National Cancer Institute 

CARCINOGENESIS 

Technical Report Series 

No. 158 

1979 

BIOASSAY OF 

(2-CHLOROETHYL) 

TRIMETHYLAMMONIUM CHLORIDE (CCC) 

FOR POSSIBLE CARCINOGENICITY 

CAS No. 999-81-5 

NCI-CG-TR-158 

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

Public Health Service 

National Institutes of Health 

BIOASSAY OF
 

(2-CHLOROETHYL)TRIMETHYLAMMONIUM CHLORIDE
 

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) 79-1714
 
ii

 
BIOASSAY OF
 

(2-CHLOROETHYL)TRIMETHYLAMMONIUM CHLORIDE
 

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
 

(2-chloroethyl)trimethylammonium chloride 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. A negative result, in
 

which the test animals do not have a greater incidence of cancer
 

than control animals, does not necessarily mean that the test
 

chemical is not a carcinogen, inasmuch as the experiments are
 

conducted under a limited set of circumstances. A positive
 

result demonstrates that the test chemical is carcinogenic for
 

animals under the conditions of the test and indicates that
 

exposure to the chemical is a potential risk to man. The actual
 

determination of the risk to man from chemicals found to be
 

carcinogenic in animals requires a wider analysis.
 

CONTRIBUTORS; This bioassay of (2-chloroethyl)trimethylammonium
 

chloride was conducted at the NCI Frederick Cancer Research
 

Center (FCRC) (1), Frederick, Maryland, operated for NCI (2) by
 

Litton Bionetics, Inc.
 

The manager of the bioassay at FCRC was Dr. B. Ulland, the
 

toxicologist was Dr. E. Gordon, and Drs. R. Cardy and D. Creasia
 

compiled the data. Ms. S. Toms was responsible for management of
 

data, Mr. D. Cameron for management of histopathology, Mr. L.
 

Callahan for management of the computer branch, and Mr. R. Cypher
 

for management of the facilities. Mr. A. Butler performed the
 

computer services. Histopathologic evaluations for rats and mice
 

were performed by Dr. D. G. Fairchild (1). The diagnoses
 

included in this report represent his interpretations.
 

Animal pathology tables and survival tables were compiled at EG&G
 

Mason Research Institute (3). Statistical analyses were
 

performed by Dr. J. R. Joiner (4) and Ms. P. L. Yong (4), using
 

methods selected for the bioassay program by Dr. J. J. Gart (5).
 

ill
 
The chemicals used in this bioassay were analyzed at FCRC by Dr.
 

W. Zielinsky, and the chemical analyses were reviewed and
 

approved by Dr. W. Lijinsky.
 

This report was prepared at Tracer Jitco (4) under the direction
 

of NCI. Those responsible for the report at Tracer Jitco were
 

Dr. C. R. Angel, Acting Director of the Bioassay Program; Dr. S.
 

S. Olin, Deputy Director for Science; Dr. J. F. Robens,
 

toxicologist; Dr. R. L. Schueler, pathologist; Dr. G. L. Miller,
 

Ms. L. A. Owen, Ms. M. S. King, and Mr. W. D. Reichardt, 

bioscience writers; and Dr. E. W. Gunberg, technical editor, 

assisted by Ms. Y. E. Presley. 

The following scientists at NCI were responsible for evaluating
 

the bioassay experiment, interpreting the results, and reporting
 

the findings: Dr. Kenneth C. Chu, Dr. Cipriano Cueto, Jr., Dr.
 

J. Fielding Douglas, Dr. Richard A. Griesemer, Dr. Thomas E.
 

Hamm, Dr. William V. Hartwell, Dr. Morton H. Levitt, Dr. Harry A.
 

Milman, Dr. Thomas W. Orme, Dr. A. R. Patel, Dr. Sherman F.
 

Stinson, Dr. Jerrold M. Ward, and Dr. Carrie E. Whitmire.
 

(1) Frederick Cancer Research Center, P.O. Box B, Frederick,
 

Maryland.
 

(2) Carcinogenesis Testing Program, Division of Cancer Cause and
 

Prevention, National Cancer Institute, National Institutes of
 

Health, Bethesda, Maryland.
 

(3) EG&G Mason Research Institute, 1530 East Jefferson Street,
 

Rockville, Maryland.
 

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

Maryland.
 

(5) Mathematical Statistics and Applied Mathematics Section,
 

Biometry Branch, Field Studies and Statistics, Division of
 

Cancer Cause and Prevention, National Cancer Institute,
 

National Institutes of Health, Bethesda, Maryland,
 

iv
 
SUMMARY

 

A bioassay of (2-chloroethyl)trimethylammonium chloride for 

possible carcinogenicity was conducted by administering the test 

chemical in feed to F344 rats and B6C3F1 mice. 

Groups of 50 rats of each sex were administered either 1,500 or 

3,000 ppm of the compound for 108 weeks, and 50 mice of each sex 

were administered 500 or 2,000 ppm for 102 weeks. Matched 

controls consisted of 20 untreated rats and 20 untreated mice of 

each sex. All surviving animals were killed at the end of the 

period of administration of the test chemical. 

Mean body weights of dosed rats and mice were lower than those of 

corresponding controls for part or all of the bioassay, except 

for the dosed male mice, whose mean body weights were essentially 

the same as those of the corresponding controls. Survival was 

not affected significantly in any of the dosed groups of rats or 

mice and was at least 64% in every dosed or control group of each 

species at the end of the bioassay. Sufficient numbers of dosed 

and control rats and mice of each sex were at risk for the 

development of late-appearing tumors. Since there was virtually 

no decrease in mean body weight in dosed male mice and only a 

slight decrease in female mice, and since there were no other 

toxic signs and no dose-related mortality, the animals may have 

been able to tolerate higher doses. 

No tumors occurred in the rats or mice of either sex at 

incidences that could be associated with administration of the 

test chemical. 

It is concluded that under the conditions of this bioassay, 

(2-chloroethyl)trimethylammonium chloride was not carcinogenic 

for F344 rats or B6C3F1 mice of either sex. 
vi

 
TABLE OF CONTENTS
 

Page
 

I. Introduction 1
 

II. Materials and Methods 3
 

A. Chemical 3
 

B. Dietary Preparation 3
 

C. Animals 4
 

D. Animal Maintenance 5
 

E. Subchronic Studies 7
 

F. Chronic Studies 10
 

G. Clinical and Pathologic Examinations 10
 

H. Data Recording and Statistical Analyses 13
 

III. Results - Rats 19
 

A. Body Weights and Clinical Signs (Rats) 19
 

B. Survival (Rats) 19
 

C. Pathology (Rats) 22
 

D. Statistical Analyses of Results (Rats) 23
 

IV. Results - Mice 25
 

A. Body Weights and Clinical Signs (Mice) 25
 

B. Survival (Mice) 25
 

C. Pathology (Mice) 28
 

D. Statistical Analyses of Results (Mice) 29
 

V. Discussion 31
 

VI. Bibliography 35
 

APPENDIXES
 

Appendix A Summary of the Incidence of Neoplasms in Rats
 

Administered CCC in the Diet 37
 

Table Al Summary of the Incidence of Neoplasms in Male
 

Rats Administered CCC in the Diet 39
 

Table A2 Summary of the Incidence of Neoplasms in Female
 

Rats Administered CCC in the Diet 43
 

vli
 
Page 

Appendix B Summary of the Incidence of Neoplasms in Mice 

Administered CCC in the Diet 47 

Table Bl Summary of the Incidence of Neoplasms in Male 

Mice Administered CCC in the Diet 49 

Table B2 Summary of the Incidence of Neoplasms in Female 

Mice Administered CCC in the Diet 52 

Appendix C Summary of the Incidence of Nonneoplastic 

Lesions in Rats Administered CCC in the Diet.. 57 

Table Cl Summary of the Incidence of Nonneoplastic 

Lesions in Male Rats Administered CCC 

in the Diet 59 

Table C2 Summary of the Incidence of Nonneoplastic 

Lesions in Female Rats Administered 

CCC in the Diet 66 

Appendix D Summary of the Incidence of Nonneoplastic 

Lesions in Mice Administered CCC 

in the Diet 73 

Table Dl Summary of the Incidence of Nonneoplastic 

Lesions in Male Mice Administered CCC 

in the Diet 75 

Table D2 Summary of the Incidence of Nonneoplastic 

Lesions in Female Mice Administered CCC 

in the Diet 79 

Appendix E Analyses of the Incidence of Primary Tumors in 

Rats Administered CCC in the Diet 83 

Table El Analyses of the Incidence of Primary Tumors in 

Male Rats Administered CCC in the Diet 85 

Table E2 Analyses of the Incidence of Primary Tumors in 

Female Rats Administered CCC in the Diet 91 

Appendix F Analyses of the Incidence of Primary Tumors in 

Mice Administered CCC in the Diet 95 

viii
 
Page 

Table Fl Analyses of the Incidence of Primary Tumors in 

Male Mice Administered CCC in the Diet 97 

Table F2 Analyses of the Incidence of Primary Tumors in 

Female Mice Administered CCC in the Diet 100 

TABLES 

Table 1 CCC Subchronic Feeding Studies 

in Rats and Mice 9 

Table 2 CCC Chronic Feeding Studies in Rats 11 

Table 3 CCC Chronic Feeding Studies in Mice 12 

FIGURES 

Figure 1 Growth Curves for Rats Administered 

CCC in the Diet 20 

Figure 2 Survival Curves for Rats Administered 

CCC in the Diet 21 

Figure 3 Growth Curves for Mice Administered 

CCC in the Diet 26 

Figure 4 Survival Curves for Mice Administered 

CCC in the Diet 27 

ix
 

I. INTRODUCTION 

CH, 

I 

CICH-CH- -N®-CH, •CI® 2 Z I 3 

CH3 

(2-Chloroethyl) trimethylammonium chloride
 

(2-Chloroethyl)trimethylammonium chloride (CAS 999-81-5; NCI
 

C02960) is a plant growth regulator, or dwarfing agent, used on
 

poinsettias and azaleas in the United States (Meister, 1977), and
 

on several food crops, specifically cereal grains, grapes, and
 

pears in Europe (Vettorazzi, 1977; WHO/FAO, 1973). It has been
 

marketed as Cyclocel since 1959, and is known by the common
 

names chlormequat, chlorocholine chloride, or CCC (Spencer,
 

1973). The term CCC will be used in this report.
 

The acute oral LD of CCC, has been reported to be in the
 

range of 215 to 1,020 mg/kg in mice and 330 to 750 mg/kg in rats
 

(WHO/FAO, 1973). Hennighausen and Tiefenbach (1975) found that
 

500 mg/kg of CCC given orally killed 21/40 male mice (strain not
 

specified).
 
Acutely toxic doses of CCC cause lacrimation, salivation, and
 

intestinal motility, and although these signs of toxicity of CCC
 

in mammals resemble those of anticholinesterase agents,, the
 

chemical does not inhibit cholinesterase. These effects are
 

produced by stimulation at muscarinic receptors and are partially
 

antagonized by low doses of atropine, a cholinergic blocking
 

agent which specifically blocks muscarinic receptors. Lethal
 

doses cause respiratory failure that is due to neuromuscular
 

blockage and that is unaffected by atropine treatment
 

(Hennighausen et al., 1974; Hennighausen and Tiefenbach, 1975).
 

NCI initiated long-term tests with CCC in the early 1960*s as
 

part of an effort to assess the carcinogenic potential of
 

chemicals that were of concern to public health because of their
 

industrial importance or widespread use in the environment. In
 

these chronic tests, some animals developed hepatomas, but these
 

could not clearly be associated with the administration of the
 

test chemical (innes et al., 1969). Because these studies were
 

preliminary, the chemical was selected for study in the
 

Carcinogenesis Testing Program using expanded protocols.
 
II. MATERIALS AND METHODS
 

A. Chemical
 

CCC (C..H Cl N) was obtained as technical-grade nonformulated material from American Cyanamid Co. The material was a 

yellow-white crystalline solid made by reacting ethylene 

dichloride with trimethylamine. The compound had a stated 

technical-grade purity of 97 to 98%. The effluent from highpressure liquid chromatography using a refractive index detector 

contained three components of which 90% was CCC. Elemental 

analysis showed 36.4% carbon, 8.5% hydrogen, and 8.2% nitrogen 

(theoretical: 38.0% carbon, 8.2% hydrogen, and 8.9% nitrogen). 

The test material had a melting point of 240 C with decomposition (literature: 245 C with decomposition). 

The CCC was stored at 7°C until used. 

B. Dietary Preparation 

Test diets containing CCC were prepared fresh every 1 to 1-1/2 

weeks in 6- to 12-kg batches at appropriate doses. A known 
weight of the chemical was first mixed with an equal weight of 

autoclaved Wayne® Sterilizable Lab Meal with 4% fat (Allied 

Mills, Inc., Chicago, 111.), using a mortar and pestle. The 

mixing was continued with second and third additions of feed, and 

final mixing was performed with the remaining quantity of feed 

for a minimum of 15 minutes in a Patterson-Kelly twin-shell 

blender with an intensifer bar. 

The diets were stored at 7 C in plastic bags until used. 

C. Animals 

Male and female F344 (Fischer) rats and B6C3F1 mice were obtained 

as 4-week-old weanlings, all within 3 days of the same age, from 

the NCI Frederick Cancer Research Center animal farm (Frederick, 

Md.). The animals were housed within the test facility for 2 

weeks and then were assigned four rats to a cage and five mice to 

a cage on a weight basis for each cage of animals of a given 

species and sex. For use in the chronic study, the male rats 

were required to weigh 90 to 105 g, averaging at least 100 g; the 

female rats, 80 to 95 g, averaging at least 90 g; the male mice, 

18 to 22 g, averaging at least 19.5 g; and the female mice, 17 to 
21 g, averaging at least 18.5 g. Individual animals were
 

identified by ear punch.
 

D. Animal Maintenance
 

The animals were housed in polycarbonate cages (Lab Products, 

Inc., Garfield, N.J.), 19 x 10-1/2 x 8 inches for the rats and 

11-1/2 x 7-1/2 x 5 inches for the mice. The cages were suspended 

from aluminum racks (Scientific Cages, Inc., Bryan, Tex.) and 

were covered by nonwoven polyester-fiber 12-mil-thick filter 

paper (Hoeltge, Inc., Cincinnati, Ohio). The bedding used was 

Absorb-dri® hardwood chips (Northeastern Products, Inc., 

Warrenburg, N.Y.). The feed was presterilized Wayne® 

Sterilizable Lab Meal, provided ad_ libitum in suspended stainless 

steel hoppers and replenished at least three times per week. 

Water, acidified to pH 2.5, was supplied ad libitum from glass 

bottles. Sipper tubes (Lab Products, Inc.) were suspended 

through the tops of the cages. 

The contaminated bedding was disposed of through an enclosed 

vacuum line that led to a holding tank from which the bedding was 

fed periodically into an incinerator. The cages were sanitized 

twice per week and the feed hoppers twice per month at 82 to 
88 C in a tunnel-type cagewasher (Industrial Washing Corp., 

Mataway, N.J.), using the detergents, Clout (Pharmacal 

Research Laboratories, Greenwich, Conn.) or Oxford D'Chlor 

(Oxford Chemicals, Atlanta, Ga.). The glass bottles and sipper 

tubes were sanitized at 82 to 88 C in a tunnel-type bottle 

washer (Consolidated Equipment Supply Co., Mercersburg, Pa.) 

three times per week, using a Calgen Commercial Division 

detergent (St. Louis, Mo.). The racks for the cages were sanitized at or above 82 C in a rack washer (Consolidated Equipment 

Supply Co.) once per month, using the Calgen Commercial Division 

detergent, and the filter paper was changed at the same time. 

