 10
 TOTAL PP.IMAEY TUMORS 11
 

TOTAL ANIMALS WITH BENIGN TUMORS 5
 TOTAL BENIGN TUMORS 5
 

TOTAL ANIMAL S WITH MALIGNAN T TUHOBS 6
 TOTAL MALIGNAN T TUMORS 6
 

TOTAL ANIMAL S WITH SECONDARY TUMORS*
 TOTAL SECONDARY TUMOES
 

TOTAL ANIMALS WITH TUMORS USCFPTAIN­ BENIGN OR MALIGNAN T
 

TOTAL UNCERTAIN TUMORS
 

TOTAL ANIMALS WITH TUMORS UNCERTAIN­ PPTMAR Y 08 BETASTATIC
 

TOTAL UNCERTAIN TUMORS
 

PRIMARY TUMORS: ALL TUMOES EXCEPT SECONDARY TUMORS
 

L OW DOSE HIGH DOSE 01-MOU 7 01-MOU8 

50 50
 33 17
 

1
 

17 32
 

20 20
 30 26
 

1 1 13
 15 15
 

12 10
 14 11
 

1
 1
 

1
 

1
 

SECONDARY TUMORS: METASTATIC TUMOES OR TUMORS INVASIVE INTO AN ADJACENT ORSAN
 

A-5
 
TABLE A2 

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

CONTHOL(VEH ) LOW DOSE HI3H DOSE 01-FOU6 01-FOU7 01-FOU3 

ANIMAL S INITIALL Y IN STUD Y 20 50 50 

ANIMAL S NSCROPSIED 20 50 50 

ANIMAL S EXAMINE D HISTOPATHOLOGICALLY* * 20 <t 2 US . 

INTE'3'IMESTA3 Y SYSTE M 

*SKT» J (20) (50) (50) 

SQUAMOU S CEL L CARCINOM A 1 (2%) 

"I?ROSA!>COM A 1 (2%) 

*SU3CU T TISSU E (20) (50) (50) 

FIBROM A 1 (5%) 3 (6X) 

FIBROSA3COM A 2 (U% ) 

T IS ROIIS HISTIOCYTOMA , MALIGNAN T 1 (2S) 

tlPOH A 1 (2«) 

RESPIRATOR Y SYSTE M 

*LUS G (20) (it 2) (15) 

CARCINOMA , NOS , SETASTATI C 2 (US) 

AEStiOCARCISOMA . NOS , METASTIiTIC 1 (2%) 2 (U*) 

FIBROSARCOMA , METASTATI C 1 (2X) 1 (2%) 

HE?1ANGIOSAHCOM A 1 (5%) 

*.1ULTIPLF ORGAN S (20) (50) (50) 

MAII3.LYMPHOMA , HISTIOCYTIC TYP E 1 (2%) 

GRAN'ILOCYTI C LEUKEMI A 1 (2%) 

• SPLEE N (20) (U1 ) (U5 ) 

CAPCINOMA , NOS , METASTATI C 1 (2%) 

H'SAN3IOSARCOM A 1 (2%) 1 (2*) 

ICFSVICUl LYMPH NODE (20) (39) C*3)
 

CR3CINOMA , NOS, MSTASTATIC 1 (2%)
 

• MESENTE3IC L. NODE (20) (39) (U3)
 

CARCINOMA , NOS, MSTASTATIC 1 (2%)
 

HEMAXGIOSAPCOHA
 

# NIHBE" 0 ? ANIMAL S WIT H TISSU E EXAMINE D MICROSCOPICALL Y 

* NUIEE P O F ANIMAL S NECROPSIE C 

"EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

A-6 
TABLE A2 (CONTINUED)
 

CONTROL (VSH) LOW COSE HIG H DOSE 

01-FOtS 01-F047 01-F0118 

CIRCULATOR Y SYSTE M 

3IOHE 

DIGESTIV E SYSTE M 

#LIVE R (20) (*2) (15) 

CARCINOMA ,

NEOPLASTIC

 NOS, METASTATI C 

 NODUL E 1 (2%) 

1

1

 (2«) 

 (2S) 

HEPATOCELLULA R CARCINOM A 1 (2*) 

COPTICAL CARCINOMA, METASTATIC 1 (2%) 

((PANCREA S (19) C«0) (45) 

CARCINOMA , NOS, METASTATI C 2 (4X) 

tESOPHASD S

CARCINOMA , NOS, METASTATIC 

 (20) (HI ) (11)

1 (2%) 

tSTOHACH (20) C*2) (15) 

CAPCINOMA,NO S 2 (<U) 

ISMAI L INTESTIN E (19) (tl) (13) CAECINOSA , NOS, METASTATI C 1 (2%) 

tlARG E INTESTIN E (20) (12) (15) CARCINOMA , NOS, METASTATI C 1 <2X ) 

UBI1IAP Y SYSTE M 

fKIDNE Y (20) (12) (15) CARCINOMA , NOS, NETASTATI C 1 (2X) 

T03ULAR-CEL L ADENOCARCINOM A 1 (2*) 

MIXE D TOMOR , MALIGNAN T 2 (5%) 

HBMARTOMA + 1 (5%) 1 (2%) 

*DHIN'AS Y 31ADDEB (19) (38) (15) CAPCINOMA , NOS, METASTATI C 1 (2S) 

ENDOCRINE SYSTEM
 

»I'ITUITAR Y (20) (U2 ) (tl) 

CHROMOPHOBE_ADENgilA 5__[252]_ J_J1Z*L J2_ 

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

* NUMBE R OF ANIMAL S NSCPOPSIE D 

+ THIS IS CONSIDERED TO BE A BENIGN FORM OF THE MALIGNANT MIXED TUMOR OF THE KIDNEY AND CONSISTS OF PROLIFERATIVE LIPOCYTES, TUBULAR STRUCTURES, FIBROBLASTS, AND VASCULAR SPACES IN 

VARYING PROPORTIONS. 

