National Cancer Institute 

CARCINOGENESIS 

Technical Report Series 

No. 2 

February 1976 

CARCINOGENESIS BIOASSAY OF
 

TRICHLOROETHYLENE
 

CAS No. 79-01-6 

NCI-CG-TR-2 

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

Public Health Service 

National Institutes of Health 

NCI-CG-TR-2

 

February 1976 

CARCINOGENESIS BIOASSAY OF 

TRICHLOROETHYLENE 

CAS No. 79-01-6 

These studies were conducted by Hazleton Laboratories, 

Inc., initially under direct contract to the National 

Cancer Institute and subsequently under subcontract 

with Tracor Jitco, Inc., Prime Contractor for the 

Carcinogenesis Bioassay Program, National Cancer Institute. 

CARCINOGEN BIOASSAY AND PROGRAM RESOURCES BRANCH 

CARCINOGENESIS PROGRAM 

DIVISION OF CANCER CAUSE AND PREVENTION 

NATIONAL CANCER INSTITUTE 

BETHESDA, MARYLAND 

DHEW Publication No. (NIH) 76-802 

For sale by the Superintendent of Documents, U.S. Government Printing Office 

Washington, D.C. 20402 - Price $3.30 

Stock Number 017-042-00117-0 

FOREWORD
 

One of the major goals of the National Cancer Institute is to
 

determine the causative factors responsible for human cancer as
 

a basis for preventive measures, both at the environmental and
 

at the host levels. The identification of chemical and physical
 

agents which represent carcinogenic hazards has been recognized
 

as an essential task. The Carcinogenesis Program of the
 

Division of Cancer Cause and Prevention includes among its
 

responsibilities that of testing chemicals for carcinogenic
 

activity.
 

Methods for carcinogenesis bioassay have continued to evolve in
 

the last few decades. While progress was initially slow, and
 

bioassay methods were crude, methodology has greatly improved
 

within the past decade. With better bioassay testing procedures
 

and more extensive efforts in this direction, more chemicals
 

capable of causing cancer in humans will be detected first
 

through experimental tests rather than exclusively through
 

epidemiological studies. More than ever the experimentalists
 

and epidemiologists can now interact to provide direction and
 

leads to the enormous task of exploring the complex problems
 

associated with chemicals and cancer in our society.
 

Technological developments of the last few decades have
 

resulted in thousands of chemicals being introduced into the
 

environment. A number of these chemicals can be expected to be
 

found carcinogenic. The number of chemicals which have been
 

shown to have carcinogenic activity has continued to increase.
 

Two of the major goals of the Carcinogenesis Program are to
 

identify carcinogenic chemicals and to develop improved
 

methodology for testing. The most reliable test method available at this time is the long term bioassay study using laboratory rodents. This is the primary procedure used currently by
 

the bioassay program in its systematic testing for carcinogenicity.
 

Due to the lack of in-house facilities adequate for conducting
 

carcinogenesis bioassay studies, the NCI Carcinogenesis Program
 

has implemented this activity through collaborative research
 

contracts.
 

Several hundred chemicals have been selected for bioassay in
 

recent years. Included among these is a series of chlorinated
 

organic compounds. The bioassay of trichloroethylene, the
 
subject of the present report, is one of the first of this
 

series to have been completed.
 

The selection of this test dates back to decisions made in the
 

early phases of implementation of the Carcinogenesis Program
 

during the development of research on screening methods for
 

carcinogenicity testing. Trichloroethylene was one of 18
 

chemicals tested under a contract awarded to Hazleton Laboratories, Incorporated. Vienna, Virginia, on May 1, 1971, as a
 

result of a Request for Proposals advertised in the C£mmerc_e
 o n March 15, 1969.
 

This bioassay was initiated in 1972 according to the methods
 

used and widely accepted at that time; it represents a valid
 

carcinogenesis test. The design of carcinogenesis bioassays has
 

evolved since then in some respects and several improvements
 

have been developed. The currently recommended procedures are
 

described in detail in the first volume of this series (NCI-CGTR-1) entitled "Guideline for Carcinogenesis Testing in Small
 

Rodents" (1976). The most notable changes pertain to preliminary toxicity studies, number of controls used, and extent
 

of pathological examination.
 

The present report, the first of its kind, provides a detailed
 

documentation of all the aspects of the bioassay, including all
 

the individual animal data and diagnoses.
 

The publication of such detailed reports fulfills a commitment
 

made in 1968 when NCI developed its ''Plan for Chemical Carcinogenesis and the Prevention of Cancers". Methods and capabilities for a fully detailed documentation and publication of well
 

defined and relatively large bioassay studies had to be
 

developed by the Program. They include the development of the
 

Carcinogenesis Bioassay Data System, guidelines for bioassay
 

protocols and procedures, and a network of bioassay resources
 

for chemicals, animals, testing facilities, experimental
 

design, pathology, data processing, and statistical analysis.
 

Methods and criteria for pathological diagnosis and
 

classification were developed.
 

This type of publication also fulfills a recommendation made by
 

the International Union Against Cancer (UICC) in 1969 (Berenblum, 1969). In fact, the definition and exhaustive documentation of carcinogenesis bioassays was recommended by the UICC
 

international expert panels. Those recommendations served as an
 

essential basis for the development of the present reporting 

and publication system. A workshop on "Data Dissemination in 

Carcinogenesis" held by the NCI Carcinogenesis Program in 

January 19 74 endorsed the Technical Report serie^s as a means
 

for disseminating carcinogenesis test results.
 

The present bioassay is clearly the result of team effort, Many
 

people contributed to the selection of the test, the
 

ii
 
development and design of the protocols and the diagnostic and
 

analytical procedures, the establishment and monitoring of
 

facilities, the conduct of the animal tests, and the analysis
 

of results. All of them share in the authorship of this study.
 

Their names and credit for their contribution to the study are
 

given under the heading of "Contributors".
 

The interpretation of carcinogenesis bioassay results. in
 

relation to the complex task of assessing human hazards, is
 

beyond the scope of the present study. This report is designed
 

to provide a factual basis for the interpretative efforts by
 

giving a full, open, and detailed account of the observations
 

made during this bioassay.
 

Norbert P. Page D.V.M
 

Chief, Carcinogen Bioassay Umberto Saffiotti, M.D.
 

and Program Resources Branch Associate Director,
 

Carcinogenesis Program Carcinogenesis Program
 

Division of Cancer Cause Division of Cancer Cause
 

and Prevention and Prevention
 

National Cancer Institute National Cancer Institute
 

1 11
 
CONTRIBUTORS
 

Many individuals and institutions have participated in the
 

planning and conduct of this study. The selection of the
 

chemical and test animals, design of the experiments, and much
 

of the monitoring of progress was the responsibility of Dr.
 

John Weisburger, previously of the National Cancer Institute,
 

now with the American Health Foundation, and Dr. Elizabeth
 

Weisburger, National Cancer Institute (NCI). Dr. John Weisburger served as project officer from inception of the contract
 

until the fall of 1972. Drs. Elizabeth Weisburger and Norbert
 

Page served in that capacity from that time until the present.
 

The actual animal experiments were conducted under contract to
 

the Hazleton Laboratories, Incorporated (HLI), Vienna, Virginia. Principal Investigators for HLI were Drs. Willard Weatherholtz, William Olson, Marcelina Powers, and Richard Voelker.
 

Dr. Robert Habermann conducted the microscopic examination of 

the tissues. Ms. Klara Petrovics was responsible for much of 

the routine technical aspects of the study. Tracor Jitco, 

Incorporated, Rockville, Maryland, as Prime Contractor for
 

Bioassay Operations, with the assistance of the Hazleton Laboratories, Incorporated and National Cancer Institute staffs,
 

has prepared this report. Dr. Jane Robens was responsible for
 

the coordination and major effort required in its preparation.
 

Dr. Charlie Barron conducted a review and confirmation of the
 

histopathologic diagnoses as submitted by Hazleton Laboratories. Dr. Miles Davis conducted the statistical analysis, Dr.
 

Stephen Olin prepared the chemical sections of this report, and
 

Ms. Nancy Palmer functioned as Technical Editor.
 

The Biomedical Information Sciences Department,, EG&G/Mason
 

Research Institute, Bethesda, Maryland, operations contractor
 

for the Carcinogenesis Bioassay Data System (CBDS), under the
 

direction of Mr. Dalton Tidwell, was responsible for the compilation of the individual animal pathology tables and some of
 

the summary tables. Drs. Norbert Page, Cipriano Cueto, and
 

Umberto Saffiotti of the National Cancer Institute outlined the
 

format of this report, worked closely with the Tracor Jitco and
 

Hazleton Laboratories staffs in preparing it, reviewed its content, and contributed to the discussion and interpretation of
 

the findings. Dr. John Gart, Head, Mathematics and Statistics
 

Section, Field Studies and Statistics Program, Division of
 

Cancer Cause and Prevention, NCI, and his staff wc»re responsible for verifying the accuracy of the data, tables, and the
 

statistical analysis of the data.
 

iv
 
Advisory Groups to the Carcinogenesis Program have provided 

guidance in the further development of the carcinogenesis 

bioassay methodology and the review of the Carcinogenesis 

Bioassay Program and its contracts. Members include Dr. 

Richard Adamson, NCI; Dr. Clyde Dawe, NCI; Dr. William 

Deichmann, University of Miami; Dr. Leo Friedman (deceased),
 

Food and Drug Administration; Dr. John Gilbert, Harvard
 

Computing Center; Dr. Harold Grice, Canadian Food and Drug
 

Directorate; Dr. Paul Harris, Indianapolis, Indiana; Dr.
 

Charles Irving, Memphis Veterans' Administration Hospital; Dr.
 

Gerhard Krueger, NCI; Dr. Bernard McNamara, Edgewood Arsenal;
 

Dr. Paul Newberne, Massachusetts Institute of Technology; Dr.
 

Norbert Page, NCI; Dr. Lionel Poirier, NCI; Dr. William
 

Priester, NCI; Dr. James Sontag, NCI; Dr. Robert Squire, NCI;
 

Dr. Elizabeth Weisburger, NCI; Dr. John Weisburger, American
 

Health Foundation; Dr. Harry Wood, NCI; and Mr. Samuel Poiley,
 

NCI.
 

We are especially grateful for the contributions and valuable
 

constructive criticism provided by the reviewers of this
 

report. NCI staff reviewers were Drs. Thomas Cameron, John
 

Cooper, Kenneth Chu, Cipriano Cueto, Herman Kraybill, Umberto
 

Saffiotti, Sidney Siegel, James Sontag, Robert Squire, and
 

Elizabeth Weisburger. Consultants who reviewed the report and
 

provided valuable advice were Dr. Norman Breslow, University of
 

Washington; Dr. Herbert Blumenthal, Food and Drug Administration; Dr. William D'Aguanno, Food and Drug Administration; Dr.
 

Harold Grice, Canadian Food and Drug Directorate; Dr. Elton
 

Homan, U.S. Environmental Protection Agency; Dr. Philip Issenberg, University of Nebraska Medical Center; Dr. William Lloyd,
 

National Institute for Occupational Safety and Health; Dr.
 

Roscoe Moore, National Institute for Occupational Safety and
 

Health; Dr. Verald Rowe, Dow Chemical Company; Mr. Sheldon
 

Samuels, American Federation of Labor/Congress of Industrial
 

Organizations; Dr. Raymond Shapiro, Food and Drug Administration; Dr. Zeb Bell, Jr., PPG Industries, Inc.; Mr. Larry
 

Sargert, PPG Industries, Inc.; Dr. John Weisburger, American
 

Health Foundation; Dr. Jerome Wesolowski, California Department
 

of Health.
 

In addition to those mentioned, appreciation is given to the
 

numerous other staff personnel of Tracor Jitco, Inc., the
 

National Cancer Institute's contractors, and the NCI Carcinogenesis Program for their contributions to these studies.
 

While this Technical Report documents in detail the design,
 

conduct, and results of the study, any further inquiries
 

regarding the study should be directed to the Carcinogenesis
 

Program of the National Cancer Institute.
 

SUMMARY
 

Trichloroethylene (TCE), a halogenated chemical, has been
 

tested for carcinogenicity in the National Cancer Institute's
 

Carcinogenesis Bioassay Program. Trichloroethylene has been
 

used primarily as a solvent in industrial degreasing operations. Other uses have been as a solvent in dry cleaning and
 

food processing, as an ingredient in printing inks, paints,
 

etc., and as a general anesthetic or analgesic.
 

Industrial grade (>99% pure) trichloroethylene was tested using
 

50 animals per group at 2 doses and with both sexes of Osborne-


Mendel rats and B6C3F1 mice. Twenty of each sex and species
 

were maintained as matched controls, in addition to colony and
 

positive carcinogen controls. Animals were exposed to the compound by oral gavage 5 times per week for 78 weeks. At the end
 

-of treatment, animals were observed until terminal sacrifice at
 

110 weeks for rats and 90 weeks for mice. A complete necropsy
 

and microscopic evaluation of all animals (except 7 of the
 

original 480) was conducted.
 

Two doses were used with animals started on test at approximately 6 weeks of age. The initial doses used in this test
 

were the estimated maximum tolerated dose (MTD) and 1/2 MTD, as
 

predicted from data obtained in a 6-week toxicity study. For
 

rats, the initial doses were 1300 and 650 mg/kg body weight.
 

These were changed, based upon survival and body weight data,
 

so that the "time-weighted average" doses were 549 and 1097
 

mg/kg for both male and female rats. For mice, the initial
 

doses were 1000 and 2000 mg/kg for males and 700 and 1400 mg/kg
 

for females. The doses were increased so that the "timeweighted average" doses were 1169 and 2339 mg/kg for male mice
 

and 869 and 1739 mg/kg for female mice.
 

Clinical signs of toxicity, including reduction in weight, were
 

evident in treated rats. These, along with an increased mortality rate, necessitated a reduction in doses during the test. In
 

contrast, very little evidence of toxicity was seen in mice, so
 

doses were increased slightly during the study. The increased
 

mortality in treated male mice appears related to the presence
 

of liver tumors.
 

A variety of neoplastic lesions were observed in rats with no
 

significant difference between trichloroethylene-treated and
 

control animals. The only lesion that might be attributed to
 

vii
 
the treatment was a chronic nephropathy found in both sexes and
 

at both dose levels.
 

With both male and female mice, primary malignant tumors of the
 

liver, i.£.» hepatocellular carcinoma, were observed in high
 

numbers. For males, 26/50 low dose and 31/48 high dose animals
 

had hepatocellular carcinomas as compared with 1/20 matched
 

controls and bill colony controls. The differences between
 

treated and matched control males at both doses were highly
 

significant (P < 0.01). For females, hepatocellular carcinomas
 

were observed in 4/50 low dose and Wlkl high dose animals as
 

compared with 0/20 matched controls and 1/80 colony controls.
 

While the difference between the high dose female mice and
 

matched controls was also highly significant (P < 0.01), the
 

difference at the low dose was less (P = 0.09). For both male
 

and female mice, age-adjusted tests for linear trend (dose
 

response) were highly significant for hepatocellular carcinoma
 

(P < 0.001 for males and P = 0.002 for females).
 

In male mice at the high doses, hepatocellular carcinomas were
 

observed early in the study. The first was seen at 27 weeks; 9
 

others were found in male mice dying by the 78th week. The
 

tumor was not observed so early in low dose male or female
 

mice. The diagnosis of hepatocellular carcinoma was based on
 

size, histologic appearance, and presence of metastasis,
 

especially to the lung. No other lesion was significantly
 

elevated (P < 0.05) in treated mice. The incidence of hepatocellular carcinomas in the trichloroethylene-matched controls
 

was typical of that observed in colony controls.
 

Carbon tetrachloride (CCI4) was used as a positive control for
 

the series of chlorinated chemicals which included trichloroethylene. While virtually all male and female mice developed
 

hepatocellular carcinomas following carbon tetrachloride treatment, the response in the Osborne-Mendel rat was considerably
 

less. Only about 5% developed hepatocellular carcinomas. Thus,
 

there appears to be a marked difference in sensitivity to
 

induction of carcinomas by chlorinated compounds between the
 

B6C3F1 mouse and the Osborne-Mendel rat.
 

The results of this carcinogenesis test of trichloroethylene
 

clearly indicate that trichloroethylene induced a hepatocellular carcinoma response in mice. While the absence of a
 

similar effect in rats appears most likely attributable to a
 

difference in sensitivity between the Osborne-Mendel rat and
 

the B6C3F1 mouse, the early mortality of rats due to toxicity
 

must also be considered.
 

viii
 
TABLE OF CONTENTS
 

Page
 

1.0	 INTRODUCTION 1
 

2 .0	 MATERIALS 2
 

2.1	 Name and Synonyms 2
 

2.2	 Formula, Molecular Weight, Identifying Numbers, and
 

Characteristics 2
 

2 .3	 Procurement 2
 

2.4	 Chemical Analysis 3
 

2.5	 Preparations Used for All Bioassays 3
 

2.6	 Safety Procedures 3
 

3 .0	 TEST ANIMALS AND ENVIRONMENT 5
 

4.0	 PRECHRONIC PHASES: METHODS AND RESULTS 7
 

4.1 Acute Study	 7
 

4.2 Eight-Week Subchronic Study	 7
 

4.2.1 Methodology	 7
 

4.2.2 Results - Rats	 8
 

4.2.3 Results - Mice	 8
 

4.2.4 Selection of MTD	 8
 

5.0	 CHRONIC TESTING: METHODOLOGY 9
 

5 .1 Experimental Design	 9
 

5.1.1 Experimental Groups	 9
 

5.1.2 Dates of Study	 9
 

5.1.3 Preparations and Doses	 9
 

5.1.4 Treatment Schedule	 10
 

5 . 2 Observations	 10
 

5.3	 Necropsy 10
 

5.4	 Histological Preparation and Microscopic Examination.. 13
 

5.5	 Data Processing and Confirmation 14
 

5.6	 Positive Controls 15
 

6.0	 CHRONIC TESTING: RESULTS - RATS 16
 

6.1	 Body Weights 16
 

6.2	 Clinical Observations 16
 

6 .3	 Survival 16
 

6.4	 Pathology 17
 

6.5	 Tumor Probabilities 21
 

6.6	 Controls 22
 

6.6.1 Survival	 22
 

6.6.2 Tumors	 22
 

IX
 
7 .0 CHRONIC TESTING: RESULTS - MICE. . . 24
 

7 .1 Body Weights 24
 

7 .2 Clinical Observations 24
 

7.3 Survival 24
 

7.4 Pathology 27
 

7.5 Tumor Probabilities 34
 

7 .6 Colony Controls 35
 

7.7 Positive Controls 39
 

8.0 DISCUSSION 41
 

8.1 Design of Bioassay 41
 

8.1.1 Selection of Animal Species 41
 

8.1.2 Route of Exposure 41
 

8.1.3 Use of High Doses 42
 

8.1.4 Methodology 42
 

8.2 Test Compound Purity 43
 

8.3 Metabolism, Distribution, and Excretion 43
 

8 .4 Toxicology 44
 

8.5 Pathology and Survival 45
 

8.5.1 Rats 45
 

8.5.2 Mice 45
 

8.6 Effect of Various Compounds in the Same Room 46
 

8.7 Relationship to Toxicity of Carbon Tetrachloride...... 46
 

8 .8 Conclusions 47
 

9 .0 BIBLIOGRAPHY 48
 

APPENDIX A: CHEMISTRY 53
 

Chemical and Physical Characteristics 54
 

Technical Product and Impurities 54
 

Manufacturing Processes 54
 

Chemical Analysis 55
 

Identification of Trace Components 61
 

Feed Analysis 66
 

Water Analysis 68
 

APPENDIX B: WEIGHTS AND SURVIVAL 69
 

Mean Body Weights and Survival - Subchronic Study 70
 

Mean Body Weights and Survival - Chronic Study 74
 

APPENDIX C: STATISTICS 87
 

Statistical Methodology 88
 

Data for Statistical Analysis 93
 

Survival Probabilities 97
 

Tumor Probabilities 102
 
APPENDIX D: PATHOLOGY 117 

Tissues Examined - Trichloroethylene-Treated Animals

Tumor Summary Tables by Site of Origin and by Anatomic 

Site - Trichloroethylene-Treated Animals

Individual Pathology - Trichloroethylene-Treated Animals...

 118 

 120 

 134 

APPENDIX E: POSITIVE CONTROLS 

Individual Liver Pathology - Positive Control 

(Carbon Tetrachloride) Animals 17 7 

XI
 

LIST OF TABLES
 

Page
 

Table I. Identification of Trichloroethylene 

Used in Study 2 

Table II. Design and Survival Results -

Trichloroethylene Subchronic Study 7 

Table III. Dosage and Observation Schedule -

Trichloroethylene Chronic Study 11 

Table IV. Dosage and Observation Schedule ­

Carbon Tetrachloride Study 15 

Table V. Tumor Incidence - Rats with Tumors 17 

Table VI. Comparison of Survival of Colony Controls and 

Trichloroethylene- and Carbon TetrachlorideTreated Rats 22 

Table VII. Incidence of Liver Tumors - Colony Control and 

Carbon Tetrachloride-Treated Rats 22 

Table VIII. Incidence of Hepatocellular Carcinoma ­

Trichloroethylene-Treated Mice 35 

Table IX. Probability of Observing Hepatocellular Carcinoma 

in Trichloroethylene-Treated Male Mice 36 

Table X. Incidence of Hepatocellular Carcinoma ­

Colony Control Mice 39 

Table XI. Comparison of Survival in Colony Controls ­

Vehicle-Treated and Trichloroethyleneand Carbon Tetrachloride-Treated Mice 39 

Table Xlla. Comparison of Hepatocellular Carcinoma Incidence 

in Colony Controls - Vehicle Treated and 

Trichloroethylene- and Carbon TetrachlorideTreated Mice 40 

Table Xllb. Comparison of Time to Liver Tumor Detection 

in Colony Controls - Vehicle Treated and 

Trichloroethylene- and Carbon TetrachlorideTreated Mice 40 

Table Xllla.

Table Xlllb.

 Mean Body Weights and Survival -

Trichloroethylene Subchronic Study - Male Rats....

 Mean Body Weights and Survival -

Trichloroethylene Subchronic Study - Female Rats.,

 70 

 71 

xiii 
Table XlVa. Mean Body Weights and Survival -

Trichloroethylene Subchronic Study - Male Mice.... 72 

Table XlVb. Mean Body Weights and Survival -

Trichloroethylene Subchronic Study - Female Mice.. 73 

Table XVa. Mean Body Weights, Food Consumption, and 

Survival - Trichloroethylene Chronic 

Study - Male Rats 74 

Table XVb. Mean Body Weights, Food Consumption, and 

Survival - Trichloroethylene Chronic 

Study - Female Rats 76 

Table XVIa. Mean Body Weights, Food Consumption, and 

Survival - Trichloroethylene Chronic 

Study - Male Mice 78 

Table XVIb. Mean Body Weights, Food Consumption, and 

Survival - Trichloroethylene Chronic 

Study - Female Mice 80 

Table XVII. Identity of Tumor Marks . 92 

Table XVIIIa. Data for Statistical Analysis ­

Trichloroethylene-Treated Male Rats 93 

Table XVIIIb. Data for Statistical Analysis ­

Trichloroethylene-Treated Female Rats 94 

Table XlXa. Data for Statistical Analysis ­

Trichloroethylene-Treated Male Mice . . 95 

Table XlXb. Data for Statistical Analysis ­

Trichloroethylene-Treated Female Mice 96 

Table XXa. Product-Limit Estimates of Probability of 

Survival - Trichloroethylene-Treated Male Rats.... 97 

Table XXb. Product-Limit Estimates of Probability of 

Survival - Trichloroethylene-Treated Female Rats.. 98 

Table XXc. Statistical Tests Comparing Estimated 

Probability of Survival among Control 

and Trichloroethylene-Treated Rats 99 

Table XXIa. Product-Limit Estimates of Probability of 

Survival - Trichloroethylene-Treated Male Mice.... 100 

Table XXIb. Product-Limit Estimates of Probability of 

Survival - Trichloroethylene-Treated Female Mice.. 100 

xiv
 
Table XXIc. Statistical Tests Comparing Estimated 

Probability of Survival among Control 

and Trichloroethylene-Treated Mice 101 

Table XXII. Tumor Incidence - Trichloroethylene-Treated Rats.. 102 

Table XXIIIa. Statistical Tests Comparing Estimated Probability 

of Observing Reticulum-cell Sarcoma, Lymphosarcoma, or Malignant Lymphoma (Mark b) among 

Control and Trichloroethylene-Treated Rats 103 

Table XXIIIb. Statistical Tests Comparing Estimated Probability 

of Observing Fibroadenoma of the Mammary Glands 

(Mark g) among Control and TrichloroethyleneTreated Rats 104 

Table XXIIIc. Statistical Tests Comparing Estimated Probability 

of Observing Hemangioma of Any Site (Mark h) 

among Control and Trichloroethylene-Treated Rats.. 104 

Table XXIIId. Statistical Tests Comparing Estimated Probability 

of Observing Follicular Adenocarcinoma of the 

Thyroid (Mark p) among Control and Trichloroethylene-Treated Rats 105 

Table XXIIIe. Statistical Tests Comparing Estimated Probability 

of Observing Chromophobe Adenoma of the Pituitary 

(Mark t) among Control and TrichloroethyleneTreated Rats 105 

Table XXIVa. Exact Statistical Tests Comparing Proportions 

of Animals (Not Adjusted for Age) with Observed 

Hepatocellular Carcinoma of the Liver among 

Pooled Control and Trichloroethylene-Treated Rats. 106 

Table XXIVb. Exact Statistical Tests Comparing Proportions 

of Animals (Not Adjusted for Age) with Observed 

Hepatocellular Carcinoma or Neoplastic Nodule of 

the Liver among Pooled Control and Trichloroethylene-Treated Rats 107 

Table XXIVc. Exact Statistical Tests Comparing Proportions 

of Animals (Not Adjusted for Age) with Observed 

Hepatocellular Carcinoma of the Liver among 

Pooled Control and Carbon TetrachlorideTreated Rats 108 

Table XXIVd. Exact Statistical Tests Comparing Proportions 

of Animals (Not Adjusted for Age) with Observed 

Hepatocellular Carcinoma or Neoplastic Nodule of 

the Liver among Pooled Control and Carbon 

Tetrachloride-Treated Rats 109 

xv
 
Table XXV. Tumor Incidence - Trichloroethylene-Treated Mice.. Ill 

Table XXVIa. Statistical Tests Comparing Estimated Probability 

of Observing Hepatocellular Carcinoma (Mark a) 

among Control and Trichloroethylene-Treated Mice.. 112 

Table XXVIb. Statistical Tests Comparing Estimated Probability 

of Observing Reticulum-cell Sarcoma, Lymphosarcoma, or Malignant Lymphoma (Mark b) among 

Control and Trichloroethylene-Treated Mice 112 

Table XXVIc. Statistical Tests Comparing Estimated Probability 

of Observing Carcinoma or Adenocarcinoma of the 

Lung or Alveoli (Mark c) among Control and 

Trichloroethylene-Treated Mice 113 

Table XXVId. Statistical Tests Comparing Estimated Probability 

of Observing Adenoma of the. Lung (Mark d) among 

Control and Trichloroethylene-Treated Mice 113 

Table XXVle. Statistical Tests Comparing Estimated Probability 

of Observing Carcinoma, Adenocarcinoma, or 

Adenoma of the Lung or Alveoli (Mark c or d) 

among Control and Trichloroethylene-Treated Mice.. 114 

Table XXVII. Exact Statistical Tests Comparing Proportions 

of Animals (Not Adjusted for Age) with 

Observed Hepatocellular Carcinoma among 

Pooled Control and Trichloroethylene-Treated Mice. 115 

Table XXVIIIa. Numbers of Tissues Examined ­

Trichloroethylene-Treated Rats 118 

Table XXVIIIb. Numbers of Tissues Examined ­

Trichloroethylene-Treated Mice 119 

Table XXIXa. Primary Tumors by Site of Origin ­

Trichloroethylene-Treated Rats 121 

Table XXIXb. Tumors by Anatomic Site ­

Trichloroethylene-Treated Rats 124 

Table XXXa. Primary Tumors by Site of Origin ­

Trichloroethylene-Treated Mice 127 

Table XXXb. Tumors by Anatomic Site ­

Trichloroethylene-Treated Mice 130 

Table XXXIa. Individual Pathology - TrichloroethyleneTreated Male Rats 135 

Table XXXIb. Individual Pathology - TrichloroethyleneTreated Female Rats 147 

xvi 
Table XXXIla. Individual Pathology •- TrichloroethyleneTreated Male Mice 158 

Table XXXIIb. Individual Pathology - TrichloroethyleneTreated Female Mice 168 

Table XXXIIIa. Liver Histopathology for Positive Controls 

(Carbon Tetrachloride) - Low Dose Male Rats 178 

Table XXXIIIb. Liver Histopathology for Positive Controls 

(Carbon Tetrachloride) - High Dose Male Rats 181 

Table XXXIIIc. Liver Histopathology for Positive Controls 

(Carbon Tetrachloride) - Low Dose Female Rats 184 

Table XXXIIId. Liver Histopathology for Positive Controls 

(Carbon Tetrachloride) - High Dose Female Rats.... 187 

Table XXXIVa. Liver Histopathology for Positive Controls 

(Carbon Tetrachloride) - Low Dose Male Mice 190 

Table XXXIVb. Liver Histopathology for Positive Controls 

(Carbon Tetrachloride) - High Dose Male Mice 192 

Table XXXIVc. Liver Histopathology for Positive Controls 

(Carbon Tetrachloride) - Low Dose Female Mice 194 

Table XXXIVd. Liver Histopathology for Positive Controls 

(Carbon Tetrachloride) - High Dose Female Mice.... 196 

xvii
 
LIST OF FIGURES
 

Page 

Figure la. Dosage Schedule - Chronic Study ­

Trichloroethylene-Treated Rats 12 

Figure lb. Dosage Schedule ­ Chronic Study ­

Trichloroethylene-Treated Mice 12 

Figure 2a. Product-Limit Estimates of Probability of 

Survival - Trichloroethylene-Treated Male Rats.... 18 

Figure 2b. Product-Limit Estimates of Probability of 

Survival - Trichloroethylene-Treated Female Rats.. 19 

Figure 3a. Product-Limit Estimates of Probability of 

Survival - Trichloroethylene-Treated Male Mice.... 25 

Figure 3b. Product-Limit Estimates of Probability of 

Survival - Trichloroethylene-Treated Female Mice.. 26 

Figure 4. Primary hepatocellular carcinoma, mouse (high 

dose male #32). The rather well differentiated 

tumor of trabecular pattern has a recognizable 

boundary (—>) with the pre-existent hepatic 

tissue. Hematoxylin and eosin, x75 28 

Figure 5 . Same as Figure 4, x300 28 

Figure 6. Primary hepatocellular carcinoma, mouse (high 

dose male #26). The rather anaplastic solid 

tumor has a recognizable boundary (—>) with 

the pre-existent hepatic parenchyma. Mitoses 

are abundant. Hematoxylin and eosin, x240 30 

Figure 7. Primary hepatocellular carcinoma, mouse (high 

dose male #15) with both solid trabecular 

pattern and papillary pattern. Most cells 

are large and resemble normal hepatocytes. 

There are also smaller cells with less cytoplasm and smaller, more basophilic nuclei. 

These cells usually occur in clusters and are 

especially prominent in the larger papillary 

structures 30 

XVlll
 
Figure 8. Primary hepatocellular carcinoma, mouse (high 

dose male #32). There is an area of highly 

anaplastic cells in a papillary pattern 

contiguous with an area of well differentiated 

cells in a trabecular pattern. Hematoxylin and 

eosin, xl20 32 

Figure 9. Secondary hepatocellular carcinoma, lung, 

mouse (high dose male #15). The metastatic hepatocellular carcinoma has invaded and extended 

into a bronchiole. The bulk of the tumor consists 

of rather well differentiated hepatocytes but 

there are scattered foci of smaller, more 

anaplastic, basophilic, neoplastic cells 

(—>) also. Hematoxylin and eosin, xl20 32 

Figure 10. Metastatic hepatocellular carcinoma, lung, mouse 

(high dose male #15). Well differentiated hepatocytes comprise a large nodule (edge at upper left) 

and invade perivascularly at lower center. 