The animal rooms were maintained at 22 to 24 C and 45 to 55% 

relative humidity. Incoming air was passed through a filter of 

65% efficiency and a bag filter of 95% efficiency at the intake 

and was expelled without recirculation through a "Z"-type 

roughing filter of 30% efficiency and a bag system of 90 to 95% 

efficiency at the exhaust (American Air Filters, Louisville, Ky., 

Mine Safety Appliances, Pittsburgh, Pa.). The room air was 

changed 15 times per hour. The air pressure was maintained 

negative to a clean hallway and positive to a return hallway. 

Fluorescent lighting was provided automatically on a 

12-hour-per-day cycle. 
Rats administered CCC and their corresponding controls were
 

housed in the same room as rats on feeding studies of the
 

following chemicals:
 

(CAS 86-06-2) 2,4,6-trichlorophenol
 

(CAS 51-03-6) piperonyl butoxide
 

Mice administered CCC and their corresponding controls were
 

housed in the same room as mice on feeding studies of the
 

following chemicals:
 

(CAS 156-62-7) calcium cyanamide
 

(CAS 95-80-7) 2,4 diaminotoluene
 

(CAS 19010-66-3) lead dimethyldithiocarbamate
 

(CAS 86-30-6) N-nitrosodiphenylamine
 

(CAS 88-96-0) phthalamide
 

(CAS 120-62-7) piperonyl sulfoxide
 

(CAS 137-17-7) 2,4,5-trimethylaniline 

E. Subchronic Studies
 

Subchronic feeding studies were conducted to estimate the maximum
 

tolerated doses (MTD's) of CCC, on the basis of which two
 

concentrations (referred to in this report as "low" and "high"
 

doses) were selected for administration in the chronic studies.
 

Groups of five rats and five mice of each sex were administered
 

feed containing CCC at one of several doses for 7 weeks followed
 

by 1 week of observation, and groups of five control animals of
 
each species and sex were administered basal diet only. Each 

animal was weighed twice per week. 

Table 1 shows the survival of animals in each dosed group at the 

end of the course of chemical administration, and the mean body 

weights of dosed animals at week 7 expressed as percentages of 

mean body weights of the controls. 

At the end of the subchronic studies, all animals were killed 

using CO. and necropsied. Clinical observations exclusive of 

weight and microscopic examination, showed no dose-related 

changes for male or female rats dosed at 3,150 or 6,800 ppm nor 

for male or female mice dosed at 10,000 ppm. 

Ten percent depression in body weight was the major criterion for 

the estimation of MTD's. The doses required to produce this 

response were determined by the following procedure: first, 

least squares regressions of mean body weights versus days on 

study were used to estimate mean body weights of each of the 

dosed groups at day 49. Next, probits of the percent weights of 

the dosed groups at day 49 relative to weights of corresponding 

control groups were plotted against the logarithms of the doses, 

and least squares regressions fitted to the data were used to 

estimate the doses required to induce 10% depression in weight. 

8 
Table 1. CCC Subchronic Feeding Studies in Rats and Mice
 

Male Female
 

Mean Weight Mean Weight
 

at Week 7 at Week 7
 

Dose Surviv- as % of Surviv- as % of
 

(ppm) al (a) Control al (a) Control
 

RATS
 

3,150 5/5 85 5/5 88
 

4,600 5/5 83 5/5 81
 

6,800 5/5 79 5/5 68
 

10,000 4/5 45 4/5 49
 

14,700 0/5 0/5
 

MICE
 

1,200 5/5 86 5/5 79
 

2,500 5/5 78 5/5 85
 

4,000 5/5 81 5/5 66
 

5,000 5/5 77 5/5 56
 

7,000 5/5 74 5/5 68
 

10,000 5/5 70 5/5 53
 

20, 000 2/5 60 2/5 59
 

(a) Number surviving/number in group.
 
The low and high doses for rats were set at 1,500 and 3,000 ppm;
 

for mice, 500 and 2,000 ppm.
 

F. Chronic Studies
 

The test groups, doses administered, and durations of the chronic
 

feeding studies are shown in tables 2 and 3.
 

G. Clinical and Pathologic Examinations
 

All animals were observed twice daily. Observations for sick,
 

tumor-bearing, and moribund animals were recorded daily.
 

Clinical examination and palpation for masses were performed each
 

month, and the animals were weighed at least once per month.
 

Moribund animals and animals that survived to the end of the
 

bioassay were killed using C0_ and necropsied.
 

The pathologic evaluation consisted of gross and microscopic
 

examination of major tissues, major organs, and all gross
 

lesions. The tissues were preserved in 10% neutral buffered
 

formalin, embedded in paraffin, sectioned, and stained with
 

hematoxylin and eosin. The following tissues were examined
 

10
 
Table 2. CCC Chronic Feeding Studies in Rats
 

Sex and Initial CCC Time on 

Test No. of in Diet (b) Study 

Group Animals (a) (ppm) (weeks) 

Male
 

Mat ched-Contro 1 20 0 108
 

Low-Dose 50 1,500 108
 

High -Dose 50 3,000 108
 

Female
 

Matched-Control 20 0 108
 

Low-Dose 50 1,500 108
 

High -Dose 50 3,000 108
 

(a)	 Al animal l s were approximately 6 weeks of age when placed on
 

study.
 

(b)	 Test and control diets were provided ad_ libitum 7 days
 

per week.
 

11
 
Table 3. CCC Chronic Feeding Studies in Mice
 

Sex and Initial CCC Time on 

Test No. of in Diet (b) Study 

Group Animals (a) (ppm) (weeks) 

Male
 

Matched-Control 20 0 102
 

Low-Dose 50 500 102
 

High-Dose 50 2,000 102
 

Female
 

Matched-Control 20 0 102
 

Low-Dose 50 500 102
 

High-Dose 50 2,000 102
 

(a)	 Al animal l s were approximately 6 weeks of age when placed on
 

study.
 

(b)	 Test and control diets were provided £d_ libitum 7 days
 

per week.
 

12
 
microscopically: skin, lungs and bronchi, trachea, bone marrow
 

(femur), spleen, lymph nodes (mesenteric and submandibular),
 

thymus, heart, salivary glands (parotid, sublingual, and
 

submaxillary), liver, pancreas, esophagus, stomach (glandular and
 

nonglandular), small and large intestine, kidney, urinary
 

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

prostate, mammary gland, uterus, ovary, brain (cerebrum and
 

cerebellum), and all tissue masses. Peripheral blood smears also
 

were made for all animals, whenever possible.
 

Necropsies were also performed on all animals found dead, unless
 

precluded in whole or in part by autolysis or cannibalization.
 

Thus, the number of animals from which particular organs or
 

tissues were examined microscopically varies and does not
 

necessarily represent the number of animals that were placed on
 

study 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
 

13
 
design, clinical observations, survival, body weight, and individual pathologic results, as recommended by the International
 

Union Against Cancer (Berenblum, 1969). Data tables were generated for verification of data transcription and for statistical
 

review.
 

These data were analyzed using the appropriate 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 section.
 

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) for testing two groups for equality and Tarone's (1975)
 

extensions of Cox's methods for testing for 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.
 

14
 
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 is examined (denominator). In most instances,
 

the denominators included only those animals for which that site
 

was examined histologically. However, when macroscopic
 

examination was required to detect lesions prior to histologic
 

sampling (e.g., skin or mammary tumors), or when lesions could
 

have appeared at multiple sites (e.g., lymphomas), the
 

denominators consist of the numbers of animals necropsied.
 

The purpose of the statistical analyses of tumor incidence is to
 

determine whether animals receiving the test chemical developed a
 

significantly higher proportion of tumors than did the control
 

animals. As a part of these analyses, the one-tailed Fisher
 

exact test (Cox, 1970) was used to compare the tumor incidence of
 

a control group with that of a group of dosed animals at each
 

dose level. When results for a number of dosed 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) 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
 

15
 
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), was also used. Under the
 

assumption of a linear trend, this test determines 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 is a positive dose relationship. This method also provides a two-tailed test of departure
 

from linear trend.
 

A time-adjusted analysis was applied when numerous early deaths
 

resulted from causes that were not associated with the formation
 

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

first tumor was observed were excluded by basing the statistical
 

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

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

such an early tumor was found, comparisons were based exclusively
 

on animals that survived at least as long as the animal in which
 

the first tumor was found. Once this reduced set of data was
 

obtained, the standard procedures for analyses of the incidence
 

of tumors (Fisher exact tests, Cochran-Armitage tests, etc.) were
 

followed.
 

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

incidence of tumors. Curves of the proportions surviving without
 

an observed tumor were computed as in Saffiotti et al. (1972).
 

The week during which an animal died naturally or was 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 less than 0.05, two-


tailed test) were also noted.
 

The approximate 95 percent confidence interval for the relative
 

risk of each dosed group compared with its control was calculated
 

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

relative risk is defined as p./p where p is the true
 

binomial probability of the incidence of a specific type of tumor
 

in a dosed group of animals and PC is the true probability of
 

the spontaneous incidence of the same type of tumor in a control
 

group. The hypothesis of equality between the true proportion of
 

a specific tumor in a dosed group and the proportion in a control
 

group corresponds to a relative risk of unity. Values in excess
 

17
 
of unity represent the condition of a larger proportion in the
 

dosed 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% of a large number of identical experiments, the
 

true ratio of the risk in a dosed 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 (P less than 0.025 one-tailed test when the
 

control incidence is not zero, P less than 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. RESULTS - RATS
 

A. Body Weights and Clinical Signs (Rats)
 

Mean body weights of dosed male and female rats were lower than
 

those of corresponding controls and were dose related throughout
 

the bioassay, although differences between dosed and control
 

males were slight (figure 1). Other clinical signs, such as
 

corneal opacity and tissue masses, were observed at comparable
 

incidences in dosed and control groups.
 

B. Survival (Rats)
 

The Kaplan and Meier curves estimating the probabilities of
 

survival for male and female rats administered CCC in the diet at
 

the doses of this bioassay, together with those of the matched
 

controls, are shown in figure 2. The result of the Tarone test
 

for dose-related trend in mortality is not significant in either
 

sex.
 

In male rats, 32/50 (64%) of the high-dose group, 37/50 (74%) of
 

the low-dose group, and 14/20 (70%) of the control group lived to
 

19
 
60 

TIME ON STUDY (WEEKS) 

Figure 1. Growth Curves for Rats Administered CCCin the Diet 

20 
TIME ON STUDY (WEEKS) 

Figure 2. Survival Curves for Rats Administered CCC in the Diet 

21 
the end of the bioassay. In females, 41/50 (82%) of the 

high-dose group, 35/50 (70%) of the low-dose group, and 13/20 

(65%) of the control group lived to the end of the bioassay. 

Sufficient numbers of rats of each sex were at risk for the 

development of late-appearing tumors. 

C. Pathology (Rats) 

Histopathologic findings on neoplasms in rats are summarized in
 

Appendix A, tables Al and A2; findings on nonneoplastic lesions
 

are sunmarized in Appendix C, tables Cl and C2.
 

From an inspection of the numerical differences in the incidences
 

of leukemia or malignant lymphoma in the female rats (controls
 

3/20, low-dose 11/50, high-dose 14/50), one could infer an
 

increase in neoplasia in the animals receiving CCC. There was
 

also an apparent dose-related increase in the incidence of
 

islet-cell adenomas of the pancreas of the male rats (controls
 

0/18, low-dose 1/47, high-dose 7/45). No islet-cell adenomas of
 

the pancreas were seen in any of the female rats.
 

Several chronic inflammatory, degenerative, or proliferative
 

22
 
lesions frequently seen in aged F344 rats occurred with 

approximately equal frequency and severity in each sex of the 

dosed and control animals. 

Based on the histopathologic examination, there was no clear 

evidence of carcinogenicity in F344 rats due to the 

administration of CCC under the conditions of this bioassay. 

D. Statistical Analyses of Results (Rats) 

Tables El and E2 in Appendix E contain the statistical analyses 

of the incidence of those primary tumors that occurred in at 

least two animals of one group and at an incidence of at least 5% 

in one or more than one group. 

In male rats, the result of the Cochran-Armitage test for 

positive dose-related trend in the incidence of islet-cell 

adenoma is significant (P = 0.023), but the results of the Fisher 

exact test are not significant. The historical records of this 

laboratory show an incidence of 16/416 (4%) in male controls, 

with incidences in individual control groups- as high as 3/16 

(19%) or 2/19 (11%) to as low as 0/20. 

23 
The incidences of female rats with lymphoma or leukemia are 3/20 

(15%) in the control group, 11/50 (22%) in the low-dose group, 

and 14/50 (28%) in the high-dose groups. The results of the 

Cochran-Armitage test and the Fisher exact test are not 

significant. The historical records of this laboratory show an 

incidence of 42/420 (10%) in female controls with incidences in 

individual control groups as high as 4/20 (20%) or 3/20 (15%) to 

as low as 0/20. 

Significant results in the negative direction are observed in the 

incidence of C-cell tumors of the thyroid in male rats and in the 

incidence of fibroadenomas of the mammary gland in female rats. 

In each of the 95% confidence intervals for relative risk, shown 

in the tables, the value of one or less than one is included; 

this indicates the absence of significant positive results. It 

should also be noted that each of the intervals, except that for 

the incidence of C-cell tumors of the thyroid in the high-dose 

male rats, has an upper limit greater than one, indicating the 

theoretical possibility of the induction of tumors by CCC, which 

could not be detected under the conditions of this test. 

24 
IV. RESULTS - MICE
 

A. Body Weights and Clinical Signs (Mice)
 

Mean body weights of the dosed male mice were essentially
 

unaffected by administration of the test chemical throughout the
 

bioassay. Mean body weights of the female mice were unaffected
 

during the first 40 weeks, but thereafter were slightly lower
 

than those of the corresponding controls (figure 3). Other
 

clinical signs, such as tissue masses, were observed at
 

comparable incidences in the dosed and control groups.
 

B. Survival (Mice)
 

The Kaplan and Meier curves estimating the probabilities of
 

survival for male and female mice administered CCC in the diet at
 

the doses of this bioassay, together with those of the matched
 

controls, are shown in figure 4. In male mice, the result of the
 

Tarone test for dose-related trend in mortality is not
 

significant. An indicated departure from linear trend (P =
 

0.014) is observed, because the control animals did not survive
 

as long as the dosed animals. The result of the Cox test between
 

25
 
8 D a B 8 8 s\ 7> 

8 A 

§ B a 

ill 

S n 

Q

O 

a> 

2 

ui 

5 

MALE MICE 

D MATCHED CONTROL 

O LOW DOSE 

A HIGH DOSE 

10 20 30 40 50 60 70 90 100 

TIME ON STUDY (WEEKS) 

D D 

a o a 

D 

^ .* £ 8 D 

fi 

a 

o n 

8 

r

e .6 

FEMALE MICE 

D MATCHED CONTROL 

O LOW DOSE 

A HIGH DOSE 

20 40 50

 100 1 0 

TIME ON STUDY (WEEKS) 

Figure 3. Growth Curves for Mice Administered CCC in the Diet 

26 
"S-" •&-+" A~ -O 

I *A... 

(L A-i^ 

D 

CO 

u. 

t 

m 

O 

cc 

MALE MICE 020­ D MATCHED CONTROL 

O LOW DOSE 

0 10­ /1 HIGH DOSE 

45 60 75 

TIME ON STUDY (WEEKS) 

^ 

^>-\ A _ o—. ""A: 

h A-­ A A 

H^ 

CO 

° 060­

§ 040­

m 

O 

cc 

FEMALE Ml CE 020-

D MATCHED CONTROL 

O LOW DOSE 

010- L\ HIGH DOSE 

c 15 30 45 60 75 90 1C 5 12 

TIME ON STUDY (WEEKS) 

Figure 4. Survival Curves for Mice Administered CCCin the Diet 

27 
the control and the low-dose groups is significant (P = 0.034),
 

but in the negative direction. In females, the result of the
 

Tarone test is not significant.
 