A-7 
TABLE A2 (CONTINUED) 

CONTROL(VEH ) LOW DOSE HIGH DOSE 01-F046 01-F047 01-F048 

*ADHENA L (20) (42) (44) 

CARCINOMA , NOS, METASTATIC 1 (2X) 

CORTICAL ADENOM A 2 (5X) 

CORTICAL CARCINOM A 2 (5X) 1 (2X) PHEOCHROMOCYTOMA 1 (2X) 

tTHYBOID (20) (41) (44) 

FOLLICULAR-CELl ADENOM A 1 (5X) 2 (5X) 1 (2X) FOLLICULAR-CEL L CARCINOM A 2 (5X) 2 (5X) 

C-CELL ADENOM A 1 (2X) 3 (7X) 

C-CELL CARCINOM A 2 (5X) 

REPRODUCTIVE SYSTEM
 

*MA«MAR Y 3LAND (20) (50) (50)
 

UDENOCARCINOMA, NOS 1 (53S) 12 (2HX) 11 (22X)
 

FIBHOADENOHA U (20*) 13 (26X) 11 (22X)
 

*PRKPUTIAL 3LAND (20) (50) (50)
 

CARCINOMA,NOS 1 (2X)
 

tOTEHU S (20) (M1 ) (Ult) 

CARCINOMA , NOS, HETASTATIC 1 (21) 

SQUAHOU S CELL CARCINOM A 1 (2X) 

ENDOHETRIA L STROMAL POLYP 2 (5*) 6 (ItX) 

5NDOMFTRIA L STROMA L SARCOMA 1 (2X) 

HEMANGIOSARCOM A 1 (2X) 

#OVAR Y (19) (H1) (43) 

CARCINOMA , NOS, METASTATIC 1 (2X) 

GRANULOSA-CEL L TUSOR 1 (5%) 1 (2X) 

NERVOU S SYSTE M 

NONF _ _ _ 

SPECIAL S'NSE ORGANS
 

NONE
 

MUSCULOSKELETAL SYSTEM
 

__IONE
 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 » NUMBER OF ANIMALS NECROPSIED
 

A-8
 
TABLE A2 (CONCLUDED) 

CONTROLLED) LOW DOSE HIGH DOSE 01-FOH6 01-FOU7 01-FOU8 

BODY CAVITIES
 

*ABDOMINAL CAVITY (20) (50) (50)
 

PIBROSARCOMA 1 (5X)
 

'"MESENTERY (20) (50) (50) 

ENDOSETRIAL. STROMAL SARCOMA, MET 1 (2%) 

ALL OTHEF SYSTEMS 

' DIAPHRAG M 

CARCINOMA, NOS, METASTATIC 1
 

ANIMA L DISPOSITION SUMMARY
 

ANIMALS INITIALLY IN STUDY 20 50 50
 NATURAL DEATHS 6 12 17
 

MORIBUND SACRIFICE 1
 

SCHEDULED SACRIFICE
 

ACCIDENTALLY KILLED
 

TERMINA L SACRIFICE 1 <J 38 32 

ANIMAL MISSING
 

3 INCLUDES AUTOLYZED ANIMALS
 

TUMOR SUMMARY
 

TOTAL ANIMALS WITH PBIMARY TUaOHS* 11 33 37
 TOTAL PRIMARY TUMORS 16 59 61
 

TOTAL ANIMALS WITH BENIGN TUMORS 10 22 2U
 TOTAL BENIGN TUMORS 12 31 33
 

TOTAL ANIMALS WITH MALIGNANT TUMOPS 3 2« 23
 TOTAL MALIGNANT TUMORS 3 26 27
 

TOTAL ANIMALS WITH SECONDARY TUMORS* 3 5
 TOTAL SECONDARY TUMORS . 3 21
 

TOTAL ANIMALS WITH TUMORS UNCEBTAIN­ BENIGN OR MALIGNANT 1 2 1
 

TOTAL UNCERTAIN TUMORS 1 2 1
 

TOTAL ANIMALS WITH TUMORS UNCERTAIN­ PEIMARY OR METASTATIC
 

TOTAL UNCERTAIN TUMORS
 

* PSIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS
 * SECONDARY TUMO8S: METASTATIC TUMORS OB TUMORS INVASIVE INTO AN ADJACENT ORGAN
 

A-9 

APPENDIX B
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS
 

IN MICE TREATED WITH CLONITRALID
 

TABLE Bl 

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

CONTROL (VEH) LOW DOSE HIGH DOSE 

02-M05 2 02-M053 02-M054 

ANIMAL S INITIALL Y IN STCDY 20 50 50 

ANIMAL S MISSIN G 1 3 

ANIMAL S NEC80PSIED 16 143 13 

ANIMAL S EXAMINE D HISTOPATHOLOGICALLY ** 16 ' 43 13 

iNTESUMENTAR Y SYSTE M 

*SUBCBT TISSUE (16) (13) (43) 

FIBROSARCOM A 2 (5X) 3 (7%) 

SESPIRATOS Y SYSTE M 

*LUSG (16) (43) (43) AlVEOLAR/BEONCHIOLA R ADENOM A 1 (2X)

 

HEHATOPOIETIC SYSTEM 

*MULTTPL E ORGAN S (16) (43) (43) 

MALIG.LYMPHOMA , LYMPHOCYTI C TYP E 1 (2X) 1 (2X) 

tKIDNEY (16) (43) (43) 

1ALIS.LYBPHOMA , LYMPHOCYTI C TYP E 1 (6X) 

CIRCULATORY SYSTEM
 

NONE
 

DIGESTIVE SYSTEM
 

*LIVEH (16) (43) (43)
 

HEPATOCELLULAR CARCINOMA 1 (2X)
 

URINARY SYSTEM
 

NONE
 

ENDOCRINE SYSTEM
 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECEOPSIED
 

**EXCLUDES PARTIALLY AUTOLYZED ANIMALS
 

B-3
 
REPRODUCTIV E SYSTE M 

*TESTIS


INTERSTITIAL-CELL TUMOB


NEBVOUS SYSTEM
 

NONF
 

SPECIAL SENSE ORSANS
 

NONE
 

MUSC'JLOSKELETAL SYSTEM
 

NOSE
 

BODY CAVITIES
 

NONE
 

ALL OTHER SYSTEMS
 

NOME
 

ANIMAL DISPOSITION SUMKAEY
 

ANIMALS INITIALLY IN STUDY
 NATURA L DEATHS


MORIBUND SACRIFICE
 

SCHEDULED SACRIFICE
 

ACCIDENTALLY KILLED


TERMINAL SACRIFICE


ANIMA L MISSING


# NUMBE R O F ANIMAL S WIT H TISSU E

* NUMBE R OF ANIMAL S NECEOPSIE C 

TABLE Bl (CONTINUED) 