Numerous foci of smaller, more anaplastic, 

basophilic, neoplastic cells occur in vessels 

(—>) and alveolar capillaries, and invade 

perivascularly. Hematoxylin and eosin, x96 32 

Figure 11. Product-Limit Estimates of Probability of 

Observing Hepatocellular Carcinoma ­

Trichloroethylene-Treated Male Mice 37 

Figure 12. Comparison of Incidence of Hepatocellular 

Carcinoma in Trichloroethylene-Treated 

Male and Female Mice 38 

Figure 13a. Mean Body Weight vs. Time on Study ­

Trichloroethylene-Treated Male Rats 82 

Figure 13b. Mean Body Weight vs. Time on Study ­

Trichloroethylene-Treated Female Rats 83 

Figure 14a. Mean Body Weight vs. Time on Study ­

Trichloroethylene-Treated Male Mice 84 

Figure 14b. Mean Body Weight vs. Time on Study ­

Trichloroethylene-Treated Female Mice 85 

xix
 

1.0 INTRODUCTION
 

In the late 1960s, scientists at the National Cancer Institute noted that a
 

group of halogenated compounds extensively used as solvents in industrial
 

processes had not been adequately tested for chronic toxicity. A related
 

compound, carbon tetrachloride, however, had already been found carcinogenic in mice (Eschenbrenner and Miller, 1944, 1946), hamsters (Delia Porta
 

et al. , 1961), and rats (Reuber and Glover, 1970). Thus, carcinogenesis
 

bioassays of a group of these solvents were initiated.
 

Trichloroethylene was one of the chemicals selected to be tested and this
 

report describes the conduct and results of its bioassay. Production of
 

trichloroethylene was reported as 609, 514, and 427 million pounds in 1970,
 

1971, and 1972, respectively, in the Chemical Economics Handbook (1972).
 

The primary use (about 90%) of trichloroethylene is in the vapor degreasing
 

of metals and equipment. It has also been used as a solvent in dry cleaning, in the processing of certain medicines and foods, and in other
 

processes, as an ingredient in printing inks, paints, lacquers, varnishes,
 

and adhesives, as a chemical intermediate, and in a variety of other
 

applications such as a grain fumigant (Wiseman, 1972; Frear, 1969).
 

A pharmaceutical grade of trichloroethylene has also been used as a general
 

anesthetic in surgical and obstetrical procedures, administered by inhalation. It is a potent analgesic but will not produce appreciable skeletal
 

muscle relaxation at the concentrations used. As an analgesic it has been
 

used for minor procedures such as cleaning and debridement of burns,
 

orthopedic manipulations, cystoscopy, incision of abscesses, surface
 

biopsy, changing painful dressings, and treating trigeminal neuralgia
 

(Price and Dripps, 1965).
 

Trichloroethylene has been identified in low concentrations in certain
 

municipal water supplies as reported by the Environmental Protection Agency
 

(Dowty et al., 1975). Residues may result from the use of trichloroethylene as a solvent in the processing of foods. Tolerances for
 

trichloroethylene of 25 ppm in decaffeinated ground coffee, 10 ppm in
 

decaffeinated soluble (instant) coffee extract, and 30 ppm in spice
 

oleoresins have been established by the Food and Drug Administration (Code
 

of Federal Regulations, Title 21) . Thus, exposure may occur indirectly to
 

the general population through residues in water and food.
 

The National Institute of Occupational Safety and Health (1973) has issued
 

a comprehensive review of the uses, exposure, and known biological effects
 

of trichloroethylene. Criteria for a recommended standard for occupational
 

exposures are given in this document. It recommends that occupational
 

exposure to trichloroethylene be controlled so that no worker shall be
 

exposed to a peak concentration of trichloroethylene in excess of 150 ppm
 

as measured over a maximum sampling time of 10 minutes, or to a concentration in excess of 100 ppm determined as a time-weighted average exposure
 

for an 8-hour workday as measured over a minimum sampling time of 10
 

minutes. The Occupational Safety and Health Administration has recently
 

proposed changes in their regulations to reflect these recommendations.
 
2.0 MATERIALS
 

2.1 Name and Synonyms
 

Chemical Abstracts and IUPAC Name: Trichloroethene
 

Synonyms and Common Name: Trichloroethylene
 

Acetylene trichloride
 

Ethinyl trichloride
 

1,1,2-Trichloroethylene
 

TCE
 

(Christensen and Luginbyhl, 1974;
 

Deichmann and Gerarde, 1969)
 

2.2 Formula, Molecular Weight, Identifying Numbers, and Characteristics
 

Formula: C2HCI3
 

Molecular Weight: 131.40
 

Wiswesser Line Notation: GYGU1G
 

Chemical Abstracts Service Registration Number: 79-01-6
 

NCI Number: CO4546
 

For chemical and physical characteristics, technical product and impurities,
 

and manufacturing processes, see Appendix A.
 

2.3 Procurement
 

Four batches of trichloroethylene were procured from Aldrich Chemical Company
 

as given in Table I:
 

Table I. Identification of Trichloroethylene Used in Study
 

Batch Manufacturer's Received by Analysis Report 

No. Lot No. Hazleton (date) (date) Use 

1 050191 7/23/71 1/9/73 Prechronic study 

and first 2 weeks 

of chronic study. 

2 061891 7/23/71 Weeks 3-15 in 

chronic study. 

3 063017 5/5/72 2/16/74 Weeks 16-36 in 

chronic study. 

4 063014 10/12/72 7/8/74 Weeks 37-78 in 

chronic study. 
Each batch was received in one or more large amber bottles. Containers
 

were stored in the dark at room temperature.
 

2.4 Chemical Analysis
 

The purity of the trichloroethylene used in the bioassay was determined by
 

gas chromatography and infrared spectroscopy. Minor components subsequently were identified by gas chromatography-mass spectrometry and confirmed with reference standards.
 

Analyses of gas chromatographic total area data showed the major component
 

to be at least 99% in each batch. Infrared spectra compare well with
 

trichloroethylene reference spectra. The minor components comprise a
 

mixture of stabilizers routinely added to commercial formulations of
 

trichloroethylene. They include 1,2-epoxybutane (0.19%), ethyl acetate
 

(0.04%), epichlorohydrin (0.09%), N-methylpyrrole (0.02%), and
 

diisobutylene (0.03%) . Percentages were determined by FID gas
 

chromatography with standards after completion of the bioassay. No detectable quantities of 1, 1, 2, 2-tetrachloroethane (<5 ppm) or 1,1,1,2-tetrachloroethane (<2 ppm) were indicated by gas chromatography, using reference
 

standards. (For details of chemical analysis, jse_e Appendix A.)
 

2•5 Preparations Used for All Bioassays
 

Fresh solutions of trichloroethylene in corn oil were prepared weekly in
 

amounts sufficient to treat all animals for one week, sealed, and
 

refrigerated until use. Concentrations for the chronic test are given in
 

Table III. The corn oil (purchased from the distributor, C. F. Sauer
 

Company, Richmond, VA) was not analyzed for impurities or reaction products
 

during this study.
 

2.4 Safety Procedures
 

Laboratory personnel working with undiluted experimental compound were
 

required to wear the following protective gear: safety goggles, latex
 

gloves, disposable full-body protective suit with attached feet and
 

open-face hood, half-face Welsh respirator fitted with a dust and mist
 

filter over a chemical cartridge for organic vapors. All work with these
 

materials was conducted in a negative-pressure room and under a hood.
 

Personnel involved with compound administration, animal weighing, and
 

feeding wore a disposable laboratory coat, head covering, the Welsh
 

respirator, and disposable latex gloves. Personnel involved with animal
 

care, i-e., cage changing and washing, wore heavy duty gloves, safety
 

shoes, half-face surgical mask (3M), and head covering. Any person entering the animal rooms was required to wear a head covering and 3M mask. As
 

an additional personnel safety measure and to minimize cross contamination
 

within animal rooms, actual intubation procedures were performed within the
 

confines of a fume hood. Test solutions were kept in an ice bath during the
 

dosing procedures to minimize evaporation. In the rooms housing rats, hoods
 

were located in the corner of the room and each rack was wheeled to the
 

hood each time the animals were dosed. Mouse racks were transported
 

through the hall to another room with a large hood and each cage was placed
 

directly under the hood during the intubation process. The testing
 
laboratory's health and safety officer in conjunction with laboratory 

personnel made on-site inspections to insure compliance with the above 

safety precautions. 
3.0 TEST ANIMALS AND ENVIRONMENT
 

Random-bred Osborne-Mendel rats (Battelle Memorial Institute, Columbus, OH)
 

and B6C3F1 (C57BL/6 x C3H/He) hybrid mice (Charles River, Wilmington, MA)
 

in the chronic study were obtained at 35 and 25 days of age, respectively,
 

from suppliers under contract to NCI. Trichloroethylene-treated rats and
 

their controls were born within 6 days of each other, with a median birth
 

date of February 23, 19 72. Trichloroethylene-treated mice and their controls were born within 6 days of each other, with a median birth date of
 

July 17, 19 72. Upon arrival at the laboratory, all animals were isolated
 

for at least 10 days. They were observed at arrival and weighed immediately before being placed on study. Weight ranges of trichloroethylenetreated animals and their controls in the chronic study were: male rats,
 

168-229 g; female rats, 130-170 g; male mice, 11-22 g; female mice, 11-18
 

g. Animals were randomly* assigned to treatment groups, so that initially
 

the average weight in each group was approximately the same.
 

The rats were individually housed in hanging galvanized steel cages, 25.4 x
 

17.8 x 17.8 cm (Wahmann) with wire mesh fronts and floors. There were 72
 

cages per rack placed to allow 2 racks per 100 square feet of floor space.
 

Paper collection trays (National Paper Products) were placed under the
 

cages and were changed twice weekly. The rats were transferred to freshly
 

cleaned cages weekly. The soiled cages and racks were washed under
 

pressure at 80°C in water containing Super Soilax detergent (Economics
 

Laboratory, Inc.), rinsed at 80°C, and steamed in a Matawan 375 gallon
 

automatic cycle industrial washer. The water bottles with stainless steel
 

sipper tubes inserted in rubber stoppers were changed twice weekly; the
 

dirty bottles and tubes were washed, rinsed, and steamed in a 220 gallon
 

industrial washer similar to that used for cage washing. Feed was supplied
 

in glass jars within each cage. Trichloroethylene-treated rats and their
 

controls were maintained in a room housing other rats being treated with
 

one of the following compounds: dibromochloropropane, ethylene dichloride,
 

1,1- dichloroethane, and carbon disulfide. Four groups of vehicle-treated
 

controls were in the same room.
 

The mice in the chronic phase were housed in polypropylene cages (Lab
 

Products), 47 x 24.1 x 15.2 cm, which contained 10 animals of one sex per
 

cage. (In the prechronic phases mice were housed individually in hanging
 

wire mesh steel cages, 17.8 x 12.7 x 10.2 cm.) Animal rooms contained 40
 

cages per rack to allow 1.5 racks per 100 square feet of floor space. The
 

cover for each unit was welded stainless steel wire over which was placed a
 

non-woven polyester fiber filter bonnet. Each cage contained a galvanized
 

iron, compartmentalized lfgangif feeder (Dixie Sheet Metal Co.) with a screen
 

top. All mice were transferred to clean cages containing fresh bedding
 

(Sanichips, a heat-treated hardwood product, Shurfire Products, Inc.) twice
 

weekly. The soiled cages were washed and rinsed at 80°C and steamed. The
 

steel wire bar covers of the cages were washed on a weekly basis and the
 

racks on a monthly basis. The filter bonnets were washed and autoclaved
 

weekly. The water bottles and stainless steel drinking tubes were changed
 

^•Animals were not distributed according to a table of random numbers.
 

5 
3 times a week and cleaned by washing, rinsing, and steaming as stated
 

ab o ve.
 

Mice treated with trichloroethylene were maintained in a room housing other
 

mice being treated with one of the following 17 compounds: 1,1,2,2-tetrachloroethane, chloroform, 3-chloropropene, chloropicrin, 1,2-dibromochloropropane, 1,2-dibromoethane, ethylene dichloride, 1,1-dichloroethane,
 

3-sulfolene, iodoform, methyl chloroform, 1,1,2-trichloroethane, tetrachloroethylene, hexachloroethane, carbon disulfide, trichlorofluoromethane,
 

and carbon tetrachloride. Nine groups of vehicle controls and 9 groups of
 

untreated controls were also housed in this room.
 

All feeders for both rats and mice were changed weekly for the first 10
 

weeks and every 4 weeks thereafter and were cleaned by the same process of
 

washing, rinsing, and steaming.
 

Following changing, the clean cages for all animals were placed on the
 

racks in the same manner as before; however, the racks were repositioned
 

within the room on a daily basis for the first 78 weeks, and weekly
 

thereafter. The floors of each room were cleaned daily using Mikro-Bac, a
 

phenolic detergent-disinfectant (Economics Laboratory, Inc.).
 

The total air in each room was changed 10-15 times per hour with all
 

incoming air filtered through 2-inch thick fiberglass disposable filters
 

which were changed at least once weekly. The relative humidity of the room
 

air was maintained between 45 and 55% and the temperature range was 20 to
 

24°C. Rooms were illuminated by fluorescent lighting 12 hours per day.
 

There was no communication between rooms, _i.j^.> there were no connecting
 

doors, separate groups of technicians handled the rats and mice, each room
 

had individual air ducts, and rooms were under negative pressure. Samples
 

of ambient air were not tested for presence of volatile materials.
 

Wayne Lab-Blox meal was fed to the animals aid libitum. (Appendix A
 

contains a feed ingredient list, analysis of protein, fat, and fiber by the
 

manufacturer, and analyses for pesticide residues in selected feed
 

batches.) Drinking water, from a local artesian well, was supplied ad
 

libitum in glass bottles. (Appendix A presents a water analysis.)
 

Hazleton Laboratories, Inc., was certified as a research facility in August
 

1967 under the Animal Welfare Act by the USDA, Animal and Plant Health
 

Inspection Service. The animal care facilities were fully accredited by
 

the American Association for Accreditation of Laboratory Animal Care
 

beginning June 4, 1971.
 

6 
4.0 PRECHRONIC PHASES: METHODS AND RESULTS
 

4•1 Acute Study
 

Single-dose range-finding studies were conducted with male rats and female
 

mice to determine the highest dose to be used in the 8-week subchronic
 

study. Groups of 2 animals each were administered a single dose of
 

trichloroethylene in corn oil by gavage by oral intubation and observed for
 

14 days. Ten dosages were used: 100, 178, 316, 562, 1000, 1420 (rats
 

only), 1780 (mice only), 3160, 5620, 10,000, and 17,800 mg/kg. The lowest
 

doses causing death, 5620 mg/kg for rats and 10,000 mg/kg for mice, were
 

selected as the highest doses to be used in the 8-week subchronic study.
 

4.2 Eight-Week Subchronic Study
 

The objective of this study was to estimate the Maximum Tolerated Dose
 

(MTD) for trichloroethylene to be used in rats and mice in the bioassay for
 

carcinogenicity. In this context the MTD is defined as the highest dose 

that can be administered during the chronic study, which will not be 

expected to alter the animals' survival rate from effects other than 

carcinogenicity. 

4.2.1 Methodology
 

Animals were placed into 6 groups each of 5 males and 5 females so that
 

initially the average weight per animal in each treatment group was the
 

same. Five groups received the test compound at varying dosages; one group
 

served as the control and received only the vehicle (corn oil).
 

Trichloroethylene was dissolved in corn oil and animals were dosed by
 

gavage under a hood for 5 consecutive days per week for 6 weeks on the
 

basis of milligrams trichloroethylene per kilogram body weight. Doses
 

ranged from 562 to 5620 mg/kg in rats and from 1000 to 10,000 mg/kg in mice
 

(Table II).
 

Table II. Design and Survival Results - Trichloroethylene Subchronic Study
 

Rats Mice
 

Group Dose Concn Survival0 Dose Concn Survival0
 

No. (mg/kg)a (mg/ml)b Males Females (mg_/kg)a (mg/ml)b Males Females
 

1 0 0 5/5 5/5 0 0 5/5 5/5
 

2 562 562 5/5 5/5 1000 100 5/5 5/5
 

3 1000 1000 5/5 5/5 1780 178 5/5 5/5
 

4 1780 1780 5/5 5/5 3160 316 5/5 3/5
 

5 3160 3160 5/5 5/5 5260 562 1/5 1/5
 

6 5620 5620 0/5 0/5 10,000 1000 0/5 0/5
 

amg Trichloroethylene/kg body weight,
 b

mg Trichloroethylene/ml corn oil.
 cAt 8 weeks.
 
Animals were weighed weekly and the most recent weight was used as a guide
 

for the dosage. All animals of one sex within a treatment group received
 

the same dosage, that is, the volume administered to all animals was based
 

on the mean body weight for the group.
 

Dosing was stopped after 6 weeks and the animals were maintained for an
 

additional 2 weeks under control conditions to detect delayed toxicity.
 

Body weight was recorded on day 0 and weekly thereafter. Food consumption
 

and observations of appearance, behavior, and signs of toxic effects were
 

recorded weekly. Observations of mortality were made daily. Each animal
 

that died and all animals killed at termination at 8 weeks were gross
 

necropsied. No histopathology was performed. Mean body weights for each
 

group, including standard deviations, and survival by week were determined
 

(Tables XIIIa, Xlllb, XlVa, and XlVb, Appendix B) .
 

4.2.2 Results - Rats
 

While body weight gains of all treated groups were below those of controls,
 

the reduction exceeded 20% for doses above 1780 for females and 3160
 

mg/kg/day for males.
 

No abnormal clinical signs were evident for doses of 1780 mg/kg/day and
 

below. Hunching, discoloration of the fur due to urine stains, alopecia,
 

and labored respiration were observed at 3160 and 5620 mg/kg/day. Kidney
 

lesions were observed in 2 males at 1780 mg/kg/day, one a dilated kidney
 

pelvis and the other a dark red renal medulla. Incidental findings
 

included large abscessed areas in all lobes of the lungs of 2 test animals.
 

No other gross lesions were noted.
 

4.2.3 Results - Mice
 

Body weight gains in all surviving groups were not significantly affected
 

in a dose-related manner. Except for death at the higher doses, there were
 

no signs attributable to the compound. All survivors appeared normal at
 

termination. No lesion was noted in any mouse at necropsy.
 

4.2.4 Selection of MTD
 

Based on body weight gains and survival rates, the initial high doses
 

(estimated maximum tolerated dose) were selected as 1300 mg/kg for both
 

male and female rats, 2000 mg/kg for male mice, and 1400 mg/kg for female
 

mice.
 

8 
5.0 CHRONIC TESTING: METHODOLOGY
 

5.1 Experimental Design
 

5.1.1 Experimental Groups
 

Trichloroethylene was administered at 2 doses to both sexes of Osborn-


Mendel rats and B6C3F1 mice, in groups of 50 animals each. Therefore, a
 

total of 400 treated animals divided into 8 groups was used. Groups of 20
 

matched-*- vehicle-treated controls were used for each sex of each species.
 

Ninety-nine male and 98 female rats and 77 male and 80 female mice were
 

used as vehicle-treated "colony controls*', and 70 male and 76 female mice
 

were used as untreated colony controls. They served as matched controls to
 

trichloroethylene and to other compounds that were tested simultaneously. A
 

group of positive control animals treated with carbon tetrachloride was
 

also studied; see section 5.6. Treated and control animals came from the
 

same source and were otherwise comparable. Animals were randomly^
 

assigned to treatment and control groups, so that initially the average
 

weight in each group was approximately the same.
 

5.1.2 Dates of Study
 

Rats receiving trichloroethylene and their controls were placed on study at
 

48 days of age on April 11, 1972 and killed after 110 weeks on May 23,
 

19 74. Mice receiving trichloroethylene and their controls were placed on
 

study at 35 days of age on August 21, 1972 and killed after 90 weeks on May
 

15, 1974.
 

5.1.3 Preparations and Doses
 

Trichloroethylene was dissolved in corn oil at concentrations of 60% w/v
 

for rats and 10-24% w/v for mice and administered by gavage for 5 consecutive days per week. The amount of solution to be administered was calculated weekly on the basis of the animal body weight, using the following
 

factor: dose (mg/kg)/concentration (mg/ml) = F (ml/kg). For instance, a
 

group of mice scheduled to receive trichloroethylene at a dose of 1000
 

mg/kg/day and weighing an average of 20 g at the end of week 2 actually
 

received 0.13 ml of a 15% solution during week 3 (1000/15 = F = 6.6 ml/kg;
 

6.6 x 20/1000 = 0.13 ml/20 g) . Later in the study when weighing was done
 

only once monthly, each newly calculated dose was administered for 4 weeks.
 

All animals of one sex within a treatment group received the same dosage,
 

that is, the volume of trichloroethylene solution administered to all
 

animals was based on the mean body weight for the group. Controls received
 

by gavage a dose of vehicle (corn oil) based on the factor calculated for
 

the high dose group.
 

*• Vehicle-treated controls will be assumed to be matched, and will not be
 

referred to as such in the remainder of this report.
 

^Animals were not distributed according to a table of random numbers.
 
5.1.4 Treatment Schedule
 

At the beginning of the chronic study, the high dose groups received the
 

estimated maximum tolerated dose as determined in the 8-week subchronic
 

study. The low dose was one-half of the high dose in all cases. In order
 

to maintain the animals at the maximum doses that could be actually
 

tolerated, body weight changes and survival were monitored, and,
 

accordingly, doses were changed for the rats after 7 and 16 weeks of
 

treatment, and for the mice after 12 weeks. To help assure survival until
 

planned termination the dosing schedule was changed for rats to a cycle of
 

1 week of no treatment followed by 4 weeks of treatment. Seventy-eight
 

weeks after the start of the test, dosing of rats and mice was stopped and
 

observation of the animals continued until the test was terminated after
 

110 weeks for rats and 90 weeks for mice. This dosing schedule is outlined
 

in Table III and Figures la and lb.
 

5.2 Observations
 

Individual body weights and food consumption per cage were recorded weekly
 

for the first 10 weeks and monthly thereafter (Tables XVa, XVb, XVIa, and
 

XVIb, and Figures 13a, 13b, 14a, and 14b, Appendix B) . Records of
 

appearance, behavior, and signs of toxic effects were maintained at the
 

same intervals as above. Animals were checked daily for mortality, and
 

moribund animals were killed.
 

5.3 Necropsy
 

A gross necropsy was performed on each animal that died or was killed and
 

on all survivors at termination. The weight of any discrete subcutaneous
 

tissue mass was recorded. The dissection of rats and mice followed the same
 

systematic technique whether the animals died or were killed. However,
 

their blood smears for microscopic evaluation were prepared from the tail
 

of each living animal immediately prior to injecting it with Diabutal^
 

intraperitoneally (0.3 to 0.5 ml for rats and 0.05 to 0.1 ml for mice).
 

After induction of a state of anesthesia, the spinal cord and blood vessels
 

at the back of the neck were severed with sharp-pointed scissors and the
 

animal was exsanguinated and immediately necropsied.
 

The cervical lymph nodes, salivary glands, and thyroid with attached
 

parathyroids, trachea, larynx, and esophagus (en bloc) were removed. The
 

eyes, brain, pituitary, and nasal turbinates were removed, examined, and
 

fixed. Thigh muscle and accompanying sciatic nerve and the femur were then
 

excised, followed by abdominal skin (with mammary gland) and subcutaneous
 

masses. The thoracic and abdominal cavities were then opened and the
 

sternum was removed by cutting through the costochondral junctions. The
 

thymus, heart (with small attached length of aorta), and lungs were
 

removed. The lungs were fixed in their entirety. The thoracic spinal cord
 

was removed. All lobes of the liver were taken including the free margin
 

of each lobe. Any nodule or mass was represented in a block 10 x 5 x 3 mm
 

cut from the liver and fixed in a marked capsule. The spleen was removed
 

with a small piece of pancreas attached. The stomach was separated from
 

the small intestines and esophagus and opened. Following removal of the
 

stomach contents, its lining was examined. The mesenteric lymph nodes were
 

10
 
Table III. Dosage and Observation Schedule - Trichloroethylene Chronic Study
 

Dosage
 

Group
 

Low dose
 

males and
 

females
 

High dose
 

males and
 

females
 

Low dose
 

males
 

High dose
 

males
 

Low dose
 

females
 

High dose
 

females
 

Dose (mg Percent of
 

TCE/kg TCE in
 

Body Wt) Corn Oil
 

650 60.0
 

750 60.0
 

500 60.0
 

500 60.0
 

no treatment
 

1300 60.0
 

1500 60.0
 

1000 60.0
 

1000 60.0
 

no treatment
 

1000 15.0
 

1000 10.0
 

1200 24.0
 

no treatment
 

2000 15.0
 

2000 20.0
 

2400 24.0
 

no treatment
 

700 10.0
 

900 18.0
 

no treatment
 

1400 10.0
 

1400 20.0
 

1800 18.0
 

no treatment
 

Age at
 

Dosinga
 

(weeks)
 

Rats
 

7
 

14
 

23
 

37
 

85
 

7
 

14
 

23
 

37
 

85
 

Mice
 

5
 

11
 

17
 

83
 

5
 

11
 

17
 

83
 

5
 

17
 

83
 

5
 

11
 

17
 

83
 

Treatment
 

Period
 

(weeks)
 

7C
 

9c
 

14C
 

48d
 

32
 

7C
 

9c
 

14c
 

48d
 

32
 

6C
 

6C
 

66C
 

12
 

6C
 

6C
 

66C
 

12
 

1 2 c
 

66C
 

12
 

6C
 

6C
 

66C
 

12
 

Time-Weighted
 

Av. Dose" (mg
 

TCE/kg Body Wt)
 

549
 

1097
 

1169
 

2339
 

869
 

1739
 

Matched controls received doses of corn oil by gavage calculated on the
 

basis of the factor for the high dose animals (see section 5.1.3).
 aAge at initial dose or dose change.
 

^Time-weighted average dose = £ (dose in mg/kg x no. of days at that
 

dose)/I (no. of days receiving any dose). In calculating the time-weighted
 

average dose, only the days an animal received a dose are considered.
 c

Dosing 5 days per week each week.
 

^Dosing 5 days per week, cycle of 1 week of no treatment followed by 4
 

weeks of treatment. (Animals were treated for 38 weeks of the 48 week
 

period.)
 

11
 
Figure la. Dosage Schedule - Chronic Study ­ Figure lb. Dosage Schedule - Chronic Study ­ Trichloroethylene-Treated Rats
 Trichloroethylene-Treated Mice
 
excised along with a small amount of surrounding fatty tissue. The
 

intestines were removed and straightened by cutting through their
 

mesenteric attachment and were examined externally for abnormalities.
 

Portions of duodenum, jejunum, ileura, and colon (about 1.5 cm long) were 

excised and placed in a capsule for fixation. If any gross lesion was 

noted from the serosal surface, the intestines were opened. After the 

selected portions were excised for separate preservation, the remaining
 

portions of intestines and cecum were fixed in the container with the bulk
 

of the tissues. The adrenals were removed with surrounding fatty tissue.
 

After the kidneys were removed and their capsular surface freed of other
 

tissues, they were bisected longitudinally and placed in tissue capsules.
 

The urinary bladder was removed, a small amount of 10% formalin was
 

injected if contracted, and it was opened slightly to examine the lining.
 

The prostate and seminal vesicles were removed together. The testes with
 

epididymides attached were fixed en bloc after the attached fat pad was
 

removed. The ovaries, uterus, and vagina were removed and fixed en. bloc.
 

The posterior 2 cm of rectum was removed with surrounding connective tissue
 

and fixed en. mass.
 

Subcutaneous and other masses were removed and weighed. Location, size
 

(dimensions or weight), color, consistency, and general appearance were
 

recorded. Representative sections, 5 mm thick, were excised and fixed. If
 

an animal had more than one mass or nodule, sections of each were placed in
 

a marked capsule. If a mass involved the head, the entire skull was fixed
 

after the cranial cavity had been opened.
 

The above protocol applied to animals necropsied after February 28, 1974,
 

when the carcasses were no longer retained. Before that time, similar
 

procedures were used, but fewer tissues were taken, and the carcasses were
 

preserved.
 

5•4 Histological Preparation and Microscopic Examination
 

The necropsy was conducted by a dissector under the supervision of a
 

pathologist. Each time a tissue was removed the presence of any gross
 

lesion was recorded. All tissues were fixed in 10% buffered formalin
 

(Appendix D, Tables XXVIIIa and XXVIIIb). The histologic technician
 

trimmed the tissues under a fume hood to a thickness of 3-6 mm; any missing
 

tissues or unusual lesions were brought to the attention of the
 

pathologist. Tissues were dehydrated, cleared, and infiltrated on an
 

Autotechnicon tissue processor, using a series of Technicon reagents (S-29
 

and UC-670) and Paraplast. Tissues were placed in a vacuum oven and
 

subsequently embedded in Paraplast, using a Tissue Tek system. All blocks
 

were sectioned at 5 microns; one slide was prepared from each block and
 

stained with hematoxylin and eosin. Special staining procedures such as
 

PAS, Trichrome, and PTAH were used occasionally as requested by the
 

pathologist to diagnose a specific tumor cell type. All paraffin blocks
 

were sealed with Paraplast and stored in plastic cabinets. Slides were
 

boxed and delivered with all necropsy forms and work sheets to the
 

pathologist at Hazleton Laboratories, Inc., for examination. Seven to 9
 

slides were prepared per mouse, and 7-10 per rat.
 

13
 
5.5 Data Processing and Confirmation
 

Summaries of the numbers of tissues examined for rats and mice are given in
 

Tables XXVIIla and XXVIIlb. The pathologist recorded his findings into a
 

dictaphone for the typist to transcribe on a computer data form, the
 

Individual Animal Data Record (IADR). Information was included on the
 

death of the animal, details of the necropsy, including physical
 

abnormalities and tumors. IADRs were transmitted to EG&G/Mason Research
 

Institute, Bethesda, MD, NCI operations contractor for the computerized
 

system for collection, maintenance, and analysis of bioassay data. This
 

system is known as the Carcinogenesis Bioassay Data System (CBDS) (Linhart
 

et al. , 1974). Diagnoses of tumors and other animal abnormalities were
 

coded using the coding system described in the "Systematized Nomenclature
 

of Pathology" (SNOP), prepared by the Committee on Nomenclature and the
 

Classification of Disease, College of American Pathologists, Chicago, 1965.
 

The SNOP code has been modified for use in the Carcinogenesis Bioassay Data
 

System. One code is entered for the topography or site of the lesion, and
 

another for the diagnosis. Output from CBDS is in the form of the
 

Individual Animal Pathology Report, which presents the complete pathology
 

data for the animals within a group, including both tumor and non-tumor
 

diagnoses. The initial output was reviewed and corrected by the
 

pathologist at Hazleton. A further review of the findings was conducted by
 

pathologists at Tracor Jitco, Inc., and the National Cancer Institute with
 

special attention given to liver lesions. The differences in opinion were
 

minor, and, in general, supported the diagnoses as presented. The final
 

data are presented in the form as seen in Tables XXXIa, XXXIb, XXXIIa, and
 

XXXIIb.
 

14
 
5.6 Positive Controls
 

In this bioassay carbon tetrachloride was administered as a positive
 

control to both rats and mice, obtained from the same source and maintained
 

under the same environmental conditions as the animals receiving trichloroethylene. Solutions were prepared and administered by gavage in the manner
 

described for trichloroethylene. Dosing was 5 times per week thoughout the
 

study according to the dosage schedule in Table IV:
 

Table IV. Dosage and Observation Schedule - Carbon Tetrachloride Study
 

Time-Weighted 

Dose Percent of Age at Treatment Av. Dose" (mg 

Dosage (mg CCl^kg CCI4 in Dosinga Period CCl4/kg 

Group Body Wt) Corn Oil (weeks) (weeks) Body Wt) 

Rats 

Low dose males 25 2.5 6 10c
 

50 5.0 16 6 8C
 

no treatment 84 32 47
 

High dose males 50 2.5 6 10c
 

100 5.0 16 68C
 

no treatment 84 32 94
 

Low dose females 100 10.0 6 i 4c
 

75 7.5 20 6 4C
 

no treatment 84 32 80
 

High dose females 14c
 200 10.0 6
 

150 7.5 20 64C
 

no treatment 84 32 159
 

Mice
 

Low dose males 1250 25.0 5 78C
 

and females no treatment 83 12 1250
 

High dose males 2500 25.0 5 78C
 

and females no treatment 83 12 2500
 

Matched controls received doses of corn oil by gavage calculated on the
 

basis of the factor for the high dose animals (see section 5.1.3).
 aAge at initial dose or dose change.
 