In male mice, 42/50 (84%) of the high-dose group, 49/50 (98%) of
 

the low-dose group, and 16/20 (80%) of the control group lived to
 

the end of the bioassay. In females, 46/50 (92%) of the
 

high-dose group, 41/50 (82%) of the low-dose group, and 16/20
 

(80%) of the control group lived to the end of the bioassay.
 

Sufficient numbers of mice of each sex were at risk for the
 

development of late-appearing tumors.
 

C. Pathology (Mice)
 

Histopathologic findings on neoplasms in mice are summarized in
 

Appendix B, tables Bl and B2; findings on nonneoplastic lesions
 

are summarized in Appendix D, tables Dl and D2.
 

There was a slightly increased incidence of hemangiomas and
 

hemangiosarcomas in the dosed females (controls 1/20, low-dose
 

4/50, high-dose 5/50).
 

28
 
Several chronic inflammatory, degenerative, or proliferative
 

lesions frequently seen in aged B6C3F1 mice occurred with
 

approximate equal frequency and severity in the dosed and control
 

an ima1 s.
 

Based on the histopathologic examination, there was no clear
 

evidence of carcinogenicity in B6C3F1 mice due to administration
 

of CCC under the conditions of this bioassay.
 

D. Statistical Analyses of Results (Mice)
 

Tables Fl and F2 in Appendix F contain the statistical analyses
 

of the incidences of those primary tumors that occurred in at
 

least two animals of one group and at an incidence of at least 5%
 

in one or more than one group.
 

In male mice, the result of the Cochran-Armitage test for
 

dose-related trend in the incidence of hepatocellular carcinoma
 

is significant (P = 0.036), but the results of the Fisher exact
 

test are not significant. In female mice, a slightly increased
 

incidence of hemangiomas and hemangiosarcomas is not significant.
 

Significant trends in the negative direction are observed in the
 

29
 
incidences of lymphoma and of cortical adenoma of the adrenal in
 

male mice and of adenoma of the pituitary in females.
 

In each of the 95% confidence intervals for relative risk, shown
 

in the tables, the value of one is included; this indicates the
 

absence of significant positive results. It should also be noted
 

that each of the intervals has an upper limit greater than one,
 

indicating the theoretical possibility of the induction of tumors
 

by CCC, which could not be detected under the conditions of this
 

test.
 

30
 
V. DISCUSSION
 

Mean body weights of the dosed rats and mice were lower than 

those of corresponding controls for part or all of the bioassay, 

except for the dosed male mice, whose mean body weights were 

essentially unaffected by administration of the test chemical. 

Survival was not affected significantly in any of the dosed 

groups of rats or mice and was 64% or greater in both dosed and 

control groups of each species at the end of the bioassay. 

Sufficient numbers of rats and mice of each sex were at risk for 

the development of late-appearing tumors. Since there was 

virtually no decrease in mean body weight in dosed male mice and 

only a slight decrease in female mice and since there were no 

other toxic signs and no dose-related mortality, the mice may 

have been able to tolerate higher doses. 

Islet-cell adenomas of the pancreas occurred in the male rats at 

incidences that were dose related (P = 0.023), but in direct 

comparisons the incidences in the individual dosed groups were 

not significantly higher than those in the control group 

(controls 0/19, low-dose 2/47, high-dose 7/49)s In female rats, 

lymphoma or leukemia occurred in a higher percentage of dosed 

than control animals (controls 3/20, or 15%, low-dose 11/50, or 

31 
22%, and high-dose 14/50, or 28%). The results of the 

statistical analyses were not, however, significant. Hepatocellular carcinomas occurred in the male mice at incidences that 

were dose related (P = 0.036), but in direct comparisons the 

incidences in the individual dosed groups were not significantly 

higher than that in the control group (controls 7/20, low-dose 

13/50, high-dose 23/49). Thus, the occurrence of pancreatic 

tumors in the dosed male rats, lymphoma or leukemia in the dosed 

female rats, and liver tumors in the dosed male mice cannot 

clearly be related to administration of the test chemical. No 

tumors occurred in the female mice at incidences that were 

significant either for positive dose-related trend or for greater 

incidences in dosed groups than in control groups. 

In previous long-term feeding studies of CCC, administration of 

1,000 ppm for 78 weeks to CFLP mice caused no adverse effect on 

the survival and only about 6% decrease in body weight gained; an 

incidence of benign lung tumors of 20/52 in the dosed males was 

higher than that of 10/51 in the controls, but was considered to 

be within the normal range under the conditions of the test 

(WHO/FAO, 1973). In other long-term feeding studies in mice, 

administration of CCC at 21.5 mg/kg by stomach tube for 4 weeks, 

then in the diet at 65 ppm for 18 months, to B6C3F1 and B6AKF1 

hybrids led to incidences of hepatomas in 5/18 males of each 

32 
hybrid compared with incidences of 6/257 and 7/240 in the
 

corresponding controls (Innes et al., 1969; WHO/FAO 1973). When
 

rats of unidentified strain were administered 500 or 1,000 ppm
 

CCC in the diet for 2 years, they showed no signs of toxicity or
 

histopathologic abnormalities attributable to the test chemical
 

(WHO/FAO, 1973).
 

It is concluded that under the conditions of this bioassay, CCC
 

was not carcinogenic for F344 rats or B6C3F1 mice of either sex.
 

33
 
34

 
VI. BIBLIOGRAPHY
 

Armitage, P., Statistical Methods in Medical Research, John Wiley
 

& Sons, Inc., New York, 1971, pp. 362-365.
 

Berenblum, I., ed., Carcinogenicity Testing, A Report of the
 

Panel on Carcinogenicity £f the Cancer Research Commission £f_ the
 

UICC, Vol. "L, International Union Against Cancer, Geneva, 1969.
 

Cox, D. R., Regression models and life tables. £._ R._ Statist.
 

^ B 34(2): 187-220, 1972.
 

Cox, D. R., Analysis £f Binary Data, Methuen & Co., Ltd., London,
 

1970, pp. 48-52.
 

Gart, J. J., The comparison of proportions: A review of
 

significance tests, confidence limits and adjustments for
 

stratification. Rev. Int. Statist. Inst. 39(2) ;148-169, 1971.
 

Hennighausen, G. and Tiefenbach, B., The toxicological and
 

pharmacological properties of chlorocholine chloride. In:
 

Developmental and Genetic Aspects £f Drug and Environmental
 

Toxicity. Proceedings of the European Society of Toxicology,
 

Vol. 16, American Elsevier Publishing Co., Inc., New York, 1975,
 

pp. 300-302.
 

Hennighausen, G., Tiefenbach, B., and Dietrich, C. ,
 

Untersuchungen fiber toxikologische und phannakologische
 

Eigenschaften von Chlorcholinchlorid und N,N-Dimethyl-(2­

broma*thyl)-hydraziniumbromid. Acta biol. med. germ. 33:89-98,
 

1974.
 

Innes, J. R. M., Ulland, B. M., Valeric, M. G., Petrucelli, L. ,
 

Fishbein, L., Hart, E. R., Pallotta, A. J., Bates, R. R., Falk,
 

H. L., Gart, J. J., Klein, M., Mitchell, I., and Peters, J.,
 

Bioassay of pesticides and industrial chemicals for
 

tumorigenicity in mice: a preliminary note. J._ Natl. Cancer
 

Inst. 42(6):1101-1114.
 

Kaplan, E. L., and Meier, P., Nonparametric estimation from
 

incomplete observations. J^ Amer. Statist. Assoc. 53:457-481,
 

1958.
 

35
 
Linhart, M. S., Cooper, J. A., Martin, R. L., Page, N. P., and
 

Peters, J. A., Carcinogenesis bioassay data system. Comp. and 

Biomed. Res. 7:230-248, 1974. 

Meister, R. T., ed., Chlormequat chloride. In: 1977 Farm 

Chemicals Handbook, Meister Publishing Co., Willoughby, Ohio,
 

1977, p. D60.
 

Miller, R. G., Jr., Simultaneous Statistical Inference,
 

McGraw-Hill Book Co., New York, 1966, pp. 6-10.
 

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

Kaufman, D. G., Respiratory tract carcinogenesis in hamsters
 

induced by different numbers of administrations of benzo (a)
 

pyrene and ferric oxide. Cancer Res. 32:1073-1081, 1972.
 

Spencer, E. Y., Chlormequat. In: Guide to the Chemicals Used in
 

Crop Protection, University of Western Ontario, London, Ontario,
 

1973, p. 104.
 

Tarone, R. E., Tests for trend in life table analysis.
 

Biometrika 62:679-682, 1975.
 

Vettorazzi, G., State of the art of the toxicological evaluation
 

carried out by the Joint FAO/WHO Expert Committee on Pesticides
 

Residues. III. Miscellaneous pesticides used in agriculture and
 

public health. Residues Review 66:137-184, 1977.
 

WHO/FAO, Chlormequat. In: 1972 Evaluations of_ Some Pesticide
 

Residues in Food ^ The Monographs, World Health Organization,
 

Geneva, 1973, pp.103-130.
 

36
 
APPENDIX A
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN RATS
 

ADMINISTERED CCC IN THE DIET
 

37
 
38

 
TABLE A1.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS 

ADMINISTERED CCC IN THE DIET 

MATCHED 

CONTROL LOW DOSE HIGH DOSE 

ANIMALS INITIALLY IN STUDY 20 50 50
 

ANIMALS NECROPSIED 20 50 50
 

ANIMALS EXAMINED HISTOPATHOLOGICALLY 20 49 50
 

INTEGUMENTARY SYSTEM
 

*SKIJI (20 ) (50) (50)

 

BASAL-CELL TUHOR 1 (2X)

 

TRICHOEPITHELIOMA 2 (4X) 

FIBROMA 

*SUBCOT TISSUE (20) (50) (50)
 

SARCOMA, NOS 1 (5X) 1 (2X)
 

FIBROMA 1 (5X) 1 (2X) 3 (6X)
 

LIPOMA 2 (4X)
 

BISPIRATORY SYSTEM 

*LUNG (20) (49) (50)
 

CARCINOHA, NOS, METASTATIC 1 (2X)
 

ALVEOLAR/EEONCHIOLAR ADENOMA 3 (6X)
 

ALVEOLAR/EFONCHIOLAR CARCINOMA 2 (4*)
 

HEMATOPOIETIC SYSTEM
 

*MOLTIPLE ORGANS (23) (50) (50) 

MALIGNANT 1YHEHOMA, NOS 5 (25X) 5 (10X) 11 (22%) 

MALIG.LYMPHOMA , UNDIFFER-TYPE 1 (5X) 

MALIG.LYMPHOMA , LYMPHOCYTIC TYPE 2 (4X) 

MALIG.LYMPHOMA, HISTIOCYTIC TYPE 2 1 (2X) 

MONOCYTIC IEUKEMIA 1 

#SPLtEN (20) (49) (50) 

MALIGNANT IYMPHOMA, NOS 1 (2X) 

*THYMUS (7) (36) (43) 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

39
 
TABLE A1. MALE RATS: NEOPLASMS (CONTINUED) 

MATCHED 

CONTROL LOW DOSE HIGH DOSE 

FIBEOSAECCBA 1 (2%) ____.___„•__ _ ____.______._______.» _ _ — «__ —— ___ _ — _— — __^_ _._.» — —____.«.— ——_,..»— — — — — —•— — ___• — __. 

CIRCUiATORY SYSTEM 

NONE 

DIGESTIVE SYSTEfl 

tLIVER

HiPATOCEIlULAR CARCINOMA

 (20)

 1 (5S)

 (49)

 2 (HI)

 (50) 

2 (431) 

• PANCREAS

ACINAR-CEL1 ADENOMA

 (19) (47)

 1 (2X)

 (49) 

1 (2X) 

ISTOMACH

SQUAMOUS CILL CARCINOMA

 (20) (49)

 1 (2%) 

 (50) 

URINASY SYSTEH
 

4KIDNEY (20) (49) (50)
 

TDBULAR-CEIL ADENOMA 1 (2X)
 

LIPOSARCCHA 1 (5X)
 

«KIDNEY/CAPSUIE (20) (49) (50) 

SARCOMA, NCS, METASTATIC 1 

* URINAR Y BLACtE R (19) (49) (47) 

THANSITICNAL-CELL CARCINOMA 1 (2%) 

ENDOCRINE SYSTEM
 

#PITUITARY (20) (49) (47)
 

CHROMOPHOEH AEENOMA 6 (30X) 11 (22X) 16 (34S)
 

CHROMOPHOEE CARCINOMA 4 (8X)
 

lADR^NAL (20) (49) (50)
 

CARCINOMA, NCS, METASTATIC 1 (2%)
 

CORTICAL CARCINOMA 1 (2%)
 

PHEOCHRCMCCYTOMA 1 (5X) 3 (6X)
 

tTHYBOID (20) (50)
 

EOLLICOLAR=CII1_A£ENCMA
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMttER OF AKIMALS NECROPSIED
 

40
 
TABLE A1. MALE RATS: NEOPLASMS (CONTINUED) 

MATCHED 

CONTROL LOW DOSE HIGH DOSE 

FOLLICULAB-CELL CARCINOH A 2 (4*) 

C-CELL ABOCHA 3 (15X) 7 (15X) 

C-CELL CARCINOMA 1 (2X) 

CYSTADENCMA , NO S 1 (2«) 

#PANCREATIC ISLE1S (19) C*7) (49) 

ISLET-CELL ADENOMA 2 (<i%) 7 (14%) 

REPBODUCTIVE SYSTE M 

*MAMMAR Y GLAK I (20) (50) (50)

 

FIBROADENOM A 1 (2*)

 

*PHEPUTIAL GLAND (20) (50) (50) 

ADENOMA , NCS 1 (2«)

 

#TESXIS (20) (49) (49 ) 

INTERSTITIAL-CELL TUMOR 17 (95%) 42 (86%) 38 (78X) 

LIPOMA 1 (2X) 

NERVOU S SYSTEM 

IBRAIN (20) (49) (4S) 

OSTEOSABCCHA 1 (2%) 

OLIGODENDROGLIOM A 1 (2*) 

SPECIAL SENSE ORGANS 

NONE 

MUSCULOSKELETAL SYSTEM
 

NO Hi
 

BODY CAVITIES 

*BODY CAVITIES (20) (50) (50) 

MriSOTHELICHA, NOS 1 (2X) 

*PEBITONEUM (20) (50)
 

FIBBOSABCCBA
 

t NUHflER OF ASIHALS HITH TISSUE EXAMINED HICROSCOPICALLY
 

* NUHBER OF ANIMALS NECROPSIED
 

41
 
TABLE A1. MALE RATS: NEOPLASMS (CONTINUED) 

*PLEURA 

CARCINOMA, NOS 

MESOTHELIOMA, NOS 

SLL OTHER SYSTEMS 

*HULTIPLE ORGANS 

FIBROSABCCKA 

MiSOTHELICMA, MALIGNAN T 

ANIMAL DISPOSI1ICN SUMMAR Y 

ANIMALS INITIALLY IN STUDY 

NATURA L EEATH3 

MORIBUND' SACRIFICE 

SCHEDULED SACRIFICE
 

ACCIDENT&LLY KILLED
 

TERMINAL SACRIFICE
 

ANIMAL MISSING
 

INCLUDES AC1CLYZED ANIMALS
 

1UMOH SUMMARY
 

TOTAL ANIMALS WITH PRIMARY TDHORS*
 

TOTAL PRIMARY TUMORS
 

TOTAL ANIMALS BITH BENIGN TUMORS
 

TOTAL BENIGN TUMORS
 

TOTAL ANIMALS WITH MALIGNANT TUMORS
 

TOTAL MALIGNANT TUMORS
 

TOTAL ANIMALS HITH SECONDARY TUMOBS*


TOTAL SECONDARY TUMORS


TOTAL ANIMALS WITH TUMORS UNCERTAINEENIGN OR MALIGNANT


TOTAL UNCERTAIN TUMORS


TOTAL ANIMAIS WITH TUMORS UNCERTAIN­ PRIMARY OR METASTATIC
 

TOTAL UNCERTAIN TUMORS
 

MATCHED 

CONTROL LOW DOSE HIGH DOSE 

(20) (53) (50) 