CONTROL(VEH ) LOW DOSE HIGH DOSE 02-M052 02-M053 02-M054 

 (16)	 (42) (43)
 

1 (2S)
 

 20 50 50
 19 46 43
 

2
 

1 3 2
 

1 3
 

 EXAMINE D MICROSCOPICALL Y 

B-4
 
TABLE Bl (CONCLUDED)
 

CONTRt 3LJVEH ) LOW DO: HIGH DOSE 0?-M < 02-Mo:if 02-M054 

TUMOR SUMMAR Y
 

TOTAL ANIMAL S HITH PRIMARY TUMORS* 1 5 5
 TOTAL PRIMARY TUMORS 1 5 5
 

TOT HI. ANIMALS WITH BENIGN TUMORS 1 1
 TOTAL BENIGN TUMORS 1 1
 

TOTAI ANIMAL S WITH MALIGNAN T TUMORS 1 1 4
 TOTAL MALIGNANT TUMORS 1 it 4
 

TOTAL ANIMAL S SITH SECONDARY TUMORS*
 TOTAL SECONDARY TUMORS
 

TOTAL ANIMAL S WITH TUMOR S UNCERTAIN­ BENIGN OP MALIGNAN T
 

TOTAL UNCERTAIN TUMORS
 

TOTAL ANIMAL S WITH TUMOSS UNCERTAIN­ PRIMARY OR METASTATIC
 

TOTAL UNCERTAIN TUMORS
 

* PRIMAR Y TUMORS: ALL TUrtOSS OCF?T SECONDARY TUMORS
 # SECONDARY TUMORS: 1FTR3TATIC TUMOR S OE TUMORS INVASIVE INTO AN ADJACENT ORGAN
 

B-5
 
TABLE B2

 

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

CONTROL (VEH ) 02-F052 LOW DOSE 02-F055 HIGH DOSE 02-F056 

ANIMAL S INITIALL Y IN STUD Y 20 50 50 

ANIMAL S MISSIN G 1 

ANIMAL S NSCPOPSIE D 20 49 47 

ANIMAL S EXAMINE D HISTOPATHOLOGICALLY* * 2 0 4 9 4 7 

INTFGOMENTAR Y SYSTEM 

NON E 

RESPIRATOR Y SYSTE M 

tLUN G (20) (<*9) (47) 

ALVEOLA3/8RONCHIOLA R ADENOM A 2 (10%) 1 (2)1) 

ALVEOLAa/BRONCHIOLA H CARCINOM A 1 (5% ) 

HEMATOPOIETI C SYSTEM 

*KULTIPL E ORGAN S .(20) (49) (47) 

MALIG.LYMPHOMA , LYMPHOCYTI C TYP E 4 (20%) 1 (2%) 

MALIG.LYMPHOP1A , HISTIOCYTIC TYPE 3 (6%) 1 (2X) 

CIRCilLATOH Y SYSTE M 

NON F 

DIGESTIV E SYSTE M 

NOS E 

URINAP Y SYSTE M 

NON F 

ENDOCRIN E SYSTE M 

NON F 

* NUM9E E O F ANIMAL S HIT H TISSU E EXAMINE D MICROSCOPICALL Y 

* ND.1»5'S 0? ANIMAL S NECROPSIE D 

"EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

B-6 
TABLE B2 (CONTINUED) 

CONTROL(VEH ) ' LOW DOSE HIGH DOSE 02-F052 02-F055 02-F056 

REPRODUCTIVE SYSTEM
 

*MASKAR Y GLAND (20) (49) (47)
 

AD5NOCARCINOMA, NOS 1 (5%) 1 (2*)
 

IUTEBU S (20) (49) (46) 

ENDOMETBIA L STHOMA L POLYP 1 (2X) 1 (2X) 

tOVAR Y (18) (48) (45) 

CYSTADENOMA , NOS 1 (6%) 

ilERVOU S SYSTE M 

SPECIAL SENSE ORGANS
 

*HAHDEBI»N GLAND (20) (49) (47)
 

ADENOMA, NOS 1 (2S)
 

?IUSC'ILOSKFLETA L SYSTEM 

NON F 

BODY CAVITIES
 

NONE
 

ALL OTHER SYSTEMS
 

NONE
 

ANIMA L DISPOSITION S'JNBAFY
 

ANIMAL S INITIALLY IN STUDY 20 50 50
 NATURAL DEATHS 2 4 4
 

MORIBUND SACRIFICE
 

SCHEDULED SACRIFICE
 ACCIDENTALLY KILLED
 

T^BBINA L SACRIFICE 18 46 45 

ANIMA L MISSING 1
 

3>_!!£!2DES_iaT.OL.YZED_ANI.!SALS
 

•I NUMBE R OP ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIEE
 

B-7 
TABLE B2 (CONCLUDED)
 

CONTROL (VEH) LOW DOSE HIGH DOSE
 02-F05Z 02-F055 02-F056
 

TUM0 3 SUMMAR Y 

TOTAL ANIMAL S WITH PRIMAR 9 Y TUKORS* 5 5 TOTA L PRIMAR Y TUMOR S 9 5 5 

TOTA L ANIMAL S WIT H BENIG N TUROR S 3 1 3 TOTA L BENIG N TUMORS 3 1 3

 

TOTSL SNIKAL S WIT H "IALI3NAN T TUMOR S 6 It 2 TOTAL MALIGNAN T TUMORS 6 It 2 

TOTAI ANIMAL S WITH SECONDARY TUMORS*
 TOTAL SECONDARY TUMORS
 

TOTAL ANIMAL S WITH TUMORS UNC?RTMN­ PENI3N OP MALIGNAN T
 TOTAL UNCERTAIN T'IMOPS
 

TOTAL ANIMAL S WITH TUMORS UNCERTAIN­ PRIMAR Y OR METASTATIC
 TOTAL UNCERTAIN TUMORS
 

* PRIMAR Y TUMOBS: SLL TUMORS EXCEPT SECONDARY TURORS
 * SECONDARY TUMORS: METASTATIC TUMOR S OH TUMORS INVASIVE INTO AN ADJACENT ORGAN
 