^Time-weighted average dose = £ (dose in mg/kg x no. of days at that
 

dose)/Z (no. of days receiving any dose). In calculating the time-weighted
 

average dose, only the days an animal received a dose are considered.
 c

Dosing 5 days per week each week.
 

15
 
6.0 CHRONIC TESTING: RESULTS - RATS
 

Sections 6.1 - 6.5 refer to trichloroethylene-treated rats and their
 

vehicle-treated controls.
 

6.1 Body Weights
 

The range of the mean body weights for male rats was 193-194 g, and for
 

female rats, 144-146 g, when placed on experiment (Tables XVa and XVb,
 

Appendix B) . Weights in male rats peaked at 622 g at 46 weeks in control,
 

575 g at 46 weeks in low dose, and 535 g at 38 weeks in high dose animals.
 

At these times there were, respectively, 20 control, 43 low dose, and 39
 

high dose animals. The average weights of surviving males in all groups
 

were much lower than this at the termination of the test at 110 weeks. At
 

this time the surviving 2 controls averaged 382 g, the 8 low dose males,
 

383 g, and the 3 high dose males, 423 g.
 

Average group weights of female rats peaked at 404 g at 62 weeks in
 

control, 322 at 70 weeks in low dose, and 326 g at 94 weeks in high dose
 

animals. At these times there were, respectively, 17 control, 23 low dose,
 

and 20 high dose animals. The average weights of female rats were also
 

lower at termination at 110 weeks. At this time, 8 surviving controls
 

averaged 326 g, and 13 low dose and 13 high dose females both averaged 311
 

g. These decreases in average weights may reflect weight losses in 

individual animals and/or a mortality pattern where larger animals died 

sooner. 

6 .2 Clinical Observations
 

During the first year of the study, the appearance and behavior of the
 

treated rats were generally comparable with the controls except that
 

occasionally hunched appearance and discoloration of the fur of the lower
 

abdomen by urine stains were noted in a few test animals as early as week
 

2. Respiratory involvement characterized by labored breathing, wheezing,
 

and/or reddish nasal discharge was noted in both treated and control
 

groups, and increased as the animals aged.
 

Adverse clinical signs in all treatment groups were noted at a low or
 

moderate incidence during the first year, and with gradually increasing
 

frequency in the treated animals during the second year of the study. These
 

signs included hunched appearance; roughening of the haircoat; eyes
 

squinted or showing a reddish discharge; localized alopecia on extremities
 

or body; sores, particularly on the tail; and stains on the haircoat.
 

6.3 Survival
 

Data for rats are given in the individual pathology tables in Appendix D.
 

The methodology for statistical analysis is described in Appendix C. Data
 

for statistical analysis are summarized in Tables XVIIIa and XVIIIb,
 

Appendix C. Tables of results are also in Appendix C.
 

A high proportion of rats died during the experiment. For males, 17/20
 

control, 42/50 low dose, and 47/50 high dose animals died prior to
 

16
 
scheduled termination. For females, 12/20 control, 35/48 low dose, and
 

37/50 high dose animals died before scheduled termination. (Two low dose
 

females were missing and were not counted in the denominator for that
 

group.)
 

Survival probabilities were estimated by the product-limit procedure of
 

Kaplan and Meier (1958) (Tables XXa and XXb and Figures 2a and 2b). The
 

estimated probabilities (standard errors) of survival to 110 weeks were
 

0.100 (0.067) for male control, 0.140 (0.049) for male low dose, 0.060
 

(0.034) for male high dose, 0.400 (0.110) for female control, 0.252 (0.061)
 

for female low dose, and 0.260 (0.062) for female high dose.
 

The survival times of vehicle control, low dose, and high dose groups of
 

rats were compared (Table XXc). Among male rats, the age-adjusted test for
 

linear trend (Tarone, 19 75) is significant at P = 0.001, and the high dose
 

vs. control test and the high dose vs. low dose test are significant at P =
 

0.001, indicating that high dose male rats died earlier than low dose and
 

control male rats and that earlier death is associated with higher dose.
 

Among female rats, the low dose group died earlier than the control group,
 

as shown by a test with P = 0.028. The high dose female rats died earlier
 

than control female rats (P = 0.049), but slightly, and not significantly,
 

later than the low dose female rats. The dose-response test, therefore,
 

shows only P = 0.117.
 

6.4 Pathology
 

A variety of neoplastic and non-neoplastic lesions were recorded among
 

control, low dose, and high dose rats. Tumors in specific organ systems by
 

site of origin and by anatomic site are summarized in Tables XXIXa (page
 

121) and XXIXb (page 124), and pathologic observations for individual
 

animals are listed in Tables XXXIa (page 135) and XXXIb (page 147) in
 

Appendix D.
 

There are no significant differences In t_he incidences o_f total j^ujnorjs or
 

of a. specific ^umoj: type .between treated and control rats. As seen in
 

Table V, there is no indication of a treatment-related effect. While the
 

percentage of tumor-bearing animals is actually lower in treated animals,
 

this is likely related to the decrease in their survival.
 

Table V. Tumor Incidence - Rats with Tumors
 

Males Females 

Low High Low High 

Control Dose Dose Control Dose Dose 

Before 110 weeks 4/17 5/42 4/47 4/12 6/35 4/37
 

At 110 weeks 1/3 2/8 1/3 3/8 6/13 8/13
 

Total 5/20 7/50 5/50 7/20 2/48 12/50
 

Percent 20% 14%- 10% 35% 25% 24%
 

17
 
TIME ON STUDY (WEEKS)
 

Figure 2a. Product-Limit Estimates of Probability of
 

Survival - Trichloroethylene-Treated Male Rats
 

18
 
Figure 2b. Product-Limit Estimates of Probability of
 

Survival - Trichloroethylene-Treated Female Rats
 

19
 
Incidences of the most common tumor types according to those dying before
 

and at 110 weeks are presented in Table XXII (page 102) in Appendix C. The
 

tumors observed are described in the following paragraphs.
 

Primary malignant renal tumors were observed in 4 rats, 2 of which were
 

unilateral malignant mixed tumors of the kidney in male controls (010,
 

019). One of these (010) also had a hamartoma in the outer medulla of the
 

opposite kidney. A similar hamartoma affected the outer renal medulla of
 

one of the low dose males (002) . The hamartomas contained an admixture of
 

adipose tissue, spindle cells, and tubular structures with ciliated
 

columnar epithelium. The malignant mixed tumors consisted primarily of
 

proliferating fat cells and spindle cells mushrooming from the renal
 

medulla through the cortex to form a large extrarenal protuberance. Mitoses
 

were abundant and frequently abnormal. The proliferating tissues were
 

invasive and contained as an integral component small tubular nests of
 

proliferating epithelial tissue. Thus, the 2 renal hamartomas and the 2
 

malignant mixed tumors contained the same cellular elements. They appeared
 

to arise in or near the renal medulla. The only other primary renal tumor
 

was an adenocarcinoma arising from nephronic epithelium in a low dose male
 

(015).
 

The relationship between the renal hamartomas (2 rats) and renal malignant
 

mixed tumors (one of the same rats and another) remains to be clarified.
 

The hamartomas clearly arose near the corticomedullary junction, the mixed
 

tumors apparently so. The similarities of site of origin and tissue
 

components suggest that they were developmentally related. The hamartoma
 

was probably congenital and may have undergone malignant transformation to
 

the malignant mixed tumor. The mixed tumor had structural similarities
 

comparable to nephroblastoma, a well known tumor in many species including
 

human beings, swine, and others. Because the mixed tumors in these older
 

rats had both epithelial and nonepithelial components and were obviously
 

highly aggressive, the term malignant mixed tumor is considered suitable.
 

Primary tumors of the thyroid were found in 5 animals. Of these, 2 were
 

benign follicular-cell adenomas found in a control male and a low dose
 

male. Three malignant follicular-cell adenocarcinomas were also found in a
 

low dose male, a high dose male, and a high dose female.
 

No pituitary or mammary tumors were seen in the male rats. Among the
 

females, however, chromophobe adenomas and a benign tumor of the pituitary
 

were found in 4/20 controls, 2/47 low dose, and 6/49 high dose groups.
 

Mammary fibroadenomas were found in 3/20 controls, 5/45 low dose, and 7/48
 

high dose female rats. Multiple mammary fibroadenomas were found in one
 

low dose female (050), and 3 high dose females (023, 030, and 033). The
 

only other primary mammary tumor was an adenocarcinoma in a control female
 

(014) . Three other tumors of the reproductive system were found, an
 

ovarian granulosa-cell carcinoma in a control female (009) and 2 sarcomas
 

of the endometrium in a low dose female (026) and a high dose female (037) .
 

The sarcomas were rather poorly differentiated and a more specific
 

classification is not considered feasible.
 

There were several other miscellaneous malignant neoplastic entities of
 

epithelial origin, each occurring in a different animal. One control male
 

20
 
(007) had a primary pulmonary carcinoma with both glandular and squamous
 

differentiation; multiple metastases with similar biphasic differentiation
 

were present in the lungs, cervical lymph node, and kidneys. This was a
 

very aggressive tumor. A low dose male (Oil) had a squamous cell carcinoma
 

in the axillary region. A high dose male (034) had a pilomatrixoma of the
 

skin and another (014) had a massive aortic body tumor at the base of the
 

heart. One low dose female (023) had an adrenal cortical carcinoma.
 

Two rats had fibromas of the subcutis, a low dose male (025) and a low dose
 

female (013). Malignant tumors included a fibrosarcoma of the subcutis in
 

a low dose male (004) and a malignant anaplastic giant-cell tumor in the
 

abdomen of a control male (002) . The latter is the only tumor of this type
 

in any rat on the study. A low dose female (032) had a subcutaneous
 

liposarcoma. Hemangiosarcomas were recognized in a control male, a low
 

dose male, 2 high dose males, and a low dose female.
 

Tumors of hematopoietic type were limited to reticulum-cell sarcoma, a
 

malignant tumor which affected 1 control female (004), 1 low dose female
 

(016), and 1 high dose female (006).
 

No non-neoplastic lesions appeared to be related to treatment with the
 

exception of renal changes. Slight to moderate degenerative and
 

regenerative tubular alterations, primarily affecting proximal tubular
 

epithelium, were common in treated rats but lacking in controls. However,
 

chronic renal disease occurred frequently among aged treated and control
 

rats. A high incidence of chronic respiratory disease was observed among
 

the rats without any apparent difference in type, severity, or morbidity as
 

to sex or group. No significant toxic hepatic changes were observed.
 

6.5 Tumor Probabilities
 

For the purpose of statistical analysis, letters, or marks, were assigned
 

to sets of pathologic diagnoses (Table XVII). Frequencies of the more
 

commonly observed marks are summarized in Table XXII, Appendix C.
 

The probabilities of observing histopathologic diagnoses among control, low
 

dose, and high dose groups of rats were estimated. See Appendix C for a
 

description of methods of estimation and statistical testing. Tests were
 

performed for reticulum-cell sarcoma, lymphosarcoma, or malignant lymphoma
 

(mark b), fibroadenoma of the mammary gland (mark g), hemangioma of any
 

site (mark h), follicular adenocarcinoma of the thyroid (mark p), and
 

chromophobe adenoma of the pituitary (mark t). Since neither hepatocellular carcinoma of the liver (mark a) nor adenoma and carcinoma of the
 

lung (mark c and d) was observed in any rat, as they were in mice, tests
 

comparing these marks were not performed. Test results are shown in Tables
 

XXIIIa-e. None of the tests demonstrates increases in the probability of
 

observing a^ tumor for dosed rats over control rats.
 

21
 
6.6 Controls
 

6.6.1 Survival
 

Table VI shows a comparison of the survival of the rats receiving carbon
 

tetrachloride and trichloroethylene with pooled colony controls at 78 and
 

110 weeks:
 

Table VI. Comparison of Survival of Colony Controls and
 

Trichloroethylene- and Carbon Tetrachloride-Treated Rats
 

Trichloroethylene Carbon Tetrachloride 

Controls Low Dose High Dose Low Dose High Dose 

Interval M F M F M F M F M F 

Initial 100 100 50 50 50 50 50 50 50 50
 

78 weeks 67 75 31 20 12 23 34 38 34 21
 

110 weeks 26 51 8 13 3 13 14 26 7 14
 

At both time periods a slightly greater number of rats survived which had
 

received carbon tetrachloride than those which had received trichloroethylene. Survival among the controls was generally greater.
 

6.6.2 Tumors
 

The incidence of both hepatocellular carcinoma and neoplastic nodule in
 

colony controls and in rats receiving carbon tetrachloride is given in
 

Table VII:
 

Table VII. Incidence of Liver Tumors ­

Colony Control and Carbon Tetrachloride-Treated Rats
 

Hepatocellular Neoplastic
 

Animal Group Carcinoma Nodule
 

Males controls 1/99 0/99
 

low dose 2/50 2/50
 

high dose 2/50 1/50
 

Females controls 0/98 2/98
 

low dose 4/49 2/49
 

high dose 1/49 3/49
 

A low incidence of both hepatocellular carcinoma and neoplastic nodule was
 

found in both colony controls and carbon tetrachloride-treated rats.
 

Neither of these lesions was found in any of the rats receiving trichloroethylene. Statistical tests show no difference between control and
 

trichloroethylene-treated animals (Tables XXIVa and XXIVb). However, the
 

comparison of observed hepatocellular carcinomas in carbon tetrachloride-


treated low dose female rats compared with pooled vehicle-treated female
 

controls is significant•by a one-tailed Fisher exact test (P = 0.011) (see
 

Table XXIVc). When observed hepatocellular carcinomas and neoplastic
 

nodules are analyzed together by the same test, there are significantly
 

22
 
more lesions among both the carbon tetrachloride-treated male rats (P =
 

0.033) and female rats (P = 0.009) than among respective male and female
 

pooled controls (see Table XXIVd). Individual animal data for carbon
 

tetrachloride-treated rats are given in Tables XXXIIIa-d, Appendix D.
 

In the carbon tetrachloride-treated rats marked hepatotoxicity with
 

resultant fibrosis, bile duct proliferation, and regeneration was observed.
 

The majority of liver nodules observed were diagnosed as being regenerative
 

rather than neoplastic. These regenerative nodules were composed of
 

hepatocytes which were generally larger and paler staining than the
 

adjacent hepatic parenchyma. These nodules were multiple and circumscribed
 

by mature fibrous connective tissues.
 

The lesions diagnosed as neoplastic nodules contained hepatocytes which
 

varied in appearance from large pale-staining cells to smaller, more
 

basophilic cells with a disorganized pattern, poorly defined sinusoids, and
 

essential absence of portal triads. The hepatocytes composing the neoplastic nodules were much more variable in appearance than those of the
 

regenerative nodules.
 

The diagnosis of hepatocellular carcinoma was based on the presence of less
 

organized architecture and more variability in the cells comprising the
 

neoplasms. Often the neoplastic cells were arranged in thickened cell
 

plates or occasionally in an acinar pattern. Several of the carcinomas had
 

a prominent vascular supply as opposed to the neoplastic and regenerative
 

nodules.
 

23
 
7.0 CHRONIC TESTING: RESULTS - MICE
 

Sections 7.1 - 7.6 refer to trichloroethylene-treated mice and their
 

vehicle-treated controls.
 

7.1 Body Weights
 

Weights of male mice averaged 17 g, and female mice, 14 g, when placed on
 

experiment (Tables XVIa and XVIb). Average weights in male mice peaked at
 

34 g at 34 weeks in all groups and average weights in female mice peaked at
 

27 g at 34 weeks. Survivors of both sexes maintained approximately these
 

respective weights until termination of the experiment after 90 weeks.
 

7.2 Clinical Observations
 

During the first year of the study, the appearance and behavior of the
 

treated and control mice were generally comparable. Alopecia (generalized
 

and/or localized), sores on the tail and other parts of the body, and a
 

hunched appearance were noted in an increasing number of mice, mostly
 

males, in all groups beginning on week 14 and persisting during the study.
 

After 50 weeks of treatment, bloating or abdominal distention was the
 

predominant observation in the high dose males. By week 74, approximately
 

50% of all treated males had a bloated appearance which persisted until
 

they died or were killed after 90 weeks. A few treated females also showed
 

abdominal distention prior to termination. Subsequent necropsy of the
 

animals confirmed the presence of liver tumors.
 

7.3 Survival
 

Data for mice are given in the pathology tables in Appendix D. Data for
 

statistical analysis are summarized in Tables XlXa and XlXb, Appendix C.
 

Tables of results are also in Appendix C.
 

Some mice died before the end of the experiment at 90 weeks from other than
 

accidental causes. For males, 12/20 controls, 14/50 low dose, and 28/50
 

high dose died before termination. For females, 0/20 controls, 8/50 low
 

dose, and 8/47 high dose died. (Three high dose females were missing and
 

were not counted in the denominator for that group.)
 

Survival probabilities were estimated by the product-limit procedure of
 

Kaplan and Meier (1958) (Tables XXIa and XXIb, Figures 3a and 3b). The
 

estimated probabilities (standard errors) of survival of mice to the end of
 

the chronic test at 90 weeks were 0.400 (0.110) for male control, 0.715
 

(0.064) for male low dose, 0.409 (0.070) for male high dose, 1.000 (0.000)
 

for female control, 0.835 (0.053) for female low dose, and 0.830 (0.055)
 

for female high dose.
 

The survival of control, low dose, and high dose groups of mice was
 

compared (Table XXIc). The age-adjusted test for linear trend (Tarone,
 

1975) among male mice is marginally significant (P = 0.096), but high dose
 

is not significantly different from control. High dose male mice lived
 

24
 
40 50 60 70 

TIME ON STUDY (WEEKS) 

Figure 3a. Product-Limit Estimates of Probability of
 

Survival - Trichloroethylene-Treated Male Mice
 

25
 
30 40 50 60 70
 

TIME ON STUDY (WEEKS)
 

Figure 3b, Product-Limit Estimates of Probability of
 

Survival - Trichloroethylene-Treated Female Mice
 

26
 
significantly shorter lives than low dose male mice (P = 0.001), but
 

control male mice also had significantly shorter lives than low dose male
 

mice (P = 0.004). Among female mice, low dose and high dose groups are not
 

significantly different, but mice in both groups had significantly shorter
 

lives than the control group (P = 0.035). The age-adjusted test for linear
 

trend yields P = 0.068, only marginally significant.
 

7.4 Pathology
 

A variety of neoplastic and non-neoplastic lesions were observed among
 

control and treated mice. Tumors in specific organ systems by site of
 

origin and by anatomic site are summarized in Tables XXXa (page 127) and
 

XXXb (page 130), and individual animal pathology is listed in Tables XXXIIa
 

(page 158) and XXXIIb (page 168) in Appendix D.
 

As presented and discussed in section 7.5, highly significant differences
 

in the incidences of primary malignant tumors of the liver, i.e., hepatocellular carcinomas, were found between treated and control grjou_ps. Hepatocellular carcinoma was observed in 1/20 control males, 26/50 low dose
 

males, 31/48 high dose males, 0/20 control females, 4/50 low dose females,
 

and 11/47 high dose females (Table XXV). Metastasis of the hepatocellular
 

carcinoma to the lung, looked for on the basis of single sections., occurred
 

in 4/50 low dose males, and in 3/48 high dose males. One control male (019)
 

with hepatocellular carcinoma died during week 72 of the study. Among the
 

low dose males the first hepatocellular carcinoma was observed in a mouse
 

(029) that died during week 81 and the first metastasis was in one (007)
 

that died during week 88. Among the high dose males the first hepatocellular carcinoma was observed in a mouse (046) that died during week 27;
 

10 mice that died on or before week 78 had hepatocellular carcinoma and the
 

first metastasis was in a mouse (035) that died during week 83. Hepatocellular carcinoma was found only in females killed at termination at 90
 

weeks. Thus, the incidence <of hepatocellular carcinoma was higher _in dosed
 

than in control mice of each sex ajid much higher in males than females. The
 

major difference between low and high dose males _is the earlier detection
 

of these tumors in high dose mice.
 

The hepatocellular carcinoma varied in size and number among the affected
 

mice. The diagnosis was based on size of neoplasm, histologic appearance,
 

and the presence of metastasis. The tumors varied from those composed of
 

well differentiated hepatocytes in a relatively uniform trabecular arrangement to rather anaplastic lesions in which mitotic figures occurred in
 

cells which varied greatly in size and tinctorial characteristics. Many of
 

the tumors were characterized by the formation of relatively discrete areas
 

of highly anaplastic cells within the tumor proper which were, in turn,
 

surrounded by relatively well differentiated neoplastic cells. In general,
 

various arrangements of hepatocellular carcinoma occurred, as described in
 

the literature, including those with an orderly cord-like arrangement of
 

neoplastic cells, those with a pseudoglandular pattern resembling adenocarcinoma, and those composed of sheets of highly anaplastic cells with
 

minimal cord or gland-like arrangement. Multiple metastatic lesions
 

were observed in the lung, including several neoplasms which were
 

differentiated and relatively benign in appearance. The morphology of
 

these tumors is illustrated in Figures 4-10.
 

27
 
Figure 4.	 Primary hepatocellular carcinoma, mouse (high dose male
 

#32). The rather well differentiated tumor of trabecular
 

pattern has a recognizable boundary (->) with the preexistent hepatic tissue. Rematoxylin and eosin, x75
 

Figure 5. Same as Figure 4, x300
 

28
29
 
Figure 6.	 Primary hepatocellular carcinoma, mouse (high dose
 

male #26). The rather anaplastic solid tumor has a
 

recognizable boundary (->) with the pre-existent
 

hepatic parenchyma. Mitoses are abundant.
 

Hematoxylin and eosin, x240
 

Figure 7.	 Primary hepatocellular carcinoma, mouse (high dose
 

male #15) with both solid trabecular pattern and
 

papillary pattern. Most cells are large and
 

resemble normal hepatocytes. There are also smaller
 

cells with less cytoplasm and smaller, more
 

basophilic nuclei. These cells usually occur in
 

clusters and are especially prominent in the larger
 

papillary structures.
 

30
31
 
Figure 8.	 Primary hepatocellular carcinoma, mouse (high dose
 

male #32). There is an area of highly anaplastic cells
 

in a papillary pattern contiguous with an area of well
 

differentiated cells in a trabecular pattern.
 

Hematoxylin and eosin, x120
 

Figure 9.	 Secondary hepatocellular carcinoma, lung, mouse (high
 

dose male #15). The metastatic hepatocellular
 

carcinoma has invaded and extended into a bronchiole.
 

The bulk of the tumor consists of rather well
 

differentiated hepatocytes but there are scattered
 

foci of smaller, more anaplastic, basophilic,
 

neoplastic cells (->) also. Hematoxylin and eosin,
 

x120
 

Figure 10. Metastatic hepatocellular carcinoma, lung, mouse (high
 

dose male #15). Well differentiated hepatocytes
 

comprise a large nodule (edge at upper left) and
 

invade perivascularly at lower center. Numerous foci
 

of smaller, more anaplastic, basophilic, neoplastic
 

cells occur in vessels (->) and alveolar capillaries,
 

and invade perivascularly. Hematoxylin and eosin, x96
 

32
33
 
In addition to hepatocellular carcinomas, malignant lymphoid tumors and
 

pulmonary tumors appeared somewhat elevated although not significantly at
 

the 0.05 level. Malignant lymphoid tumors (reticulum-cell sarcoma and
 

lymphosarcoma) were recognized in 1/20 control males, 4/47 low dose males,
 

2/45 high dose males, 1/19 control females, 5/49 low dose females, and 6/47
 

high dose females. The number of tissues involved varied among the
 

different cases.
 

Benign fibrous tumors consisted of fibroma of the subcutis in a low dose
 

male (002) and neurofibroma in skeletal muscle of the back of a high dose
 

male (044) . Malignant tumors included fibrosarcoma of the skin or subcutis
 

in 3 of 20 control males. A high dose male (034) that died after 90 weeks
 

on study had a primary fibrosarcoma within the abdominal cavity with
 

multiple metastases. Among the females, one low dose mouse (003) killed
 

terminally had uterine fibrosarcoma. A control female (008) had a highly
 

vascular osteosarcoma of the soft tissues of the back and a high dose male
 

(043) had a hemangiosarcoma of the lung.
 

Of the respiratory tumors, benign pulmonary adenomas were diagnosed in 5/50
 

low dose males, 1/48 high dose males, 3/50 low dose females, and 5/47 high
 

dose females. Alveolar adenocarcinoma, a malignant tumor, was diagnosed in
 

1/48 high dose males, 2/49 low dose females, and 2/47 high dose females.
 

The one in the high dose male (035) had metastasized to regional lymph
 

nodes, periaortic tissues, and the skin of the chest.
 

Other benign tumors included adenoma of the Harderian gland in 1 low dose
 

male, 1 low dose female, and 2 high dose females. An adenoma of tubular
 

origin was found in the kidney of a high dose male (013), and an ovarian
 

cystadenoma in a low dose female (043) . Other carcinomas included an
 

endometrial adenocarcinoma in a control female (003), an ovarian granulosacell carcinoma in a low dose female (033), and a mammary adenocarcinoma in
 

a low dose female (038).
 

7.5 Tumor Probabilities
 

See Appendix C for a description of the statistical tests and tables of the
 

results.
 

Frequencies of the more commonly observed marks are summarized in Table
 

XXV. Estimated probabilities of observing histopathologic diagnosis among
 

control, low dose, and high dose groups of mice were compared. Tests were
 

conducted for hepatocellular carcinoma (mark a), reticulum-cell sarcoma,
 

lymphosarcoma, or malignant lymphoma (mark b) , carcinoma or adenocarcinoma
 

of the lung or alveoli (mark c), adenoma of the lung (mark d), and
 

carcinoma, adenocarcinoma, or adenoma of the lung (mark c or d) . Test
 

results are shown in Tables XXVIa-e. No mouse was observed to have
 

fibroadenoma of the mammary gland (mark g) or chromophobe adenoma of the
 

pituitary (mark t), so tests comparing these marks were not performed as
 

they had been for the rats.
 

34
 
The age-adjusted tests for linear trend (Tarone, 1975) were highly
 

significant for hepatocellular carcinoma in male mice (P < 0.001) and
 

female mice (P = 0.002). The tests of high dose vs. control are also
 

highly significant in both male mice (P < 0.001) and female mice (P =
 

0.008). A similar test of low dose vs. control shows significant
 

differences for male mice (P = 0.004) and female mice (P = 0.090). Estimated probabilities of observing hepatocellular carcinoma in male mice,
 

depending on the week on study in which they died, are displayed in Table
 

IX and are graphed in Figure 11. In male mice, the tumors were observed
 

earlier in the high dose group than in the low dose group, which may result
 

from earlier mortality among high dose male mice. At the end of the study
 

at 90 weeks, the estimated probability (standard error) of observing
 

hepatocellular carcinoma in male mice was 0.938 (0.043) for high dose,
 

0.683 (0.076) for low dose, and 0.077 (0.074) for control. In female mice,
 

no tumors were observed before terminal sacrifice at 90 weeks; therefore,
 

no graphic presentation is included. At 90 weeks the estimated
 

probabilities (standard errors) of observing hepatocellular carcinoma in
 

female mice were 0.282 (0.072) for the high dose, 0.100 (0.047) for the low
 

dose, and 0.000 (0.000) for the controls. Figure 12 shows a comparison of
 

the percentage of animals of either sex with observed hepatocellular
 

carcinoma. These data are summarized in Table VIII:
 

Table VIII. Incidence of Hepatocellular
 

Carcinoma - Trichloroethylene-Treated Mice
 

Males Females
 

Controls 1/20 0/20
 

Low dose 26/50 (P = 0.004) 4/50 (P = 0.090)
 

High dose 31/48 (P < 0.001) 11/47 (P = 0.008)
 

Test for significant difference of each group
 

from controls. In addition, age-adjusted tests
 

for linear trend were also highly significant for
 

male mice (P < 0.001) and female mice (P = 0.002).
 

None of the tests of other marks showed significance at levels of 0.05 or
 

less. However, the age-adjusted tests for linear trend for carcinoma or
 

adenocarcinoma of the lung (mark c) may indicate a relationship with treatment at a level P = 0.109 for male mice and P = 0.225 for female mice. Similarly, the comparison of mark b, sarcoma, lymphosarcoma, and lymphoma,
 

between high dose and control female mice may indicate a relationship (P =
 

0.172).
 

7.6 Colony Controls
 

The incidence of hepatocellular carcinomas for other control groups maintained in the same room as trichloroethylene is given in Table X. Groups
 

A-D, which include the trichloroethylene-matched control group, were
 

treated with corn oil. The incidence of hepatocellular carcinomas in the
 

trichloroethylene-matched control group was typical of that observed in
 

35
 
Table IX. Estimated Probabilities of Observing Hepatocellular
 

Carcinoma - Trichloroethylene-Treated Male Mice
 

Control Low Dose High Dose 

j n n' P j n n' P j n n' P 

0 

32 

20 

20 

20 

20 

.000 

.000 

0 

16 

50 

50 

50 

50 

.000 

.000 

0 

13 

48 

48 

48 

48 

.000 

.000 

39 19 19 .000 18 49 49 .000 14 47 47 .000 

60 18 18 .000 31 48 48 .000 15 46 46 .000 

61 17 17 .000 51 47 47 .000 24 44 44 .000 

64 16 16 .000 53 46 46 .000 25 42 42 .000 

66 15 15 .000 58 45 45 .000 27 40 39 .025 

68 14 14 .000 63 44 44 .000 28 39 38 .050 

72 13 12 .077 65 43 43 .000 29 37 37 .050 

76 12 12 .077 77 41 41 .000 30 36 35 .076 

77 10 10 .077 81 40 39 .025 36 34 33 .104 

78 9 9 .077 86 38 37 .051 42 33 33 .104 

90 8 8 .077 88 37 36 .076 53 32 31 .132 

90 35 12 .683 60 31 31 .132 

j = Week on study 61 

70 

29 

28 

28 

27 

.162 

.191 n = No, of animals 71 27 26 .221 

alive at beginning 74 26 25 .251 

of the week 75 25 25 .251 

78 24 23 .283 n' = No. of animals 83 23 22 .314 

without observed 88 22 21 .345 

tumor at the end 

of the week 

90 21 2 .938 

P = Kaplan-Meier estimate of the 

probability of 

observed tumor 
No comparable figure is presented
 

for female mice because no tumors
 

were observed before terminal
 

sacrifice at 90 weeks.
 

TIME ON STUDY (WEEKS)
 

Figure 11. Product-Limit Estimates of Probability of Observing
 

Hepatocellular Carcinoma - Trichloroethylene-Treated
 

Male Mice 

37 
Figure 12. Comparison of Incidence of Hepatocellular Carcinoma
 

in Trichloroethylene-Treated Male and Female Mice
 
other control groups, thus the significance of heptocellular carcinoma in
 

trichloroethylene-treated mice remained similar when compared with the
 

pooled colony controls.
 

Table X. Incidence of Hepatocellular Carcinoma - Colony Control Mice
 

Control Median Males Females
 

Group Birth Date Vehicle Untreated Vehicle Untreated
 

Aa 7-17-72 1/20 0/20
 

B 3-20-72 1/18 0/20
 

C 5/31/72 1/19 1/20
 

D 10/12/72 2/20 0/20
 

E 3/27/72 1/19 0/19
 

F 6/15/72 2/18 0/19
 

G 10/27/72 2/18 2/20
 

H 10/04/72 0/15 0/18
 

Total 5/77 5/70 1/80 2/76
 

a

Matched controls for trichloroethylene.
 

7.7 Positive Controls
 

Few mice receiving carbon tetrachloride survived until the planned termination of the test, compared with a considerable number in each of the
 

trichloroethylene-treated groups as shown in Table XI:
 

Table XI. Comparison of Survival in Colony Controls - Vehicle-Treated and
 

Trichloroethylene- and Carbon Tetrachloride-Treated Mice
 

Trichloroethylene Carbon Tetrachloride 

Controls Low Dose High Dose Low Dose High Dose 

Interval M F M F M F M F M F 

Initial 77 80 50 50 48 47 50 50 50 50
 

78 weeks 53 71 40 41 23 21 11 10 2 4
 

90 weeks 38 65 35 40 40 39 0 0 0 1
 

Hepatocellular carcinomas were found in practically all mice receiving
 

carbon tetrachloride, including those dying before termination of the test.
 

The incidence of liver tumors was considerably greater in carbon tetrachloride-treated mice than in trichloroethylene-treated mice as shown in
 

Table Xlla. Individual animal data for carbon tetrachloride-treated mice
 

are given in Tables XXXIVa-d, Appendix D.
 