1 (5X) 

1 (5X) 

(20) (50) (50) 

1 (2X) 

1 (5X) 

20 50 50 

3 11 11 

3 2 7 

37 32 

20 47 49 

41 98 98 

18 1*5 47 

29 76 72 

9 17 20 

11 22 25 

1 1 

1 2 

1 1 

1 1 

* PRIMARY TUMORS: ALL TUHORS EXCEPT SECONDARY TUMORS
 

# SECONDARY TUMORS: METASTATIC TUHORS OR TUMORS IN7ASIVE INTO AN ADJACENT ORGAN
 

42
 
TABLE A2.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE RATS 

ADMINISTERED CCC IN THE DIET 

MATCHED
 

CONTROL LOW DOSE HIGH DOSE
 

ANIMALS INITIALLY IN STUEY 20 50 50
 

ANIMALS NECROESIED 20 50 50
 

ANIMALS EXAMINEE HISTOPATHOiOGICALLY 20 50 50
 

INTEGUMENTARY SYSTEM
 

*SKIN (20) (50) (50)
 

PAPILLOMA, NOS 1 (2%)
 

SQUAMOUS CILL CARCINOMA 1 (2%)
 

TRICHOEPITHELIOMA 1 (2%)
 

KERATOACANTHOMA 1 (2%)
 

*SUBl.DT TISSUE (20) (50) (50)
 

FIBROMA 1 (2X)
 

FIBROSAECCKA 1 (2%)
 

HEMANGIOSAECCHA 1 (2%)
 

RESPIRATORY SYSTEM
 

tTRACHEA (20) (US) (49)
 

ADENOCARCINOMA, NOS 1 (5X)
 

#LUNG (20) (50) («9)
 

ADENOCARCINOMA, NOS, METASTATIC 1 (5X)
 

ALVEOLAR/EBONCHIOLAR ADENOMA 2 (4X) 1 (2*)
 

HEMATOPOIETIC SYSTEM
 

*MOLTIPLE ORGANS (20) (50) (50)
 

MALIGNANT LYMPHOMA, NOS 5 (10X) 13 (26%)
 

HALIG.LYMPHOMA, HISTIOCYTIC TYPE 1 (2X)
 

L£UKEMIA,NCS 1 (5«)
 

BONOCYTIC IEUKEHIA 1 (5«) 1 (2%)
 

*BLOOD (20) (50) (50)
 

LEUKEMIA, NCS 2 (<**)
 

LYMPHOCYTIC _LEUKEHIA 1__L2XJ
 

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

43
 
TABLE A2. FEMALE RATS: NEOPLASMS (CONTINUED) 

*SPLfcEN

MALIGNAN T LYMPHOMA , NOS

MALIG.LYMPHOMA , HISTIOCYTIC TYPE

*MANDIBULA R I. NOD E

ADENOCARCINOMA , NOS, METASTATIC

#THYHUS


CABCINOMfl,NOS
 •_ __________ _ ___________________ _ — — _ — — — —. —


CIBCULATOEY SYSTEM
 

NONE
 

C1GESTIVE SYSTEM
 

ILIVEE


HEPATOCELLULAR CARCINOMA


ftCECUN

ADENOMATOU S POLYP, NOS

URINAR Y SYSTE K 

*URINAR Y BLAEDER

TRANSITIONAL-CELL CARCINOM A

ENDOCRINE SYSTEM 

#PITUITARY
 

CARCINOMA,NOS
 

AOENOMA, NCS
 

CHROMOPHOBE ADENOMA
 

CHROMOPHOEI CARCINOMA
 

#ADRENAL
 

CORTICAL iEENOMA
 

PHEOCHRCMCCYTCMA
 

ITHYKOID
 

MATCHED 

CONTROL LOW DOSE HIGH DOSE 

 (20) (50) (50) 

1 (5«) 1 (2X) 

1 (2%) 

 (20) (49) (4S) 

1 (5%) 

 (12)	 (38) (38)
 

1 (3%)
 ___«.____ -._____ ___-—-_ — _——.— __ — — — — — — — •._— — —--,- — .
 

 (20)	 (50) (50)
 

1 (2X)
 

 (18) (50) (SO) 

1 (2%) 

 (19) (50) (50) 

1 (2%) 

(20)	 (49) (49)
 

1 (2%)
 

1	 (2*)
 

5 (25*) 22 (45X) 20 (41X)
 

1 (5«) 2 1 (2X)
 

(20)	 (50) (50)
 

1 (2%)
 

1 (2X)
 

(20) (49)	 (49)
 

# NOMUER OF ANIMALS HITH TISSUE	 EXAMINED MICROSCOPICALLY
 

*	 NUMBER OF ANIMALS NECROPSIED
 

44
 
TABLE A2. FEMALE RATS: NEOPLASMS (CONTINUED) 

MATCHED 

CONTROL LOW DOSE HIGH DOSE 

ADENOMA , NOS 1 (2%) 

C-CELL ADifcCM A 3 (6%) 2 

C-CELL CABCINOM A 1 (5X) 1 (2%) 

REPRODUCTIVE SYSTEM
 

*MAMMARY GLANI (20) (50) (50) 

AD3NOMA, NOS 1 (2%) 

CXSTADENCMA, NOS 1 (5X) 

FIBROMA 1 (555) 

FIBROADENOMA 4 (205E) 7 (14X) 2 (4S) 

dUTEROS (20) (50) (50) 

ADENOCARCINOMA , NOS 1 (2%) 

LEIOMYOM A 1 (2%) 

LEIOMYOSAFCOHA 1 (5X) 1 (2%) 

NERVOUS SYSTEM
 

IBRAIN (20) (50) (50)
 

CHROMOPHOEE CARCINOMA, INVASIVE 1 (2%)
 

CHROMOPHOEE CARCINOMA, METASTATI 1 (2%)
 

ASTROCYTCMA 1 (5%)
 

SPECIAL SENSE CRGANS
 

NONE
 

MUSCULOSKELETAL SYSTEM 

NONfi 

EOCY CAVITIES 

NONE 

ALL OTHER SYSTEMS 

NONi 

* NUMbER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

45
 
TABLE A2. FEMALE RATS: NEOPLASMS (CONTINUED) 

JNIMAL DISEOSI1ION SUMMABY
 

ANIHALS INITIALLY IN STUDY
 

NATURAL DEATHS
 

MORIBUND SACRIFICE
 

SCHEDULED SACRIFICE
 

ACCIDENTALLY KILLED
 

TERMINAL SACRIFICE
 

ANIMAL MISSING
 

3 INCLUDES AUTCLYZED ANIMALS
 

TUMOB SUMMARY
 

TOTAL ANIMALS WITH PRIMARY TUMORS*
 

TOTAL PRIMARY TUMORS
 

TOTAL ANIMALS WITH BENIGN TUMORS
 

TOTAL BENIGN TUMORS
 

TOTAL ANIMALS WITH MALIGNANT TUMORS
 

TOTAL MALIGNANT TUMORS
 

TOTAL ANIMALS WITH SECONDARY TUMORS*


TOTAL SECCKDARY TUMORS


TOTAL ANIHALS WITH TUMORS UNCERTAIN­ BENIGN OR MAIIGNANT
 

TOTAL UNCERTAIN TUMOES
 

TOTAL ANIMALS WITH TUMORS UNCERTAIN­ PRIHAHY OB METASTATIC
 

TOTAL UNCERTAIN TDMOES
 

MATCHED
 

CONTROL
 

20
 

13
 

19
 

8
 

1 1
 

1


 2


LOW DOSE HIGH DOSE 

5) 50 

35 U1 

41 37 

55 

30 27 

37 33 

17 19 

17 22 

1 1 

1 1 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUHORS
 * SECONDARY TUMORS: METASTATIC TUMORS OR TUMORS INVASIVE INTO AN ADJACEKT ORGAN
 

46
 
APPENDIX B
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MICE
 

ADIMINISTEREO CCC IN THE DIET
 

47
 
48

 
TABLE B1.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE 

ADMINISTERED CCC IN THE DIET 

ANIMALS INITIALLY IN STUDY
 

ANIMALS NECEOESIED
 

ANIMALS EXAMINID HISTOPATHC10GICALIY
 

INTEGUMENTARY SYSTEM
 

*SUBCUT TISSUE
 

HEMANGIOMA
 

RESPIRATORY SYSTEM
 

*IUNG
 

ALVEOLAR/ERCNCHIOLAH ADENOMA
 

ALVEOLAH/EEONCHIOLAE CAECINOMA
 

HEMATOPOIETIC SYSTEM
 

*MULTIPLE OBGANS
 

MALIGNANT IYMPHOMA, NOS
 

SMESiNTERIC 1. NODE
 

HEMANGIOMA
 

MALIGNANT IYMFHOMA, NOS
 

IKIDNEY
 

MALIGNANT LYMPHOMA, NOS
 

tTHYMUS
 

THYMOMA, MALIGNANT
 

MALIGNANT LYMPHOMA, NOS
 

CIRCULATORY SYSTEM
 

NONE
 

DIGESTIVE SYSTEM
 

*LIVER
 

HEPATOCELLULAR CARCINOMA
 

MATCHED
 

CONTROL
 

20
 

20
 

20
 

(20)
 

(20)
 

2 (1055)
 

2 (10*)
 

(20)
 

3 (15*)
 

(20)
 

(20)
 

(18)
 

(20)
 

7 (35*)
 

LOW DOSE
 

50
 

50
 

50
 

(50)
 

(50)
 

3 (6%)
 

7 (14%)
 

(50) 

7 (14%) 

(50) 

1 (2%) 

1 (2%) 

(50) 

1 (2%) 

(43) 

1 (2«) 

1 (2«) 

(50)
 

13 (26%}
 

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

49
 

HIGH DOSE
 

50
 

49
 

49
 

(49)
 

1 (2%)
 

(49)
 

2 (4*)
 

3 (6%)
 

(49)
 

2 (4*)
 

(49)
 

(49)
 

(44)
 

(49)
 

23_J47X.l
 
TABLE B1. MALE MICE: NEOPLASMS (CONTINUED) 

MATCHED 

CONTROL LOW DOSE HIGH DOSE 

HEMANGIOSAICCM A 1 (2%) 

•ESOPHAGUS (20) (41) (44) PAPILLOMA, NOS 1 (5%)

 

ISTOHACH (20) (50) (49) SUOAMOUS CILL CARCINOM A 1 (2%) ADENOMATOU S POLYP, NOS 1 (2S) 

UBINAHY SYSTEE 

HONE
 

ENDOCRINE SYSTIM
 

• ADRENAL (20) (48) (49)
 

CORTICAL ADENOM A 2 (10X) 1 (2%)
 

REPRODUCTIVE SYSTEM
 

NON£
 

NERVOUS SXSTEH
 

NONB
 

SPECIAL SENSE ORGAN S
 

NONE
 

BUSCUiOSKELETAL SYSTEM
 

NONE
 

EODY CAVITIES
 

NONe
 

ALL OTHER SYSTEMS
 

• NDBBEE OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUHBER OP ANIMALS NECROPSIED
 

50
 
TABLE B1. MALE MICE: NEOPLASMS (CONTINUED)
 

MATCHED
 

CONTROL
 

ANIMAL DISEOSITICN SUMMAR Y 

ANIflALS INITIALLY IN STUDY 

NATURAL DEATHS 

MORIBUND SACRIFICE 

SCHEDULED SACRIFICE 

ACCIDENTALLY KILLED 

TERMINAL SACRIFICE 

ANIMAL MISSING 

20 

16 

3 INCLUDES AU1CLYZED ANIMALS 

TUMOR SUMMARY
 

TOTAL ANIMALS WITH PRIMARY TUMORS* 12
 

TOTAL PRIMARY TUMORS 17
 

TOTAL ANIMALS WITH BENIGN TUMORS 4
 

TOTAL EENIGN TUMORS 5
 

TOTAL ANIMALS WITH MALIGNANT TUMORS 10
 

TOTAL MALIGNANT TUMORS 12
 

TOTAL ANIMALS WITH SECONDARY TUMORS*
 

TOTAL SECCKDARY TUMORS
 

TOTAL ANIMALS WITH TUMORS UNCEBTAIN­ EENIGN OR MALIGNANT
 

TOTAL UNCERTAIN TUMORS
 

TOTAL ANIMALS WITH TUMOBS UNCERTAIN­ PRIMARY OF KITASTATIC
 

TOTAL UNCERTAIN TUMORS
 

LOW DOSE HIGH DOSE 

50 

1 

12 

29 

38 

29 

32 

28 

32 

26 

29 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS
 

# SECONDARY TUKORS: METASTATIC TUMORS OR TUMORS INVASIVE INTO AN ADJACENT ORGAN
 

51
 
TABLE B2.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICE 

ADMINISTERED CCC IN THE DIET 

ANIMALS INITIAILY IN STUDY
 

ANIMALS NECROPSIED
 

ANIMALS EXftMINID HISTOPATHCLOGICALLY
 

INTEGUMENTARY SYSTEM
 

*SKIN
 

HEMANGIOSARCOMA
 

*SUBoUT TISSUE
 

RHABDOMYOSARCOMA
 

HEMANGIOMA
 

HEMANGIOSARCOMA
 

RESPIRATORY SYSTEK
 

#LUNG
 

ALVEOLAR/ERONCHIOLAR ADENOMA
 

ALVEOLAR/EBONCHIOLAH CARCINOMA
 

HEMATOPOIETIC SYSTEM
 

*MULTIPLE OEGANS
 

MALIGNANT IYMPHOHA, NOS
 

* BLOOD
 

Lx.UKEMIA,NCS
 

IBONE MARROW
 

HEMANGIOSARCOMA
 

ISPLiEN
 

HEMANGIOSARCOMA
 

MALIGNANT IYMPHOMA. NOS
 

ILUNG
 

MALIGNANT IYMPHOMA, NOS
 

MATCHED
 

CONTROL LOW DOSE
 

20 50
 

20 50
 

2) 50
 

(20) (50)
 

(2)) (50)
 

1 (5X)
 

1 (2%)
 

(20) (49)
 

1 (2X)
 

1 (5X) 2 (4X)
 

(20)	 (50)
 

7 (35%) 8 (16X)
 

(20)	 (50)
 

1 (5%)
 

(19)	 (50)
 

1 (2X)
 

(19)	 (48)
 

2 (4X)
 

(20) (49)
 

f	 NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

*	 NOMaER OF ANIMALS NECROPSIED
 

52
 

HIGHE IOSE
 

SO
 

50
 

50
 

(50)
 

1 (2X)
 

(50)
 

1 (2X)
 

(50)
 

1 [2*)
 

1 (2*)
 

(50)
 

1C (20%)
 

(50)
 

(4X)
 ^
 

(50)
 

(50)
 

1 (2X)


1	 (2X)
 

(50)
 

1 (2»)
 
TABLE B2. FEMALE MICE: NEOPLASMS (CONTINUED) 

MATCHED 

CONTROL LOW DOSE HIGH DOSE 

*LIV£R (19) (49) (50) 

MALIGNAN T IYMPHOMA , NOS 1 (2%) 

*MES£NTERY (20) (50) (50)
 

MALIGNANT LYMPHOMA, NOS 1 (2%)
 

IKIDHEY (20) (49) (50)
 

MALIGNANT LYMPHOMA, NOS 1 (2X)
 

tTHYMUS	 (16) (41) (46)
 

MALIGNANT IYMPHOHA, NOS	 1 (2%) 1 (2%)
 

CIRCULATORY SYSTEM
 

NONE
 

EIGESTIVE SYSTIM
 

«SALIVABY GLAND (19) (47) (50)
 

CYSTADENCKi, NOS 1 (255)
 

• LIVSR (19) (49) (50) 

HEPATOCELLULAR CARCINOM A 4 (21S) 7 (14X) 4 (855) 

ItUODENUH (18) (45) (50)
 

HEMANGIOMA 1 (2%)
 

DHINAHY SYSTIfl
 

NONE
 

INEOCKINE SYSTHM
 

•PITUITARY	 (16) (49) (48)
 

ADENOMA, NOS 2 (13X) 2 (4X)
 

*ADRENAL (19) (49) (50)
 

CORTICAL AEINCM A 1 (2«)
 

«THYBOID (19) (47) (5C)
 

FOLLICULAR-CELL ADENOMA 1_J5XI_. 