B-8
 
APPENDIX C
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN RATS TREATED WITH CLONITRALID
 

TABLE Cl 

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

CONTROL(VEH) LOW DOSE HIGH DOSE
 01-M046 01-M047 01-M048
 

ANIMAL S INITIALLY IN STUDY 20 50 50
 

ANIMALS NECROPSIED 20 49 49
 

ANIMALS EXAMINED HIS70PATHOLOGICA LLY** 20 1*3 36
 

INTEGUMENTARY SYSTEM
 

*SKIN (20) (49) (49)
 

EPIDERMAL INCLUSION CYST 1 (2%)
 

*SUBCUT TISSUE (20) (49) (49)
 ULCER, NOS 1 (2%)
 

NECROSIS, FAT 1 (2S)
 

RESPIRATORY SYSTEM
 

*NASAL CAVITY (20) (49) (49)
 

INFLAMMATION, NOS 2 (4X)
 

HONG/BRONCHUS (20) (43)' (36)
 

ABSCESS, NOS 8 (19%)
 

ILIJNG (20) (43) (36)
 

MINERALIZATION 1 (2%)
 

ATELECTASIS 1 (3%)
 

CONGESTION, NOS 5 (25%) 17 (40%)
 

EDEMA, NOS 2 (5S)
 

INFLAMMATION, NECROTIZING 1 (251)
 

PNEUMONIA, CHRONIC MUHINE 18 (90%) 40 (93*) 24 (67S)
 

HEMATOPOIFTIC SYSTEM
 

#SPLEEN (18) (43) (35)
 

INFLAMMATION, NOS 1 (3%)
 

PEHIARTBSITIS 1 (6%)
 

HEMATOPOIESIS 3 (7%) 1 (3X)
 

fMESSNTSRIC L. NODE (18) (36) (33)
 

INFLAMMATION^ NOS 1 .US1__
 

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

* NUMBE R OF ANIMAL S NECROPSIE D 

**EXCLUD£S PARTIALLY AUTOLYZED ANIMALS 

C-3 
TABLEC1 (CONTINUED) 

CONTRO L (VEH ) LOW DOSE 01-MO 46 01-M047 

CIRCULATOR Y SYSTE M 

IHEftS T 

DILATATION, NOS 

?IBH03IS 

?E?IARTSRITIS 

CALClnCl DEPOSIT 

(20) 

1 (5«) 

(U3) 

1

1

 (2%) 

 (2%) 

tMYOCARDI'IM 

MINERALIZATION 

(20) (U3) 

INFLAMMATION , NOS 

INFLAMMATION, FOCAL 

INFLAMMATION, DIFFUSE 

INFLAMMATION, INTERSTITIAL 

FIBFOSIS 

"TBBOSIS, DIFFUSE 

DEGENERATION, NOS 

CALCIFICATION, NOS 

3 (15%) 

3 (15X) 

1 (5%) 

1 (554) 

U

2

1

 (9%) 

 (5%) 

 (2%) 

»ENDOCA3DIUM 

INFLAMMATION , NOS 

FIBROSIS 

(20) (43) 

*AOPTA 

INFLAMMATION , NOS 

ARTERIOSCLEROSIS, NOS 

MEDIAL CALCIFICATION 

CALCIUM DEPOSIT 

CALCIFICATION, NOS 

(20) (49) 

2

1

 (4%) 

 (2X) 

"CORONARY ARTER Y 

MI3ESALI3ATION 

(20) (49) 

1 (2%) 

DI3ESTIVF SYSTEM
 

*S»LIV6SY GLAND (11) (22)
 

'1YPEPPLASIH, INTRADUCTAL 1 (9%)
 

#LIVEB (20) (43)
 

COSHESTIOS, HOS 4 (20%) 12 (28%)
 

INFLAMMATION, NOS 1 (2X)
 

NFCPOSIS, NOS 1 (5%)
 

METAMOEPHOSI3 FATTY 2 (10%) 7 (16%)
 

tPCAL_CELLULAR_CHANGE
 

* NUMBE R OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALLY
 

* NUMBER OF ANIMAL S NECROFSIEC
 

C-4
 

HIGH DOSE 01-MOua 

(36)
 

1 (3%)
 

(36)
 

1 (3X)
 

2 (6*)
 

1 (3%)
 

6 (17%)
 

(36)
 

1 (3J)
 

1 (3%)
 

(49)
 

1 (2«)
 2 (14%)
 

1 (2X)
 

(49)
 

(23)
 

(36)
 

1 (3*)
 
TABLE Cl (CONTINUED)
 

CONiaOL(VEH) LOW DOSE HIGH DOSE
 01-MOU6 01-MOU7 01-BQ1J8
 

HYPERPLASTIC NODULE	 1 (5%)
 

HYPERPLASIA, NOS	 1 (2%) 2 (6X)
 

ILIVES/CENTRILOBULAS (20) (13) (36)
 

DEGENERATION, NOS 1 (2%)
 

NECROSIS, NOS 1 (2%)
 

METAMORPHOSIS FATTY 2 <5»)
 

•LIVKR/HEPATOCYTES	 (20) (13) (36)
 

INFLAMMATION, NOS 2 (5%)
 

INFLAMMATION, DIFFUSE 1 (2%)
 HYPERPLASIA, NOS 1 (5%) 1 (2%)
 

*BILE DUCT (20) (19) (19)
 

INFLAMMATION, NOS 5 (25%) 3 (6%)
 

HYPEBPLASIA, NOS 6 (30%) 19 (39%) 6 (12%)
 

•PANCREAS	 (18) (11) (32)
 

THROMBOSIS, NOS 1 (3%)
 

PESIARTPP.ITIS 2 (11%) U (10%)
 

ATROPHY, NOS 2 (11%) 3 (7S) 2 (618)
 

t STOMACH (20) (13) (35)
 

MINERALIZATION 2 (10%) U (9%)
 