39
 
Table Xlla. Comparison of Hepatocellular Carcinoma Incidence
 

in Colony Controls - Vehicle-Treated and
 

Trichloroethylene- and Carbon Tetrachloride-Treated Mice
 

Trichloroethylene Carbon Tetrachloride
 

Animals Controls Low Dose High Dose Low Dose High Dose
 

Males 5/11 26/50 31/48 49/49 47/48
 

Females 1/80 4/50 11/47 40/40 43/45
 

These liver tumors in carbon tetrachloride-treated mice varied greatly in
 

appearance from lesions which contained well differentiated hepatic cells
 

that had a relatively uniform arrangement of the cords to very anaplastic
 

liver cells having large hyperchromatic nuclei, often with inclusion
 

bodies, and with vacuolated, pale cytoplasm. Arrangement of the neoplastic
 

liver cells varied from short stubby cords to nests of hepatic cells and
 

occasionally acinar arrangements. Mitotic figures were often present. Some
 

of the tumors were characterized by discrete areas of highly anaplastic
 

cells surrounded by relatively well differentiated tumor cells. The
 

neoplasms occurring in the treated mice were similar in appearance to those
 

noted in the trichloroethylene-treated mice.
 

The test week at which the first animal died in which a hepatocellular
 

carcinoma was observed in each group is given in Table Xllb:
 

Table Xllb. Comparison of Time (weeks) to Liver Tumor Detection
 

in Colony Controls - Vehicle-Treated and Trichloroethyleneand Carbon Tetrachloride-Treated Mice
 

Trichloroethylene Carbon Tetrachloride
 

Animals Controls Low Dose High Dose Low Dose High Dose
 

Males 72 81 27 48 26
 

Females 90 90 91 16 19
 

In addition to the higher incidence, hepatocellular carcinomas were
 

observed much earlier in carbon tetrachloride-treated mice than in the
 

trichloroethylene-treated mice. Tumors in control mice were observed much
 

later.
 

40
 
8.0 DISCUSSION
 

Trichloroethylene is one among a series of halogenated chemicals tested in
 

the National Cancer Institute carcinogenesis bioassay program. The results
 

clearly indicate that exposure to trichloroethylene has resulted in 

hepatocarcinogenicity in both sexes of the B6C3F1 mouse. No evidence of 

such activity was apparent in the rat tests. 

8•1 Design of Bioassay
 

8.1.1 Selection of Animal Species and Strain
 

Since it is well known that species vary in their carcinogenesis response
 

to chemicals, two species are routinely used in the carcinogenesis bioassay
 

program to decrease the possibility of false negative results based on
 

tests in a resistant strain of animals. A clearly positive result in only
 

one test species is a valid indication of the carcinogenic activity of the
 

test compound while failure to detect carcinogenicity in another species
 

only indicates a lower susceptibility of the latter under the conditions of
 

the test.
 

Rats and mice were selected as the test species because they are genetically standardized, readily available species that reproduce well and can be
 

easily, successfully, and economically maintained. They have been the most
 

extensively used species for carcinogenesis testing.
 

The Osborne-Mendel strain of rats was chosen because of the experience
 

gained by the Food and Drug Administration where the strain was used for
 

many years as a general purpose test animal. In addition, it had been
 

found by Reuber and Glover (1970) to be sensitive to the carcinogenicity of
 

carbon tetrachloride by subcutaneous administration. The B6C3F1 strain of
 

mice, an Fl hybrid cross of the C57BL/6 female with the C3H/He male, was
 

chosen because it had been extensively and satisfactorily used in the NCI
 

carcinogenesis bioassays.
 

8.1.2 Route of Exposure
 

Trichloroethylene was administered by gavage in this study while the main
 

human exposures are by inhalation of vapors and by ingestion through
 

contaminated water and food products. The selected route of exposure is
 

considered relevant to all modes of human exposure because trichloroethylene is readily absorbed and distributed to all organs following
 

ingestion or inhalation. This has been shown in several species, including
 

the rabbit (Gasq, 1936), the dog (Barrett and Johnston, 1939), and the
 

guinea pig (Fabre and Truhaut, 1952). The fact that in this study tumor
 

induction occurred in the liver indicates that trichloroethylene was
 

absorbed and that systemic exposure of tissues occurred.
 

41
 
8.1.3 Selection of Doses and Duration of Exposure
 

An attempt was made to select the highest dose that could be administered
 

for most of the animal's lifetime without altering the animal's normal
 

longevity from effects other than carcinogenicity (estimated maximum
 

tolerated dose). A lower dose, corresponding to one half of the high dose,
 

was also tested to assure that at least one group would survive well
 

through the test period if unpredicted toxic effects occurred in the high
 

dose group.
 

The selection of doses for bioassay is made to assure the greatest
 

probability of detecting a carcinogenic effect within an experimental
 

protocol which must use relatively small numbers of animals compared with
 

the large number of human beings exposed.
 

8.1.4 Methodology
 

The protocol and methodology of this bioassay generally conformed to
 

recognized methodology for assessing carcinogenicity as adopted by the NCI.
 

While the NCI has revised in several respects its test procedures since the
 

time the present tests were designed (Guidelines for Carcinogenesis
 

Bioassay in Small Rodents, 1975), these changes do not reduce the meaningfulness of this study. The major changes in current NCI protocols as
 

compared with the test procedures used in this study are: (a) More complete
 

and longer prechronic studies are carried out in an effort to predict the
 

MTD more accurately. Prechronic studies are now routinely conducted for 90
 

days. Histopathological studies are performed on animals from all dose
 

groups, necessary to establish a maximum tolerated dose. Clinicopathological tests are also performed when indicated. Experience has shown that
 

more extensive prechronic tests reduce or eliminate the need to change dose
 

levels during the course of the chronic study. In the past, survival and
 

slight decrease in weight gain were used as the main indicators of the MTD.
 

(b) Larger matched control groups are now also included with each test so
 

that pooling of controls from several compounds is not necessary. In this
 

bioassay 20 matched controls for each sex and species were started. However, 18 chemicals in the case of the mice and 5 chemicals in the case of
 

rats were tested concurrently, and pooling of the controls for the various
 

chemicals, in order to have a larger number of colony control animals, was
 

anticipated.
 

It has been recommended by several panels on carcinogenesis testing
 

(Berenblum, 1969; FDA Advisory Committee, 1971; Golberg, 1974) that
 

administration of a food additive in a carcinogenesis test might begin
 

prior to conception and continue in the offspring because such treatment
 

would provide a more thorough examination of the carcinogenic potential of
 

test compounds. The present bioassay makes no attempt to measure either
 

transplacental or neonatal carcinogenic effects.
 

Commercial laboratory diets were fed to the test animals. Recommendations
 

have been made to use semi-synthetic diets of known and constant composition as a step toward uniformity in interpretation of experimental results
 

from different laboratories and to decrease contamination with mycotoxins,
 

pesticides, or other agents. However, problems in preparation and storage,
 

42
 
nutritional adequacy, palatability, and handling have precluded so far
 

their adoption for practical use in these large scale lifetime studies.
 

8.2 Test Compound Purity
 

The purity of the trichloroethylene used in the chronic bioassay was
 

greater than 99%, as determined by gas chromatographic total area data and
 

infrared spectra (see Appendix A) . In the gas chromatographic analysis,
 

the percentage of each component was determined from the relative area of
 

its gas chromatographic peak. The sum of the area comprising the peaks was
 

considered to total 100%. In the final analytical work, in which the minor
 

components were actually identified, standard samples of each component
 

were used to correct for differences in gas chromatographic detector
 

response. Nonvolatile materials, such as polymers and inorganic salts,
 

would not have been detected by gas chromatography but there is no reason
 

to suspect their presence. Any significant quantity of such impurities
 

would have been detected in the infrared spectra, which, in fact, compare
 

well with reference trichloroethylene spectra.
 

Inhibitors and stabilizers are commonly added to trichloroethylene used for
 

vapor degreasing. Some of the trace components, identified in Batch #4 by
 

a combination of gas chromatography and mass spectrometry in subsequent
 

analyses, are 1,2-epoxybutane (0.19%), ethyl acetate (0.04%), epichlorohydrin (0.09%), N-methylpyrrole (0.02%), and diisobutylene (0.03%).
 

According to a major manufacturer, these are typical inhibitors in
 

commercial formulations of trichloroethylene used for vapor degreasing.
 

It is reported that trichloroethylene in the mid-1960s contained
 

substantial quantities of 1,1,2,2-tetrachloroethane, a highly toxic
 

precursor in the acetylene-based manufacturing process, but current
 

technology and the switch to ethylene-based processes now result in a
 

highly pure commercial product (NIOSH, 1973). That conclusion is
 

consistent with the analytical data on the batches used for the bioassay.
 

Analytical work on bioassay Batch #4 confirmed the absence of detectable
 

quantities of 1,1,2,2,-tetrachloroethane (<5 ppm) and 1,1,1,2-tetrachloroethane (<2 ppm).
 

While the results obtained in the present bioassay could possibly have been
 

influenced by an impurity in the trichloroethylene used, the extremely low
 

amounts of impurities found make this improbable.
 

8.3 Metabolism, Distribution, and Excretion
 

The metabolism, distribution, and excretion of trichloroethylene in humans,
 

rats, and mice was reviewed in Criteria for a Recommended Standard...
 

Occupational Exposure to Trichloroethylene (1973), issued by the National
 

Institute for Occupational Safety and Health.
 

The data describing evidence for the metabolic pathways of trichloroethylene in mice are not as extensive as that available for man and rats.
 

However, the data do indicate similar metabolic pathways for this compound
 

in the 3 species and do not support the concept that metabolic differences
 

among these species might explain the lack of carcinogenic response of the
 

43
 
rat to trichloroethylene as compared with the mouse. Recent data on
 

trichloroethylene metabolism in man are consistent with earlier findings in
 

experimental animals (Ertle et a1., 1972; Ikeda et al., 1971, 1972; Ikeda
 

and Ohtsuji, 1972; Ikeda and Inamura, 1973; Kimmerle and Eben, 1973;
 

Nomiyama and Nomiyama, 1971; Parkhouse, 1969; Soucek and Vlachova, I960;
 

Stewart and Dodd, 1964; Vignoli et al., 1970).
 

8.4 Toxicology
 

Depression of the central nervous system is the primary acute toxic effect
 

of trichloroethylene in both humans and animals. This effect was
 

demonstrated in rats by Adams et al. (1951) in short term toxicity studies.
 

In humans, nausea and vomiting, headache, vertigo, dizziness, tinnitis,
 

unsteady walk, fatigue, sleepiness, and even excitement have all been
 

reported in patients and in those persons inadvertently or occupationally
 

exposed. This and other acute and subchronic toxicity of the compound are
 

extensively reviewed in Criteria for a Recommended Standard...Occupational
 

Exposure to Trichloroethylene (1973) issued by the National Institute for
 

Occupational Safety and Health.
 

Although data on chronic or carcinogenesis studies were not available in
 

the published literature prior to this report, data from subchronic studies
 

indicated that the liver and kidney were target sites of the toxic effects
 

of trichloroethylene.
 

The effects of this chemical on liver and kidney were noted by Adams et al.
 

(1951) in subchronic inhalation studies using rats as an experimental
 

model. Liver and kidney weights of both male and female rats were increased
 

but no histopathological abnormalities were reported. Increased liver
 

weights were also observed in both guinea pigs and rabbits. In mice, trichloroethylene is not as nephrotoxic as several other chlorinated methane,
 

ethane, and ethylene derivatives (Plaa and Larson, 1965). However, the
 

hepatotoxicity of trichloroethylene has been reported by several authors.
 

It is much less severe than that of carbon tetrachloride, an intensively
 

studied chlorinated methane that was used as a positive control in this
 

bioassay. In a review of trichloroethylene von Oettingen (1955) described
 

hepatic damage which was reported as early as 1932 in dogs. He also quotes
 

Fiessinger and Laur in 1936 as observing "centrolobar degeneration and
 

vacuolization, and in some animals, a picture similar to yellow atrophy of
 

the liver". Clinical dysfunction of the liver in dogs was reported by
 

Seifter (1944). Mice have shown hepatic dysfunction when exposed to
 

trichloroethylene by inhalation for up to 8 weeks (Kylin et al. , 1965) or
 

by intraperitoneal injection (Klaassen and Plaa, 1966). In other studies,
 

the hepatotoxicity of trichloroethylene was enhanced by pretreatment with
 

acetone or isopropyl alcohol (Traiger and Plaa, 1974).
 

Thus, hepatic damage of varying degrees has been found following repeated
 

exposures of short duration to trichloroethylene. These observations
 

establish the need for chronic studies.
 

44
 
8.5 Pathology and Survival
 

8.5.1 Rats
 

In the bioassay various neoplastic entities, both benign and malignant,
 

occurred in all rat dosage groups and controls. Some are rather rare types
 

of tumors in rats, such as pilomatrixoma of skin and aortic body tumor, but
 

none was significantly related to treatment with trichloroethylene. No
 

toxic hepatic change nor primary hepatic tumor was observed in the rats.
 

Chronic respiratory disease occurred without regard to sex or treatment
 

group in rats. The only treatment-related lesion was a chronic nephropathy
 

encountered among most rats of both sexes and at both high and low doses of
 

the compound. This nephropathy was characterized by degenerative and
 

regenerative changes in tubular epithelium with questionable interstitial
 

response. The lesion is dissimilar from the chronic nephropathy so
 

commonly encountered in rats with advancing age and recognized in some rats
 

on this experiment. No treatment-related lesions severe enough to appear
 

responsible for death were detected in the rats. Nevertheless, decreased
 

survival was generally dose-related.
 

Only a low incidence of both neoplastic nodule and hepatocellular carcinoma
 

was observed in rats receiving carbon tetrachloride, the positive control
 

compound. Survival was slightly better among the carbon tetrachloride-


treated than among the trichloroethylene-treated rats.
 

In this bioassay as in other previous carcinogenic bioassays of chlorinated
 

organic compounds, some chemicals have been identified as liver carcinogens
 

in the mouse, but have produced no observed carcinogenic effect in the rat.
 

The difference in susceptibility to the induction of tumors by certain
 

chemicals in the rat as compared to the mouse may be attributed to many
 

factors and is beyond the scope of these studies.
 

8.5.2 Mice
 

In the mice, there was a significant increase in the incidence of hepatocellular carcinomas in both low and high dose males and high dose females
 

(P < 0.05) and low dose females (P = 0.09). The tumors were found in 12 of
 

27 mice dying during the experiment and in 19 of 21 high dose male mice
 

killed at termination of the experiment. Low dose male mice as well as
 

high and low dose female mice had good survival rates. With the exception
 

of 3 low dose male mice dying at 81 to 88 weeks, hepatocellular carcinoma
 

was found only in those animals killed at the end of the experiment. The
 

lower probability of tumor observation in female mice may reflect either
 

the lower doses they received or an actual sex difference in response.
 

Thus, trichloroethylene was found to have a dose-related carcinogenic
 

effect on the liver of both sexes of mice. No other tumor was related to
 

treatment.
 

Only 20 matched vehicle control mice were started for each species and sex
 

in this bioassay; however, the significance of hepatocellular carcinoma in
 

trichloroethylene-treated mice remains similar when compared with data from
 

control mice for other halogenated solvents tested concurrently.
 

45
 
In this study the doses of trichloroethylene administered to the test
 

animals resulted in a greater survival of mice than of rats. The increased
 

mortality in rats may in part have resulted from intercurrent disease as
 

well as from a greater sensitivity to the toxic effects of trichloroethylene.
 

In the positive control group, hepatocellular carcinoma was observed in
 

practically all mice receiving carbon tetrachloride. Only one of these
 

animals survived until planned termination of the test. Except for high
 

dose male mice, hepatocellular carcinomas were observed at an earlier age
 

among carbon tetrachloride-treated than among trichloroethylene-treated
 

mice. Thus, carbon tetrachloride was a much stronger hepatocarcinogen than
 

was trichloroethylene, under the conditions of these tests.
 

8.6 Effect of Various Compounds in the Same Room
 

Several halogenated solvents were being tested simultaneously in the same
 

room, i. e., 5 in the room housing rats and 18 in the room housing mice.
 

This is not expected to change the results significantly; however, no
 

experimental studies of cross contamination or simultaneous administration
 

are available. A protective effect from simultaneous exposure to other,
 

and halogenated solvents would not be expected, and it is highly unlikely
 

that an interaction of possible airborne contaminant amounts of solvents
 

would bring about false positives, considering the high doses of trichloroethylene used.
 

The species in which tumors were found, i.e., mice, were housed in a room
 

where 17 other chemicals were being tested; however, stringent precautions
 

against cross contamination were employed. The mice were kept in cages
 

with filter tops which limit the amount of expired chemical in the air
 

available for inhalation by other animals, the total air in each room was
 

changed 10 to 15 times per hour, and the mouse racks were transported to
 

another room with a large hood for the daily intubations. Furthermore, the
 

hepatocarcinomas in mice were present at a greater than P = 0.01 level of
 

significance and were produced by doses of trichloroethylene of 700 to 2400
 

mg/kg, which are several thousand-fold greater than any possible contamination could have been. A dose-related effect was observed and, any
 

possible chemical in the general room air did not affect controls. Thus,
 

although this room arrangement is not desirable as is stated in the
 

Guidelines for Carcinogen Bioassay in Small Rodents, there is no evidence
 

the results would have been different with a single compound in a room.
 

8.7 Relationship to the Toxicity of Carbon Tetrachloride
 

Carbon tetrachloride was used as a positive control because of its
 

demonstrated ability to produce liver tumors in rats, hamsters, and mice
 

(Reuber and Glover, 1970, Delia Porta et al., 1961, Eschenbrenner and
 

Miller, 1946). The doses used were approximately 10-fold less than for
 

trichloroethylene in rats, but were only slightly higher for males and 50%
 

higher for females than for trichloroethylene in mice.
 

Pathology of the liver was evident in carbon tetrachloride-treated rats.
 

Hepatocellular carcinomas and neoplastic nodules were found in a few rats
 

46
 
of both dose groups and sexes in contrast to the results with

trichloroethylene where no tumors and very little non-tumor pathology of

the liver were reported. This is particularly significant since the dose

of trichloroethylene used was approximately 10 times greater than for

carbon tetrachloride.

Hepatocellular carcinomas were found in practically all mice including

those dying before termination of the test. The incidence was considerably

greater than for trichloroethylene-treated mice (see section 7.7).

Although in mice the MTD values of carbon tetrachloride and trichloroethylene were similar, all except one carbon tetrachloride-treated animal

(of both sex and dose groups) died prior to termination at 90 weeks. The

survival of the trichloroethylene-treated female mice was excellent, and

even 40% of the high dose males survived until termination of the test.

Death in the carbon tetrachloride-treated animals could have resulted

either from toxicity or carcinogenicity since tumors were observed in

practically all animals. These results confirm previous work. Both

Klaassen and Plaa (1966) and Gehring (1968) have shown that in mice the

hepatotoxicity of carbon tetrachloride is much greater than that of

trichloroethylene, both on an absolute basis and in relation to anesthetic

effects and to the LD5 0 value.

8.8 Conclusions

The administration of trichloroethylene under the experimental conditions

described in this report induced a high incidence of hepatocellular

carcinoma in B6C3F1 mice of both sexes. The test in rats is inconclusive:

large numbers of rats died prior to planned termination; in addition, the

response of this rat strain to the hepatocarcinogenicity of the positive

control compound, carbon tetrachloride, appeared relatively low. Although

direct extrapolation to man is not possible, the identification, using this

methodology, of trichloroethylene as a carcinogen in animals serves as a

warning of its possible carcinogenicity in humans.

47
9.0 BIBLIOGRAPHY
 

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(1951) Vapor Toxicity of Trichloroethylene Determined by Experiments
 

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Armitage, P. (1971) Statistical Methods in Medical Research, Chapter 14,
 

John Wiley and Sons, New York, NY.
 

Barrett, H. M. ; Johnston, J. H. (1939) The Fate of Trichloroethylene in the
 

Organism, J^ Biol^ ChenK_ 127:765-770.
 

Berenblum, I. (Ed.) (1969) Carcinogenicity Testing; A Report of the Panel
 

on Carcinogenicity of the Cancer Research Commission of the UICC, UICC
 

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

Geneva.
 

Chemical Economics Handbook (1972) Chemical Information Services, Stanford
 

Research Institute, Menlo Park, CA.
 

Christensen, H. E. ; Luginbyhl, T. T. (Eds.) (1974) The Toxic Substances
 

List, 1974 Ed., USDHEW, National Institute for Occupational Safety and
 

Health, Rockville, MD.
 

Cochran, W. G. (1954) Some Methods for Strengthening the Common Chi-square
 

Tests, Biometrics jLO:417-454.
 

Cox, D. R. (1972) Regression Models and Life-Tables (with discussion), J^
 

Roy. Statist. Soc. B34:184-220.
 

Deichmann, W. B.; Gerarde, H. W. (1969) Toxicology of Drugs and Chemicals,
 

Academic Press, New York, NY.
 

Delia Porta, G.; Terracini, B; Shubik, P. (1961) Induction with Carbon
 

Tetrachloride of Liver Cell Carcinomas in Hamsters, JN_ Natl. Cancer
 

Inst. 2^6:855-863.
 

Dowty, B. ; Carlisle, D.; Laseter, J. L.; Storer, J. (1975) Halogenated
 

Hydrocarbons in New Orleans Drinking Water and Plasma, Science
 

187:75-77.
 

Dreisbach, R. H. (1974) Handbook of Poisoning: Diagnosis and Treatment,
 

8th Ed., Lange Medical Publications, Los Altos, CA.
 

Ertle, T.; Henschler, D.; Mueller, G.; Spassowski, M. (1972) Metabolism of
 

Trichloroethylene in Man - I. The Significance of Trichloroethanol in
 

Long-term Exposure Conditions, Arch. Toxicol. 29:171-188.
 

48
 
Eschenbrenner, A. B.; Miller, E. (1944) Studies on Hepatomas. I. Size and
 

Spacing of Multiple Doses in the Induction of Carbon Tetrachloride
 

Hepatomas, J^ Natl^ Cancer Inst. 4:385-388.
 

Eschenbrenner, A. B.; Miller, E. (1946) Liver Necrosis and the Induction of
 

Carbon Tetrachloride Hepatomas in Strain A Mice, J^ Natl. Cancer Inst,
 

6:325-341.
 

Fabre, R. ; Truhaut, R. (1952) Contribution a I/etude de la Toxicologie du
 

Trichlorethylene - II. Resultats des Etudes Experimentales chez
 

L'animal, Brit. J^ Ind._ Med. 9^:39-43.
 

Food and Drug Administration Advisory Committee on Protocols for Safety
 

Evaluation: Panel on Carcinogenesis Report on Cancer Testing in the
 

Safety Evaluation of Food Additives and Pesticides (1971) Toxicol.
 

Appl. Pharma^ol^ 2^:419-438.
 

Food and Drug Administration (1974) Code of Federal Regulations, Title 21,
 

121.1041.
 

Frear, D. E. H. (Ed.) (1969) Pesticide Index, 4th Ed., College_Science
 

Publishers, State College, PA, p. 377.
 

Gasq, M. (1936) Etudes Toxicologie et Hygienic des Solvents Chlores
 

Acethyleniques, Delmar, Bordeaux.
 

Gehring, P. J. (1968) Hepatotoxic Potency of Various Chlorinated Hydrocarbon Vapors Relative to their Narcotic and Lethal Potencies in Mice,
 

Toxicol. Appl. Pharmacol. U:287-298.
 

Golberg, L. (1974) Carcinogenesis Testing of Chemicals, Proceedings,
 

Conference on Carcinogenesis Testing in the Development of New Drugs,
 

May 23-25, 1973, Washington, DC, CRC Press, Inc., Cleveland, OH.
 

Ikeda, M. ; Imamura, T. (1973) Biological Half-life of Trichloroethylene and
 

Tetrachloroethylene in Human Subjects, Int. AL^L L Arbetsmed.
 

31--2Q9-224.
 

Ikeda, M.; Ohtsuji, H. (1972) A Comparative Study of the Excretion of
 

Fujiwara Reaction-Positive Substances in Urine of Humans and Rodents
 

Given Trichloro- and Tetrachloro-Deri'vatives of Ethane and Ethylene,
 

Brit. J^ Ind. Med. 29:99-104.
 

Ikeda, M.; Ohtsuji, H.; Imamura, T.; Komoike, Y. (1972) Urinary Excretion
 

of Total Trichloro-Compounds, Trichloroethanol and Trichloroacetic
 

Acid as a Measure of Exposure to Trichloroethylene and Tetrachloroethylene, Bjrit^ J^_ Ind. Med. 29:328-333.
 

49
 
Ikeda, M.; Ohtsuji, H.; Kawai, H.; Kuniyoshi, M. (1971) Excretion Kinetics 

of Urinary Metabolites in a Patient Addicted to Trichloroethylene, 

Brit. J^ Ind. Med^ 28:203-206. 

International Labour Office (1972) Encyclopedia of Occupational Health and
 

Safety, Vol. II, McGraw-Hill, New York, NY.
 

Kaplan, E. L.; Meier, P. (1958) Nonparametric Estimation from Incomplete
 

Observations, J^ Amer^ Statist. Ass. 53:457-481.
 

Kimmerle, G.; Eben, A. (1973) Metabolism, Excretion and Toxicology of Trichloroethylene after Inhalation, Arch. Toxicol. 30:127-138.
 

Klaassen, C. D.; Plaa, G. L. (1969) Comparison of the Biochemical
 

Alterations Elicited in Livers from Rats Treated with Carbon
 

Tetrachloride, Chloroform, 1,1,2-Trichloroethane, and 1,1,1-Trichloroethane, Biochem. Pharmacol. 18:2019-2027.
 

Klaassen, C. D.; Plaa, G. L. (1966) Relative Effect of Various Chlorinated
 

Hydrocarbons on Liver and Kidney Function in Mice, Toxicol. Appl.
 

Pharmacol. 9^:139-151.
 

Kylin, B. ; Sumegi, I.; Yllner, S. (1965) Hepatotoxicity of Inhaled Trichloroethylene and Tetrachloroethylene, Long-term Exposure, Acta
 

Pharmacol. Toxicol. 2^2:379-385.
 

Linhart, M. S.; Cooper, J.; Martin, R. L.; Page, N.; Peters, J. (1974)
 

Carcinogenesis Bioassay Data System, Computers Biomed. Research
 

7_:230-248.
 

Mantel, N. (1963) Chi-square Tests with One Degree of Freedom: Extensions
 

of the Mantel-Haenzel Procedure, J^ Amei^ Statist. Ass. 58:690-700.
 

Mantel, N. (1966) Evaluation of Survival Data and Two New Rank Order
 

Statistics Arising in Its Consideration, Cancer Chemother. Rep.
 

5(0:163-170.
 

National Cancer Institute (March 1975) Guidelines for Carcinogen Bioassay
 

in Small Rodents, TR-1, Division of Cancer Cause and Prevention,
 

National Institutes of Health, Bethesda, MD.
 

National Institute for Occupational Safety and Health (1973) Occupational
 

Exposure to Trichloroethylene, USDHEW, Public Health Service,
 

Washington, DC.
 

Nomiyama, K.; Nomiyama, H. (1971) Metabolism of Trichoroethylene in Humans.
 

Sex Difference in Urinary Excretion of Trichloroacetic Acid and
 

Trichloroethanol, Int. Arch. Arbeitsmed. 28:37-48.
 

50
 
Parkhouse, J. (1969) Uptake and Metabolism of Trichloroethylene. Can.
 

Anaesthetists* Soc. J^ 16:113-118.
 

Patty, F. A. (Ed.) (1963) Industrial Hygiene and Toxicology, Vol. II, D. W.
 

Fassett and D. D. Irish (Eds.), Interscience Publishers, New York, NY.
 

Plaa, G. L.; Larson, R. E. (1965) Relative Nephrotoxic Properties of
 

Chlorinated Methane, Ethane, and Ethylene Derivatives in Mice,
 

Toxicol. Appl. Pharmacol. 2:37-44.
 

Price, H. L.; Dripps, R. D. (1965) General Anesthetics, II. Volatile
 

Anesthetics: Diethyl Ether, Divinyl Ether, Chloroform, Halothane,
 

Methoxyflurane, and other Halogenated Volatile Anesthetics, The Pharmacological Basis of Therapeutics, 3rd Ed., L. S. Goodman and A.
 

Gilman (Eds.), MacMillan, New York, NY, pp. 83-99.
 

Rechnagel, R. 0.; Glende, E. A.; Jr. (1973) Carbon Tetrachloride
 

Hepatotoxicity: An Example of Lethal Cleavage, Crit. Rev. Toxicol.
 

2:263-297.
 

Reuber, M. D.; Glover, E. L. (1970) Cirrhosis and Carcinoma of the Liver in
 

Male Rats Given Subcutaneous Carbon Tetrachloride, J^_ Natl. Cancer
 

Inst. 44:419-423.
 

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

(1972) Respiratory Tract Carcinogenesis in Hamsters Induced by
 

Different Numbers of Administrations of Benzo(a)pyrene and Ferric
 

Oxide, Cancer Res. 32:1073-1081.
 

Sax, N, I. (1968) Dangerous Properties of Industrial Materials, 3rd Ed.,
 

Van Nostrand Reinhold, New York, NY, p. 1180.
 

Seifter, J. (1944) Liver Injury in Dogs Exposed to Trichloroethylene, J^
 

Ind. Hyg. Toxicol. 2^6:250-252.
 

Soucek, B.; Vlachova, D. (I960) Excretion of Trichloroethylene Metabolites
 

in Human Urine, Brit. J^_ Ind. Med. 17:60-64.
 

Stecher, P. G. (Ed.) (1968) The Merck Index, 8th Ed., Merck & Co., Inc.,
 

Rahway, NJ, p. 1069.
 

Stewart, iL D.; Dodd, H. C. (1964) Absorption of Carbon Tetrachloride,
 

Trichloroethylene, Tetrachloroethylene, Methylene Chloride, and
 

1,1,1-Trichloroethane through the Human Skin, Amer. Ind. Hyg. Ass. J.
 

25:439-446.
 

Tarone, R. E. (1975) Test for Trend in Life-Table Analysis, Biometrika,
 

62:679-682.
 

51
 
Traiger, G. J.; Plaa, G. L. (1974) Chlorinated Hydrocarbon Toxicity:
 

Potentiation by Isopropyl Alcohol and Acetone, Arch. Environ. Health
 

2^:276-278.
 

Vignoli, L.; Jouglard, J.; Vignoli, P.; Terrasson de Fougeres, P. (1970)
 

Acute Intoxication by Trichloroethylene Ingestion. Metabolism of this
 

Poison, Med_^ Leg. Assicuorazioni 1_8:789-798.
 

von Oettingen, W. F. (1955) The Halogenated Hydrocarbons: Toxicity and
 

Potential Dangers, PHS Publication No. 414, U. S. Government Printing
 

Office, Washington, DC, pp. 203-227.
 

Wiseman, P. (1972) Introduction to Industrial Organic Chemistry, Applied
 

Science Publishers Ltd., Essex, England.
 

52
 
APPENDIX A: CHEMISTRY 

53 
Chemical and Physical Characteristics
 

Physical state: Liquid, nonflammable (Patty, 1963; Sax, 1968)
 

Odor: Resembling chloroform (Patty, 1963)
 

Melting point: -73°C (Patty, 1963)
 

Boiling point: 86.7°C (760 mm Hg) (Stecher, 1968)
 

Solubility: 0.1 g/100 ml water at 20°C; soluble in ether, ethyl alcohol,
 

and chloroform (Patty, 1963); dissolves most fixed and volatile oils
 

(Stecher, 1968)
 

Odor threshold: 24.1 ppm
 

Specific gravity: 1.45560 (25°/4°C) (Patty, 1963)
 

Vapor density: 4.54 (air = 1) (Patty, 1963)
 

Vapor pressure: 77 mm Hg (25°C) (International Labour Office, 1972)
 

Refractive index: 1.4777 (20°C) (Patty, 1963)
 

Percent in "saturated" air: 10.2 (25°C) (Patty, 1963)
 

Handling precautions: Use with adequate ventilation (Stecher, 1968). Must
 

be stored in sealed, light-resistant containers.
 

Technical Product and Impurities
 

Tetrachloroethane has been reported as an impurity in technical trichloroethylene, particularly in that produced by the acetylene-based process
 

(Dreisbach, 1974; NIOSH Criteria Document, 1973). The chemical used for
 

this bioassay contained no detectable quantity of tetrachloroethane.
 