* NUMBER OF ANIMALS HITH TISSUI EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

53
 
TABLE B2. FEMALE MICE: NEOPLASMS (CONTINUED) 

REPRODUCTIVE SYSTEM
 

*MAMMARY GLANI
 

ADENOCARCINOMA, NOS
 

*UTEHUS
 

ADENOCARCINOMA, NOS
 

LEIOMYOMA
 

#OVAiiY

CYSTADENCEi, NOS

GRANULOSA-CEIL TUMOR __._.__ ____^_«._^.^«._ _.^_.— —._


NERVOUS SYSTEH
 

NONE
 

SPECIAL SENSE CRGANS
 

NONE
 

MUSCUiOSKEIETAI SYSTEM
 

NON£
 

EODY CAVITIES 

*MES£NTERY


LEIOMYOSARCOMA


ALL OTHER SYSTEMS
 

*BULTIPLE ORGANS


 ^^_..«._ _^_«._


MATCHED 

CONTROL LOW DOSE HIGH DOSE 

(20)	 (50) (50)
 

1 (2%)
 

(20)	 (U6) (50)
 

1 (2%)
 

1 (2X)
 

 (20) (47) (5C) 

1 (55S) 

2 (4«) _ v_ ~_ *.___ _ ..•_ ^_— _„— —.-•,— ——— ____«.I_.V.B«.«I. —— — «•»•••
 

 (20)	 (50) (50)
 

1 (2X)
 

 (20)	 (50) (50)
 

# NUMi3ER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 * NUMBER OF ASIMALS NECROPSIED
 

54
 
TABLE B2. FEMALE MICE: NEOPLASMS (CONTINUED) 

MATCHED
 

CONTROL LOW DOSE HIGH DOSE
 

ANIMAL DISEOSI1ICN SUMMAEY
 

ANIHALS INITIALLY IN STUDY 5J 50
 

NATURAL DEATHS)
 

MORIBUND SACRIFICE
 

SCHEDULED SACRIFICE
 

ACCIDENTALLY KILLED
 

TERMINAL SACRIFICE 16
 

ANIMAL MISSING
 

INCLUDES AUTCLYZED ANIMALS
 

luMOR SUBMABY
 

TOTAL ANIMALS WITH PRIMARY TUMORS* 14 25 26
 

TOTAL PRIMARY TUMORS 18 3J 34
 

TOTAL ANIMALS WITH BENIGN TUKORS 3 5 4
 

TuTAL BENIG N TUMORS 5 5 4
 

TOTAL ANIMAL S HITH MALIGNANT TUMORS 1 1 19 24
 

TOTAL MALIGNANT TUMORS 13 23 30
 

TOTAL ANIMAL S WIT H SECONDARY TUMOBS# 

TOTAL SECONDAR Y TUMOflS 

TOTAL ANIMALS WITH TUHOBS UNCERTAIN­ BENIGN OB MAIIGNANT 2
 

TOTAL UNCERTAIN TUMORS 2
 

TOTaL ANIMAI S HITH TUMORS UNCERTAIN­ EBIBARY OR KITASTATIC
 

TOTAL UNCERTAIN TUMORS
 

* PRIMARY TUMCRS: ALL TUMORS EXCEPT SECONDARY TUMORS
 

# SECONDARY TDMOES: METASTATIC TUMORS OR TUMORS INVASIVE INTO AN ADJACENT ORGAN
 

55
 
56

 
APPENDIX C
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN RATS
 

ADMINISTERED CCC IN THE DIET
 

57
 
58

 
TABLE C1.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS 

ADMINISTERED CCC IN THE DIET 

MATCHED 

CONTROL LOW DOSE 

ANIMALS INITIAILY 111 STUDY 

ANIMALS NECROPSIED 

ANIMAi.3 EXAMINED HISTOPATHOLOGICALLY 

20 

20 

20 

50 

50 

49 

INTEGUMENTARY SYSTEM
 

*SKIN (20) (50)
 

EPIDERMAL INCLUSION CYST 1 (2%)
 

*SUBCUT TJSSOI (20) (50)
 

H&MATOMA, KOS
 

HEMORHHAGIC CYST
 

STEATITIS 1 (2%)
 

FIBROSIS 1 (2%)
 

RESPIRATORY SYSTEM
 

#TRACHEA (20) (46)
 

INFLAMMATICN, CHBONIC
 

#LUNG/BRONCHUS (20) (49)
 

LYMPHOCYTIC INFLAMMATORY INFILTR 11 (55«) 13 (21%)
 

#LUNG (20) (49)
 

LYMPHOCYTIC INFLAMMATOHY INFILTR 24 (49X)
 

INFLAMMATICN, INTERSTITIAL 1 (2»)
 

I&FLAMMATICN, FOCAL GRANULOMATOU 1 (2%)
 

HEMATOPOIETIC SYSTEM
 

*BLOOD (20) (50)
 

MONOCYTOSIE 2 (4*)
 

LjiUKOPENIA, NOS 1 (5X) 1 (2«)
 

HYPERPLASIA, NEOTBOPHILIC 2 (4«)
 

#BON£ MAREOH (20) (49)
 

HYPJHPLASIAj GRANOLOCYTIC 1_J2«I_
 

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

HIGH DOSE
 

50
 

50
 

50
 

(50)
 

1 (2X)
 

(50)
 

1 (2X)
 

1 (2%)
 

(50)
 

1 (2%)
 

(50)
 

(50)
 

39	 (76%)
 

1 (2%)
 

(50)
 

1 (2%)
 

(50)
 

59
 
TABLE C1. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

MATCHED 

CONTROL LOW DOSE HIGH DOSE 

•SPLiEN	 (23) (49) (50)
 

CONGESTION, NOS 2 (1035) 2 (4%)
 

INFARCT, NCS 1 (2%)
 

HhBOSIDEFCSIS 3 (635) 3 (635)
 

HYPERPLASIA, BETICDLUB CELL 1 (235)
 

HYPERPLASIA, LYBPHOID 1 (5X)
 

HKBATOPOIISIS 10 (5031) 29 (59X) 20 (40*)
 

MYELOPOIESIS 1 (235)
 

fLYBPH NODE (20) (49) (50)
 

INFLABBATICN, NECROTIZING 1 (235)
 

HYPERPLASIA, DIFFUSE 1 (5*)
 

HYPERPLASIA, LYMPHOID 1 (2%)
 

•MANDIBULAR	 L. NODE (20) (49) (50)
 

CYST, NOS 1 [2%) 2 (455)
 

CONGESTION, NOS 1 (2*)
 

EDEMA, NOS 1 (2X)
 

HEMORRHAGE 1 (255)
 

FIBROSIS 1 (2%)
 

P£RIARTEBI1IS 1 (2X)
 

HYPERPLASIA, NOS 1 (2*)
 

HYPERPLASIA, DIFFUSE 2 (1036)
 

PLASMACYTCSIS 6 (30*) 12 (24X)
 

EBYTHROPHAGOCYTOSIS 1 (2X)
 

HYPERPLASIA, HETICULUH CELL 1 (2X)
 

HYPERPLASIA, LYMPHOID 1 (235) 1 (2X)
 

•MEDIASTINAL L.NCDE (20) (49) (50)
 

P1GBENTATICN, NOS 1 (5*)
 

•MESENTERIC I. NODE (20) (49) (50) .
 

CYST, NOS 2 (4X)
 

EDEMA, NOS 1 (536) 1 (231) 1 (2X)
 

PIGMENTA1ICN, NOS 1 (5*)
 

HYPERPLASIA, RETICULUM CELL 1 (2*)
 

•THYMUS	 (7) (36) (43)
 

CONGESTIOK, NOS 1 (335)
 

CIRCULATORY SYSTEM
 

IHEART (20) (49) (50)
 

IBHQMBOS. MURAL !_J2SJ 1 12%)
 

* NUMBER OF ANIBALS WITH TISSUE EXAMINED MICROSCOPICALLY
 * NUBBER OF ANIBALS NECROPSIED
 

60 
TABLE C1. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

" 

INFLAMMATION, SUPPURATIVE 

FIBROSIS 

FIBROSIS, FOCAL 

iHEAST/ATRIOfl 

THROMBOSIS, NOS 

fMYOCARDIO B 

INFLAMMATION , StJPPORATIVE 

INFLAMMATION, CHRONIC 

*PULi1ONARY AETER Y 

HYPERTROPHY, NOS 

DIGESTIVE SYSIBfl 

ISALIVARY GLAN D 

INFLAMMATION, CHRONIC 

#LIVER 

CONGESTION, NOS 

LYMPHOCYTIC INFLAMMATOR Y INFILTR 

INFLAMMATICN , NECROTIZING 

CHOLANGICFIBROSIS 

CIRRHOSIS, NOS 

CIRRHOSIS, PORTAL 

DiGENERATICN, HYDROPIC 

NECROSIS, NOS 

NECROSIS, POCAL 

AflYLOIDOSIS 

METAMORPHOSIS FATTY 

LIPOIDOSIS 

BASOPHILIC CYTO CHANG E 

CLEAR-CELL CHANGE 

MEGALOCYTCSIS 

LnUKEMOID FEACTION 

JtLIVER/CENTRIIOBOLAR 

DEGENERATION , HYDROPIC 

ILIVER/HEPATCCYTES 

METAMORPHOSI S FATTY 

#BILfi DOCT 

INFLABMASICN^CHEONIC FCCA.1 

MATCHED 

CONTROL LOW DOSE HIGH DOSE 

1 (5X) 27 (55X) 40 (80*) 

11 (55«) 16 (33X) 

(20) (49) (50) 

1 (5X) 

(20) (49) (50) 

1 (2X) 

1 (2X) 

(20) (50) (50) 

8 (16X) 

(20) (49) (50) 

1 (2*) 

(20) (49) (50 ) 

1 (2X) 

1 (5X) 

11 (55 X) 

2 (10X) 

1 (2X) 

1 (2X) 

41 (84*) 37 (74*) 

3 (6X) 

1 (2X) 

1

5

 (2*) 

 (10*) 

1 (2X) 

1 <5X) 

2 (10X) 9 (18*) 9 (18X) 

1 (5X) 

1 (5X) 

13 (27*) 

2 (4S) 

5 (10X) 

2 (4*) 

1 (2X) 

3 (6X) 

1 (2X) 

2 (4*) 

1 (2X) 

(20) (49) (50) 

1 (2X) 

120) (49) (50) 

1 (5X) 

(20) 

1 (5X) 

(49) (50) 

# NUMriEH OF ANIMALS HITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

61
 
TABLE C1. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

MATCHED
 

CONTROL LOW DOSE
 

HYPERPLASIA, NOS 1 (5%) 

SPANCREAS 

CONGESTION, NCS 

FIBROSIS 

FIBROSIS, FOCAL 

PtRIARTEFJIJS 

ATROPHY, NCS 

(19) 

1

1

1

 (5«) 

 (5X) 

 (5*) 

(47) 

U

6

 (9S) 

 (13*) 

IPANCREATIC ACINUS 

FIBROSIS, FOCAL 

HYPERPLASIA, NOS 

(19) (47) 

3 (6J5) 

tSTOMACH 

ULCER, NOS 

ULCER, FCCAL 

INFLAMMATION, SUPPUHATIVE 

INFLAMMATION, NECROTIZING 

(20) 

1 (5*) 

(49) 

1

1

 (2%) 

(2%) 

INFLAMMATION, ACUTE SOPPDRATIVE 1 (2%)
 

GRANULOftA, FOREIGN BOEY 1 (2%)
 

ULCER, PERFORATED
 

FIBROSIS, FOCAL
 

tGASlRIC MUCOSA (20) (49)
 

DILATATION, NOS
 

#ILEUM (19) (49)
 

FIBROSIS, BOCAL
 

URINABY SYSTEM
 

#KIDNEY	 (20) (49)
 

CAST, NOS 13 (65%) 42 (86*)
 

PYELONEPHRITIS SUPPURATIVE
 

ABSCESS, f«CS 1 (535) 1 (2»)
 

INFLAMMATION, CHRONIC 43 (88X)
 

INFLAMMATION, CHRONIC FOCAL 9 (U5%)
 

INFLAMMATION, CHRONIC DIFFUSE 1 (5%)
 

INFLAMMATION, PYOGR ANULCM ATQUS 1 (2X)
 

PERIARIERITIS
 

DEGENERATION, HYALINE 1 (5%)
 

HYPERPLASIA, 1UBULAR CELL
 

#KIDNEY/TUBULE (20) (49)
 

METAMORPHOSIS FATTY 1_J2Xj_
 

« NUHBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

HIGH DOSE
 

(49)
 

1 (2X)
 

3 (6*)
 

3 (6X)
 

1 (2*)
 

3 (6*)
 

(49)
 

1 (2X)
 

(50)
 

1 (2*)
 

1 (2%)
 

1 (2%)
 

1 (2%)
 

(5C)
 

1 (2%)
 

(49)
 ' 1 [2%)
 

(50)
 

38	 (76%)
 

2 (4%)
 

40 (80%)
 

1 (2%)
 

1 (2%)
 

(50)
 

62
 
TABLE C1. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

PIGMENTATION, SOS
 

H£MOSIDEBCSIS
 

SURINARY ELAEIER
 

CALCULUS, NOS
 

INFLAMMATION, NECROTIZING
 

INFLAMMATION, CHRONIC FCCAL
 

ENDOCRINE SYSTEM
 

#PITUITARY
 

CYST, NOS
 

ANGIECTASIS
 

# ADRENAL
 

LYMPHOCYTIC INFLAMMATORY


NiCROSIS, FOCAL
 

Hi-TAMORPHCSIS FATTY
 

ANGIECTASIS
 

(tADRENAi CORIIX
 

METAMORPHOSIS FATTY
 

((ADRENAL MEEUILA
 

HYPERPLASIA, NODULAR
 

ITHYROID
 

HYPERPLASIA, C-CELL
 

IPANCREATIC ISLETS
 

HYPERPLASIA, NOS
 

REPRODUCTIVE SYSTEM
 

+MAMHARY GLANE
 

DILATATICN/CUCTS
 

CYST, NOS
 

*PREi?UTIAL GLAND
 

CYST, NOS
 

IPRObTATE
 

INFLAMMATION.,. SOPPURATIVE
 

 INFILTR
 

MATCHED
 

CONTROL
 

3 (15*)
 

(19)
 

(20)
 

2 (10%)
 

2 (10%)
 

[20)
 

1 (5»)
 

1 (5%)
 

4 (20?!)
 