HEMORRHAGE 2 (5%)
 

ULCER, NOS 3 (15%) 3 (7%)
 ULCER, FOCAL 1 (5%) 1 (2%)
 

INFLAMMATION, CHRONIC 1 (3*)
 CALCIUM DEPOSIT 3 (9%)
 

ACANTHOSIS 1 (5X)
 

•GASTRIC	 MUCOSA (20) (13) (35)
 

MINERALIZATION 1 (2%)
 

•LARGE	 INTESTINE (18) (13) (31)
 

SFHATODIASIS 1 (2%) 2 (6X)
 

URINAR Y SYSTEM
 

•KIDNEY	 (20) (13) (35)
 

.1IHEPALIZATION 1 (5%) 1 (2%)
 

MYD80NEPHROSIS 1 (3*)
 CONGESTION, NOS 5 (25%) 18 (U2%)
 

INFLAMMATION, CHRONIC 17 (35R) 34 (79%) 22 (63S)
 

•KIDNEY/CORTEX	 (20) (13) (35)
 

CYSTX_N05
 

* NUM3EB OF ANIMAL S »I1H TISSUE EXAMINED MICROSCOPICALLY
 

* NII13ER OF ANIMALS NECROPSIED
 

C-5
 
TABLE Cl (CONTINUED) 

CONTROL(VEH ) LOB DOSE HIGH DOSE 

01-M046 01-BOK7 01-M048 

#KIt>NEY/PELVI S (20) (13) (35)

 

INFLAMMATION , NOS 2 (5%)

 

tORINAR Y BLADDE R (18) («1) (31)

 INFLAMMATION , NOS 1 (6%) 2 (5X) 2 (6%)

 

ENDOCRIN E SYSTEM 

*PITUITAR Y (19) (10) (33) CYST, NOS 2 (5X)

 HYPEHPLASIA , CH80MOPHOBE-CELL 2 (11%) 2 (5X)

 

*ADRENA L (19) (13) (36)

 

CONGESTION, NOS 2 (11)S)

 

ANGIF.CTASIS 2 (11%) 3 (7%) 3 (8X)

 

IADRENA L CORTE X (19) (13) (36)

 

DEGENERATION , NOS 7 (37S) 10 (23%) 3 (8X)

 

#THYROI D (20) (12) (35) 

COLLOID CYST 1 (5*) 1 (2*) 

HYPF.RPLASIA , C-CELL 2 (10%) 1 (2%) 1 (3%) 

HYPERPLASIA , FOLLICULAP-CELL 1 (5%) 1 (2%) 1 (3%) 

IPARATHYROI D (1) (1) (3) HYPERPLASIA , NOS 1 (100%) H (100%) 3 (100%) 

REPRODUCTIV E SYSTEM 

tPBOSTATE (13) (30) (26) 

INFLAMMATION , NOS 2 (15%) U (13%) 2 (8X) 

• SKMINA L VESICLE (20) (19) (19)

 INFLAMMATION, NOS 1 (2%)

 

tTESTIS (20) (13) (36) 

PFRIARTESITIS 1 (5*) 

ATROPHY, NOS 5 (25%) 16 (37%) 4 (11%) 

ASPERMATOGENESI S 2 (10%) 9 121X) 2 (61) 

HYPOSPERMATOGENESI S 1 (20%) 7 (16%) 1 (3%) 

*EPIDIDYHI S (20) (19) (19) 

NECROSIS, FAT 3 (6%) 

NERVOU S SYSTEM 

NONE 

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

* NUMBE R OF ANIMAL S NECROPSIE S 

C-6 
TABLE Cl (CONCLUDED)
 

CONTROL(VEH) LOB DOSE
 01-N04S 01-H047
 

SPECIAL SENSE ORGANS
 

*EYE (20) (49)
 INFLAMMATION, NOS 1 (2*)
 

*EYE/CORNEA (20) (49)
 INFLAMMATION, NOS 2 (4X)
 

MUSCULOSKELFTAL SYSTEM
 

*SKELETAL MUSCLE (20) (49)
 MINERALIZATION 1 (5%)
 INFLAMMATION, NOS 1 (5%) 1 (2X)
 

FI3BOSIS 1 (2%)
 

BODY CAVITIES
 

*ABDONINAl CAVITY (20) <»9)
 INFLAMMATION, CHRONIC 1 (5)1)
 

PERIARTF.RITIS 1 (2X)
 NECROSIS, FAT 1 (5%)
 

*PE3ITONEt'M (20) (U9)
 INFLAMMATION, NOS
 

'PERICARDIUM (20) (49)
 INFLAMMATION, NOS
 

INFLAMMATION, SUPPURATIVE 1 (2%)
 

*MESENTSBY (20) (49)
 PERIARTERITIS 1 (5%) 6 (12X)
 

1ECROSIS, FAT 1 (2%)
 

ALL OTHER SYSTEMS
 

NONE
 

SPECIAL MORPHOLOGY SUMMAR Y
 

NECROPSY PERF/NO HISTO PERFORMED 6
 

AUTOLYSIS/NO NECROPSY 1
 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

HIGH DOSE
 01-M048
 

(49)
 

(49)
 

(49)
 

(49)
 

1 (2*)
 

(49)
 

2 (4*)
 

(49)
 

1 (2*)
 

(49)
 

5 (10X)
 

1 (2X)
 

13
 

1
 

C-7
 
TABLE C2

 

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

CONTROL (VEH) LOW DOSE HIGH DOSE
 01-F046 01 -FOU7 01-FOU3
 

ANIMALS INITIALLY IN STUDY 20 50 50
 

ANIMALS NECROPSIED 20 50 50
 

ANIMALS "XANISED HISTOPATHO LO GICA LLY** 20 1*2 45
 

INTFTHMENTBRY SYSTEM
 

*SUBCDT TISSUE (20) (50) (50)
 

ABSCESS, NOS 2 (4?,)
 

ABSCESS, -CHBONIC 1 (2*)
 

RESPIRATORY SYSTEM
 

*SASAL CAVITY (20) (50) (50)
 

INFLAMMATION, NOS 1 (2%)
 

INFLAMMATION, SUPPURAT1VE 1 (2%)
 