Manufacturing Processes
 

In one industrial process, trichloroethylene is produced from acetylene.
 

The process involves addition of chlorine to acetylene to give 1,1,2,2­

tetrachloroethane. The more common process since 1972 involves the
 

addition of chlorine to ethylene to give ethylene dichloride and then
 

further chlorination to 1,1,1,2-tetrachloroethane, followed by HC1
 

elimination to yield trichloroethylene (Wiseman, 1972).
 

54
 
Chemical Analysis
 

Trichloroethylene Batch #1 - January 9, 1973*
 

Gas Chromatography (Table Al)
 

Detector: Flame ionization
 

Recorder range: 1 mv full scale
 

Column: 3' x 1/4" od, aluminum, 80-100 mesh Porapak Q
 

Temperatures (°C): Injection port 205, detector 250, column oven programmed from 60 (2 min) to 205 (20 min) at 6°/min
 

Flow rates (ml/min): Nitrogen carrier 45, hydrogen 45, air 475 

Attenuation: From 1 x 16 to 103 x 32 

Remarks: About 0.8 yl sample was injected 

*Conducted by Hazleton Laboratories, Inc. 

Table Al. Analysis of Total Area Data - Batch #1 

Component By 

Retention Time 

(min) 

Area 

(cm2) 

Total Areaa 

(cm2) 

Percent 

(A/At x 100) Av. (%) 

2.8 

3.4 

3.3 

0.1 

0.1 

0.1 

A 

B 

C 

0.00067 

0.00082 

0.00075 

0.001 

13.3 

13.6 

13.8 

0.54 

0.34 

0.41 

A 

B 

C 

0.0036 

0.0031 

0.0034 

0.003 

18.0 

18.0 

18.4 

118 

72 

83 

A 

B 

C 

0.79 

0.65 

0.69 

0.7 

19.2 

19.4 

19.8 

8 

6 

7 

A 

B 

C 

0.053 

0.054 

0.058 

0.06 

21. lb 

20.9b 

21.3b 

14,840 

10,920 

11,900 

A 

B 

C 

99.1 

99.2 

99.1 

99.1 

26.0 

26.0 

25.5 

13 

9 

12 

A 

B 

C 

0.087 

0.082 

0.10 

0.09 

Total 100.0 

aA
 = 14,980
 

B = 11,010
 

C = 12,000
 

^Trichloroethylene
 

Infrared Spectroscopy (Figure Al)
 

55
 
14 l-ijl J jtl>?|tliUi t-M|^ M I Mllljiiljifj 

Figure Al. Infrared Spectrum of Trichloroethylene
 
Trichloroethylene Batch #3 - February 15, 1974*
 

Gas Chromatography (Table A2) 

Same as for Batch #1, except as follows: 

Flow rates (ml/min): Hydrogen 60, air 450 

Attenuation: From 1 x 16 to 10^ x 128 

Remarks: About 1.0 yl sample was injected 

^Conducted by Hazleton Laboratories, Inc. 

Table A2. Analysis of Total Area Data - Batch #3 

Component By 

Retention Time 

(min) 

Area 

(cm2) 

Total Areaa 

(cm2) 

Percent 

(A/At x 100) Av. (%) 

12.7 

12.6 

12.7 

3. 

3. 

3. 

6 

3 

9 

A 

B 

C 

0.01 

0.01 

0.01 

0.01 

16.8 

16.8 

17.0 

305 

284 

343 

A 

B 

C 

0.63 

0.62 

0.68 

0.6 

18.2 55
 

18.1 52
 

18.2 63
 

A

B

C
 

0.11
 

0.12 0.1
 

0.13
 

20.2b 48,200
 

20.2b 45,400
 

20.2b 49,800
 

24.2 23
 

24.2 21
 

24.2 25
 

26.0 86
 

26.0 79
 

26.1 81
 

Total
 

aA
 = 48,670
 

B = 45,840
 

C = 50,320
 

^Trichloroethylene
 

Infrared Spectroscopy (Figure A2)
 

A
 

B
 

C
 

A
 

B
 

C
 

A
 

B
 

C
 

99.0
 

99.0
 

99.0
 

0.05
 

0.05
 

0.05
 

0.18
 

0.17
 

0.16
 

99.0
 

0.05
 

0.2
 

100.0
 

57
 
00 

Figure A2* Infrared Spectrum of Trichloroethylene
 
Trichloroethylene Batch #4 - July 8, 1974*
 

Gas Chromatography (Table A3)
 

Same as for Batch #1, except as follows:
 

Temperatures (°C): Injection port 215, detector 240
 

Flow rates (ml/min): Hydrogen 65, air 450
 

Attenuation: From 1 x 16 to 10^ x 16
 

Remarks: About 1.2 yL sample was injected
 

^Conducted by Hazleton Laboratories, Inc.
 

Table A3. Analysis of Total Area Data - Batch #4
 

Component By 

Retention Time Area 

(min) (cm2

) 

20.7 1.7 

20.7 1.3 

20.6 1.7 

22.8 440 

22.7 370 

22.7 410 

24.1 280 

23.9 230 

23.9 250 

26.4b 115 ,500 

26.2b 104 ,300 

26.2b 111 ,300 

34.8 37 

34.8 33 

34.7 34 

Total 

a A
 = 116,250
 

B = 104,930
 

C = 112,000
 

^Trichloroethylene
 

Infrared Spectroscopy (Figure A3)
 

Total Areaa
 

(cm2)
 

A
 

B
 

C
 

A
 

B
 

C
 

A
 

B
 

C
 

A
 

B
 

C
 

A
 

B
 

C
 

Percent
 

(A/At x 100)
 

0.001
 

0.001
 

0.001
 

0.38
 

0.36
 

0.37
 

0.24
 

0.22
 

0.22
 

99.4
 

99.4
 

99.4
 

0.03
 

0.03
 

0.03
 

Av. (%)
 

0.001
 

0.4
 

0.2
 

99.4
 

0.03
 

100.0
 

59
 
Figure A3, Infrared Spectrum of Trichloroethylene 
Identification of Trace Components
 

July 10, 1975
 

Bioassay Batch #4*
 

Analysis Techniques
 

Temperature-programmed vapor-phase chromatography (vpc) was performed on
 

trichloroethylene in order to obtain relative areas and retention times of
 

the major component and impurities present in the compound. Vapor-phase
 

chromatography-mass spectrometry was performed to obtain the mass peaks of
 

the components present. Vapor-phase chromatography was then repeated on
 

the sample spiked with compounds which had the correct mass and fragmentation properties to confirm enhancement of the impurity peaks. In addition, vpc with electron capture detection was used to analyze for trace
 

amounts of the tetrachloroethanes.
 

Results - Vapor-Phase Chromatography
 

Survey System 1: Experimental conditions were as follows: Bendix 2500,
 

1.8 x 4 mm id Chromosorb column, flame ionization detection, oven temperature programmed from 100 to 250°C at 10°/min.
 

A major peak and 5 impurities were detected.
 

Retention Retention Time Area (Relative Possible 

Peak Time (min) (Relative to TCE Peak) to TCE Peak) Identity 

1 7.0 0.70 0.003 unknown 

2 7.9 0.79 0.4 1,2-epoxybutane 

3 8.6 0.85 0.3 ethyl acetate 

4 10.1 1.00 100 trichloroethylene 

5 11.5 1.14 0.05 N-methylpyrrole 

6 12.9 1.28 0.06 diisobutylene 

*Conducted by Midwest Research Institute, Kansas City, MO.
 

Survey System 2: Experimental conditions: Tracor MT 220, 5% Carbowax 20M
 

TRA on 60-80 mesh Gas-Chrom Q, 1.8 m x 4 mm column, flame ionization detection, oven temperature 40°C.
 

A major peak and 2 impurites were detected.
 

Peak 

Retention 

Time (min) 

Retention Time

(Relative to TCE Peak)

 Area (Relative 

 to TCE Peak) 

Possible 

Identity 

4 

5 

7 

2.4 

6.0 

10.9 

1.00 

2.50 

4.54 

100.00 

0.27 

0.80 

trichloroethylene 

N-methylpyrrole 

epichlorohydrin 

61
 
Quantitation and Identity System 1 (Epoxybutane): Experimental conditions:
 

Tracor MT 220, Chromosorb 102, 1.8 m x 4 mm id column, flame ionization
 

detection, oven temperature 100°C.
 

By addition of known standards to the trichloroethylene sample, it was
 

determined that peak 2 was not due to ethyl vinyl ether, ethyl acetate,
 

tetrahydrofuran, methyl ethyl ketone, or 1,2-epoxyisobutane. However,
 

addition of an authentic sample of 1,2-epoxybutane enhanced the peak. The
 

epoxybutane in trichloroethylene was quantitated against a known standard
 

of epoxybutane.
 

Retention Quantitation
 

Peak Time (min) Identity (%)
 

1 - unknown
 

2 18.11 1,2-epoxybutane 0.19
 

3 22.52 ethyl acetate
 

Quantitation and Identity System 2 (Ethyl Acetate): Experimental conditions: Bendix 2500, Chromosorb 102, 1.8 m x 4 mm id, flame ionization
 

detection, oven temperature 135°C.
 

The vpc-mass data indicated that peak 3 could be ethyl acetate. Addition
 

of an authentic sample of ethyl acetate to the trichloroethylene enhanced
 

this peak. Ethyl acetate in the trichloroethylene was quantitated using an
 

ethyl acetate standard.
 

Retention Quantitation
 

Peak Time (min) Identity (%)
 

1 5.7 unknown
 

2 7.8 1,2-epoxybutane
 

3 9.8 ethyl acetate 0.04
 

Quantitation and Identity System 3 (N-Methylpyrrole and Trimethylpentene):
 

Experimental conditions: Tracor MT 220, Chromosorb 102, 1.8 m x 4 mm id,
 

flame ionization detection, oven temperature 175°C.
 

The vpc-mass data indicated that peak 5 could be N-methylpyrrole. Addition
 

of an authentic sample of ltf-methylpyrrole to the trichloroethylene enhanced
 

this peak.
 

The vpc-mass data indicated that peak 6 could be diisobutylene (2,2,4-trimethylpentene) . Addition of an authentic sample of 2,2,4-trimethylpentene
 

enhanced this peak.
 

Peaks 5 and 6 were quantitative against the authentic standards.
 

62
 
Retention Quantitation
 

Peak Time (min) Identity (%)
 

4 4.0 trichloroethylene
 

5 6.5 N-methylpyrrole 0.02
 

6 10.0 trimethylpentene 0.03
 

Quantitation and Identity System 4 (Epichlorohydrin); Experimental conditions: Tracor MT 220, 5% Carbowax 20M TRA on 60-80 mesh Gas-Chrom Q, 1.8 m
 

 4 mm, flame ionization detection, oven temperature 40°C.
 

The vpc-mass data indicated that peak 7 could be epichlorohydrin. Addition
 

of an authentic sample of epichlorohydrin enhanced this peak. The epichlorohydrin present in the trichloroethylene sample was quantitated using
 

an epichlorohydrin standard. On the Chromosorb 102 column, epichlorohydrin
 

has the same retention time as trichloroethylene.
 

Retention Quantitation
 

Peak Time (min) Identity (%)
 

4 trichloroethylene 

5 8.19 N-methylpyrrole 

7 11.18 epichlorohydrin 0.09 

Quantitation and Identity System 5 (Tetrachloroethane): Experimental conditions: Tracor MT 220, Chromosorb 102, 1.8 m χ 4 mm id, 63 Ni electron
 

capture detection, oven temperature 190°C.
 

On injection of a 6 μ neat sample of trichloroethyene, Lot No. 063014, no
 

peaks were detected at the retention times of either tetrachloroethane
 

isomer. The concentration of tetrachloroethane in the trichloroethylene
 

(Lot No. 063014) is therefore less than 2 ppm for the 1-, 1,1,2-isomer and
 

less than 5 ppm for the 1,1,2,2-isomer.
 

Note: A fresh sample of trichloroethylene (Aldrich Lot No. 090947) did
 

contain a peak at the same retention time as 1,1,1,2-tetrachloroethane.
 

Quantitation against known 1,1,1,2-tetrachloroethane indicated a concentration of 3 ppm.
 

Minimum
 

Retention Detectable
 

Standard Time (min) Amount (ng)
 

1,1,1,2-Tetrachloroethane 12.0 0.1
 

1,1,2,2-Tetrachloroethane 16.7 0.3
 

63
 
Vapor-Phase Chromatography-Mass Spectrometry: Experimental conditions:
 

Varian MAT CH-4B interfaced via a Watson-Biemann helium separator to a
 

Micro-Tek 2000 MF gas chromatograph, data processed by Varian 620/i computer, Chromosorb 102 column, 1.8 m x 4 mm id. See Table A4.
 

Conclusions
 

Six small impurities were detected in trichloroethylene (Lot No. 063014) by
 

vapor-phase chromatography. Mass spectrometry data indicated that the 5
 

larger impurities were 1,2-epoxybutane, ethyl acetate, Nhmethylpyrrole,
 

diisobutylene, and epichlorohydrin. Trichloroethylene samples spiked with
 

authentic samples of the impurities showed enhancement of the corresponding
 

peaks. Quantitation studies indicated the following impurity concentrations: 1,2-epoxybutane, 0.19%; ethyl acetate, 0.04%; _N-methylpyrrole,
 

0.02%; diisobutylene, 0.03%; epichlorohydrin, 0.09%. The first impurity
 

peak (0.003%, relative area) was not detected by mass spectrometry.
 

1,1,2,2- and 1,1,1,2-tetrachloroethane were not detected by electron
 

capture detection or mass spectrometry; the levels of minimum detection on
 

the electron capture detector for the 2 tetrachloroethane isomers were 5
 

and 2 ppm, respectively.
 

64
 
Table A4. Vapor-Phase Chromatography-Mass Spectrometry

a

Peak notation the same as previously indicated.

Eight Peak Index of Mass Spectra, Vol. I (1970), Mass Spectrometry

Data Centre, AWRE, Aldermaston, Reading, RG7 4PR, United Kingdom.

Peaka

2

3

5

6

7

Mass

28 (N )

42 2

41

27

72

29

39

71

57

43

29

27

45

61

42

70

26

88

81

80

39

42

53

28

55

27

57

55

97

29

56

112

27

69

57

27

29

49

31

62

28

51

Intensity Relative

to Base Peak

100

92

92

58

28

32

35

36

20

100

43

23

49

49

16

30

8

15

93

59

24

18

27

100 (N2)

13

52

100

88

47

51

51

31

17

24

100

100

79

88

65

52

100 (N2)

25

Mass

42

41

27

72

29

39

71

57

43

29

27

45

61

42

70

26

81

80

39

42

53

28

55

27

57

55

97

29

56

112

27

69

57

27

29

49

31

62

28

51

Literature Values for:

1,2-Epoxybutanek

Intensity Relative

to Base Peak

100

93

39

34

30

28

28

20

Ethyl Acetate*5

100

25

13

13

10

6

5

4

N-Methylpyrrole

100

80

37

30

28

17

17

15

Diisobutylene

100

42

29

22

19

15

11

9

Epichlorohydrin

100

39

31

25

22

18

16

8

65
Manufacturer's Feed Analysis (Wayne Lab-Blox Meal)
 

(Allied Mills Inc., Chicago, IL 60606) 

Crude protein (Min.) 24 .0% 

Crude fat (Min.) 4.0% 

Crude fiber (Max.) 4.5% 

Ingredients
 

Animal liver meal
 

Fish meal
 

Dried whey
 

Corn and wheat flakes
 

Ground yellow corn
 

Soybean meal
 

Wheat middlings
 

Cane molasses
 

Soybean oil
 

Brewer's dried yeast
 

Vitamin A palmitate
 

Irradiated dried yeast (source of vitamin D)
 

D-Activated animal sterol
 

Vitamin E supplement
 

Menadione sodium bisulfite (source of vitamin K activity)
 

Riboflavin supplement
 

Niacin
 

Calcium pantothenate
 

Choline chloride
 

Thiamine
 

Ground limestone
 

Dicalcium phosphate
 

Salt
 

Manganous oxide
 

Copper oxide
 

Iron carbonate
 

Ethylene diamine dihydriodide
 

Cobalt carbonate
 

Zinc oxide
 

66
 
Basal Feed Analysis
 

A study of apparent pesticide residues in basal feed samples was conducted
 

at Gulf South Research Institute under contract to NCI on November 10,
 

19 72. Three samples of feed used at Hazleton were analyzed. No lindane,
 

heptachlor, aldrin, heptachlor epoxide, endrin, DDD, chlordane, methoxychlor, toxaphene, or organophosphate was found. Two of the samples
 

contained 0.00392 and 0.00682 ppm DDE, and 2 contained 0.00629 and 0.02016
 

ppm DDT. One sample contained 0.03955 ppm Aroclor 1254. The 3 samples
 

contained 0.00249, 0.00141, and 0.00167 ppm dieldrin. The method of
 

analysis was as follows:
 

A 20 g sample was extracted with 150 ml 6% diethyl ether in hexane on a 25
 

mm od x 40 cm chromatographic column containing 1" Na2S(>4. With the
 

stopcock closed, 25 ml extracting solvent was added. The feed sample was
 

added and allowed to settle. The column was filled with extracting solvent
 

and the sample again was allowed to settle. The stopcock was opened and
 

150 ml extracting solvent was collected in a 500 ml standard taper round-


bottom flask. A Synder column was placed on the flask and the extracting
 

solvent was evaporated on a 65-70°C heating mantle. The Snyder column was
 

rinsed with 30 ml hexane and the extract was reduced to about 10 ml.
 

Evaporation continued and the Snyder rinse was repeated a second time. When
 

the extract again was reduced to 10 ml, the heat was removed and the
 

extract was reduced to about 2 ml under vacuum.
 

The extracted sample was cleaned up on a 25 mm od x 40 cm chromatographic
 

column containing 5" Florisil (activated 15 hours at 135°C) and topped with
 

1/2" Na2SO4. The sample extract was quantitatively transferred to the 

column with a small portion of hexane. The following extracting solvents 

were passed through the column and collected: 

Fraction A: 175 ml hexane
 

Fraction B: 200 ml 6% diethyl ether in hexane
 

Fraction C: 225 ml 15% diethyl ether in hexane
 

Fraction D: 200 ml 30% diethyl ether in hexane
 

Each fraction was evaporated as before almost to dryness and the residue
 

was quantitatively transferred to a 10 ml volumetric flask and diluted to
 

volume using hexane. An aliquot was taken for gas-liquid chromatographic
 

analysis. Compounds were identified according to chromatographic retention
 

times only and, therefore, should be considered tentative.
 

67
 
Water Analysis (local artesian veil)
 

(mg/liter except pH)
 

Calcium
 

Magnesium
 

Potassium
 

Sulfate
 

Nitrate
 

Nitrite
 

Ammonia
 

Phenol
 

Chlorine
 

Chloride
 

Fluoride
 

3.8
 

1.3
 

1.05
 

0.01
 

2.77
 

0.05
 

0.06
 

0.001
 

0.001
 

1.90
 

0.01
 

Cyanide
 

Iron
 

Copper
 

Zinc
 

Cadmium
 

Chromium
 

Lead
 

Alkalinity
 

Hardness
 

pH
 

0.005
 

0.02
 

0.005
 

0.024
 

0.006
 

0.05
 

0.001
 

63.1
 

10.6
 

6.35 units
 

68
 
APPENDIX B: WEIGHTS AND SURVIVAL
 

69
 
Table Xllla. Mean Body Weights and Survival - Trichloroethylene Subchronic Study - Male Rats 

Time
 

Interval
 

(weeks)
 

0
 

1
 

2
 

3
 

4
 

5
 

6
 

7
 

8
 

Mean av. wt gain (g)
 

o % of control wt gain
 

0
 

1
 

2
 

3
 

4
 

5
 

6
 

7
 

8
 

Mean av. wt gain (g)
 

% of control wt gain
 

Body
 

Mean
 

(g)
 

GrouD
 

230
 

263
 

295
 

323
 

358
 

373
 

384
 

406
 

417
 

187
 

—
 

Group
 

230
 

246
 

278
 

310
 

342
 

355
 

364
 

388
 

404
 

174
 

93
 

Weight
 

Std Dev. Surv.
 

(g)
 

1 (0 mg/kg/day)
 

19.4
 

21.0
 

19.2
 

29.4
 

26.9
 

26.3
 

27.2
 

28.7
 

22.1
 

5/5
 

5/5
 

5/5
 

5/5
 

5/5
 

5/5
 

5/5
 

5/5
 

5/5
 

4 (1730 mg/kg/day)
 

21.0
 

14.4
 

19.4
 

21.1
 

27.1
 

33.2
 

36.0
 

37.5
 

37.6
 

5/5
 

5/5
 

5/5
 

5/5
 

5/5
 

5/5
 

5/5
 

5/5
 

5/5
 

Body Weight Body Weight 

Mean Std Dev. Surv. Mean Std Dev. Surv. 

(R) (g) (g) (g) 

Group 2 (562 mg/kg/day) Group 3 (1000 mg/kg/day) 

230 21.4 5/5 231 21.2 5/5 

255 34.5 5/5 259 27.8 5/5 

284 40.4 5/5 294 29.8 5/5 

319 39.9 5/5 327 30.6 5/5 

344 36.7 5/5 359 33.9 5/5 

357 36.7 5/5 374 33.1 5/5 

362 39.4 5/5 376 31.3 5/5 

384 42.0 5/5 406 33.9 5/5 

400 39.2 5/5 418 34.1 5/5 

170 187 

90.9 100 

Group 5 (3160 mg/kg/day) Group 6 (5620 mg/kg/day) 

230 18.4 5/5 228 20.5 5/5 

218 18.8 5/5 193 32.6 3/5 

240 23.4 5/5 226 — 1/5 

273 30.0 5/5 242 - 1/5 

299 34.9 5/5 279 - 1/5 

298 40.0 5/5 - - 0/5 

304 43.2 5/5 

344 45.7 5/5 

370 48.4 5/5 

140 51 

74.9 27.3 
Table Xlllb. Mean Body Weights and Survival - Trichloroethylene Subchronic Study - Female Rats
 

Time Body Weight Body Weight Body Weight
 

Interval Mean Std Dev. Surv. Mean Std Dev. Surv. Mean Std Dev. Surv.
 

(weeks) (g) (g) (g) (g) (g) (g)
 

Group 1 (0 mg/kg/day) Group 2 (562 mg/kg/day) Group 3 (1000 mg/kg/day)
 

0 168 14.1 5/5 167 10.9 5/5 170 10.6 5/5
 

1 183 12.6 5/5 178 7.8 5/5 182 12.1 5/5
 

2 193 26.0 5/5 195 8.8 5/5 197 12.0 5/5
 

3 218 29.6 5/5 214 9.1 5/5 218 12.7 5/5
 

4 235 28.5 5/5 230 10.2 5/5 237 16.8 5/5
 

5 244 29.8 5/5 236 12.1 5/5 238 18.0 5/5
 

6 249 31.5 5/5 234 12.1 5/5 239 17.5 5/5
 

7 266 31.6 5/5 248 9.3 5/5 252 18.9 5/5
 

8 276 28.9 5/5 258 10.5 5/5 260 20.3 5/5
 

Mean av. wt gain (g) 108 91 90
 

% of control wt gain — 84.3 83.3
 

Group 4 (1730 mg/kg/day) Group 5 (3160 mg/kg/day) Group 6 (5620 mg/kg/day)
 

0 170 8.3 5/5 170 8.8 5/5 169 14.0 5/5
 

1 111 9.2 5/5 171 13.3 5/5 178 9.a 2/5
 

2 183 7.1 5/5 173 19.4 5/5 169 — 1/5
 

3 202 9.7 5/5 188 24.7 5/5 184 - 1/5
 

4 227 9.5 5/5 217 23.3 5/5 204 - 1/5
 

5 226 7.5 5/5 212 21.4 5/5 - - 0/5
 

6 223 8.0 5/5 217 19.9 5/5
 

7 239 11.2 5/5 229 16.0 5/5
 

8 245 14.5 5/5 241 21.7 5/5
 

Mean av. wt gain (g) 75 71 35
 

% of control wt gain 69.4 65.7 32.3
 
Table XlVa. Mean Body Weights and Survival - Trichloroethylene Subchronic Study — Male Mice
 

Time Body Weight Body Weight Body Weight 

Interval Mean Std Dev. Surv. Mean Std Dev. Surv. Mean Std Dev. Surv. 

(weeks) (g) (g) (g) (g) (g) 

Group 1 (0 mg/kg/day) Group 2 (1000 mg/kg/day) Group 3 (1780 mg/kg/day) 

0 15 0.9 5/5 15 0.9 5/5 15 0.9 5/5 

1 16 0.3 5/5 15 1.4 5/5 16 1.7 5/5 

2 18 0.5 5/5 19 1.5 5/5 18 0.8 5/5 

3 19 0.3 5/5 20 1.5 5/5 19 0.9 5/5 

4 20 0.1 5/5 21 1.4 5/5 20 1.2 5/5 

5 20 0.5 5/5 21 1.4 5/5 21 1.3 5/5 

6 20 0.4 5/5 22 1.2 5/5 21 1.7 5/5 

7 20 0.4 5/5 22 0.9 5/5 21 1.2 5/5 

8 21 2.1 5/5 23 1.1 5/5 21 1.5 5/5 

Mean av. wt gain (g) 6 8 6 

% of control wt gain — 133 100 

Group 4 (3160 mg/kg/day) Group 5 (5620 mg/kg/day) Group 6 (10,000 mg/kg/day) 

0 15 0.9 5/5 15 1.0 5/5 15 1.0 5/5 

1 16 1.0 5/5 14 0.3 2/5 - - 0/5 

2 18 0.7 5/5 13 3.5 2/5 

3 20 1.0 5/5 18 0.5 2/5 

4 21 1.3 5/5 18 1.3 2/5 

5 22 1.3 5/5 20 1.4 2/5 

6 22 1.5 5/5 20 - 1/5 

7 22 1.4 5/5 20 - 1/5 

8 23 1.2 5/5 20 - 1/5 

Mean av. wt gain (g) 8 5 -

% of control wt gain 133 83.3 
Table XlVb. Mean Body Weights and Survival Trichloroethylene Subchronic Study Female Mice
 

Time Body Weight Body Weight Body Weight
 

Interval Mean Std Dev. Surv. Mean Std Dev. Surv. Mean Std Dev, Surv.
 

(weeks) 1E) (g) (g) (g) (g) (g)
 

Group 1 (0 mg/kg/day) Group 2 (1000 mg/kg/day) Group 3 (1780 mg/kg/day)
 

0 11 2.2 5/5 11 2.2 5/5 11 2.2 5/5
 

1 13 1.6 5/5 11 1.8 5/5 13 1.6 5/5
 

2 16 1.6 5/5 15 1.1 5/5 14 1.0 5/5
 

3 17 1.0 5/5 17 1.2 5/5 16 1.1 5/5
 

4 19 0.6 5/5 18 1.1 5/5 18 0.9 5/5
 

5 19 0.9 5/5 18 1.4 5/5 18 0.5 5/5
 

6 20 0.9 5/5 19 1.4 5/5 19 1.0 5/5
 

7 20 0.6 5/5 18 1.6 5/5 19 0.9 5/5
 

8 21 0.8 5/5 19 2.0 5/5 20 0.9 5/5
 

Mean av. wt gain (g) 10 8 9
 

% of control wt gain — 80.0 90.0
 

Group 4 (3160 rng/kg/day) Group 5 (5620 mg/kg/day) Group 6 (10,000 ι ng/kg/day)
 

0 11 2.4 5/5 11 2.5 5/5 11 2.6 5/5
 

1 13 1.5 3/5 13 0.5 2/5 - - 0/5
 

2 15 0.5 3/5 13 4.0 2/5
 

3 17 1.7 3/5 13 6.9 2/5
 

4 20 1.5 3/5 20 — 1/5
 

5 20 1.7 3/5 20 — 1/5
 

6 20 1.5 3/5 20 — 1/5
 

7 21 1.3 3/5 20 — 1/5
 

8 21 1.0 3/5 20 1/5
 

Mean av. wt gain (g) 10 9 ­

% of control wt gain 100 90.0
 
Table XVa. Mean Body Weights, Food Consumption, and Survival -


Trichloroethylene Chronic Study - Male Rats
 

Vehicle Controls Low Dose High Dose 

Time Body Weighta No. of Body Weight No. of Body Weight No. of 

Interval Mean Std Dev. Foodb Animals Mean Std Dev Food Animals Mean Std Dev Food Animals 

(weeks) (g) (g) (g) Weighed (g) (g) (g) Weighed (g) (g) (g) Weighed 

0 193 15.0 0 20 193 15.8 0 50 194 16.7 0 50 

1 242 18.7 139 20 240 21.8 145 50 237 22.7 128 50 

2 288 20.9 155 20 281 26.4 162 50 269 26.5 157 49 

3 312 22.2 162 20 309 21.2 166 50 296 27.9 148 48 

4 334 22.6 164 20 329 23.5 164 50 313 34.2 156 48 

5 360 24.0 161 20 352 25.4 166 50 340 26.2 161 47 

6 387 35.2 162 20 377 27.8 168 50 360 28.7 163 47 

7 402 27.6 189 20 382 28.8 196 50 375 27.5 170 46 

8 412 31.8 157 20 399 30.6 158 50 381 27.8 152 46 

9 437 32.8 155 20 423 32.5 157 50 406 31.2 139 46 

10 458 34.2 154 20 433 34.7 162 50 406 32.2 156 46 

14 504 39.4 153 20 474 40.4 150 50 446 45.3 140 45 

18 535 38.9 161 20 503 41.7 163 48 481 42.5 152 44 

22 552 38.0 161 20 519 46.3 158 48 493 43.9 148 43 

26 570 43.3 156 20 533 47.1 158 47 500 47.2 150 43 

30 587 41.5 155 20 544 48.8 164 46 503 51.3 159 42 

34 607 40.3 157 20 564 49.3 159 45 528 49.3 148 40 

38 617 46.6 153 20 566 50.5 155 45 535 52.1 143 39 

42 612 45.2 175 20 565 47.0 177 44 528 57.2 178 38 

46 622 47.7 158 20 575 49.7 151 43 526 52.1 151 36 

50 618 64.5 152 20 582 49.0 153 42 528 51.3 142 34 

54 616 50.2 149 20 573 47.6 147 40 513 48.3 134 30 

58 618 45.2 164 20 576 53.5 158 40 508 52.4 137 27 

62 628 49.6 167 20 586 47.5 161 38 507 56.7 165 25 

66 615 53.6 156 20 562 46.8 156 37 493 57.2 146 19 

70 611 59.8 139 19 567 60.8 139 35 506 52.4 137 17 

74 581 65.8 151 18 539 62.7 153 33 485 45.8 151 14 

78 559 76.7 153 16 523 67.3 158 31 462 47.2 148 12 

82 519 92.4 144 15 503 78.3 159 29 450 55.6 169 11
 
Table XVa. Mean Body Weights, Food Consumption, and Survival -


Trichloroethylene Chronic Study - Male Rats (continued)
 

86 500 106.7 136 13 490 88.6 152 24 415 52.2 157 10 

90 517 94.6 114 9 482 84.5 124 20 422 53.0 120 9 

94 515 69.5 138 8 467 96.3 129 18 415 73.7 136 8 

98 459 80.7 120 7 449 97.2 144 16 403 64.7 149 7 

102 423 42.2 137 4 420 98.6 141 15 389 72.5 141 6 

106 394 37.6 94 3 401 88.9 109 11 418 46.9 142 4 

110 382 26.9 91 2 383 101.5 114 8 423 45.3 255 3 

a

Calculated using individual animal weight.
 

^Average weight per animal per week.
 