(20)
 

(20)
 

1 (5X)
 

(20)
 

(19)
 

1 (5*)
 

(2J)
 

(20)
 

(9)
 

1 -Q1«I
 

LOW DOSE
 

1 (235)
 

(49)
 

1 (2%)
 

(49)
 

(49)
 

2 (4»)
 

18 (37%)
 

(49)
 

2 (4%)
 

(49)
 

(48)
 

4 (8»)
 

(47)
 

(50)
 

8 (16%)
 

(50)
 

1 (2*)
 

(37)
 

2_J5%1_
 

HIGH DOSE
 

(47)
 

1 (2%)
 

1 (2%)
 

(47)
 

4 (9%)
 

1 (2%)
 

(50)
 

1 (2?)
 

28 (56%)
 

(50)
 

1 (2%)
 

(50)
 

(50)
 

1 (2%)
 

(49)
 

1 (2%)
 

(50)
 

12	 (24X)
 

1 (2%)
 

(5C)
 

(47)
 

_2_J4»1
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

63
 
TABLE C1. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

"
 MATCHED
 

CONTROL LOW DOSE HIGH DOSE
 

INFLAMMATION, ACUTE 1 (2X)
 

IMFLABBATICN, ACDTE SOPPDRATIVE 1 (3X)
 

FIBROSIS 1 (2X)
 

HYPEBPLASIA, NODULAR 1 (11*)
 

HYPERPLASIA, NOS 2 (4*)
 

*SEBINAL VESICLE (20) (50) (50)
 

INFLAMMATION, SUPPURATIVE 2 (4X) 1 (2X)
 

INFLABBATICN, ACDTE SOPPUBATIVE 1 (2X)
 

FIBROSIS, IOCAL 1 (2X)
 

fTESIIS (20) (49) (OS)
 

GfiANULOHA, SPERMATIC 1 (5X) 1 (2X)
 

CYTOMEGAIY 1 (2X) 1 (2X)
 

ATROPHY, DCS 1 (5X) 10 (20%) 13 (21%)
 

ASPEHMATCGINESIS 1 (2X)
 

tTESTIS/TOBULI (20) (49) (49)
 

MINERALIZATION 1 (2X)
 

DEGENERATION, HYALINE 1 (5X)
 

*EPIDIDYBIS (20) (50) (50)
 

INFLABHATICN, CHRONIC 1 (2%)
 

FIBROSIS, EIFFOSE 1 (2X)
 

N£CROSIS, PAT 1 (2X)
 

NIRVOUS SYSTEH
 

#CER£BRDB (20) (49) (49)
 

ABSCESS, NCS 1 (2X)
 

*BRAIN (20) (49) (49)
 

HYDROCEPHALUS, NOS 1 (2X)
 

HiDROCEPHAIUS, INTERNAL 1 (2X)
 

HEMORRHAGE 1 (25!)
 

NECROSIS, NOS 2 (4S)
 

#EHA1N/THALABCS (20) (49) (49)
 

H£BORRHAGE 1 (2%)
 

*SPINAL CORD (20) (50) (5C)
 

HiBOREHAGE 1 <2X)
 

NECROSIS,, NOS 1 (2X)
 

* NUMBER OF AKIBALS WITH TISSUE EXABINED MICROSCOPICALLY
 * NUMBER OF AN3BALS NECROPSIED
 

64
 
TABLE C1. MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

SPECIAL SENSE ORGAN S 

*EYE

CATARACT


MUSCULOSKELETAI SYSTEM
 

*SKELETAL MUSCLE


INFLAMMATION, FOCAL


BODY CAVITIES
 

*ABDOMINAL CAVITY


SIEATITIS


*PERITONEUM


INFLAMMATION, NOS


INFLAMMATION, SUPPUBATIVE


INFLAMMATION, GRANULOMATOUS


*MESjiNTERY


SIEATITIS


PiRIABTEBITIS


NriCBOSIS, JAT


ALL OTHER SYSTEMS
 

^MULTIPLE ORGANS


LEUKEMOIE fEACTION


SPECIAL MORPHC10GY SUMMARY
 

AUTO/NECBOESY/NO HISTO


MATCHED 

CONTROL LOW DOSE HIGH DOSE 

 (20)

1 (5%) 

 (50) (5C) 

 (20)

1 (5X) 

 (50) (50) 

 (20) (50)

1 (2%) 

 (50) 

 (20) (50) (50) 

1 (23t) 

1 (5X) 

1 (2») 

 (20) (50) (50) 

1 (2%) 

6 (12X) 

1 (2%) 

 (20) (50) (50) 

1 (2%) 

1 

« NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

65
 
TABLE C2.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS 

ADMINISTERED CCC IN THE DIET 

MATCHED 

CONTROL LOW DOSE 

ANIMALS INITIAILY IN STUDY

ANIMALS NECROPSIED

ANIMALS EXAMINED HISTOPATHOIOGICALLY

 20 

 20 

 20 

50 

50 

50 

INTEGUMENTA8Y SYSTEM
 

*SKIN (20) (50)
 

EPIDERMAL INCLUSION CYST
 

INFLAMMATION, CHRONIC SOPPURATIV 1 (2%)
 

*SUBCUT TISSUI (20) (50)
 

NuCROSIS, FAT
 

HYPERPLASIA, NOS
 

RESPIRATORY SYSTEM
 

ITRACHEA (20) (48)
 

INFLAMMATION, NOS
 

tLONG/BRONCHUS (20) (50)
 

LYMPHOCYTIC INFLAMMATORY INFILTR 18 (9055)
 

#LUNG (20) (5))
 

HEMORRHAGE
 

LYMPHOCYTIC INFLAMMATORY INFILTR 42 (84%)
 

INFLAMMATION, INTERSTITIAL 2 (4»)
 

INFLAMMATION, FOCAL GRANULCMATOU 1 (2X)
 

CHOLESTEEC1 DEPOSIT 1 (2X)
 

HEMATOPOIETIC SYSTEM
 

*BLOOD (20) (50)
 

CYTOPLASMIC VACUOLIZATION 1 (5%)
 

CYTOMEGALY 1 (5%)
 

LiUKOCYTOSIS, NOS
 

LEUKOCYTOSIS, NEOTROPHILIC 2 (10%)
 

LYMPHOCYTOSIS 1 (2X)
 

t NUMBER OF ANIMALS HITH TISSUE EXAMINED MICROSCOPICALLY
 * NUMBER OF ANIMALS NECROPSIED
 

HIGH DOSE
 

50
 

50
 

50
 

(50)
 

1 (2X)
 

(5C)
 

1 (2%)
 

1 (2X)
 

(49)
 

4 (3%)
 

(49)
 

(49)
 

1 (2X)
 

42 (86%)
 

(50)
 

1 (2X)
 

66
 
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

MATCHED 

CONTROL LOW DOSE HI8HDOK 

LEUKOPENIA , NC S 1 (2X) 

EHYTHHOBLASTOSI S 1 (2X) 

HYPERPLASIA , NEDTROPHILIC 1 (5%) 2 (4X) 2 <4JI) 

ISPLtEN (20) (50) (SO) 

CONGESTION , NCS 1 (2X) 

INFARCT, NCS 1 (5X) 

HuMOSIDEBCEI S 5 (255S) 39 (78X) 31 (681) 

HYPERPLASIA, RETICULUM CELL 1 (2X) 3 (6X) 

HtMATOFOIESIS 14 (70X) 38 (76X) 36 (72X) 

*LYM?H NODE (20) (49) (49) 

CONGESTION, NCS 1 (2X)

 PJ.GMENTATICN , NOS 1 (2X) 

PLASMACY1CSIS 1 (2X) 

HYPERPLASIA , LYMPHOI D 1 (2X) 

MANDIBULAR L. NODE (20) (49) (49)

CYST, NOS 2 (41) 

CONGESTION, NOS 1 (5X) 1 (2X) 1 (2S) 

EDEMA, NOS 1 (2X) 

PIGMENTATION, NOS 1 (2%) 

HiMOSIDEBCSIS 1 (2X) 

HYPERPLASIA, NOS 1 (2X) 

PLASMACYTCSIS 11 (55*) 14 (29X) 20 (41*) 

HYPERPLASIA, LYMPHOID 2 (4X) 

MEDIASTINAL I.NODE (20) (49) 

CONGESTION, NOS 1 (5X) 1 (2X) 

PIGMENTATION, NOS 1 (58) 20 (41X) 5 (10*) 

MESENTERIC L. NODE (20) (49) 

CONGESTION, NOS 2 (4X)

 

EDEMA, NOS 1 (2X)

 

PIGMENTATION, NOS 4 (8X)

 

HYPERPLASIA, LYMPHOID 1 (2X) 

*THYMUS (12) (38) (38) 

HYPERPLASIA , NOS 

CIRCULATORY SYSTEM 

((HEART (20) (50) (19)

FIBROSIS 41 (82X1 37 J76IA 

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

67
 
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

" 

MATCHED 

CONTROL LOW DOSE HIGH DOSE 

FIBROSIS, FOCAL 

BhTAMORPHOSIS FATTY 

HjiflOSIDEBCSIS 

13

1

 (65X) 

 (5«) 

1 (2X) 

*HEAfiT/ATRIUB 

THROHBOSIS, NOS 

(20) (50) 

1 {2X) 

(49) 

*PUiflOHARY ARTERY 

HYPERTROPHY, NOS 

(20) (50) (50) 

1 (2X) 

riGESTIVE SYSTEH
 

ISALIVARY GLAND (19) (50) (49)
 

CYSTIC DDC1S 1 (2X)
 

INFLAHHATICN, NOS 2 (4X)
 

FIBROSIS, FOCAL 1 (2X)
 

#LI¥£R (20) (50) (SO)
 

CONGESTICK, NOS 1 (2X)
 

INFLAHBATICN, SUPPURATIVE 1 (5X)
 

G8ANULCBA, NOS 2 (W)
 

INFLABBATICN, FOCAL GRANULOBATOU 1 (2X)
 

FIBROSIS, FOCAL 1 (2%)
 

NODULE 1 (5X)
 

CHOLANGIOFIBROSIS 6 (30%) 25 (SOX) 24 (4855)
 

BETABORPHCSIS FATTY 1 (5S) 3 (6X) 6 (12X)
 

BASOPHILIC CY10 CHANGE 14 (10%) 40 (80X) 38 (7655)
 

CLEAR-CELL CHANGE 3 (6X)
 

H£GALOCYTCSIS 6 (12X) 3 (6X)
 

HYPERPLASIA, NODULAR 1 <5S)
 

#LIVtR/CENTRIIOBULAS (20) (50) (50)
 

BjETAHORPHCSIS FATTY 1 (5»)
 

#EIL£ DOCT (20) (50) (50)
 

INFLABBATICN, CHRONIC 1 (5X)
 

HYPERPLASIA, NOS 1 (5X)
 

*PANCREAS (19) (50) (50)
 

FIBROSIS 6 (12X) 3 (6X)
 

FIBROSIS, FOCAL 2 (11S) 1 (2X)
 

FIBROSIS, EIFFUSE 1 (2X)
 

PEBIARTERI1IS 1 (2X)
 

-AIRQPHXj NCS 2 (11JI_ 4 (8X1 2_J4»1
 

f NUHBER OF ANIBALS WITH TISSUE EXABINED BICROSCOPICAILY
 * NUMBER OF ABIBALS NECROPSIED
 

68
 
- -

TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

#STOMACH
 

CYST, NOS
 

ULCER, NOS
 

INFLAMMATION, ACUTE SUPPURATIVE
 

•SHALL INTESTINE
 

INFLAMMATION, ACUTE/CHBCNIC
 

#COLON
 

NEMATODIASIS
 

UBINABY SYSTEM
 

» KIDNEY
 

CAST, NOS
 

HYDBONEPHBOSIS
 

PYELONEPHEITIS, NOS
 

INFLAMMATION, CHBONIC
 

INFLAMMATION, CHBONIC FOCAL
 

PIGMENTATION, NOS
 

HiMOSIDEBGSIS
 

#KIDNEY/TUBUIE
 

PIGMENTATION, NOS
 

•KIDNEY/PELVIS
 

CALCULUS, BOS
 

iUBINABY BLAECEB
 

HEMORRHAGE
 

ENDOCBINE SYSTEM
 

fPITOITABY
 

CYST, NOS
 

HEMORRHAGE
 

HJiMORBHAGIC CYST
 

ANGIECTASIS
 

1 ADRENAL
 

FIBBOSIS
 

NECROSIS^ CORTICAL
 

MATCHED
 

CONTROL
 

(19)
 

1 (5X)
 

(18)
 

(18)
 

(20)
 

10	 (SOX)
 

1 (5X)
 

2 (10)1)
 

12 (60%)
 

(20)
 

(20)
 

(19)
 

(20)
 

2 (1QX)
 

1 (5X)
 

2 (10X)
 

(20)
 

n -r
 

LOW DOSE
 

(50)
 

1 (2X)
 

(49)
 

(50)
 

(50)
 

22 (4KX)
 

26 (52%)
 

1 (2X)
 

(50)
 

1 (2X)
 

(50)
 

1 (2X)
 

(50)
 

1 (2X)
 

(49)
 

3 (6X)
 

1 (2X)
 

(50)
 

1 (2X)
 

HIGH DOSE
 

(50)
 

1


(50)
 

1


(50)
 

1


(50)
 

18


1


28


1


(50)
 

(50)
 

(50)
 

(49)
 

16


1


(50)
 

1


 (2X)
 

 (2X)
 

 (2%)
 

 (36X)
 

 (2X)
 

 (56X)
 

 (2X)
 

 (33X)
 

 (2X)
 

 <2X)
 

t NUMBEB OF ANIMALS HITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBEB OF ANIMALS NECROPSIED
 

69
 
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

MATCHED
 

CONTROL LOW DOSE HIGH DOSE
 

HiTABOEPHOSIS FATTY 3 (6%) 1 (2%)
 

HYPERPLASIA, NODULAR 2 (4%)
 

ANGIECTASIS 7 (35%) 28 (56%) 26 (52%)
 

#ADRENAL CORTEX (20) (50) (53)
 

MINERALIZATION 1 (2%)
 

CYST, NOS 1 (2%)
 

CONGESTION, NOS 1 (2%)
 

MiTAMORPHOSIS FATTY 1 (2%) 1 (2%)
 

HYPERPLASIA, NODULAR 2 (4%)
 

HYPERPLASIA, NOS 1 (2X) 2 (4%)
 

fADRENAL MZEUILA (20) (50) (50)
 

ANGIECTASIS 1 (2%)
 

#THY.hOID (20) (49) (49)
 

INFLAMMATION, NOS 1 (2%)
 

HYPERPLASIA, C-CELL 2 (10%) 8 (16%) 1C (20%)
 

REPRODUCTIVE SYSTEM
 

*MAMi4ARY GLANI (20) (50) (50)
 

DILATATICN/DUCTS 8 (40%) 31 (62%) 2C (40%)
 

CYST, NOS 1 (5%)
 

ANGIECTASIS 1 (2%)
 

*MAM3ARY DUCT (20) (50) (5C)
 

RETENTION OF CONTENT 1 (2%)
 

#UTEHUS (20) (50) (5C)
 

PYOMETRA 1 (2%)
 

NECROSIS, FAT 1 (2%)
 

POLYP, INFLAMMATORY 2 (10%) 4 (8%) 6 (12%)
 

tCERVIX UTERI (20) (50) (50)
 

FIBROSIS 1 (2*)
 

#UTEHUS/ENDC«ITRIUM (20) (50) (5C)
 

CYST, NOS 1 (2%) 2 (4%)
 

MULTILOCUIAR CYST 1 (5%)
 

#OVAtiY (20) (50) (50)
 

CYST, NOS 1 (5%) 2 (4%) 3 (6%)
 