*L!ING/BEONCHOS (20) (42) (45)
 

ABSCESS, NOS 2 (10%) 2 (5X)
 

*LUNG (20) (12) (45)
 

CONGESTION, NOS 3 (15%) 21 (50%) 1 (2%)
 ABSCESS, NOS 1 (5%)
 

PNEUMONIA, CHEONIC MUP.INE 18 (90S) 41 (98*) 29 (64%)
 

&ALVEOLAS WAL L (20) (42) (4bj
 

EPITHELIALIZATION 1 (5T>)
 

HEKATOPOIETIC SYSTEM
 

»BONE MARROW (20) ("D (43)
 

METAMORPHOSIS FATTY 1 (2X)
 

•SPLEEN	 (20) (41) (45)
 

HEMATOPOIESIS 2 (10%) 1 (17%) 10 (22%)
 

CIRC'JLATORY SYSTEM
 

*MYOCARDIOM (20) (42) (44)
 

INFLAMMATION^ NOS ... i-isa 1 12%1 1 J2%1_ _ _
 

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

* NMEF.R OF ANIMAL S NECROPSIE E 

**EXCLUDES PARTIALLY AUTOLYZED ANIMALS 
TABLE C2 (CONTINUED)
 

CONTROL (VEH) LOS DOSE HIGH DOSE
 01-F046 01-F047 01-F048
 

INFLAMMATION, DIFFUSE 1 (5%)
 

FIBROSIS 4 (10%)
 

FIBROSIS, DIFFUSE 1 (2%)
 

DEGENERATION, NOS 3 (7%)
 

IENDOCARDIUM (20) (t2) ("«)
 INFLAMMATION, NOS 1 (2X)
 FIBROSIS 2 (5X)
 HYPERPLASIA, NOS 1 (2%)
 

*AORTA (20) (50) (50)
 

ARTFRI05CLEHOSIS, NOS 1 (2%)
 

DIGESTIVE SYSTEM
 

tSALIVAPY GLAND (14) (34) (38)
 INFLAMMATION, CHRONIC 1 (7%)
 

ATROPHY, NOS 1 <3X)
 

•LIVES	 (20) (42) (45)
 CONGESTION, NOS 3 (15%) 4 (10%)
 NFCHOSIS, NOS 1 (5%)
 

NECROSIS, FOCAL 1 (21.)
 METAMORPHOSIS FATTY 2 (10%) 5 (12%) 1 (2%)
 FOCAL CELLULAR CHANGE 3 (7%)
 HYPERPLASIA, FOCAL 1 (5%)
 

HEMATOPOIESIS 1 (2%)
 

ILIVER/CENTRILOBULAR (20) (42) (45)
 METAMORPHOSIS FATTY 1 (5%)
 

tLIVER/HEPATOCYTES (20) (42) (45)
 

DEGENEBATION, NOS 1 12%)
 NECROSIS, NOS 1 (5%)
 

FOCAL CELLOLAP CHANGE 1 <5S)
 

HYPERPLASIA, NOS 3 (15%)
 

*BILF DUCT (20) (50) (50)
 

DILATATION, NOS 1 (2«)
 CONGFSTION, NOS 1 (2%)
 INFLAMMATION, NOS 3 (15%) 12 (24%) 2 (4%)
 

HYPERPLASIA, NOS a (20%) 17 (34%) 7 (14X)
 

tPANCREAS (19) (40) (45)
 

PFRIARTERITIS 1 J1%1
 - —3 mi
 * NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIHALS NECROPSIEC
 

C-9
 
CALCIOH DEPOSIT 

ATROPHY , NO S 

tFSOPHASIJS 

ULCER , NOS 

#STOMAC H 

HFKOPRHAS E 

INFLAMMATION , NO S 

ULCER , NO S 

ULCER , FOCAL 

CALCIU M DEPOSIT 

HYPERKERATOSI S 

SCANTHOSIS 

*LARG E ISTESTINE 

N5MATODIASI S 

URINAPY SYSTEM
 

IKIONEY
 

MINEPALIZATION
 

HYDHONEPHeOSIS
 

CONGESTION, NOS
 

IHFLAJIBATION, CHRONIC
 

tKTDSF.Y/COSTSX
 

CYST, NOS
 

#KIDNEY/PELVIS
 

CALCULUS, NOS
 

INFLANMATION, NOS
 

ENDOCPIN? SYSTEM
 

IPITHITAR Y

CYST, NOS

HSHORHHA3IC CYST
 

HYPERPLASIA, CHROMOPHOBS-CELL


tADR^NAL


CONOESTION, NOS


DFGEN'i'RATION, NOS
 

ATROPHY^.SOS
 

TABLE C2 (CONTINUED)
 

CONTHOL(VEH ) 01-P046

(20)

1 (5X) 

(20)

1 (5%) 

2 (10*)

(20)

(20)
 

1 (5%)
 

3 (15*)
 

13 (65%)
 

(20)
 

(20)
 

 (20) 

1 (5%) 

2 (10%)
 

 (20)
 

1 (5%)
 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIKALS NECROP3IEB
 

C-IO
 

 LOW DOSE 01-F047 

 (4 1) 

 (42) 

2 (5%) 

3 (7X) 

1 (2%) 

3 (7%) 

4 (10*) 

 (42 ) 

4 (10%) 

(42)
 

15 (36*)
 

19 (45X)
 

(42)
 

3 (7*)
 

(42)
 

1 (2X)
 

4 (10%)
 

(42)
 

2 (5*)
 

1 (2*)
 

3 (7X)
 

(42)
 

3 (7S)
 

HIG H DOSE 01-F048 

1 (2S) 

1 (2*) 

(44) 

(45) 

1 (2») 

1 (2*) 

1 (2X) 

4 (9%) 

3 (7*) 

(45) 

2 (4%) 

(45)
 

1 (2*)
 

14 (31%)
 

(45)
 

(45)
 

(44)
 

4 (9*)
 

(44)
 

1 (2*)
 
TABLE C2 (CONTINUED) 

CONTHOL(VEH ) 01-F016 LOW EOSE O1-F0t7 

ANGIECTHSI S 9 (15%) 14 (33%) 