Table XVb. Mean Body Weights, Food Consumption, and Survival -


Trichloroethylene Chronic Study - Female Rats 

Vehicle Controls Low Dose High Dose 

Time Body Weighta No. of Body Weight No. of Body Weight No. of 

Interval Mean Std Dev. . Foodb Animals Mean Std Dev Food Animals Mean Std Dev Food Animals 

(weeks) (g) (g) (g) Weighed (g) (g) (g) Weighed (g) (g) (g) Weighed 

0 146 11.4 0 20 144 11.0 0 50 144 9.5 0 50 

1 169 15.9 110 20 170 13.4 96 50 169 11.7 99 50 

2 201 16.1 no 20 183 19.5 122 48 177 21.1 117 50 

3 205 14.1 131 20 192 23.7 113 47 196 20.7 110 49 

4 216 16.3 133 20 209 18.7 117 46 208 19.9 117 49 

5 228 20.0 132 20 220 18.6 127 45 217 22.1 127 49 

6 241 25.1 125 20 224 19.2 122 45 226 19.7 118 48 

7 242 24.8 153 20 234 19.7 132 44 235 19.8 128 48 

8 255 27.9 119 20 240 20.3 113 44 236 18.7 108 48 

9 268 26.8 132 20 256 22.0 112 44 251 20.7 112 48 

10 280 30.8 130 20 250 21.4 128 44 247 21.0 117 47 

14 302 33.8 107 20 271 24.4 98 44 262 26.3 104 47 

18 321 37.9 122 20 283 27.9 113 41 276 27.9 112 47 

22 330 36.8 119 20 286 31.0 102 37 281 26.5 102 46 

26 351 37.6 124 19 293 28.8 110 34 286 30.8 106 45 

30 367 39.5 124 19 303 33.9 95 32 304 39.0 93 43 

34 382 34.8 128 19 309 33.8 110 31 229 36.3 105 42 

38 383 42.7 121 19 309 34.9 104 30 302 38.5 102 41 

42 378 52.2 151 19 314 30.7 127 28 306 49.2 130 41 

46 382 45.9 127 19 307 30.4 116 28 301 37.1 104 39 

50 390 50.9 126 17 312 30.0 114 28 300 42.0 126 39 

54 388 51.3 128 17 315 29.8 120 28 307 38.6 113 37 

58 396 47.3 135 17 318 34.3 118 27 310 39.7 109 35 

62 404 57.4 134 17 318 34.5 129 25 310 39.8 136 34 

66 390 54.3 135 17 311 35.7 118 25 304 40.7 116 32 

70 399 59.7 116 16 322 46.1 113 23 313 46.1 108 29 

74 385 60.4 140 16 303 42.7 127 23 300 45.6 118 24 

78 373 58.2 146 16 317 39.9 145 20 317 43.5 135 23 

82 382 47.9 143 15 315 43.2 142 20 315 48.1 136 22 

86 378 43.4 137 15 311 43.2 131 20 317 51.9 131 22 
Table XVb. Mean Body Weights, Food Consumption, and Survival -


Trichloroethylene Chronic Study - Female Rats (continued)
 

90 364 41.6 101 14 314 33.6 103 19 324 48.9 107 20 

94 366 47.7 134 14 313 37.6 123 19 326 55.7 127 20 

98 327 61.7 120 13 314 58.3 119 16 321 60.3 129 18 

102 336 63.4 134 9 314 66.4 132 15 299 74.3 143 16 

106 332 74.9 105 9 308 73.7 114 13 317 70.3 130 13 

110 326 80.1 119 8 311 86.0 114 13 311 67.7 136 13 

a

L,aicuiatea using individual animal i
 

bAverage weight per animal per week.
 
Table XVIa. Mean Body Weights, Food Consumption, and Survival -


Trichloroethylene Chronic Study - Male Mice 

Vehicle Controls Low Dose High Dose 

Time Body Weighta No. of Body Weight No. of Body Weight No. of 

Interval Mean Std Dev, . Foodb Animals Mean Std Dev Food Animals Mean Std Dev Food Animals 

(weeks) (g) (g) (g) Weighed (g) (g) (g) Weighed (g) (g) (g) Weighed 

0 17 0.5 0 20 17 2.0 0 50 17 1.1 0 50 

1 20 0.0 24 20 19 1.2 23 50 20 0.5 22 50 

2 20 1.2 25 20 21 1.0 26 50 21 0.5 25 50 

3 22 0.4 22 20 22 0.9 24 50 22 0.9 25 50 

4 23 0.1 22 20 23 0.7 24 50 23 0.7 25 50 

5 25 0.4 22 20 24 0.7 25 50 25 0.6 25 50 

6 25 0.2 22 20 24 0.5 25 50 24 0.5 24 50 

7 25 0.4 25 20 26 0.7 27 50 26 1.1 27 50 

8 27 0.3 23 20 26 0.8 26 50 26 0.3 26 50 

00 9 24 1.0 26 20 26 0.8 28 50 26 0.4 29 50 

10 27 0.4 22 20 27 1.2 26 50 28 0.4 25 50 

14 28 0.1 23 20 28 0.8 27 50 29 0.4 26 49 

18 29 1.7 20 20 30 0.9 24 49 31 0.3 23 46 

22 30 0.4 28 20 30 1.5 29 48 30 0.8 29 46 

26 31 0.2 24 20 31 0.9 28 48 32 0.7 27 42 

30 33 0.1 26 20 32 1.0 28 48 32 0.6 28 38 

34 34 0.5 26 19 34 1.2 29 47 34 1.1 31 36 

38 34 0.5 24 19 35 1.4 29 47 35 0.5 29 35 

42 33 0.1 23 18 33 1.1 27 47 35 0.9 30 35 

46 34 0.4 26 18 34 0.9 28 47 35 1.6 30 34 

50 34 0.5 23 18 34 1.5 27 47 35 0.7 30 34 

54 32 0.4 21 18 33 1.0 26 45 34 0.4 30 33 

58 33 1.3 27 18 34 1.1 28 44 35 0.6 33 33 

62 35 0.5 27 16 34 0.9 30 44 35 0.2 34 • 30 

66 33 0.5 22 15 34 1.5 27 41 34 0.5 32 30 

70 33 0.9 31 13 35 1.1 33 41 35 0.5 36 29 

74 32 2.7 27 12 34 0.4 34 41 35 0.6 36 27 

78 34 0.6 24 8 34 0.8 32 40 35 1.0 39 24
 
Table XVIa. Mean Body Weights, Food Consumption, and Survival -


Trichloroethylene Chronic Study - Male Mice (continued)
 

82 33 0.2 33 8 33 0.4 31 38 34 1.2 42 22 

86 32 0.1 32 8 32 0.3 32 38 32 1.9 41 22 

90 34 0.7 34 8 33 0.7 32 35 34 1.3 38 20 

a

Calculated using individual animal weight.
 

^Average weight per animal per week.
 
Table XVIb. Mean Body Weights, Food Consumption, and Survival -


Trichloroethylene Chronic Study - Female Mice 

Vehicle Controls Low Dose High Dose 

Time Body Weighta No. of Body Weight No. of Body Weight No. of 

Interval Mean Std Dev. Foodb Animals Mean Std Dev Food Animals Mean Std Dev . Food Animals 

(weeks) (g) (g) (8) Weighed (g) (g) (g) Weighed (g) (g) (g) Weighed 

0 14 0.0 0 20 14 0.6 0 50 14 0.7 0 50 

1 17 0.0 22 20 18 0.4 24 50 17 0.6 23 50 

2 18 0.3 28 20 18 0.5 28 50 18 0.8 27 50 

3 19 0.5 24 20 19 0.3 24 50 19 0.6 23 50 

4 19 0.5 30 20 19 0.7 24 50 19 0.7 23 50 

5 20 0.5 25 20 20 0.6 24 50 20 0.7 22 50 

6 20 0.1 25 20 20 0.3 24 50 19 0.7 23 50 

7 21 0.2 32 20 21 0.3 26 50 21 0.5 24 50 

8 22 0.9 31 20 21 0.3 27 50 22 0.2 26 50 

9 20 0.3 32 20 21 0.7 27 50 21 0.6 26 50 00 10 22 0.4 23 20 22 0.4 23 50 22 0.6 23 50 

o 14 23 0.4 27 20 23 0.3 23 49 24 0.5 24 49 

18 24 0.1 18 20 25 0.9 21 49 25 1.5 20 49 

22 25 0.1 27 19 25 0.6 25 49 24 0.6 24 48 

26 25 0.2 22 19 26 0.7 23 49 25 0.5 24 47 

30 26 0.1 20 19 26 0.4 23 49 26 0.5 24 47 

34 27 0.1 24 19 27 0.4 23 48 27 0.4 24 44 

38 27 0.6 19 18 27 0.4 22 46 27 0.4 23 43 

42 28 0.8 21 18 28 0.5 23 45 27 0.4 35 41 

46 28 0.5 21 18 29 0.5 22 45 27 0.3 22 41 

50 28 1.1 20 18 28 0.4 21 45 27 0.6 22 41 

54 28 1.1 19 18 27 0.5 21 45 26 0.3 22 41 

58 29 0.2 22 18 28 0.5 33 44 27 0.4 23 41 

62 28 0.5 25 18 28 0.2 25 44 26 0.4 23 41 

66 28 0.9 20 18 28 0.7 22 42 27 0.6 21 41 

70 29 0.7 23 18 28 0.6 24 42 28 0.7 23 40 

74 29 0.6 21 18 29 0.6 24 42 28 0.7 25 40 

78 29 0.6 23 18 29 0.7 25 41 28 0.4 26 40
 
Table XVIb. Mean Body Weights, Food Consumption, and Survival -


Trichloroethylene Chronic Study - Female Mice (continued)
 

82 28 0.5 24 18 28 0.6 25 40 27 0.6 27 40
 

86 26 0.0 24 17 27 0.6 25 40 25 1.0 25 40
 

90 28 0.6 28 17 30 0.6 27 40 28 0.7 26 39
 

a

Calculated using individual animal weight.
 

^Average weight per animal per week.
 

00
 
40 60 80 

TIME ON STUDY (WEEKS) 

Figure 13a. Mean Body Weight vs. Time on Study ­ Trichloroethylene-Treated Male Rats
 

82
 
D CONTROL 

 LOW DOSE 

 HIGH DOSE 

TIME ON STUDY (WEEKS)
 

Figure 13b. Mean Body Weight vs. Time on Study ­

Trichloroethylene-Treated Female Rats
 

83
 
40 60 80 

TIME ON STUDY (WEEKS) 

Figure 14a. Mean Body Weight vs. Time on Study ­

Trichloroethylene-Treated Male Mice 

84
 

10 
40 60 

TIME ON STUDY (WEEKS) 

Figure 14b. Mean Body Weight vs. Time on Study ­

Trichloroethylene-Treated Female Mice
 

85
 

APPENDIX C: STATISTICS
 

87
 
Organization of Chronic Data for Statistical Analysis
 

Statistical analysis begins with examination of the data on survival and
 

pathology of animals in the chronic experiment. These data are presented
 

in detail in the pathology tables in Appendix D. These tables are
 

summarized in this appendix (Tables XVII, XVIIIa, XVIIIb, XlXa, and XlXb).
 

Each animal is listed in its group, identified by its animal number, and
 

arranged in order of the week on study in which it died or was killed or
 

lost. The first column is labeled "Week on Study". An asterisk after the
 

week indicates that the animal's observed lifetime was censored by
 

scheduled or terminal sacrifice, accidental killing, or otherwise lost.
 

Absence of the asterisk indicates that the animal was found dead or was
 

killed while moribund. When mice that were scheduled to be terminally
 

sacrificed at 90 weeks were in fact killed at 91 weeks, 90 weeks is adopted
 

as the week on study of death, so they can be compared with control mice
 

killed at 90 weeks. The second column lists the animal number for
 

reference to the pathology tables. The final column, labeled "Mark",
 

contains letters assigned by the pathologist and the biostatistician in
 

cooperation to sets of pathological diagnoses solely for statistical
 

analysis. The choice of letter is arbitrary, except that more frequent
 

diagnostic sets tend to be assigned to letters early in the alphabet. Each
 

animal is marked if its pathology is included in the set of diagnoses
 

associated with the mark. An animal may have several marks. If the animal
 

has not been evaluated histopathologically, a hyphen appears in the column.
 

A blank space in the column indicates that the animal has been evaluated
 

histopathologically, but has not been marked.
 

Estimation of Survival Probabilities
 

The probability that an animal survived each week of the chronic experiment
 

is estimated for each group by the product-limit procedure of Kaplan and
 

Meier (1958). These estimates are listed in Tables XXa, XXb, XXIa, and
 

XXIb in Appendix C. The estimates are also graphed in Figures 2a, 2b, 3a,
 

and 3b. A description of the estimate follows, including a description of
 

the survival tables.
 

In the survival tables, the first column, labeled j, is the week on study
 

at which death or censoring occurs. The second column, labeled n, is the
 

number of animals at the beginning of week j. The third column, labeled
 

n', is the number of animals alive at the end of week j. The estimate of
 

the conditional probability of the animal surviving week j , given that it
 

was alive at the beginning of the week, is n'/n. If an animal dies during
 

the week, n'/n is less than one. The number of animals dying during the
 

week is n minus n' . Animals whose lifetimes are censored are removed from
 

the set of surviving animals just before the beginning of the week. This
 

removal does not decrease the estimated conditional probability of
 

survival. The number of animals thus removed is found by subtracting the n
 

of this week from the n' of the previously listed week. The fourth column,
 

labeled P, is the product of the factors n'/n for all weeks up to and
 

including the current week j . It is the Kaplan-Meier product-limit estimate of the unconditional probability of an animal surviving from the
 

beginning of the chronic experiment through the end of the current week j.
 

88
 
Comparison of Survival Among Groups
 

A statistical test described by Cox (1972, p. 197), and developed by Mantel
 

(1963, 1966) and Cochran (1954) is used to compare the survival of 2 groups
 

of animals. Table XXc shows results of the test for rats, and Table XXIc
 

shows them for mice. The test accumulates over weeks the observed number
 

of deaths in the higher dosed group minus its expected value under the null
 

hypothesis of equal probability of death in each group. This accumulated
 

statistic is denoted by U in the tables. The test also accumulates the
 

variance of the observed number of deaths. The total variance is denoted
 

by V. A statistic Ζ is computed by dividing U by the square root of V.
 

Since Ζ is distributed approximately as a standard normal random variable,
 

the probability of exceeding Ζ is listed in the column labeled P. Small
 

values of Ρ indicate that the probability of death in the higher dosed
 

group is significantly larger than in the low dosed group at significance
 

level P, indicating longer life for the lower dosed group. The calculation
 

of U and V begins with forming a two-by-two table for each week in which a
 

death has occurred in either group. A typical table is:
 

Lower Dose Higher Dose Total
 

Deaths during week χ Μ
 

Survivors of week No-Mo
 Νχ-Μχ N-Μ
 

Animals at risk during week N Ν
 0 Nl
 

The observed number of deaths in the higher dosed group is Μχ, and its
 

expected value under the null hypothesis is Εχ=ΝΜχ/Ν. Their difference,
 

Μχ-Εχ, is accumulated over weeks to form the test statistic U. The
 

variance V of the test statistic U is calculated by accumulating a contribution from each table of (Μ(Ν-Μ)Αχ(Ι-Αχ) / (N-l) , where Αχ=Νχ/Ν. Four
 

comparisons are made by this test: both dosed groups pooled vs. control,
 

low dose vs. control, high dose vs. control, and high dose vs. low dose.
 

A dose-response table developed by Tarone (1975) is also applied to the
 

two-by-three table:
 

Control Low Dose High Dose Total 

Deaths during week Mo χ M2 Μ 

Survivors of week NQ-MQ Νχ-Μχ N2 -M2 N-Μ 

Animals at risk during week NQ Ni N2 Ν Dosage do *1 d2 

The test statistic U and its variance V are accumulated over weeks. The
 

contribution to U from each table is:
 

d (M -E )+d (M -E )+d (M -E ) o o o 1 1 1 2 2 2
 , where Ej=MNj/N for j = 0, 1, or 2.
 

The contribution to V from each table is:
 

2 2 )2
 ((d A +d A +d A )-(d A +d A +d A )(M(N-M)/N-l)) 0 0 1 1 1 2 0 0 1 1 2 2
 , where Aj=Nj/N,
 

for j=0 , 1, or 2.
 

89
 
Estimation of Probabilities of Observing Tumors
 

The probabilities of observing tumors are estimated by a modification of
 

the product-limit procedure of Kaplan and Meier (1958). This modified
 

estimate is described by Saffiotti ejt al. (1972). When an animal dies or is
 

killed, it is evaluated histopathologically and marked as observed to have
 

developed the pathology associated with the mark under study or as not yet
 

having developed it. The animal being marked in this context is analogous
 

to the animal having died in the survival context, and the animal not being
 

marked is analogous to having the lifetime of the animal censored by
 

scheduled sacrifice or loss. If histopathological evaluation of the
 

pathology associated with the mark was not performed, the animal is not
 

considered in the analysis of the mark under study. The probability of
 

survival estimated by the product-limit procedure in the survival context
 

corresponds to the probability of not yet having observed a tumor. A more
 

interesting quantity is the probability of having observed a tumor, which
 

is found by subtracting the probability of not having observed a tumor from
 

one. The estimated probabilities of having observed hepatocellular
 

carcinoma of the liver (mark a) for male mice are given in Table XII, and
 

graphed in Figure 11. Figure 12 shows a comparison of incidence of
 

hepatocellular carcinoma in male and female mice.
 

Comparison of Probabilities of Observing Tumors Among Groups,
 

Adjusting for Age
 

The statistical tests of Cox (1972) and Tarone (1975) used to compare
 

survival among groups can be modified to compare the probabilities of
 

observing tumors among groups by employing the analogies described above
 

between death in the survival context and observation of a tumor in the
 

context of observing pathology. Specifically, in the two-by-two and twoby-three tables of the section on the comparison of survival among groups,
 

"Deaths during week" is replaced by "animals with observed tumor during
 

week". Tumors cannot be observed while the animals are alive. No
 

distinction is made between the natural death of an animal and the censoring of its lifetime in comparisons of tumor probabilities. Animals in
 

which tumors were not observed at histopathological examination are
 

censored from the group of surviving animals for the next week's
 

comparison.
 

Calculation and interpretation of the Cox and Tarone tests proceed just as
 

in their use in the survival context. Both tests are one-tailed in the
 

direction of increasing probability of tumor with increasing dose. Small
 

values of P indicate significantly greater probability of tumor in higher
 

dosed animals.
 

These tests compare animals at the same ages in the groups under
 

comparison, removing biases introduced by differing death rates in control
 

and dosed groups.
 

Results of these tests are shown in Tables Xllla-e for rats and XXVIa-e for
 

mice.
 

90
 
Comparison of Probabilities of Observing Tumors Among Groups, by Exact
 

Tests (Not Adjusted for Age)
 

When the ages of animals at death or sacrifice are ignored, comparison of
 

probabilities of tumors between two groups is performed by the well known
 

Fisher-Irwin exact test, which is a one-tailed test. (See, e_.£., Armitage
 

(1971) for a description.) For three groups, with dosages associated with
 

each group, the test, essentially due to Armitage (1971), employs the
 

linear contrast statistic
 

U = d0M0 + d1M1 + d2M2
 

where di = ith dosage and Mi = number of animals with the tumor in the ith
 

group.
 

The exact distribution of U is computed under the null hypothesis that all
 

groups have the same probability of tumor. The probability P that the
 

observed U is equalled or exceeded is given in tables with the observed
 

proportions and percentages of tumors.
 

To test for overall heterogeneity among three groups not necessarily
 

related to the dosages, the two degree of freedom chi-square statistic for
 

the classical Pearson test for independence is computed from the data and
 

compared with the exact distribution under the null hypothesis as above.
 

Finally, chi-square statistics with one degree of freedom each are used in
 

exact tests partitioning the two degree of freedom statistic into
 

components for a linear trend on a logistic scale and deviation from the
 

linear trend.
 

Results of these tests are shown in Tables XXIVa-d for rats and Table XXVII
 

for mice.
 

91
 
Table XVII. Identity of Tumor Marks

Mark Tumor

a Hepatocellular carcinoma

b Reticulum-cell sarcoma, lymphosarcoma, or malignant

lymphoma

c Carcinoma or adenocarcinoma of the lung or alveoli

d Adenoma of the lung

f Aortic body tumor

g Fibroadenoma of the mammary glands

h Hemangiosarcoma of any site

i Malignant giant-cell tumor of soft tissues or

fibrosarcoma of the skin or subcutis

j Malignant mixed tumor of the kidney

k Hamartoma of the kidney

1 Follicular adenoma of the thyroid

m Tubular adenocarcinoma of the kidney

n Fibroma of the subcutis

p Follicular adenocarcinoma of the thyroid

q Squamous cell carcinoma of the subcutis

r Pilomatrixoma of the skin

t Chromophobe adenoma of the pituitary

u Adenocarcinoma of the mammary glands

v Granulosa-cell carcinoma of the ovaries

w Adrenal cortical carcinoma

x Liposarcoma of any site

y Sarcoma of the endometrium or fibrosarcoma of the

uterus

z Adenoma of the Harderian gland

A Neurofibroma of any site

B Adenoma of the kidney

C Papilloma of the stomach

D Osteosarcoma

E Adenocarcinoma of the endometrium

F Cystadenoma of the ovaries

92
Table XVIIIa. Data for Statistical Analysis - Trichloroethylene-


Treated Male Rats
 

Control Low Dose High Dose
 

Week on Animal Mark Week on Animal Mark Week on Animal Hark 

Study Number Study Number Study Number 

67 3 16 28 2 12 

70 11 h 17 36 2 32 

76 14 23 10 5 14 

76 15 27 6 6 4 

82 9 34 35 12 15 

82 13 40 30 17 22 

83 18 42 34 21 6 

87 16 48 3 27 2 

88 5 50 7 31 25 

88 12 53 41 33 24 

90 2 i 60 46 35 40 

91 4 61 39 40 33 

96 19 jl 65 19 42 39 

98 20 67 32 44 3 

99 10 jk 67 44 48 18 

102 17 72 40 49 30 

103 8 74 47 52 5 

110* 1 76 18 52 41 

110* 6 76 23 1 53 37 

110 7 c 80 26 h 54 13 

80 33 54 17 

82 5 56 27 * Animal's observed 83 38 58 47 

lifetime censored 83 43 59 9 

by scheduled sacri­ 85 1 61 29 fice, accidental 86 14 62 7 killing or loss. 86 20 62 38 Absence of * means 87 29 65 11 ρ natural death or 88 12 65 28 

moribund sacrifice. 90 15 m 65 45 

94 25 η 66 49 

94 49 68 10 

 Histopathology not 96 21 70 20 

performed. 97 16 71 16 

102 9 72 34 r 

103 13 72 50 

103 27 75 44 

103 31 76 21 

104 37 82 19 f 

107 8 83 43 

107 45 88 31 

108 4 i 91 36 h 

110* 2 kp 97 42 

110* 

110* 

11 

17 

q 99 

102 

8 

35 

110* 22 103 1 

110* 24 109 48 

110 42 110* 23 

110* 48 110* 26 h 

110* 50 110* 46 

93
 
Table XVIIIb. Data foi : Statistical Analysis Tnchloroethylene-


Treated Female Rats
 

Control Low Dose Iiigh Dose
 

Week on Animal Mark Week on Animal Mark Week on Animal Mark
 

Study Number Study Number Study Number
 

25 8 2 24 3 13
 

47 16 2* 29 — 5 2
 

47 20 2 42 9 32
 

68 6 t 3 2 h 21 36
 

79 1 5 20 24 49
 

87 7 7 47 28 8 

97 11 g 15 1 28 16 

98 15 16 30 30 43
 

99 16 38 35 40
 

102 14 tu 19 11 43 38
 

104 19 t 21 12 46 45
 

108 12 21 14 50 4
 

110* 3 gt 21* 21 _ 53 7
 

110* 4 bv 22 5 54 41
 

110* 5 22 39 58 24
 

110* 9 25 17 59 17
 

110* 10 g 26 36 63 31
 

110* 13 28 35 64 22
 

110* 17 33 33 67 28
 

110* 18 34 19 69 12
 

40 45 69 27 

42 22 70 18 * Animal's observed 57 6 70 20
 

lifetime censored 60 10 72 14
 

by scheduled sacri 61 44 73 1
 

fice, accidental 68 27 73 42
 

killing or loss. 69 25 74 39
 

Absence of * means
 75 15 82 26
 

natural death or 75 16 b 86 48
 

moribund sacrifice. 77	 48	 89
 47
 

86 34 95 21
 

96 23 w 96 50 t
 

Histopathology not	 96 46 101 29 g
 performed. 98 32 X 101 35
 

100 13 η 102 19
 

102 49 103 33 g
 

104 40 g 104 34
 

110* 3 110* 3
 

110* 4 110* 5 t
 

110* 7 gt 110* 6 b
 

110* 8 110* 9 gt
 

110* 9 110* 10 g
 

110* 18 g 110* 11
 

110* 26 y 110* 15
 

110* 28 110* 23 gt
 

110* 31 t 110* 25
 

110* 37 110* 30 gp
 

110* 41 110* 37 gy
 110* 43 g 110* 44
 

110 50 g 110* 46 t
 

94
 
Table XIXa. Data for Statistical Analysis - Trichloroethylene-


Treated Male Mice
 

Control Low Dose High Dose 

Week on Animal Mark Week on Animal Mark Week on Animal Mark 

Study Number Study Number Study Number 

32 10 16 10 13 10 

39 9 18 50 b 14 40 

60 8 31* 9 15 49 

61 7 51 30 15* 50 

64 6 53 40 24 19 

66 20 58 8 24 20 

68 5 63 39 25 47 

72 19 a 65 20 25 48 

76 4 65 38 27 46 a 

76 18 b 77 19 28 18 a 

77 17 i 81 28 b 28 45 

78 3 81 29 ad 29 30 

90* 1 i 86 49 a 30 9 

90* 2 88 7 ad 30 17 a 

90* 11 i 88 18 36 8 a 

90* 12 90* 1 ad 42 7 

90* 13 90* 2 η 53 39 a 

90* 14 90* 3 a 60 5 

90* 15 90* 4 abd 60 6 

90* 16 90* 5 a 61 16 a 

90* 6 a 70 38 a 

90* 11 a 71 15 a * Animal's observed 90* 12 a 72 37 

lifetime censored 90* 13 a 74 29 a 

by scheduled sacri­ 90* 14 ζ 75 4 

fice, accidental 90* 15 ab 75 36 _ killing or loss. 90* 16 78 44 aA 

Absence of * means 90* 17 a 83 35 ac 

natural death or 90* 21 a 88 3 ab 

moribund sacrifice. 90* 22 90* 1 a 

90* 23 90* 2 a 

90* 24 a 90* 11 a 

 Histopathology not 90* 25 90* 12 a 

performed. 90* 26 a 90* 13 aB 

90* 27 90* 14 a 

90* 31 a 90* 21 a 

90* 32 a 90* 22 a 

90* 33 a 90* 23 a 

90* 34 a 90* 24 b 

90* 35 a 90* 25 a 

90* 36 a 90* 26 a 

90* 37 ad 90* 27 a 

90* 41 a 90* 28 a 

90* 42 90* 31 d 

90* 43 a 90* 32 a 

90* 44 90* 33 aC 

90* 45 90 34 ai 

90* 46 90* 41 a 

90* 47 90* 42 a 

90* 48 a 90* 43 ah 

95
 
Table XlXb. Data for Statistical Analysis - Trichloroethylene-


Treated Female Mice
 

Control Low Dose High Dose
 

Week on Animal Mark Week on Animal
 

Study Number Study Number
 

20* 10 10 20
 

37* 9 32* 10
 

83* 8 D 37 19
 

90* 1 38 18
 

90* 2 41* 9
 

90* 3 Ε 55 30
 

90* 4 63 29
 

90* 5 66 40
 

90* 6 76 17
 

90* 7 d 81 16
 

90* 11 90* 1
 

90* 12 90* 2
 

90* 13 90* 3
 

90* 14 b 90* 4
 

90* 15 90* 5
 

90* 16 90* 6
 

90* 17 90* 7
 

90* 18 90* 8
 

90* 19 90* 11
 

90* 20 90* 12
 

90* 13
 

90* 14
 * Animal's observed	 90* 15
 

lifetime censored 90* 21
 

by scheduled sacri 90* 22
 

fice, accidental
 90* 23
 

killing or loss.
 90* 24
 

Absence of * means
 90* 25
 

natural death or
 90* 26
 moribund sacrifice.	 90* 27
 

90* 28
 

90* 31
 

 Histopathology not 90* 32
 

performed.	 90* 33
 

90* 34
 

90* 35
 

90* 36
 

90* 37
 

90* 38
 

90* 39
 

90* 41
 

90* 42
 

90* 43
 

90* 44
 

90* 45
 

90* 46
 

90* 47
 

90* 48
 

90* 49
 

90* 50
 

Mark
 

b
 

b
 

b
 

ay
 

a
 

a
 

c
 

a
 

V
 

i
 

bd
 

u
 

bd
 

czF
 

Week on Animal
 

Study
 

14*
 

22*
 

26*
 

32
 

32
 

33
 

38
 

39
 

40
 

69
 

88
 

90*
 

90*
 

90*
 

90*
 

90*
 

90*
 

90*
 

90*
 

90*
 

90*
 

90*
 

90*
 

90*
 

90*
 

90*
 

90*
 

90*
 

90*
 

90*
 

90*
 

90*
 

90*
 

90*
 

90*
 

90*
 

90*
 

90*
 

90*
 

90*
 

90*
 

90*
 

90*
 

90*
 

90*
 

90*
 

90*
 

90*
 

90*
 

90*
 

Number
 

10
 

40
 

50
 

30
 

49
 

39
 

38
 

20
 

37
 

9
 

36
 

1
 

2
 

3
 

4
 

5
 

6
 

7
 

8
 

11
 

12
 

13
 

14
 

15
 

16
 

17
 

18
 

19
 

21
 

22
 

23
 

24
 

25
 

26
 

27
 

28
 

29
 

31
 

32
 

33
 

34
 

35
 

41
 

42
 

43
 

44
 

45
 

46
 

47
 

48
 

Mark
 

—
 

-


-


b
 

b
 

b
 

a
 

ac
 

a
 

a
 

d
 

b
 

b
 

d
 

a
 

ac
 

a
 

d
 

abd
 

a
 

a
 

ad
 

96
 
Table XXa. Product-Limit Estimates of Probability of Survival ­

Trichloroethylene-Treated Male Rats
 

Control
 

j n n' P 

0 20 20 1.000 

67 20 19 .950 

70 19 18 .900 

76 18 16 .800 

82 16 14 .700 

83 14 13 .650 

87 13 12 .600 

88 12 10 .500 

90 10 9 .450 

91 9 8 .400 

96 8 7 .350 

98 7 6 .300 

99 6 5 .250 

102 5 4 .200 

103 4 3 .150 

110 3 2 .100 

j - Week on study
 

n = No, of animals
 

alive at beginning


of the week
 

n' = No. of animals
 

surviving the week
 

P = Kaplan-Meier


estimate of sur­ vival probability
 

Low Dose High Dose 

j n n' P j n n' P 

0 50 50 1.000 0 50 50 1 .000 

16 50 49 .980 2 50 48 .960 

17 49 48 .960 5 48 47 .940 

23 48 47 .940 6 47 46 .920 

27 47 46 .920 12 46 45 .900 

34 46 45 .900 17 45 44 .880 

40 45 44 .880 21 44 43 .860 

42 44 43 .860 27 43 42 .840 

48 43 42 .840 31 42 41 .820 

50 42 41 .820 33 41 40 .800 

53 41 40 .800 35 40 39 .780 

60 40 39 .780 40 39 38 .760 

61 39 38 .760 42 38 37 .740 

65 38 37 .740 44 37 36 .720 

67 37 35 .700 48 36 35 .700 

72 35 34 .680 49 35 34 .680 

74 34 33 .660 52 34 32 .640 

76 33 31 .620 53 32 31 .620 

80 31 29 .580 54 31 29 .580 

82 29 28 .560 56 29 28 .560 

83 28 26 .520 58 28 27 .540 

85 26 25 .500 59 27 26 .520 

86 25 23 .460 61 26 25 .500 

87 23 22 .440 62 25 23 .460 

88 22 21 .420 65 23 20 .400 

90 21 20 .400 66 20 19 .380 

94 20 18 .360 68 19 18 .360 

96 18 17 .340 70 18 17 .340 

97 17 16 .320 71 17 16 .320 

102 16 15 .300 72 16 14 .280 

103 15 12 .240 75 14 13 .260 

104 12 11 .220 76 13 12 .240 

107 11 9 .180 82 12 11 .220 

108 9 8 .160 83 11 10 .200 

110 8 7 .140 88 10 9 .180 

91 9 8 .160 

97 8 7 .140 

99 7 6 .120 

102 6 5 .100 

103 5 4 .080 

109 4 3 .060 

110 3 3 .060 

97
 
Table XXb. Product-Limit Estimates of Probability of Survival ­

Trichloroethylene-Treated Female Rats
 

Control
 Low Dose High Dose 

j η η ' Ρ j η η ' Ρ 

0 50 50 1.000 0 50 50 1 .000 

2 50 48 .960 3 50 49 .980 

3 47 46 .940 5 49 48 .960 

5 46 45 .919 9 48 47 .940 

7 45 44 .899 21 47 46 .920 

15 44 43 .878 24 46 45 .900 

16 43 41 .837 28 45 43 .860 

19 41 40 .817 30 43 42 .840 

21 40 38 .776 35 42 41 .820 

22 37 35 .734 43 41 40 .800 

25 35 34 .713 46 40 39 .780 

26 34 33 .692 50 39 38 .760 

28 33 32 .671 53 38 37 .740 

33 32 31 .650 54 37 36 .720 

34 31 30 .629 58 36 35 .700 

40 30 29 .608 59 35 34 .680 

42 29 28 .587 63 34 33 .660 

57 28 27 .566 64 33 32 .640 

60 27 26 .545 67 32 31 .620 

61 26 25 .524 69 31 29 .580 

68 25 24 .503 70 29 27 .540 

69 24 23 .482 72 27 26 .520 

75 23 21 .441 73 26 24 .480 

77 21 20 .420 74 24 23 .460 

86 20 19 .399 82 23 22 .440 

96 19 17 .357 86 22 21 .420 

98 17 16 .336 89 21 20 .400 

100 16 15 .315 95 20 19 .380 

102 15 14 .294 96 19 18 .360 

104 14 13 .273 101 18 16 .320 

110 13 12 .252 102 16 15 .300 

103 15 14 .280 

104 14 13 .260 

110 13 13 .260 

j
 

0
 

25
 

47
 

68
 

79
 

87
 

97
 

98
 

99
 

102
 

104
 

108
 

110
 

η
 

20
 

20
 

19
 

17
 

16
 

15
 

14
 

13
 

12
 

11
 

10
 

9
 

8
 

η '
 

20


19
 

17
 

16
 

15
 

14
 

13
 

12
 

11
 

10
 

9
 

8
 

8
 

1
 

Ρ
 

.000
 

.950
 

.850
 

.800
 

.750
 

.700
 

.650
 

.600
 

.550
 

.500
 

.450
 

.400
 

.400
 

j = Week on study
 

n = No. of animals
 

alive at beginning
 

of the week
 

n' = No. of animals
 

surviving the week
 

 = Kaplan-Meier
 

estimate of sur­ vival probability
 

98
 
Table XXc. Statistical Tests Comparing Estimated Probability of
 

Survival among Control and Trichloroethylene-Treated Rats
 

Male Rats Female Rats 

Comparison U V Ζ Ρ υ V Ζ Ρ 

Dose

Response 23.60 51.47 3.29 0.001 8.48 50.64 1.19 0.117 

Dosed vs. Control 6.47 18.17 1.52 0.064 7.13 14.55 1.87 0.031 

Low Dose vs. Control 0.88 12.44 0.25 0.402 6.33 11.08 1.90 0.028 

High Dose vs. Control 11.12 15.07 2.86 0.002 5.50 11.04 1.65 0.049 

High Dose vs. Low Dose 13.94 19.86 3.13 0.001 -2.56 17.84 -0.61 0.728 

U = observed test statistic minus its expected value under the null
 

hypothesis.
 