CORPUS LU1EUH CYST 1 (2%)
 

t NUHriER OF AKIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBE R OF ANIMAL S NECROPSIE D 

70 
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

CORPUS LC1EUM


#OVARY/FOLLICLE


A1RESIA


NERVOUS SYSTEM
 

tCEREBRAL VENTRICLE


HEMORRHAGE


#BRAIN


HYDROCEPHALUS, NOS


HYDROCEPHALUS, INTERNAL


HEMORRHAGE


SPECIAL SENSE ORGANS
 

NONE
 

MUSCOLOSKELETAL SYSTEM
 

NONE
 

BODY CAVITIES
 

*MEDIASTINUM


PERIARTERITIS


*MESENTERY


FIBROSIS


NECROSIS, IAT


ALL OTHER SYSTEHS
 

ADIPOSE TISSUE
 

NECROSIS, FAT


SPECIAL MORPHOLOGY SUMMARY
 

NO LESION BEPORTED


MATCHED
 

CONTROL


 17 (55%)


 (20)


 (20)


 (20)


 (20)


 (20)


 1 (5%)
 

1 (555)
 

1
 

 LOW DOSE HIGH DOSE
 

 41 (82%) Hj (86X)
 

 (50)	 (50)
 

2 (H%)
 

 (50)	 (50)
 

1 (2X)
 

 (50)	 (50)
 

1 (2%)
 

1 (2%)
 

1 (2%)
 

 (50)	 (50)
 

1 [2%)
 

 (50) (50)
 

1
 

t	 NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

*	 NUMBER OF ANIMALS NECROPSIED
 

71
 
72

 
APPENDIX D
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MICE
 

ADMINISTERED CCC IN THE DIET
 

73
 
74

 
TABLE 01.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE MICE 

ADMINISTERED CCC IN THE DIET 

MATCHED 

CONTROL. LOW DOSE HIGHD OSE 

ANIMALS INITIA1LY IN STUDY

ANIMALS NECROPSIED

ANIMALS EXAMINED HISTOP AT HOLOGIC ALLY

 20 

 20 

 20 

50 

50 

50 

50 

49 

49 

INTEGUMENTARY SYSTEM
 

*SKIN (20) (50) (49)
 

ABSCESS, NCS 2 (4«)
 

RESPIRATORY SYSTEM
 

tlUNii (20) (50) (4'5)
 

LYMPHOCYTIC INFLAMMATORY INFILTR 2 (10%) 1 (2%)
 

INFLAMMATICN, INTERSTITIAL 1 (2%) 2 (455)
 

HEMATOPOIETIC SYSTEM
 

*BLOOD (20) (50) (49)
 

Li-UKOPENIA, NCS 1 (5%)
 

#BON£ MARRO W (20) (50) (49)
 

HYPERPLASIA, RETICULUM CELL £ (4%)
 

itSPLjiEN (20) (50) (48)
 

HYPERPLASIA, RETICULUM CELL 1 (2%) C (1 J«)
 

HYPERPLASIA, LYMPHOID 1 (5%) 1 (2X)
 

#MESENTERIC L. NODE (20) (50) (49)
 

CONGESTICh, NOS 9 (45%) 15 (3J%) 17 (35%)
 

INFLAMMATICN, CHRONIC 1 (2%)
 

FIBROSIS 2 (4%)
 

PLASMACYTCSIS 1 (2%)
 

MEGAKARYCCYTOSIS 1 (2%)
 

HYPERPLASIA, RETICULUM CELL 6 (12%) 8 (16%)
 

HYPERPLASIA, LYMPHOID 2 (4%) 1 (2%)
 

CIRCULATORY SYSTE M 

*MYOi_ARDIU M (19) (50) (49) 

INFLAMMATICN , CHRONIC fCCA L 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

75 
TABLE 01. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

MATCHED
 

CONTROL LOW DOSE HIGH DOSE
 

DIGESTIVE SYSTEM
 

#SALIVARY GLAND (20) (50) (49)
 

FIBROSIS, FOCAL 1 (555)
 

tLIVER (20) (50) (49)
 

INFLAMMATION, CHRONIC FOCAL 2 (4%)
 

NECROSIS, FOCAL 1 (5%)
 

INFARCT, NCS 1 (5X)
 

MiTAMOEPHCSIS FATTY 4 (20%) 4 (8X) 6 (1255)
 

ANGIECTASIS 1 (2«)
 

*EANCREAS (20) (48) (4£)
 

MLTAMORPKOSIS FATTY 2 (455)
 

ATROPHY, ICCAI 1 (2%)
 

((PANCREATIC ACINUS (20) (48) (46)
 

ATROPHY, NCS 2 (455)
 

ISTOHACH (20) (50) (49)
 

INFLAMMATION, ACUTE FOCAL 1 (2%)
 

IPEYiRS PATCfc (20) (5'J) (48)
 

HYPERPLASIA, RETICULUM CELL 2 (455) 1 (255)
 

HYPERPLASIA, LYMPHOID 1 (255)
 

tDOCOENUM (20) (50) (48)
 

DIVERTICDICSIS 1 (255)
 

UEINAHY SYSTEK
 

f KIDNEY (20) (50) (49)
 

HYDRONEPHROSIS 1 (235)
 

LYMPHOCYTIC INFLAMMATORY INFILTR 2 (10X) 2 (435) 7 (145S)
 

INFLAMMATION, CHRONIC 2 (4X) 3 (635)
 

IsiFLAMMATICN, CHRONIC FOCAL 1 (555) 2 (455)
 

ENDOCRINE SYSTEM
 

IPITJITARY (18) (48) (49)
 

CYST, NOS 1 (6%) 1 (2%)
 

((THYROID (19) (47) (47)
 

FIBROSIS., FOCAL 1 (235)
 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NOMBER OF ANIMALS NECROPSIED
 

76
 
TABLE D1. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

AXROPHY , NCS

HYPERPLASIA, FOLLICULAR-CELL

REPRODUCTIVE SYSTEM 

*MAMMARY GLSK E

INFLAMMATICN , NOS

*SEMINA L VESICLE

D-LLATATICti, NCS

ITESTIS

ATROPHY, FCCAL

NEBVOU S SYSTI M 

NONE 

SPECIAL SENS I CKGAN S 

NONJj 

MUSCULOSKELETAL SYSTEM
 

NONE
 

EOEY CAVITIES
 

*MESjiNTERY

NhCfiOSIS, FAT

A LL OTHER SYSTIM S 

NONE 

SPECIAL MORPHCIOGY SUMMARY
 

MO LESION EEEOBTED


MATCHED 

CONTROL LOW DOSE HIGH DOSE 

1 (2% ) 

1 (5%) 

 (23) (50) (49) 

1 (2%) 

 (20) (50) (49) 

2 (4%) 1 (2%) 

 (20) (50) (49 ) 

1 (5%) 

 (20) (50) (49) 

2 (10% ) 1 (2%) 

3 6	 6
 

#	 NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

*	 NUMBER OF ANIMA1S NECROPSIED
 

77
 
TABLE D1. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

MATCHED 

CONTROL LOW DOSE HIGH DOSE 

AU1C/NECEOESY/HIS1 0 PERF

AUTOLYSIS/NO NECJtOPSY

 1 

1 

* NUMBE R

* NDHBEE

 OF

 OF

 ANIMAL S HITH TISSUE

 ANIMAL S NECROPSIED 

 EXAMINE D MICROSCOPICALL Y 

78 
TABLE D2.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE 

ADMINISTERED CCC IN THE DIET 

MATCHED
 

CONTROL LOW DOSE HIGH DOSE
 

ANIMALS INITIALLY IN STUDY 20 50 50
 

ANIMALS NECROPSIED 20 50 50
 

ANIMALS EXAMINED HISTOPATHO LOGICALLY 20 50 50
 

INTEGUMENTARY SYSTEM
 

*SUBi-UT TISSUE (20) (50) (50)
 

HiMORRHAGIC CYST 1 (2%)
 

RESPIRATORY SYSTEf
 

#LUNG (20) (49) (50)
 

LYMPHOCYTIC INFLAMMATORY INFILTR 4 (20%) 3 (6/5) 5 (10%)
 

INFLAMMATION, INTERSTITIAL 1 (2%)
 

HLMOSIDEECSIS 1 (2%)
 

HEMATOPOIETIC SYSTEM
 

*BLOOD (2)) (50) (50)
 

HYPERPLASIA, NEUTROPHILIC 1 (2%) 1 (2%)
 

ISPLjiEN (19) (48) (50)
 

HYPERPLASIA, HEMATOPOIETIC 1 (2*) 2 (4%)
 

HYPERPLASIA, RETICULUM CELL 2 (4«)
 

HYPERPLASIA, LYMPHOID 6 (32%) 7 (15X) 1 (2%)
 

#LYMPH NODE (19) (47) (50)
 

HjiMORRHAGIC CYST 1 (2%)
 

PLASMACYTCSIS 1 (2%)
 

HYPERPLASIA, LYMPHOID 1 (2%)
 

IMANDIBULAR L. NODE (19) (47) (5C)
 

CONGESTION, NOS 1 (5%)
 

HYPERPLASIA, RETICULUM CELL 1 (2%)
 

HYPERPLASIA, LYMPHOID 1 (2%)
 

#MES£.NTSRIC I. NODE (19) (47) (50)
 

CONGESTION., NOS 2 _[4%1 1 J25SJ_
 

f NUMdE R O F AMMAL S WIT H TISSUE EXAMINE D MICROSCOPICALL Y 

* NOMdE R OF AMMAL S NECROPSIE D 

79 
TABLE 02. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

MATCHED 

CONTROL LOW DOSE HIGH DOSE 

HiMORRHAGI C CYS T •1 (2%) 

HYPERPLASIA , HEMATOPOIETI C 1 (2%) 

HYPERPLASIA , RETICULU M CELL 4 (9X) 3 (6%) 

HYPERPLASIA , LYMPHOI D 2 (11*) 2 (4% ) 

CIRCULATORY SYSTEM
 

NONE
 

EIGESTIVE -SYSTEM
 

KLIViiR (19) (49) (50)
 

LYMPHOCYTIC INFLAMMATORY INFILTR 4 (21%) 13 (2)%) 5 (1J%)
 

INFLAMMATION, ACUTE fOCAL 1 (5%)
 

AiiSCESS, NCS 1 (2X)
 

ItflFLAMMATICN, FOCAL GRANULCMATO U 2 (4%)
 

NECROSIS, FOCAL 1 (2%)
 

METAMORPHOSIS FATTY 2 (11*) 5 (10*) 2 (4%)
 

ANGIECTASIS f 1 (2%) 1 (2%)
 

HYPERPLASIA, HEMATOPOIETIC 1 (2%)
 

MYELOPOIESIS 1 (2%)
 

((PANCREAS (19) (46) (47)
 

ANGIECTASIS 1 (234)
 

#STCMACH (19) (46) (49)
 

INFLAMMATICN, ACUTE FOCAL 1 (5%)
 

ISMALL INTESTINAL SUB (18) (45) (50)
 

E1VERTICULCSIS 1 (2X)
 

#PEYERS PATCH (18) (45) (50)
 

HYPERPLASIA, BETICULUH CELL 1 (2X)
 

HYPERPLASIA, LYMPHOID 1 (6%)
 

URINARY SYSTEK
 

flKIDJiEY (20) (49) (50)
 

HYDRONEPHROSIS 2 (4%)
 

LYMPHOCYTIC INFLAMMATORY INFILTR 1 (5%) 10 (20%) 4 (83E)
 

INFLAMMATICN, INTERSTITIAL 1 (5%)
 

IltFLAMMATICNj,. CHRONIC FOCAL 3(6%)
 

# NUMriER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

80
 
TABLE D2. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

MATCHED 

CONTROL LOW DOSE HIGH D IOSE 

ItiFLAMHATICN , CHRONIC EIFFUS E 1 (2%) 

ENDOCBIN E SYSTE M 

IPITUITARY (16) (49) (48)

 

HEMORRHAGI C CYST 1 (2»)

 

#ADRENA L (19) (49) (SO ) 

HYPERPLASIA , NODULA R 1 (5%) 

#ADRfiNA L CORTI X (19) (49 ) (50)

 

METAMOBPKCSI S FATTY 1 (5%) 

REPRODUCTIV E SYSTE M 

fcUTERUS (20) (46) (50) 

H2MORRHAGIC CYST 1 (5%) 

POLYP, INFLAMMATOR Y 1 (2%) 

IUTEHUS/ENDCEF.TRIU M (20) (46) (50) DJ.LATATICN, NOS 6 (30%) 2) (43%) 23 (4JX ) 

SOVAriY (20) (47) (50 ) 

MINEBAIIZATIC N 1 (2«) 

CYST, NOS 2 (10S) 11 (23S) 13 (26%) 

HiMORRHAGIC CYST 1 (5X) 1 (2%) 2 (455) 

FIBROSIS 1 (2%) 

NECROSIS, FAT 2 (10X ) 

NERVOUS SYSTEM
 

SPECIAL SENSJ CRGANS
 

NON2
 

MUSCOLOSKELETAI SYSTEM
 

NONE
 

t NUHttER OF ANIMAL S WIT H TISSUE EXAMINE D MICROSCOPICALL Y 

* NUMBER OF ANIMAL S NECROPSIE D 

81 
TABLE D2. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

MATCHED 

CONTROL LOW DOSE 

EOE Y CAVITIE S 

*ABDOMINA L CAVIT Y (20 ) (50) 

AriSCESS, NCS 1 (2%) 

*PERITONEU M (20) (50) 

livFLAMMATICN , ACUTE DIFFUS E 1 (2%) 

INFLAMMATION , CHRONIC DIFFUS E 1 (2%) 

*«ESiiNTESY (20) (50) 

NECROSIS, FAT 4 (20% ) 6 C\2%) 

ANGIECTASI S 

A LL OIHEE SYSTEMS 

*MULTIPLE OEGAN S (20 ) (50)

 

LYMPHOCYTI C INFLAMMATOR Y INFILTR 1 (5% ) 

SPECIAL MORFHC10G Y SUMMAR Y 

NJ LESIO N IEPOETE D 2 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

HIGH DOSE 

(50) 

(5C) 

(50) 

3 (655) 

1 (2%) 

(50) 

3 

82
 
APPENDIX E
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS IN RATS
 

ADMINISTERED CCC IN THE DIET
 

83
 
84

 
Table El. Analyses of The Incidence of Primary Tumors in Male Rats
 

Administered CCC in The Diet (a)
 

Topography; Morphology
 

Integumentary System: Fibroma (b)
 

P Values (c,d)
 

Relative Risk (f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

oo Lung: Alveolar /Bronchiolar
 Ln Carcinoma or Adenoma (b)
 

P Values (c,d)
 

Relative Risk (f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Matched
 

Control
 

1/20(5)
 

N.S.
 

108
 

0/20(0)
 

N.S.
 

Low
 

Dose
 

1/50(2)
 

N.S.
 

0.400
 

0.005
 

30.802
 

108
 

4/49(8)
 

N.S.
 

Infinite
 

0.394
 

Infinite
 

108
 

High
 

Dose
 

5/50(10)
 

N.S.
 

2.000
 

0.249
 

92.596
 

95
 

2/50(4)
 

N.S.
 

Infinite
 

0.123
 

Infinte
 

108
 
(continued) 

Topography;

Table El.

 Morphology 

 Analyses of The Incidence of Primary Tumors in Male Rats 

Administered CCC in The Diet (a) 

Matched 

Control 

Low 

Dose 

High 

Dose 

Hematopoietic System:

or Leukemia (b) 

 Lymphoma 

6/20(30) 10/50(20) 13/50(26) 

P Values (c,d) N.S. N.S. N.S. 