IAD3SNA L C08TEX 

DEGENERATION ,

ftTKOPHY, NOS 

 NO S 

(20) 

12 (60*) 

(12)

18 (43*) 

1 (2«) HYPE3PLASIA , NOS 1 (5*) 

tTHYPOID 

HYPERPLASIA , C-CELL 

(20) 

2 (10%) 

(41) 

3 (7X) HYPERPLRSIA , FOLLICUUR-CELL 

#PA'.HIHYROI D 

HYP"BPL»SI» , NOS 

REPRODUCTIV E SYSTE M 

*MAMMAR Y GLAN D (20) (50) 3ALACTOCELE 1 (5S) 

MFCROSIS , SOS 1 (2X) 

ttlTSRU S (20) (41) HYDPOSJTH A 3 (15%) 2 (5S) THROMBOSIS , NOS 

H V MORSHAGI C CYST 1 <5X) 

INFLAMMATION , NO S 

INFLAMMATION , CHRONI C 1 (5X) 

*UTEPUS/ENDOMETBIU M (20) (41) INFLAMMATION , NOS 1 (5%) 2 (5*) INFLAMMATION , SUPPUFATIV E 1 (2X) HYPERPLASIA , NO S 1 (5X) HYPERPLASIA , CYSTIC 1 (5%) 10 (24*) 

tOVASY/OVIDUC T (20) (41) INFLAMMATION , SUPPURATIV E 1 (5S) 

tOVAR Y (19) (41) CYST, NO S 1 (5S) 1 (2*) FOLIICDLA R CYST, NO S 1 (2X) INFLAMMATION , NOS 

NERVOU S SYSTE M 

NON E 

SPECIA L SENS E ORGAN S 

_NON E 

* NUMBER OF ANIMALS KITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

HIGH DOSE 01-F04B 

10 (23S) 

(44) 

8 (13*) 

(44) 

2 (5*) 

(1)

1 (100X) 

(50) 

(44) 

3 (7i) 

1 (2X) 

1 (2%) 

(44) 

1 (2*) 

4 (9*) 

(44) 

(13) 

3 (7*) 

1 (2X) 

c-i
 
TABLE C2 (CONCLUDED) 

CONTROL (VEH ) 01-FOU6 LOW DOSE 01-F047 HIGH DOSE O1-F0t8 

MUSCOLOSKELETAL SYSTEM 

•SKELETAL MUSCLE

INFLAMMATION, NOS

 (20) (50) (50) 

1 (2») 

BODY CAVITIES 

*ABDOMINA L CAVIT Y

NFCBOSIS, FAT

• MESENTERY

PERIAHTE8ITIS

 (20)

 1 (5?) 

 (20)

 (50)

 (50)

 1 (2%)

 (50) 

 (50) 

3 (6*) 

ALL OTHEB SYSTEMS 

NONE 

SPECIAL MORPHOLOGY SUMMAR Y 

NECROPSY PERF/NO HISTO PERFORMED

I NUMBER OF ANIMALS HITH TISSUT EXAMINED MICROSCOPICALLY 

* NUMBER OF ANIMALS NECROESIED 

8 5 

C-I2
 
APPENDIX D
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN MICE TREATED WITH CLONITRALID
 

TABLED1.

 

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

CONTROL (VEH) LOW DOSE HI3H DOSE 02-M052 02-N053 02-M054 

itNIMAL S INITIALL Y IN STUDY 20 50 50 

ANIMAL S MISSING 1 3 

ANIMAL S NECROPSIED 16 43 43 

ANIMAL S EXAMINE D HISTOPATHOLOGICALLY ** 16 43 43 

INTEGUMENTAR Y SYSTEM 

NON E 

RESPIRATOR Y SYSTEM 

tLUNG (16) (43) (43)
 

PNEUMONIA, CHRONIC F.URINE 1 (6S) 2 (5X) 1 (2%)
 

AMYLOID05IS 1 (2»)
 

HEMATOPOIETIC SYSTEM
 

tSPLFEN (16) (42) (42)
 

AMYLOIDOSIS 9 (56%) 26 (62X) 34 (81*)
 

#MESENTERIC L. NODE (13) (33) (39)
 

INFLAMMATION, NOS 1 (3*)
 

ANGIECTASIS 1 (8%) 2 (6*)
 

CIRCULATORY SYSTEM
 

tHEAPT (16) (43) (43)
 

THROMBUS, ORGANIZED 1 (2X) 3 (7*)
 

CALCIUM DEPOSIT 1 (2*)
 

tMYOCARDIUM (16) (43) (43)
 INFLAMMATION, NOS 2 (5%)
 

•ENDOCARDIUM	 (16) (43) (43)
 

INFLAMMATION, NOS 1 (2X) 2 (5«)
 

OIQESTIVE SYSTEM
 

tLIVER (16) (43) (43)
 

_____ AMYLOIDOSIS __________________________ 2__(13£l
 

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

* NUMBE R OF ANIMAL S NECROPSIES 

**EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

D-3 
TABLE Dl (CONTINUED) 

CONTRO L (VEH ) LOW EOSE HIG H DOSE 02-M052 02-M053 02-M054 

tSTOMACH (16) (41) (43)
 

ULCER, FOCAI 1 (2%)
 

CALCIUM DEPOSIT 1 (621)
 

HYPERKERATOSI3 2 (13%)
 

ACANTHOSIS 2 (13%)
 

• COLON (16) (41) (43)
 

NEMATODIASIS 1 (2*)
 

URINAFY SYSTEM
 

#KIDNE Y (16) (13) (43)
 

PYF-LONEPHRITIS, NOS 1 (6%) 1 (2%) 3 (1%)
 

INFLAMMATION, CHRONIC 12 (75%) 30 (70%) 33 (77X)
 

AMYLOIDOSIS 1 (b%) 9 (21*) 7 (16S)
 

#ORINAHY BLADDER (16) (t2) (U3)
 

INFLAHKATIOS, Nf'S 3 (19%) 1 (2X)
 

ENDOCRINE SYSTEM
 

NONE
 

REPRODUCTIVE SYSTEM
 

tPROSTATE (11) (37) (11)
 INFLAHHATION, NOS 3 (21%) 2 (5JS)
 

*SEflIN.AL VESICLE (16) (U3) (13)
 