V = variance of U.
 

 = U divided by square root V.
 

 = probability that a standard normal random variable is greater than Z,
 

See appendix on statistical methodology for full definition.
 

99
 
Table XXIa. Product-Limit Estimates of Probability of Survival
 

Trichloroethylene-Treated Male Mice
 

Control Low Dose High Dose 

j η' Ρ j η n' Ρ j η n' Ρ 0 20 20 1.000 0 50 50 1.000 0 50 50 1.000 

32 20 19 .950 16 50 49 .980 13 50 49 .980 

39 19 18 .900 18 49 48 .960 14 49 48 .960 

60 18 17 .850 31 48 48 .960 15 48 47 .940 

61 17 16 .800 51 47 46 .940 24 46 44 .899 

64 16 15 .750 53 46 45 .919 25 44 42 .858 

66 15 14 .700 58 45 44 .899 27 42 41 .838 

68 14 13 .650 63 44 43 .878 28 41 39 .797 

72 13 12 .600 65 43 41 .837 29 39 38 .777 

76 12 10 .500 77 41 40 .817 30 38 36 .736 

77 10 9 .450 81 40 38 .776 36 36 35 .715 

78 9 8 .400 86 38 37 .756 42 35 34 .695 

90 8 8 .400 88 37 35 .715 53 34 33 .674 

90 35 35 .715 60 33 31 .633 

j ­ Week on study 61 

70 

31 

30 

30 

29 

.613 

.593 = No. of animals 71 29 28 .572 

alive at beginning 72 28 27 .552 

of the week 74 27 26 .531 

75 26 24 .490 

n' == No, of animals 78 24 23 .470 

surviving the week 83 23 22 .450 

88 22 21 .429 = Kaplan-Meier 90 21 20 .409 

estimate of sur­ vival probability 

Table XXIb. Product-Limit Estimates of Probability of Survival
 

Trichloroethylene-Treated Female Mice
 

Control Low Dose High Dose 

j η η' Ρ j η n' Ρ j η n' Ρ 0 20 20 1.000 0 50 50 1 .000 0 50 50 1.000 

20 20 20 1.000 10 50 49 .980 14 50 50 1.000 

37 19 19 1.000 32 49 49 .980 22 49 49 1.000 

83 18 18 1.000 37 48 47 .960 26 48 48 1.000 

90 17 17 1.000 38 47 46 .939 32 47 45 .957 

41 46 46 .939 33 45 44 .936 

j = Week on study 55 45 44 .918 38 44 43 .915 

63 44 43 .897 39 43 42 .894 = No. of animals 66 43 42 .877 40 42 41 .872 

alive at beginning 76 42 41 .856 69 41 40 .851 

of the week 81 41 40 .835 88 40 39 .830 

90 40 40 .835 90 39 39 .830 

n' = No. of animals 

surviving the week 

 = Kaplan-Meier 

estimate of sur­ vival probability 

100
 
Table XXIc. Statistical Tests Comparing Estimated Probability of
 

Survival among Control and Trichloroethylene-Treated Mice
 

Male Mice Female Mice 

Comparison U V Z P U V Z P 

DoseResponse 6.67 26.11 1.30 0.096 4.29 8.32 1.49 0.068 

Dosed vs. 

Control -3.02 7.42 -1.11 0.866 2.75 2.27 1.82 0.034 

Low Dose vs. 

Control -5.72 4.68 -2.64 0.996 2.32 1.65 1.81 0.035 

High Dose vs. 

Control 1.11 8.77 0.37 0.354 2.36 1.66 1.83 0.033 

High Dose vs. 

Low Dose 10.72 10.35 3.33 0.001 0.21 3.99 0.10 0.459 

U = observed test statistic minus its expected value under the null
 

hypothesis.
 

V = variance of U.
 

Z = U divided by square root V.
 

P = probability that a standard normal random variable is greater than Z,
 

See appendix on statistical methodology for full definition.
 

101
 
Table XXII. Tumor Incidence - Trichloroethylene-Treated Rats
 

Male i Rats Female Rats
 

Low High Low High
 

Control Dose Dose Control Dose Dose
 

Mark b Reticulum-cell sarcoma, lymphosarcoma, or malignant lymphoma
 

Before 110 weeks 0/17 0/42 0/47 0/12 1/35 0/37
 

At 110 weeks 0/3 0/8 0/3 1/8 0/13 1/13
 

Total 0/20 0/50 0/50 1/20 1/48 1/50
 

Mark g Fibroadenoma of the mammary glands
 

Before 110 weeks 0/17 0/42 0/47 1/12 1/35 2/37
 

At 110 weeks 0/3 0/8 0/3 2/8 4/13 5/13
 

Total 0/20 0/50 0/50 3/20 5/48 7/50
 

Mark h Hemangiosarcoma of any site
 

Before 110 weeks 1/17 1/42 1/47 0/12 1/35 0/37
 

At 110 weeks 0/3 0/8 1/3 0/8 0/13 0/13
 

Total 1/20 1/50 2/50 0/20 1/48 0/50
 

Mark p Follicular adenocarcinoma of the thyroid
 

Before 110 weeks 0/17 0/42 1/47 0/12 0/35 0/37
 

At 110 weeks 0/3 1/8 0/3 0/8 0/13 1/13
 

Total 0/20 1/50 1/50 0/20 0/48 1/50
 

Mark t Chromophobe adenoma of the pituitary
 

Before 110 weeks 0/17 0/42 0/47 3/12 0/35 2/37
 

At 110 weeks 0/3 0/8 0/3 1/8 2/13 4/13
 

Total 0/20 0/50 0/50 4/20 2/48 6/50
 

Animals with Tumors (Benign and Malignant)a
 

Before 110 weeks 4/17 5/42 4/47 4/12 6/35 4/37
 

At 110 weeks 1/3 2/8 1/3 3/8 6/13 8/13
 

Total 5/20 7/50 5/50 7/20 12/48 12/50
 

Includes tumors other than those listed above.
 

102
 
Table XXIIIa. Statistical Tests Comparing Estimated Probability of
 

Observing Reticulum-cell Sarcoma, Lymphosarcoma, or Malignant Lymphoma
 

(Mark b) Control and Trichloroethylene-Treated Rats
 

Male Rats Female Rats
 

Comparison U V Ζ Ρ υ V Ζ Ρ 

Dose

Response 0.00 0.00 -0.41 1.77 -0.31 0.620 

Dosed vs. Control 0.00 0.00 -0.27 0.54 -0.37 0.644 

Low Dose vs. Control 0.00 0.00 -0.21 0.48 -0.30 0.619 

High Dose vs. Control 0.00 0.00 -0.24 0.45 -0.36 0.639 

High Dose vs. Low Dose 0.00 0.00 0.00 0.50 0.00 0.500 

U = observed test statistic minus its expected value under the null
 

hypothesis.
 

V = variance of U.
 

 = U divided by square root V.
 

 = probability that a standard normal random variable is greater than Z,
 

See appendix on statistical methodology for full definition.
 

103
 
Table XXIIIb. Statistical Tests Comparing Estimated Probability
 

of Observing Fibroadenoma of the Mammary Glands (Mark g)
 

among Control and Trichloroethylene-Treated Rats
 

Male Rats Female Rats 

Comparison U V Ζ Ρ υ V Ζ Ρ 

Dose

Response 0.00 0.00 1.91 7.09 0.72 0.237 

Dosed vs. Control 0.00 0.00 0.64 2.16 0.44 0.330 

Low Dose vs. Control 0.00 0.00 0.15 1.55 0.12 0.451 

High Dose vs. Control 0.00 0.00 0.88 1.88 0.64 0.259 

High Dose vs. Low Dose 0.00 0.00 0.94 2.28 0.62 0.267 

Table XXIlie. Statistical Tests Comparing Estimated Probability
 

of Observing Hemangioma of Any Site (Mark h)
 

among Control and Trichloroethylene-Treated Rats
 

Male Rats Female Rats 

Comparison U V Ζ Ρ υ V Ζ Ρ 

Dose

Response 1.08 1.89 0.79 0.215 -0.26 0.54 -0.35 0.638 

Dosed vs. Control -0.01 0.74 -0.02 0.506 0.17 0.14 0.46 0.324 

Low Dose vs. Control -0.31 0.45 -0.46 0.677 0.30 0.21 0.66 0.255 

High Dose vs. Control 0.51 0.75 0.59 0.276 0.00 0.00 

High Dose vs. Low Dose 1.14 0.61 1.45 0.073 -0.52 0.25 -1.04 0.851 

U = observed test statistic minus its expected value under the null
 

hypothesis.
 

V = variance of U.
 

 = U divided by square root V.
 

 = probability that a standard normal random variable is greater than Z,
 

See appendix on statistical methodology for full definition.
 

104
 
Table XXIIId. Statistical Tests Comparing Estimated Probability
 

of Observing Follicular Adenocarcinoma of the Thyroid (Mark p)
 

among Control and Trichloroethylene-Treated Rats
 

Male Rats Female Rats 

Comparison U V Ζ Ρ υ V Ζ Ρ 

Dose

Response 0.96 0.96 0.98 0.163 0.85 0.60 1.10 0.135 

Dosed vs. Control 0.46 0.35 0.77 0.219 0.24 0.18 0.55 0.289 

Low Dose vs. Control 0.27 0.20 0.61 0.270 0.00 0.00 

High Dose vs. Control 0.47 0.25 0.93 0.176 0.38 0.24 0.78 0.216 

High Dose vs. Low Dose 0.35 0.43 0.53 0.297 0.50 0.25 1.00 0.159 

Table XXIIIe. Statistical Tests Comparing Estimated Probability
 

of Observing Chromophobe Adenoma of the Pituitary (Mark t)
 

among Control and Trichloroethylene-Treated Rats
 

Male Rats Female Rats 

Comparison U V Ζ Ρ υ V Ζ Ρ 

Dose

Response 0.00 0.00 0.30 6.54 0.12 0.454 

Dosed vs. Control 0.00 0.00 -1.08 1.98 -0.76 0.778 

Low Dose vs. Control 0.00 0.00 -1.61 1.36 -1.38 0.916 

High Dose vs. Control 0.00 0.00 -0.11 2.14 -0.08 0.531 

High Dose vs. Low Dose 0.00 0.00 1.95 1.70 1.50 0.067 

U = observed test statistic minus its expected value under the null
 

hypothesis.
 

V = variance of U.
 

 = U divided by square root V.
 

 = probability that a standard normal random variable is greater than Z.
 

See appendix on statistical methodology for full definition.
 

105
 
Table XXIVa. Exact Statistical Tests Comparing Proportions of Animals (Not
 

Adjusted for Age) with Observed Hepatocellular Carcinoma of the Liver
 

among Pooled Control and Trichloroethylene-Treated Rats
 

14ale Rats Female Rats
 

Veh. Low High Exact Veh. Low High Exact
 

Cont. Dose Dose Test Cont. Dose Dose Test
 

Comparison r/n r/n r/n P r/n r/n r/n P
 

Dose-Response 1/99 0/50 0/50 0/98 0/48 0/50
 

Veh. Control 1% 0% 0% 1.000 0% 0% 0% 1.000
 

Dosed vs. 1/99 0/100 0/98 0/98
 

Veh. Control 1% 0% 1.000 0% 0% 1.000
 

Low Dose vs. 1/99 0/50 0/98 0/48
 

Veh. Control 1% 0% 1.000 0% 0% 1.000
 

High Dose vs. 1/99 0/50 0/98 0/50
 

Veh. Control 1% 0% 1.000 0% 0% 1.000
 

High Dose vs. 0/50 0/50 0/48 0/50
 

Low Dose 0% 0% 1.000 0% 0% 1.000
 

Comparison Chi-square df P Chi-square df P
 

Among High Dose,
 

Low Dose and
 

Vehicle Control 1.02 2 1.000 1.00 2 1. 000
 

Dose-Response Trend
 

Vehicle Control 0.83 1 0.749 1.00 1 1. 000
 

Deviation from Trend
 

Vehicle Control 0.19 1 1.000 0.00 1 1. 000
 

r = Number of animals with observed mark.
 

n = Number of animals examined histopathologically for the mark.
 

P = Exact probability of exceeding or equaling observed test
 

statistic under null hypothesis,
 

df = Degrees of freedom.
 

See section on statistical methodology for explanation of tests.
 

106
 
Table XXIVb. Exact Statistical Tests Comparing Proportions of Animals (Not
 

Adjusted for Age) with Observed Hepatocellular Carcinoma or Neoplastic
 

Nodule of the Liver among Pooled Control and Trichloroethylene-Treated Rats
 

Male Rats ]Female Rats 

Veh. Low High Exact Veh. Low High Exact 

Cont. Dose Dose Test Cont. Dose Dose Test 

Comparison r/n r/n r/n Ρ r/n r/n r/n Ρ
 

Dose-Response 1/99 0/50 0/50 2/98 0/48 0/50
 

Veh. Control 1% 0% 0% 1.000 2% 0% 0% 1.000
 

Dosed vs. 1/99 0/100 2/98 0/98
 

Veh. Control 1% 0% 1.000 2% 0% 1.000
 

Low Dose vs. 1/99 0/50 2/98 0/48
 

Veh. Control 1% 0% 1.000 2% 0% 1.000
 

High Dose vs. 1/99 0/50 2/98 0/50
 

Veh. Control 1% 0% 1.000 2% 0% 1.000
 

High Dose vs. 0/50 0/50 0/48 0/50
 

Low Dose 0% 0% 1.000 0% 0% 1.000
 

Comparison Chi-square df Ρ Chi-square df Ρ
 

Among High Dose,
 

Low Dose and
 

Vehicle Control 1.02 2 1.000 2.04 2 0.371
 

Dose-Response Trend
 

Vehicle Control 0.83 1 0.749 1.67 1 0.310
 

Deviation from Trend
 

Vehicle Control 0.19 1 1.000 0.37 1 1.000
 

r = Number of animals with observed mark.
 

 = Number of animals examined histopathologically for the mark.
 

 = Exact probability of exceeding or equaling observed test
 

statistic under null hypothesis,
 

df = Degrees of freedom.
 

See section on statistical methodology for explanation of tests.
 

107
 
Table XXIVc. Exact Statistical Tests Comparing Proportions of Animals (Not
 

Adjusted for Age) with Observed Hepatocellular Carcinoma
 

of the Liver among Pooled Control and Carbon Tetrachloride-Treated Rats
 

Male Rats Female Rats 

Veh. Low High Exact Veh. Low High Exact 

Cont. Dose Dose Test Cont. Dose Dose Test 

Comparison r/n r/n r/n Ρ r/n r/n r/n Ρ
 

Dose-Response 1/99 2/50 2/50 0/98 4/49 1/49
 

Veh. Control 1% 4% 4% 0.177 0% 8% 2% 0.174
 

Dosed vs. 1/99 4/100 0/98 5/98
 

Veh. Control 1% 4% 0.187 0% 5% 0.030
 

Low Dose vs. 1/99 2/50 0/98 4/49
 

Veh. Control 1% 4% 0.261 0% 8% 0.011
 

High Dose vs. 1/99 2/50 0/98 1/49
 

Veh. Control 1% 4% 0.261 0% 2% 0.333
 

High Dose vs. 2/50 2/50 4/49 1/49
 

Low Dose 4% 4% 0.691 8% 2% 0.972
 

Comparison Chi-square df Ρ Chi-square df Ρ
 

Among High Dose,
 

Low Dose and
 

Vehicle Control 1.82 2 0.601 8.83 2 0.011
 

Dose-Response Trend
 

Vehicle Control 1.48 1 0.282 1.51 1 0.281
 

Deviation from Trend
 

Vehicle Control 0.33 1 0.635 7.31 1 0.014
 

r = Number of animals with observed mark.
 

 = Number of animals examined histopathologically for the mark.
 

 = Exact probability of exceeding or equaling observed test
 

statistic under null hypothesis,
 

df = Degrees of freedom.
 

See section on statistical methodology for explanation of tests.
 

108
 
Table XXIVd. Exact Statistical Tests Comparing Proportions of Animals (Not
 

Adjusted for Age) with Observed Hepatocellular Carcinoma or Neoplastic Nodule
 

of the Liver among Pooled Control and Carbon Tetrachloride-Treated Rats
 

Comparison
 

Dose-Response
 

Veh. Control
 

Dose-Response
 

Untr. Control
 

Dosed vs.
 

Veh. Control
 

Dosed vs.
 

Untr. Control
 

Low Dose vs.
 

Veh. Control
 

Low Dose vs.
 

Untr. Control
 

High Dose vs.
 

Veh. Control
 

High Dose vs.
 

Untr. Control
 

High Dose vs.
 

Low Dose
 

Veh. Con. vs.
 

Untr. Control
 

Untr.
 

Cont.
 

r/n
 

0/20
 

0%
 

0/20
 

0%
 

0/20
 

0%
 

0/20
 

0%
 

0/20
 

0%
 

Male Rats
 Female Rats 

Exact Untr. Veh. Low High Exact 

Test Cont. Cont. Dose Dose Test 

Ρ r/n r/n r/n r/n Ρ 

2/98 6/49 4/49 

0.070 2% 12% 8% 0.057 

0/20 6/49 4/49 

0.353 0% 12% 8% 0.325 

2/98 10/98 

0.033 2% 10% 0.016 

0/20 10/98 

0.269 0% 10% 0.144 

2/98 6/49 

0.044 2% 12% 0.017 

0/20 6/49 

0.251 0% 12% 0.117 

2/98 4/49 

0.110 2% 8% 0.096 

0/20 4/49 

0.358 0% 8% 0.245 

6/49 4/49 

0.782 12% 8% 0.841 

0/20 2/98 

0.832 0% 2% 0.689 

(continued) 

Veh.
 

Cont.
 

r/n
 

1/99
 

1%
 

1/99
 

1%
 

1/99
 

1%
 

1/99
 

1%
 

1/99
 

1%
 

Low High
 

Dose Dose
 

r/n r/n
 

4/50 3/50
 

8% 6%
 

4/50 3/50
 

8% 6%
 

7/100
 

7%
 

7/100
 

7%
 

4/50
 

8%
 

4/50
 

8%
 

3/50
 

6%
 

3/50
 

6%
 

4/50 3/50
 

8% 6%
 

109
 
Table XXIVd. Exact Statistical Tests Comparing Proportions of Animals (Not
 

Adjusted for Age) with Observed Hepatocellular Carcinoma or Neoplastic Nodule
 

of the Liver among Pooled Control and Carbon Tetrachloride-Treated Rats
 

(continued) 

Comparison Chi-square df Ρ Chi-square df Ρ 

Among High Dose, 

Low Dose and 

Vehicle Control 4.89 2 0.100 6.39 2 0.046 

Dose-Response Trend 

Vehicle Control 2.98 1 0.128 3.23 1 0.105 

Deviation from Trend 

Vehicle Control 1.90 1 0.203 3.16 1 0.079 

Among High Dose, 

Low Dose and 

Untreated Control 1.67 2 0.528 2.76 2 0.259 

Dose-Response Trend 

Untreated Control 0.46 1 0.601 0.50 1 0.511 

Deviation from Trend 

Untreated Control 1.21 1 0.333 2.26 1 0.165 

r = Number of animals with observed mark. 

 = Number of animals examined histopathologically for the mark. 

 = Exact probability of exceeding or equaling observed test 

statistic under null hypothesis, 

df = Degrees of freedom. 

See section on statistical methodology for explanation of tests. 

110
 
Table XXV. Tumor Incidence - Trichloroethylene-Treated Mice
 

Male Mice Female Mice
 

Control Low Dose High Dose Control Low Dose High Dose
 

Mark a Hepatocellular carcinoma of the liver
 

Before 90 weeks 1/12 3/15 12/27 0/3 0/10 0/8
 

At 90 weeks 0/8 23/35 19/21 0/17 4/40 11/39
 

Total 1/20 26/50 31/48 0/20 4/50 11/47
 

•Mark b Reticulum-cell sarcoma, lymphosarcoma, or malignant lymphoma
 

Before 90 weeks 1/12 2/15 1/27 0/3 2/10 2/8
 

At 90 weeks 0/8 2/35 1/21 1/17 3/40 4/39
 

Total 1/20 4/50 2/48 1/20 5/50 6/47
 

Mark c Carcinoma or adenocarcinoma of the lung or alveoli
 

Before 90 weeks 0/12 0/15 1/27 0/3 0/10 0/8
 

At 90 weeks 0/8 0/35 0/21 0/17 2/40 2/39


Total 0/20 0/50 1/48 0/20 2/50 2/47
 

Mark d Adenoma of the lung
 

Before 90 weeks 0/12 2/15 0/27 0/3 0/10 0/8
 

At 90 weeks 0/8 3/35 1/21 1/17 2/40 5/39
 

Total 0/20 5/50 1/48 1/20 2/50 5/47
 

Marks c or d Carcinoma, adenocarcinoma,or adenoma of the lung or alveoli
 

Before 90 weeks 0/12 2/15 1/27 0/3 0/10 0/8
 

At 90 weeks 0/8 3/35 1/21 1/17 4/40 7/39
 

Total 0/20 5/50 2/48 1/20 4/50 7/47
 

Animals with Tumors (Benign and Malignant)a
 

Before 90 weeks 3/12 5/15 12/27 1/3 2/10 2/8
 

At 90 weeks 2/8 25/35 21/21 3/17
 12/40 17/39
 

Total 5/20
 30/50 33/48 4/20 14/50 19/47
 

Includes tumors other than those listed above.
 

Ill
 
Table XXVIa. Statistical Tests Comparing Estimated Probability
 

of Observing Hepatocellular Carcinoma (Mark a)
 

among Control and Trichloroethylene-Treated Mice
 

Comparison
 

Dose-
 Response
 

Dosed vs.
 Control
 

Low Dose vs.
 Control
 

High Dose vs.
 Control
 

High Dose vs.
 Low Dose
 

Male Mice Female Mice
 

U V Ζ Ρ υ V Ζ Ρ 

18.57 13.34 5.08 0.000 7.56 6.79 2.90 0.002 

6.72 3.68 3.51 0.000 2.66 1.86 1.95 0.026 

4.04 2.27 2.68 0.004 1.19 0.79 1.34 0.090 

8.45 4.18 4.13 0.000 3.34 1.90 2.42 0.008 

9.27 6.08 3.76 0.000 3.59 3.08 2.05 0.020 

Table XXVIb. Statistical Tests Comparing Estimated Probability of
 

Observing Reticulum-cell Sarcoma, Lymphosarcoma, or Malignant Lymphoma
 

(Mark b) among Control and Trichloroethylene-Treated Mice
 

Male Mice Female Mice 

Comparison U V Ζ Ρ υ V Ζ Ρ 

Dose

Response -0.40 2.96 -0.23 0.591 2.26 6.07 0.92 0.180 

Dosed vs. Control -0.06 0.80 -0.07 0.527 1.13 1.67 0.88 0.190 

Low Dose vs. Control 0.06 0.82 0.06 0.476 0.80 1.21 0.72 0.234 

High Dose vs. Control -0.12 0.62 -0.16 0.563 1.12 1.40 0.95 0.172 

High Dose vs. Low Dose -0.34 1.39 -0.28 0.612 0.57 2.61 0.35 0.362 

U = observed test statistic minus its expected value under the null
 

hypothesis.
 

V = variance of U.
 

 = U divided by square root V.
 

 = probability that a standard normal random variable is greater than Z.
 

See appendix on statistical methodology for full definition.
 

112
 
Table XXVIc. Statistical Tests Comparing Estimated Probability of
 

Observing Carcinoma or Adenocarcinoma of the Lung or Alveoli
 

(Mark c) among Control and Trichloroethylene-Treated Mice
 

Male Mice Female Mice 

Comparison U V Ζ Ρ υ V Ζ Ρ 

Dose

Response 0.78 0.40 1.23 0.109 1.08 2.06 0.76 0.225 

Dosed vs. Control 0.12 0.10 0.36 0.358 0.71 0.56 0.94 0.173 

Low Dose vs. Control 0.00 0.00 0.60 0.41 0.93 0.176 

High Dose vs. Control 0.26 0.19 0.59 0.277 0.61 0.42 0.94 0.173 

High Dose vs. Low Dose 0.62 0.23 1.29 0.099 0.03 0.96 0.03 0.490 

Table XXVId. Statistical Tests Comparing Estimated Probability
 

of Observing Adenoma of the Lung (Mark d)
 

among Control and Trichloroethylene-Treated Mice
 

Male Mice Female Mice
 

Comparison U V Ρ υ
 V Ρ


Dose-
 Response -0.23 2.37 -0.15 0.560 2.17 3.93 1.09 0.138
 

Dosed vs.
 Control 0.73 0.62 0.93 0.177 0.42 1.08 0.40 0.344
 

Low Dose vs.
 Control 0.90 0.72 1.07 0.144 -0.11 0.61 -0.14 0.554
 

High Dose vs.
 Control 0.28 0.20 0.62 0.268 0.82
 1.15 0.76 0.222
 

High Dose vs.
 Low Dose -1.24 1.35 -1.06 0.856 1.54 1.62 1.22 0.112
 

U - observed test statistic minus its expected value under the null
 

hypothesis·
 

V = variance of U.
 

 = U divided by square root V.
 

 = probability that a standard normal random variable is greater than Z.
 

See appendix on statistical methodology for full definition.
 

113
 
Table XXVIe. Statistical Tests Comparing Estimated Probability of
 

Observing Carcinoma, Adenocarcinoma, or Adenoma of the Lung or Alveoli
 

(Mark c or d) among Control and Trichloroethylene-Treated Mice
 

Male Mice Female Mice 

Comparison U V Ζ Ρ υ V Ζ Ρ 

Dose

Response 0.55 2.77 0.33 0.370 3.25 5.63 1.37 0.085 

Dosed vs. Control 0.85 0.72 1.00 0.160 1.13 1.55 0.90 0.183 

Low Dose vs. Control 0.90 0.72 1.07 0.144 0.49 0.97 0.50 0.309 

High Dose vs. Control 0.53 0.39 0.85 0.197 1.43 1.48 1.18 0.120 

High Dose vs. Low Dose -0.62 1.59 -0.49 0.688 1.57 2.40 1.01 0.156 

U = observed test statistic minus its expected value under the null
 

hypothesis.
 

V = variance of U.
 

 = U divided by square root V.
 

 = probability that a standard normal random variable is greater than Z,
 

See appendix on statistical methodology for full definition.
 

114
 
Table XXVII. Exact Statistical Tests Comparing Proportions of Animals (Not
 

Adjusted for Age) with Observed Hepatocellular Carcinoma or Neoplastic
 

Nodule of the Liver among Pooled Control and Trichloroethylene-Treated Mice
 

Male Mice
 

Comparison
 

Dose-Response
 

Veh. Control
 

Dose-Response
 

Untr. Control
 

Dosed vs.
 

Veh. Control
 

Dosed vs.
 

Untr. Control
 

Low Dose vs.
 

Veh. Control
 

Low Dose vs.
 

Untr. Control
 

High Dose vs.
 

Veh. Control
 

High Dose vs.
 

Untr. Control
 

High Dose vs.
 

Low Dose
 

Veh. Con. vs.
 

Untr. Control
 

Untr.
 

Cont.
 

r/n
 

5/70
 

7%
 

5/70
 

7%
 

5/70
 

7%
 

5/70
 

7%
 

5/70
 

7%
 

Veh.
 

Cont.
 

r/n
 

5/77
 

6%
 

5/77
 

6%
 

5/77
 

6%
 

5/77
 

6%
 

5/77
 

6%
 

Low
 

Dose
 

r/n
 

26/50
 

52%
 

26/50
 

52%
 

High
 

Dose
 

r/n
 

31/48
 

65%
 

31/48
 

65%
 

57/98
 

58%
 

57/98
 

58%
 

26/50
 

52%
 

26/50
 

52%
 

31/48
 

65%
 

31/48
 

65%
 

26/50 31/48
 

52% 65%
 

Exact
 

Test
 

Ρ
 

0.000
 

0.000
 

0.000
 

0.000
 

0.000
 

0.000
 

0.000
 

0.000
 

0.145
 

0.686
 

Exact
 

Test
 

Ρ
 

0.000
 

0.000
 

0.001
 

0.004
 

0.072
 

0.169
 

0.000
 

0.000
 

0.034
 

0.887
 

Untr.
 

Cont.
 

r/n
 

2/76
 

3%
 

2/76
 

3%
 

2/76
 

3%
 

2/76
 

3%
 

2/76
 

3%
 

Veh.
 

Cont.
 

r/n
 

1/80
 

1%
 

1/80
 

1%
 

1/80
 

1%
 

1/80
 

1%
 

1/80
 

1%
 

(continued)
 

Female Mice
 

Low
 

Dose
 

r/n
 

4/50
 

8%
 

4/50
 

8%
 

High
 

Dose
 

r/n
 

11/47
 

23%
 

11/47
 

23%
 

15/97
 

15%
 

15/97
 

15%
 

4/50
 

8%
 

4/50
 

8%
 

11/47
 

23%
 

11/47
 

23%
 

4/50 11/47
 

8% 23%
 

115
 
Table XXVII. Exact Statistical Tests Comparing Proportions of Animals (Not
 

Adjusted for Age) with Observed Hepatocellular Carcinoma or Neoplastic
 

Nodule of the Liver among Pooled Control and Trichloroethylene-Treated Mice
 

(continued) 

Comparison Chi~square df Ρ Chi-square df Ρ 

Among High Dose, 

Low Dose and 

Vehicle Control 52.02 2 0.000 17.76 2 0.000 

Dose-Response Trend 

Vehicle Control 47.85 1 0.000 16.96 1 0.000 

Deviation from Trend 

Vehicle Control 4.16 1 0.041 0.80 1 0.367 

Among High Dose, 

Low Dose and 

Untreated Control 47.31 2 0.000 14.41 2 0.001 

Dose-Response Trend 

Untreated Control 43.43 1 0.000 13.41 1 0.000 

Deviation from Trend 

Untreated Control 3.89 1 0.049 0.99 1 0.344 

r = Number of animals with observed mark. 