Relative Risk (f) 

Lower Limit 

Upper Limit 

0.667 

0.264 

1.989 

0.867 

0.372 

2.463 

Weeks to First Observed Tumor 108 82 92 

oo 

Pituitary:

Carcinoma

 Chromophobe 

 (b) 0/20(0) 4/49(8) 0/47(0) 

P Values (c,d) N.S. N.S. — 

Departure From Linear Trend (e) P = 0.022 

Relative Risk (f) 

Lower Limit 

Upper Limit " 

Infinite 

0.394 

Infinite 

— 

— 

— 

Weeks to First Observed Tumor 108 
Table El. Analyses of The Incidence of Primary Tumors in Male Rats
 

Administered CCC in The Diet (a)
 

(continued)
 

Topography: Morphology
 

Pituitary: Chromophobe
 

Carcinoma or Adenoma (b)
 

P Values (c,d)
 

Relative Risk (f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Adrenal: Pheochromocytoma (b)
 oo
 

P Values (c,d)
 

Relative Risk (f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Matched
 

Control
 

6/20(30)
 

N.S.
 

108
 

1/20(5)
 

N.S.
 

105
 

Low
 

Dose
 

15/49(31)
 

N.S.
 

1.020
 

0.455
 

2.828
 

108
 

3/49(6)
 

N.S.
 

1.224
 

0.108
 

62.958
 

108
 

High
 

Dose
 

16/47(34)
 

N.S.
 

1.135
 

0.514
 

3.101
 

0/50(0)
 

N.S.
 

0.000
 

0.000
 

7.475
 

70 
Table El.

(continued) 

Topography; Morphology 

 Analyses of The Incidence of Primary Tumors in Male Rats 

Administered CCC in The Diet (a) 

Matched 

Control 

Low 

Dose 

High 

Dose 

Thyroid: Follicular-cell 

Carcinoma or Adenoma (b) 0/20(0) 0/48(0) 3/50(6) 

P Values (c,d) N.S. N.S. 

Relative Risk (f) 

Lower Limit 

Upper Limit 

— 

— 

— 

Infinite 

0.250 

Infinite 

Weeks to First Observed Tumor — 99 

oo 

oo Thyroid C-cell Carcinoma or 

Adenoma (b) 3/20(15) 8/48(17) 0/50(0) 

P Values (c,d) P = 0.011 (N) N.S. P = 0.021 (N) 

Relative Risk (f) 

Lower Limit 

Upper Limit 

1.111 

0.308 

6.043 

0.000 

0.000 

0.659 

Weeks to First Observed Tumor 108 105 
(continued) 

Topography:

Table El.

 Morphology

 Analyses of The Incidence of Primary Tumors in Male Rats 

Administered CCC in The Diet (a) 

Matched 

 Control 

Low 

Dose 

High 

Dose 

Pancreatic Islets:

cell Adenoma (b)

 Islet­

 0/19(0) 2/47(4) 7/49(14) 

P Values (c,d) P = 0.023 N.S. N.S. 

Relative Risk (f) 

Lower Limit 

Upper Limit 

Infinite 

0.124 

Infinite 

Infinite 

0.787 

Infinite 

Weeks to First Observed Tumor — 108 108 

00 

VO 

Testis: Interstitial-cell 

Tumor (b) 17/20(85) 42/49(86) 38/49(78) 

P Values (c,d) N.S. N.S. N.S. 

Relative Risk (f) 

Lower Limit 

Upper Limit 

1.008 

0.840 

1.337 

0.912 

0.755 

1.265 

Weeks to First Observed Tumor 94 83 86 
Table El. Analyses of The Incidence of Primary Tumors in Male Rats
 

Administered CCC in The Diet (a)
 

(continued)
 

(a) Dosed groups received 1,500 or 3,000 ppm.
 

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

(c) Beneath the incidence of tumors in the control group is the probability level for the Cochran-


Armitage test when P is less than 0.05; otherwise, not significant (N.S) is indicated. Beneath
 

the incidence of tumors in a dosed group is the probability level for the Fisher exact test for
 

the comparison of that dosed group with the matched-control group when P is less than 0.05;
 

otherwise, not significant (N.S.) is indicated.
 

(d) A negative trend (N) indicates a lower incidence in a dosed group than in a control group.
 

(e) The probability level for departure from linear trend is given when P is less than 0.05 for any
 

comparison.
 

(f) The 95 percent confidence interval of the relative risk between each dosed group and the control
 

group.
 
Table E2. Analyses of The Incidence of Primary Tumors in Female Rats
 

Administered CCC in The Diet (a)
 

(continued)
 

Topography : Morphology
 

Hematopoietic System: Leukemia
 

or Lymphoma (b)
 

P Values (c,d)
 

Relative Risk (f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pituitary: Chromophobe
 

Carcinoma or Adenoma (b)
 

P Values (c,d)
 

Relative Risk (f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Matched
 

Control
 

3/20(15)
 

N.S.
 

58
 

6/20(30)
 

N.S.
 

108
 

Low
 

Dose
 

11/50(22)
 

N.S.
 

1.467
 

0.450
 

7.594
 

89
 

24/49(49)
 

N.S.
 

1.633
 

0.799
 

4.204
 

78
 

High
 

Dose
 

14/50(28)
 

N.S.
 

1.867
 

0.609
 

9.359
 

89
 

21/49(43)
 

N.S.
 

1.429
 

0.683
 

3.757
 

99
 
Table E2. Analyses of The Incidence of Primary Tumors in Female Rats
 

Administered CCC in The Diet (a)
 

(continued)
 

Topography: Morphology
 

Thyroid: C-cell Carcinoma
 

or Adenoma (b)
 

P Values (c,d)
 

Relative Risk (f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

\o
 

NJ
 

Mammary Gland: Fibroadenoma (a)
 

P Values (c,d)
 

Relative Risk (f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Matched
 

Control
 

1/20(5)
 

N.S.
 

108
 

4/20(20)
 

P = 0.027 (N)
 

92
 

Low High 

Dose Dose 

4/49(8) 2/49(4) 

N.S. N.S. 

1.633 0.816 

0.179 0.046 

78.704 47.195 

78 108 

7/50(14) 2/50(4) 

N.S. N.S. 

0.700 0.200 

0.207 0.020 

2.994 1.297 

65 108 
Table E2. Analyses of The Incidence of Primary Tumors in Female Rats
 

Administered CCC in The Diet (a)
 

(continued)
 

(a) Dosed groups received 1,500 or 3,000 ppm.
 

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

(c) Beneath the incidence of tumors in the control group is the probability level for the Cochran-


Armitage test when P is less than 0.05; otherwise, not significant (N.S) is indicated. Beneath
 

the incidence of tumors in a dosed group is the probability level for the Fisher exact test for
 

the comparison of that dosed group with the matched-control group when P is less than 0.05;
 

otherwise, not significant (N.S.) is indicated.
 

(d) A negative trend (N) indicates a lower incidence in a dosed group than in a control group.
 

(e) The probability level for departure from linear trend is given when P is less than 0.05 for any
 

comparison.
 

(f) The 95 percent confidence interval of the relative risk between each dosed group and the control
 

group.
 
94

 
APPENDIX F
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS IN MICE
 

ADMINISTERED CCC IN THE DIET
 

95
 
96

 
Table Fl. Analyses of The Incidence of Primary Tumors in Male Mice
 

Administered CCC in The Diet (a)
 

Topography: Morphology
 

Lung: Alveolar/Bronchiolar
 

Carcinoma (b)
 

P Values (c,d)
 

Relative Risk (f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Lung: Alveolar/Bronchiolar
 VO
 

•vl	 Carcinoma or Adenoma (b)
 

P Values (c,d)
 

Relative Risk (f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Matched
 

Control
 

2/20(10)
 

N.S.
 

80
 

4/20(20)
 

N.S.
 

80
 

Low
 

Dose
 

7/50(14)
 

N.S.
 

1.400
 

0.303
 

13.138
 

102
 

9/50(18)
 

N.S.
 

0.900
 

0.294
 

3.660
 

102
 

High
 

Dose
 

3/49(6)
 

N.S.
 

0.612
 

0.078
 

6.996
 

102
 

5/49(10)
 

N.S.
 

0.510
 

0.126
 

2.367
 

102
 
Table Fl. Analyses of The Incidence of Primary Tumors in Male Mice
 

Administered CCC in The Diet (a)
 

(continued)
 

Topography : Morphology
 

Hematopoietic System: Lymphoma (b)
 

P Values (c,d)
 

Relative Risk (f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

VO
 

oo
 Liver: Hepatocellular
 

Carcinoma (b)
 

P Values (c,d)
 

Relative Risk (f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Matched
 

Control
 

3/20(15)
 

P = 0.019 (N)
 

93
 

7/20(35)
 

P = 0.036
 

72.
 

Low
 

Dose
 

10/50(20)
 

N.S.
 

1.333
 

0.398
 

7.002
 

68
 

13/50(26)
 

N.S.
 

0.743
 

0.338
 

1.927
 

102
 

High
 

Dose
 

2/49(4)
 

N.S.
 

0.272
 

0.025
 

2.233
 

94
 

23/49(47)
 

N.S.
 

1.341
 

0.693
 

3.159
 

82
 
Table Fl. Analyses of The Incidence of Primary Tumors in Male Mice
 

Administered CCC in The Diet (a)
 

(continued)
 

Topography: Morphology 

Matched 

Control 

Low 

Dose 

High 

Dose 

Adrenal: Cortical adenoma (b) 2/20(10) 1/48(2) 0/49(0) 

P Values (c,d) P = 0.048 (N) N.S. N.S. 

Relative Risk (f) 

Lower Limit 

Upper Limit 

0.208 

0.004 

3.830 

0.000 

0.000 

1.372 

Weeks to First Observed Tumor 102 102 — 

(a) Dosed groups received 500 or 2,000 ppm.
 

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

(c) Beneath the incidence of tumors in the control group is the probability level for the Cochran-


Armitage test when P is less than 0.05; otherwise, not significant (N.S) is indicated. Beneath
 

the incidence of tumors in a dosed group is the probability level for the Fisher exact test for
 

the comparison of that dosed group with the matched-control group when P is less than 0.05;
 

otherwise, not significant (N.S.) is indicated.
 

(d) A negative trend (N) indicates a lower incidence in a dosed group than in a control group.
 

(e) The probability level for departure from linear trend is given when P is less than 0.05 for any
 

comparison.
 

(f) The 95 percent confidence interval of the relative risk between each dosed group and the control
 

group.
 
Table F2. Analyses of The Incidence of Primary Tumors in Female Mice
 

Administered CCC in The Diet (a)
 

Topography ; Morphology
 

Lung: Alveloar/Bronchiolar
 

Carcinoma or Adenoma (b)
 

P Values (c,d)
 

Relative Risk (f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

o
 

o
 Hematopoietic System: Lymphoma
 

or Leukemia (b)
 

P Values (c,d)
 

Relative Risk (f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Matched
 

Control
 

1/20(5)
 

N.S.
 

101
 

7/20(35)
 

N.S.
 

74
 

Low
 

Dose
 

3/49(6)
 

N.S.
 

1.224
 

0.108
 

62.958
 

102
 

10/50(20)
 

N.S.
 

0.571
 

0.239
 

1.560
 

85
 

High
 

Dose
 

2/50(4)
 

N.S.
 

0.800
 

0.045
 

46.273
 

102
 

15/50(30)
 

N.S.
 

0.857
 

0.405
 

2.169
 

85
 
Table F2. Analyses of The Incidence of Primary Tumors in Female Mice
 

Administered CCC in The Diet (a)
 

(continued)
 

Topography: Morphology
 

All Sites: Hemangioma or
 

Hemangiosarcoma (b)
 

P Values (c,d)
 

Relative Risk (f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Liver: Hepatocellular
 

Carcinoma (b)
 

P Values (c,d)
 

Relative Risk (f)
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Matched
 

Control
 

1/20(5)
 

N.S.
 

102
 

4/19(21)
 

N.S.
 

102
 

Low
 

Dose
 

4/50(8)
 

N.S.
 

1.600
 

0.175
 

77.169
 

81
 

7/49(14)
 

N.S.
 

0.679
 

0.202
 

2.892
 

102
 

High
 

Dose
 

5/50(10)
 

N.S.
 

2.000
 

0.249
 

92.596
 

102
 

4/50(8)
 

N.S.
 

0.380
 

0.081
 

1.880
 

102
 
Table F2. Analyses of The Incidence of Primary Tumors in Female Mice
 

Administered CCC in The Diet (a)
 

(continued)
 

Matched Low High
 

Topography: Morphology Control Dose Dose
 

Pituitary: Adenoma, NOS(b) 2/16(13) 2/49(4) 0/48(0)
 

P Values (c,d) P = 0.031 (N) N.S. N.S.
 

Relative Risk (f) 0.327 0.000
 

Lower Limit 0.026 0.000
 

Upper Limit 4.287 1.118
 

Weeks to First Observed Tumor 102 102
 —
 

(a) Dosed groups received 500 or 2,000 ppm.
 

o (b) Number of tumor-bearing animals/number of animals examined at site (percent).
 to
 

(c) Beneath the incidence of tumors in the control group is the probability level for the Cochran-


Armitage test when P is less than 0.05; otherwise, not significant (N.S) is indicated. Beneath
 

the incidence of tumors in a dosed group is the probability level for the Fisher exact test for
 

the comparison of that dosed group with the matched-control group when P is less than 0.05;
 

otherwise, not significant (N.S.) is indicated.
 

(d) A negative trend (N) indicates a lower incidence in a dosed group than in a control group.
 

(e) The probability level for departure from linear trend is given when P is less than 0.05 for any
 

comparison.
 

(f) The 95 percent confidence interval of the relative risk between each dosed group and the control
 

group.
 
 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Review of the Bioassay of (2-Chloroethyl) Trimethylammonium Chloride* for
 

Carcinogenicity by the Data Evaluation/Risk Assessment Subgroup of the
 

Clearinghouse on Environmental Carcinogens
 

December 13, 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 

on the Institute's bioassay program to identify and 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, and State health officials. 

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 (2-Chloroethyl) Trimethylammonium 

Chloride. 

The reviewer for the report on the bioassay of (2-Chloroethyl) 

Trimethylammonium Chloride agreed with the conclusion that the compound 

was not carcinogenic under the conditions of test. After a brief 

description of the experimental design, he noted the lack of data on the 

stability and content of the compound in the diet mix and the inadequate 

number of matched controls. He opined that these shortcomings probably 

did not affect the conclusion reached. Based on the results of the 

bioassay, the reviewer said that (2-Chloroethyl) Trimethylammonium 

Chloride would not appear to pose a carcinogenic hazard to human beings. 

A discussion ensued on the possible significance of the lung 

infiltrates observed among treated rats. A Program staff pathologist 

mentioned that the finding was common in aged rats, although different 

nomenclature may be used to report it. 

It was moved that the report on the bioassay of (2­

Chloroethyl) Trimethylammonium Chloride be accepted as written. 

The motion was seconded and approved without objection. 

103
 
Clearinghouse Members Present;
 

Arnold L. Brown (Chairman), University of Wisconsin Medical School
 

Joseph Highland, Environmental Defense Fund
 

William Lijinsky, Frederick Cancer Research Center
 

Henry Pitot, University of Wisconsin Medical Center
 

Verne A. Ray, Pfizer Medical Research Laboratory
 

Verald K. Rowe, Dow Chemical USA
 

Michael Shimkin, University of California at San Diego
 

Louise Strong, University of Texas Health Sciences Center
 

Kenneth Wilcox, Michigan State Health Department
 

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.
 

fcU.S . GOVERNMEN T PRINTIN G OFFICE ! 1979-281-217/301 3 

104 

DHEW Publication (NIH) No. 79-1714