INFLAMMATION, NOS 1 (2%)
 

GRANULOMA , SPERMATIC
 

*TESTIS (16) (12) (13)
 CALCIFICATION, t!OS 1 (2*)
 

ATROPHY, NOS 3 (7*)
 

*EPIDIDYMIS (16) (13) (43)
 

INFLAMMATION, NOS 1 (2JS)
 

NERVOUS SYSTEM
 

f BR.MN/MENINGE S (15) (43) (43)
 

INFLJMMATIQN^NOS , 

It NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* N'JSEFH OF ANIMALS NECROPSIED
 

D-4
 
TABLE Dl (CONCLUDED) 

CONTRO L (VEH ) LOW DOSE HIGH DOSE 02-M05 2 02-M053 02-M054 

ft BR A I N (15) («3) (43)

 

INFLAMMATION , NO S 1 (2%)

 

SPECIA L SEN3 2 ORGAN S 

"OPTIC NERV E HEA D (16) (43) (43) 

INFLAMMATION , NO S 1 (2%) 

MHSC'ILOSKELETAL SYST2M
 

NONF
 

BODY CAVITIES
 

NGN''
 

ALL OTKEF SYSTEMS
 

N 0M ?
 

SPECIAL MOFPHOLOGY SUMMAR Y
 

# NUMBE R OF ANIMAL S WITH TISSUE EXAKINED MICROSCOPICALLY
 

* M'IMBES OF ANIMALS NECPOPSISE
 

D-5
 
TABLE D2

 

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

c:ONTROL(VEH) LOW DOSE HIGH DOSE
 

02-F052 02-F055 02-F056
 

ANIMALS INITIALLY IN STUDY 20 50 50
 

ANIMALS MISSING 1
 ANIMALS NSCSOPSIED 20 19 (17
 

ANIMAIS EXAMINED HISTOPATHOLOGICALLY ** 20 49 U7
 

INTEGUMENTARY SYSTEM
 

NONE
 

RESPIFATORY SYSTEM
 

f LONG (20) C*9) (47)
 

PNEUMONIA, CHRONIC KBRINE 1 (5*) 5 (10%) 3 (6%)
 

HEMATOPOIETIC SYSTEM
 

*SPLtEN (20) (149) (17)
 

ANGIECTASIS 1 (2*)
 HEMATOPOIESIS 2 (<4X) 1 (2*)
 

tCEBVICAL LYMPH NODE (20) (U8) (43)
 

HYPEPPLASIA, LYMPHOID 1 (2*)
 

*MESENTERIC L. NODE (20) (48) (43)
 

HYPEPPLASIA, NOS 1 (5*)
 

HYPERPLASIA, LYKPHOID 1 (2)1)
 

CIRCULATORY SYSTEM
 

tMYOCARDIIW (20) (49) (47)
 

INFLAMMATION, NOS 1 (2X)
 

DIGESTIVE SYSTEM
 

tLIVER (20) (49) (47)
 

INFLAMMATION, NOS 2 (t%)
 

IPANCREAS (20) (49) (47)
 

INFLAMMATION.,. NOS 1 J2X).
 

» NU1BE R O F ANIMAL S WIT H TISSUE EXAMINE D MICROSCOPICALL Y 

* NUC1EE R OF ANIMAL S NECROPSIE D 

"EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

D-6 
TABLE D2 (CONTINUED) 

CONTROL(VEH ) 02-F05Z LOW DOSE 02-F055 HIGH DOSE 02-F056 

tSTOMACH

INFLAMMATION, NOS

HYPERKERATOSIS

ACANTHOSIS

 (20)

 1 (5«) 

1 (5%) 

 (19) (47) 

1 (2%) 

UHINAPY SYSTEM 

NOME 

ENDOCRINE SYSTEM 

&PITUITAPY

ANGIECTASIS

 (18)

 1 ((,%)

 (38)

 1 (3%) 

 (36) 

tADRENA L

ANGIFCTASIS

 (20)

 1 (5%) 

 (18) (47) 

tADBENAL CORTEX

DEGENERATION, NOS

 (20)

 1 (5%) 

 (48) (47) 

REPRODUCTIVE SYSTEM 

#UTEEUS 

HYDROMETRA 

INFLAMMATION, NOS 

(20) 

4 (20%) 

(49) 

4 (8%) 

10 (20%) 

(46) 

5 (11X) 

5 (11S) 

IDTERUS/ENDOMETHIUM 

INFLAKHATION, NOS 

HYPEPPLASIA, CYSTIC 

(20) 

1 (5%) 

4 (20%) 

(49) 

11 (22%) 

(46) 

10 (22%) 

((OVARY/OVIDUCT 

INFLAMMATION, NOS 

(20) (49) 

1 (2%) 

(46) 

1 (2%) 

*OVARY 

FOLIICULAB CYST, SOS 

PAPOVARIAN CYST 

INFLAMMATION , NOS 

INFLAMMATION , SUPP'JPATIV S 

(18) 

1 (6%) 

2 (11?) 

3 (17%) 

1 (6%) 

(48) 

4 (8%) 

5 (10%) 

10 (21%) 

(45) 

10 (22%) 

1 (2%) 

10 (22%) 

NERVOUS SYSTEM 

NOMF 

# NUMSER OF ANIMALS '-IITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMDE? OF ANIMAL S NECEOPSIED 

D-7
 
TABLE D2 (CONCLUDED) 

CONTHOL(VEH) 02-F052 LOW DOSE 02-F055 HI3H DOSE 02-F056 

SPECIAL SENSE ORGANS 

*EYE/COPNEA

ULCER, NOS

 (20) (U9) (M7) 

1 (2%) 

MUSC'JLOSKELETAL SYSTEM 

NONE 

BODY CAVITIES 

NONE 

AH OTHER SYSTEMS 

NONE 

SPECIAL MORPHOLOGY SUMMARY 

NO LESION REPORTED 2 12

ANIMA L MISSING/NO NECROPSY

AOTOLYSIS/NO NECROPSY 1

* NUMBE R OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBFR OF ANIMALS NECROPSIED 

 13 

1 

2 

D-8
 

DHEW Publication No. (NIH) 78-1341