 = Number of animals examined histopathologically for the mark. 

 = Exact probability of exceeding or equaling observed test 

statistic under null hypothesis, 

df = Degrees of freedom. 

See section on statistical methodology for explanation of tests. 

116
 
APPENDIX D : PATHOLOGY

 

117 
Table XXVIIIa. Numbers of Tissues Examined - Rats
 

Males Females 

Controls Low High Low High 

Organ Males Females Dose Dose Dose Dose 

Brain 20 20 50 49 48 49 

Spinal Cord 

Pituitary 20 20 47 47 47 49 

Thyroid 20 20 50 48 46 50 

Adrenal 20 20 50 49 47 49 

Heart 20 20 50 50 48 50 

Lung 20 20 50 50 48 50 

Spleen 20 20 49 48 47 50 

Liver 20 20 50 50 48 50 

Kidney 20 20 50 50 48 50 

Stomach 20 20 49 49 47 50 

Small Intestine 18 19 49 47 46 50 

Large Intestine 19 20 50 50 46 50 

Pancreas 20 18 50 48 48 49 

Ovary/Testes 20 20 49 47 48 49 

Uterus/Prostate 16 20 37 27 48 49 

Vagina/Seminal 

Vesicle 2 4 1 1 

Salivary Gland 18 16 33 14 25 24 

Lymph Node 20 20 49 39 41 44 

Urinary Bladder 20 18 46 46 42 43 

Gallbladder 

Nerve 

Muscle 

Eye 1 1 

Bone 20 20 50 50 48 48 

Mammary Gland 20 20 50 46 45 48 

Esophagus 19 19 49 50 47 48 

Trachea 20 20 50 48 48 50 

Thymus-Cervical 

Lymph Node 17 15 31 15 22 16 

Unusual Lesion 2 1 

Tissue Mass 6 4 5 7 7 9 

Aorta 1 

Total Animals 

Examined 20 20 50 50 48 50 

118
 
Table XXVIIIb. Numbers of Tissues Examined - Mice
 

Males Females 

Controls Low High Low High 

Organ Males Females Dose Dose Dose Dose 

Brain 20 20 50 48 50 47 

Spinal Cord 

Pituitary 16 16 35 34 44 43 

Thyroid 18 20 50 47 46 45 

Adrenal 20 20 50 47 49 47 

Heart 20 20 50 48 49 47 

Lung 20 20 50 48 50 47 

Spleen 20 20 50 48 49 47 

Liver 20 20 50 48 50 47 

Kidney 20 20 50 48 50 47 

Stomach 20 20 49 48 49 46 

Small Intestine 19 20 50 48 48 47 

Large Intestine 20 20 48 47 49 47 

Pancreas 20 20 49 47 49 47 

Ovary/Testes 20 20 50 48 47 47 

Uterus/Prostate 18 20 47 47 48 47 

Vagina/Seminal 

Vesicle 3 

Salivary Gland 15 18 49 39 47 47 

Lymph Node 20 19 47 45 49 47 

Urinary Bladder 20 19 48 47 48 44 

Gallbladder 16 16 36 15 39 34 

Nerve 

Muscle 

Eye 2 1 2 

Bone 20 20 50 48 49 47 

Mammary Gland 20 20 50 48 49 47 

Thymus 19 20 43 42 46 43 

Trachea 19 20 49 48 49 46 

Esophagus 18 20 50 48 49 47 

Unusual Lesion 3 1 5 5 4 1 

Total Animals 

Examined 20 20 50 48 50 47 

119
 
Tumor Summary Tables for Rats and Mice
 

Tables XXIXa and XXXa were designed to summarize only the number of
 

primary tumors in each organ of each system. These tables delineate each
 

system, each organ within each system, and the type of tumors within each
 

organ. The counts for each of these 3 categories, that is, the numbers of
 

animals with tumors in both system and organ and the numbers of a
 

particular tumor within each organ, are indented in a hierarchal manner in
 

the same manner as are the categories in the left hand column. If an
 

animal has more than one type of tumor within a given system the total
 

number of animals with tumors in that system may be less than the sum of
 

the organ counts (the number of animals with tumors in a particular organ
 

within that system). For example, 7 high dose female rats have mammary
 

gland tumors and 1 has a tumor of the uterus/endometrium. But since the
 

latter tumor appeared in an animal that also had a mammary gland tumor, the
 

total number of high dose female rats with tumors of the reproductive
 

system is 7, not 8.
 

In the summary, animals examined represent the number of animals started on
 

test in a specific group less the number of animals with information
 

missing (animal lost or autolyzed) in the group. The number of animals
 

with tumors may be less than the number of animals with benign tumors plus
 

the number of animals with malignant tumors since an animal may have both a
 

benign and a malignant tumor.
 

Tables XXIXb and XXXb were designed to summarize the number of tumors
 

present in each anatomic site, that is, in each organ of each system
 

regardless of their origin. Thus, all sites of metastatic tumors which
 

appear in more than one organ are included. These tables are organized in
 

the same manner as are Tables XXIXa and XXXa. The counts differ only in
 

that both system and organ counts represent number of animals with tumor in
 

a specific system or organ irrespective of the origin of the tumor. For
 

example, in low dose male rats a hemangiosarcoma of the subcutaneous tissue
 

of the integumentary system is included in Table XXX but not in Table XXIX
 

because although the tumor is present in this tissue it is not a primary
 

tumor of the tissue and originated elsewhere. The tumor summaries for each
 

table are identical except that Table XXX contains the additional counts of
 

total metastatic tumors and animals with metastatic tumors.
 

120
 
Table XXIXa.


Organ System
 

INTEGUMENTARY SYSTEM
 

Skin
 

Pilomatrixoma
 

Subcutaneous Tissue
 

Liposarcoma
 

Fibroma
 Fibrosarcoma
 

Squamous-Cell Carcinoma
 

RESPIRATORY SYSTEM
 

Lung
 

Adenosquamous Carcinoma
 

CIRCULATORY SYSTEM
 

Subcutaneous Tissue
 Hemangiosarcoma
 

Multiple Organs
 

Hemangiosarcoma
 

Pancreas
 

Hemangiosarcoma
 

Spleen
 

Hemangiosarcoma
 

DIGESTIVE SYSTEM
 

None
 

 Primary Tumors by Site of Origin - Trichloroethylene-Treated Rats
 

Male Rats Female Rats
 

Control Low Dose High Dose Control Low Dose High Dose
 

3 1 2
 

1
 

1
 

3 2
 

1
 

1 1
 

1
 

1
 

1
 

1
 

1
 

1 1 2 1
 

1
 

1
 

1
 

1
 

1
 

1
 

1 1
 

1 1
 

Continued on Next Page
 
Table XXIXa (continued)% Primary Tumors by Site of Origin - Trichloroethylene-Treated Rats
 

Organ System
 

URINARY SYSTEM
 

Kidney
 

Malignant Mixed Tumor
 

Tubular Adenocarcinoma
 

Hamartoma
 

ENDOCRINE SYSTEM
 

Pituitary
 

Chromophobe Adenoma
 

Adrenal
 

Adrenal Cortical Carcinoma
 

Thyroid
 

Follicular-Cell Adenoma
 

Follicular-Cell Adenocarcinoma
 

HEMATOPOIETIC SYSTEM
 

Multiple Organs
 

Reticulum-Cell Sarcoma
 Spleen
 

Reticulum-Cell Sarcoma
 Thymus
 

Reticulum-Cell Sarcoma
 

REPRODUCTIVE SYSTEM
 

Mammary Gland
 

Fibroadenoma
 

Adenocarcinoma
 

Uterus/Endometrium
 

Sarcoma
 

Ovary
 

Granulosa-Cell Carcinoma
 

Control
 

2
 

2
 

2
 

1
 

1
 

1
 

1
 

Male Rats Female Rats
 

Low Dose High Dose Control Low Dose High Dose
 

2
 

2
 

1
 

1
 

2 1 4 3 7
 

4 2 6
 

4 2 6
 

1
 

1
 

2 1 1
 

1
 

1 1 1
 

1 1 1
 

1
 

1
 

1
 

1
 

1
 

1
 

5 6 7
 

4 5 7
 

3 5 7
 

1
 

1 1
 

1 1
 

1
 

1
 

Continued on Next Page
 
Table XXIXa (continued).


Organ System
 

Control
 

NERVOUS SYSTEM
 

Heart
 

Aortic Body Tumor
 

MUSCULOSKELETAL SYSTEM
 

None
 

ALL OTHER SYSTEMS. 1
 

Abdomen 1
 

Giant-Cell Tumor, Malignant 1
 

PRIMARY TUMOR SUMMARY
 

Animals Examined 20
 

Animals with Benign Tumors 2
 

Total Benign Tumors 2
 

Animals with Malignant Tumors 5
 

Total Malignant Tumors 5
 

Animals with Tumors 5
 

 Primary Tumors by Site of Origin - Trichloroethylene-Treated Rats
 

Male Rats Female Rats
 

Low Dose High Dose Control Low Dose High Dose
 

1
 

1
 

1
 

50 50 20 48 50
 

3 2 6 11
 

3 2 7 8 13
 

5 3 2 5 3
 

5 3 3 6 3
 

7 5 7 12 12
 
Table XXIXb. Tumors by Anatomic Site - Trichloroethylene-Treated Rats
 

Organ System Male Rats Female Rats 

Control Low Dose High Dose Control Low Dose High Dose 

INTEGUMENTARY SYSTEM 4 1 2 

Skin 1 

Pilomatrixoma 1 

Subcutaneous Tissue 4 2

 

Liposarcoma 1

 

Hemangiosarcoma 1 

Fibroma 1 1 

Fibrosarcoma 1 

Squamous-Cell Carcinoma 1 

RESPIRATORY SYSTEM 1 1 

Lung 1 1 

Hemangiosarcoma 1 

Adenosquamous Carcinoma 1 

CIRCULATORY SYSTEM 1 1 

Heart 1 1

 

Hemangiosarcoma 1

 

Aortic Body Tumor 1

 

DIGESTIVE SYSTEM 1 1 

Liver 1

 

Reticulum-Cell Sarcoma 1

 

Pancreas 1

 

Hemangiosarcoma 1

 

Continued on Next Page
 
Table XXIXb (continued).


Organ System
 

Control
 

DIGESTIVE SYSTEM
 

Liver
 

Reticulum-Cell Sarcoma
 

Pancreas
 

Hemangiosarcoma
 

URINARY SYSTEM 3
 

Kidney 3
 

Malignant Mixed Tumor 2
 

Tubular Adenocarcinoma
 

Hamartoma 1
 

Adenosquamous Carcinoma Metast. 1
 

ENDOCRINE SYSTEM 1
 

Pituitary
 

Chromophobe Adenoma
 

Adrenal
 

Adrenal Cortical Carcinoma
 

Thyroid 1
 

Follicular-Cell Adenoma 1
 

Follicular-Cell Adenocarcinoma
 

HEMATOPOIETIC SYSTEM 2
 

Spleen 1
 

Hemangiosarcoma 1
 

Reticulum-Cell Sarcoma
 

Cervical Lymph Node 1
 

Reticulum-Cell Sarcoma
 

Adenosquamous Carcinoma, Metast . 1
 

 iiy iiiub
 

Reticulum-Cell Sarcoma
 

 Tumors by Anatomic Site - Trichloroethylene-Treated Rats
 

Male Rats Female Rats
 

Low Dose High Dose Control Low Dose High Dose
 

1 1
 

1
 

1
 

1
 

1
 

2
 

2
 

1
 

1
 

2 1 4 3 7
 

4 2 6
 

4 2 6
 

1
 

1
 

2 1 1
 

1
 

1 1 1
 

1 1 1 1
 

1 1
 

1
 

1
 

1
 

1
 

1
 

j.
 

1
 

Continued on Next Page
 
Table XXIXb (continued) . Tumors by Anatomic Site - Trichloroethylene-Treated Rats
 

Organ System
 

Control
 

REPRODUCTIVE SYSTEM
 

Mammary Gland
 

Fibroadenoma
 

Adenocarcinoma
 

Uterus/Endometrium
 

Sarcoma
 

Ovary
 

Granulosa-Cell Carcinoma
 

NERVOUS SYSTEM
 

None
 

MUSCULOSKELETAL SYSTEM
 

None
 

ALL OTHER SYSTEMS 1
 

Abdomen 1
 

Giant-Cell Tumor, Malignant 1
 

TUMOR SUMMARY
 

Animals Examined 20
 

Animals with Benign Tumors 2
 

Total Benign Tumors 2
 

Animals with Malignant Tumors 5
 

Total Malignant Tumors 5
 

Animals with Metastatic Tumors 1
 

Total Metastatic Tumors 2
 

Animals with Tumors 5
 

Male Rats Female Rats
 

Low Dose High Dose Control Low Dose High Dose
 

5 6 7
 

4 5 7
 

3 5 7
 

1
 

1 1
 

1 1
 

1
 

1
 

50 50 20 48 50
 

3 2 6 7 11
 

3 2 7 8 13
 

5 3 2 5 3
 

5 3 3 6 3
 

7 5 7 12 12
 
Table XXXa· Primary Tumors by Site of Origin ­ Trichloroethylene-Treated Mice 

Organ System Male Mice Female Mice 

Control Low Dose High Dose Control Low Dose High Dose 

INTEGUMENTARY SYSTEM 3 1 1 

Skin 1 

Fibrosarcoma 1 

Subcutaneous Tissue 2 1 1 

Fibrosarcoma 2 1 

Fibroma 1 

RESPIRATORY SYSTEM 5 2 1 4 7 

Lung 5 2 1 4 7 

Adenoma 5 1 1 2 5 

Alveolar Adenocarcinoma 1 2 2 

CIRCULATORY SYSTEM 1 

Lung 1 

Hemangiosarcoma 1 

DIGESTIVE SYSTEM 1 26 31 4 11 

Stomach 1 

Papilloma 1 

Liver 1 26 31 4 11 

Hepatocellular Carcinoma 1 26 31 4 11 

URINARY SYSTEM 1 

Kidney 1 

Adenoma 1 

Continued on Next Page 
Table XXXa (continued). Primary Tumors by Site of Origin - Trichloroethylene-Treated Mice 

Organ System
 

ENDOCRINE SYSTEM
 

None
 

HEMATOPOIETIC SYSTEM
 

Thymus
 

Lymphos arcoma
 

Multiple Organs
 

Reticulum-Cell Sarcoma
 

Lymphos arcoma
 

Spleen
 Lymphosarcoma
 

Cervical Lymph Node
 

Malignant Lymphoma
 

Mesentery Lymph Node
 

Reticulum-Cell Sarcoma
 

REPRODUCTIVE SYSTEM
 

Mammary Gland
 

Adenocarcinoma
 

Uterus
 

Fibrosarcoma
 

Adenocarcinoma
 

Ovary
 

Granulosa-Cell Carcinoma
 

Cystadenoma
 

NERVOUS SYSTEM
 

Muscle of Back
 

Neurofibroma
 

Male Mice Female Mice
 

Control Low Dose High Dose Control Low Dose High Dose
 

1 4 2 1 5 6
 

1
 

1
 

1 4 2 3 4
 

1 2 1 2 2
 

2 1 1 2
 

1 1
 

1 1
 

1
 

1
 

1
 

1
 

1 4
 

1
 

1
 

1 1
 

1
 

1
 

2
 

1
 

1
 

1
 

1
 

1
 

Continued on Next Page 
Table XXXa (continued). Primary Tumors by Site of Origin - Trichloroethylene-Treated Mice
 

Organ System 

Control 

Male Mice 

Low Dose High Dose Control 

Female Mice 

Low Dose High Dose 

MUSCULOSKELETAL SYSTEM 

Soft Tissue Osteosarcoma 

1 

1 

1 

SPECIAL SENSE ORGANS 

Harderian Gland 

Adenoma 

1 

1 

1 

1 

1 

1 

ALL OTHER SYSTEMS 

Abdomen Fibrosarcoma 

1 

1 

1 

PRIMARY TUMOR SUMMARY 

Animals Examined

Animals with Benign Tumors 

Total Benign Tumors 

Animals with Malignant Tumors

Total Malignant Tumors

Animals with Tumors

 20 

5 

5 

5 

50 

7 

7 

28 

30 

30 

48 

4 

4 

32 

36 

33 

20 

1 

1 

3 

3 

4 

50 

3 

4 

14 

15 

14 

47 

5 

5 

16 

19 

19 
Table XXXb. Tumors by Anatomic Site - Trichloroethylene-Treated Mice 

Organ System Male Mice
 

Control Low Dose
 

INTEGUMENTARY SYSTEM 3 1
 

Skin 1
 

Fibrosarcoma 1
 

Alveolar Adenocarcinoma, Metast.
 

Subcutaneous Tissue 2 1
 

Fibrosarcoma 2
 

Fibroma 1
 

Soft Tissue
 

Osteosarcoma
 

RESPIRATORY SYSTEM 9
 

Lung 9
 

Adenoma 5
 

Alveolar Adenocarcinoma
 

Hemangiosarcoma
 

Reticulum-Cell Sarcoma
 

Lymphosarcoma 1
 

Hepatocellular Carcinoma, Metast. 4
 

CIRCULATORY SYSTEM
 

Aorta
 Alveolar Adenocarcinoma, Metast.
 

DIGESTIVE SYSTEM 2 28
 

Stomach
 

Papilloma
 

Reticulum-Cell Sarcoma
 Fibrosarcoma Metastatic
 

Ileum 1
 

Reticulum-Cell Sarcoma 1
 

Lymphosarcoma
 

Female Mice
 

High Dose Control Low Dose High Dose
 

1 1 1
 

1
 

1
 

1
 

1
 

1
 

1
 

5 1 4 8
 

5 1 4 8
 

1 1 2 5
 

1 2 2
 

1
 

1
 

3
 

1
 

1
 

1
 

32 7 14
 

2 1
 

1
 

1
 

1
 

1 1
 

1
 

1
 

Continued on Next Page 
Table XXXb (continued). Tumors by Anatomic Site - Trichloroethylene-Treated Mice
 

Organ System Male Mice Female Mice
 

Control Low Dose High Dose Control Low Dose High Dose
 

REPRODUCTIVE SYSTEM 1 2 1 6
 

Mammary Gland 1
 

Adenocarcinoma 1
 

Uterus
 1 ?
 

Fibrosarcoma 1
 

Reticulum-Cell Sarcoma 1
 

Adenocarcinoma 1
 

Ovary 2
 

Granulosa-Cell Carcinoma 1
 

Cystadenoma 1
 

Vagina 1
 

Reticulum-Cell Sarcoma 1
 

Epididymis 1
 

Reticulum-Cell Sarcoma 1
 

Prostate 1 2
 

Reticulum-Cell Sarcoma 1 1
 

Lymphosarcoma 1
 

Seminal Vesicle 1
 

Reticulum-Cell Sarcoma 1
 

NERVOUS SYSTEM
 

None
 

MUSCULOSKELETAL SYSTEM 1
 

Muscle of Back 1
 

Neurofibroma 1
 

SPECIAL SENSE ORGANS 1 1
 

Harderian Gland 1 1
 

Adenoma 1 1
 

Continued on Next Page
 
Table XXXb (continued). Tumors by Anatomic Site - Trichloroethylene-Treated Mice
 

Organ System
 

DIGESTIVE SYSTEM (cont.)
 

Pancreas
 Reticulum-Cell Sarcoma
 

Fibrosarcoma Metastatic
 

Liver
 

Hepatocellular Carcinoma
 

Reticulum-Cell Sarcoma
 

Lymphosarcoma
 

URINARY SYSTEM
 

Kidney
 

Adenoma
 

Reticulum-Cell Sarcoma
 

Lymphosarcoma
 

Fibrosarcoma Metastatic
 

ENDOCRINE SYSTEM
 

Adrenal
 

Lymphosarcoma
 

Fibrosarcoma Metastatic
 

HEMATOPOIETIC SYSTEM
 

Thymus
 Lymphosarcoma
 

Spleen
 

Reticulum-Cell Sarcoma
 Lymphosarcoma
 

Lymph Node
 Malignant Lymphoma
 

Lymphosarcoma
 

Reticulum-Cell Sarcoma
 

Alveolar Adenocarcinoma,
 

Fibrosarcoma Metastatic
 

Bone Marrow
 

Lvmuhosarcoma
 

Control
 

2
 

1
 

1
 

1
 

1
 

1
 

1
 

1
 

Metast.
 

Male Mice
 

Low Dose
 

28
 

26
 

2
 

1
 

1
 

1
 

4
 

1
 

1
 

3
 

1
 

2
 

2
 

2
 

2
 

1
 

1
 

High Dose
 

2
 

1
 

1
 

32
 

31
 

1
 

4
 

4
 

1
 

1
 

1
 

1
 

1
 

1
 

1
 

4
 

2
 

1
 

1
 

4
 

1
 

1
 

1
 

1
 

Female Mice
 

Control Low Dose
 

6
 

4
 

2
 

1
 

1
 

1
 

1 4
 

1
 

1
 

3
 

1
 

2
 

1 2
 

2
 

1 1
 

High Dose
 

12
 

11
 

1
 

2
 

2
 

1
 

1
 

6
 

4
 

1
 

3
 

3
 

1
 

2
 

1
 

Continued on Next Page
 
Table XXXb (continued). Tumors by Anatomic Site - Trichloroethylene-Treated Mice
 

Organ System 

Control 

Male Mice 

Low Dose 

ALL OTHER SYSTEMS 

Abdomen Fibrosarcoma 

TUMOR SUMMARY 

Animals Examined 

Animals with Benign Tumors 

Total Benign Tumors 

Animals with Malignant Tumors 

Total Malignant Tumors 

Animals with Metastatic Tumors 

Total Metastatic Tumors 

Animals with Tumors 

20 

5 

5 

5 

50 

7 

7 

28 

30 

4 

4 

30 

High Dose
 

1
 

1
 

1
 

48
 

4
 

4
 

32
 

36
 

4
 

11
 

33
 

Control 

Female Mice 

Low Dose High Dose 

20 

1 

1 

3 

3 

50 

3 

4 

14 

15 

47 

5 

5 

16 

19 

4 14 19 

133 
Tables XXXI and XXXII - Individual Pathology
 

Rats
 

Mice
 

NATD
 

 SAC
 

MISS
 

MSAC
 

ACCK
 

Index
 

Control males
 

Low dose males
 

High dose males
 

Control females
 

Low dose females
 

High dose females
 

Control males
 

Low dose males
 

High dose males
 

Control females
 

Low dose females
 

High dose females
 

Disposition
 

Page
 

135
 

137
 

142
 

147
 

149
 

153
 

158
 

160
 

164
 

168
 

169
 

173
 

Code
 

Natural Death
 

Terminal Sacrifice
 

Missing
 

Moribund Sacrifice
 

Accident
 

134
 
Table XXXIa. Individual Pathology - Trichloroethylene-Treated Male Rats 

Control Group (Vehicle)
 

WEEKS ON DISP ANIMAL TUMORS
 

STUDY CODE NUMBER
 

TOPOGRAPHY
 

67 NATD 003
 

70 NATD 011 SPLEEN
 

76 NATD 014
 

76 NATD 015
 

82 NATD 009
 

82 NATD 013
 

83 NATD 018
 

87 NATD 016
 

88 NATD 005
 

88 NATD 012
 

90 NATD 002 ABDOMEN
 

91 NATD 004
 

MORPHOLOGY


HEMANGIOSARCOMA


GIANT-CELL TUMOR, MALIGNANT


OTHER PATH0L03Y
 

 TOPOGRAPHY
 

LUNG
 

BONE MARROW
 

 PITUITARY
 

BRONCHUS
 

SPLEEN
 

LUNG
 

THYMUS
 

SALIVARY GLAND
 KIDNEY
 

LUNG
 

KIDNEY
 

LUNG
 

BONE MARROW
 

LUNG
 

BRONCHUS
 

STOMACH
 

MESENTERIC LYMPH
 PANCREAS
 

TESTIS
 

LUNG
 

TESTIS
 

LUNG
 

LUNG
 

KIDNEY
 

 LUNG
 

TESTIS
 

LUNG
 

PLEURA
 KIDNEY
 PROSTATE
 SEMINAL VESICLE
 SUBCUT TISSUE
 

MORPHOLOGY
 

INFLAMMATION CHRONIC
 

METAMORPHOSIS PATTY
 

INFLAMMATION
 BRONCHIECTASIS
 

HEMATOPOIESIS EXTRAMED.
 INFLAMMATION CHRONIC
 

INFLAMMATION
 

INFLAMMATION
 ABSCESS
 

INFLAMMATION CHRONIC
 

INFLAMMATION CHRONIC
 

INFLAMMATION CHRONIC
 

METAMORPHOSIS FATTY
 

INFLAMMATION CHRONIC
 

BRONCHIECTASIS
 

CALCIUM DEPOSITION
 NODE POLYARTERITIS NODOSA
 POLYARTERITIS NODOSA
 

ATROPHY
 

INFLAMMATION CHRONIC
 

ATROPHY
 

INFLAMMATION CHRONIC
 

INFLAMMATION CHRONIC
 

INFLAMMATION CHRONIC
 

INFLAMMATION CHRONIC
 

ATROPHY, EILATERAL
 

INFLAMMATION CHRONIC
 

INFLAMMATION
 INFLAMMATION CHRONIC
 

INFLAMMATION
 INFLAMMATION
 ABSCESS
 

Continued on next page 
Table XXXIa (continued). Individual Pathology - Trichloroethylene-Treated Male Rats
 

WEEKS ON
 OTHER PATHOLOGY
 STUDY
 

96
 

98
 

99
 

102
 

103
 

110
 

110
 

110
 

DISP ANIMAL TUMORS
 

CODE NUMBER
 

TOPOGBAPHY
 

NATD 019	 KIDNEY
 THYROID
 

NATD 020
 

NATD 010	 KIDNEY
 

KIDNEY
 

NATD 017
 

NATD 008
 

TSAC C01
 

TSAC 006
 

NATD 007	 LUNG
 

KIDNEYS, BILATERAL
 CERVICAL LYMPH NODE
 

Control Group (Vehicle)
 

MORPHOLOGY
 

MALIGNANT MIXED TUMOR
 FOLLICULAR ADENOMA
 

MALIGNANT MIXED TUMOR
 HAMARTOMA
 

CARCINOMA, GLANDULAR AND
 

SQUAMOUS, PROBABLY PRIMARY
 IN LUNG, WITH MULTIPLE PULMONARY
 

METASTASES
 

MULTIPLE METASTATIC TUMORS
 

METASTASIC TUMORS
 

TOPOGRAPHY
 

LUNG
 

EPIDIDYMIS
 

KIDNEY
 

SPLEEN
 LUNG
 

LUNG
 

KIDNEY
 MESENTERIC LYMPH
 

LIVER
 

LUNG
 

PLEURA
 BONE MARROW
 

LUNG
 KIDNEY
 SKIN
 SKIN
 SKIN
 

LUNG
 

KIDNEY
 THYROID
 BONE MARROW
 

STOMACH
 

BONE MARROW
 

MORPHOLOGY
 

INFLAMMATION CHRONIC
 

FAT NECROSIS WITH
 

ENCAPSULATION
 

INFLAMMATION CHRONIC
 

HEMATOPOIESIS EXTRAMED.
 

INFLAMMATION CHRONIC
 

INFLAMMATION CHRONIC
 

INFLAMMATION CHRONIC
 

NODE POLYARTERITIS NODOSA
 

METAMORPHOSIS FATTY
 

INFLAMMATION CHRONIC
 

INFLAMMATION
 METAMORPHOSIS FATTY
 

INFLAMMATION CHRONIC
 

INFLAMMATION CHRONIC
 

ACANTHOSIS
 

HYPERKERATOSIS
 

INFLAMMATION
 

INFLAMMATION CHRONIC
 

INFLAMMATION CHRONIC
 

INFLAMMATION CYSTIC
 

METAMORPHOSIS FATTY
 

ULCER FOCAL
 

METAMORPHOSIS FATTY
 

end of male rat control
 
Table XXXIa (continued). Individual Pathology - Trichloroethylene-Treated Male Rats
 

Low Dose Group 

WEEKS ON DISP ANIMAL TUMORS 

STUDY CODE NUMBER 

TOPOGRAPHY MORPHOLOGY 

16 NATD 028 

17 NATD 036 

23 NATD 010 

27 NATD 006 

34 NATD 035 

40 NATD 030 

42 NATD 034 

48 NATD 003 

50 NATD 007 

53 NATD 041 

60 NATD 046 

61 NATD 039 

65 NATD 019 

67 NATD 032 

OTHER
 

TOPOGRAPHY
 

KIDNEY
 

BONE MARROW
 

KIDNEY
 

KIDNEY
 LUNG
 

KIDNEY
 BONE MARROW
 

KIDNEY
 

LUNG
 

KIDNEY
 BONE MARROW
 

KIDNEY
 

PERICARDIUM
 MYOCARDIUM
 LUNG
 

PLEURA
 

PERICARDIUM
 

MYOCARDIUM
 KIDNEY
 LUNG
 

PLEURA
 

KIDNEY
 

LUNG
 

KIDNEY
 

BRONCHUS
 

BONE MARROW
 

PATHOLOGY
 

MORPHOLOGY
 

NO SIGNIFICANT DIAGNOSIS
 

NO SIGNIFICANT DIAGNOSIS
 

NO SIGNIFICANT DIAGNOSIS
 

NO SIGNIFICANT DIAGNOSIS
 

NEPHROSIS IOXIC
 

METAMORPHOSIS FATTY
 

NEPHROSIS IOXIC
 

NEPHROSIS IOXIC
 

INFLAMMATION CHRONIC
 

NEPHROSIS IOXIC
 

METAMORPHOSIS FATTY
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

NEPHROSIS TOXIC
 

METAMORPHOSIS FATTY
 

NEPHROSIS 1OXIC
 

INFLAMMATION
 INFLAMMATION
 INFLAMMATION CHRONIC
 

INFLAMMATION
 

INFLAMMATION
 INFLAMMATION
 NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

INFLAMMATION
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

NEPHROSIS IOXIC
 

BRONCHIECTASIS
 

METAMORPHOSIS FATTY
 

Continued on next page
 
Table XXXIa (continued). Individual Pathology - Trichloroethylene-Treated Male Rats
 

WEEKS ON STUDY 

67 

72 

DISP CODE 

NATD 

NATD 

ANIMAL NUMBER 

TOPOGRAPHY 

040 

 TUMORS 

MORPHOLOGY 

Low Dose Group 

74 NATD 047 

76 NATD 018 

00 76 NATD 023 THYROID FOLLICULAR ADENOMA 

80 NATD 026 SUBCUT TISSUE/NECK HEMANGIOSARCOMA 

80 NATD 033 

82 

83 

NATD 

NATD 

005 

038 

83 NATD 043 

OTHER
 

TOPOGRAPHY
 

KIDNEY
 LUNG
 

LEFT ADRENAL
 KIDNEY
 LUNG
 

BONE MARROW
 

KIDNEY
 

LUNG
 

BONE MARROW
 

LIVER
 KIDNEY
 

LUNG
 

BONE MARROW
 

PARATHYROID
 AORTA
 

KIDNEY
 LUNG
 

STOMACH
 

PANCREAS
 

BONE MARROW
 

ADRENAL
 KIDNEY
 KIDNEY
 LUNG
 

STOMACH
 

KIDNEY
 LUNG
 

MESENTERY
 PANCREAS
 

KIDNEY
 LUNG
 

KIDNEY
 

LUNG
 

TESTIS
 

BONE MARROW
 

KIDNEY
 

LUNG
 

PATHOLOGY
 

MORPHOLOGY
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

INFLAMMATION CYSTIC
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

METAMORPHOSIS FATTY
 

NEPHROSIS IOXIC
 

INFLAMMATION CHRONIC
 

METAMORPHOSIS FATTY
 

INFLAMMATION CYSTIC
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

METAMORPHOSIS FATTY
 

HYPERPLASIA
 ARTERIOSCLEROSIS
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

CALCIUM DEPOSITION
 POLYARTERITIS NODOSA
 METAMORPHOSIS FATTY
 

AN3IECTASIS
 

NEPHROSIS TOXIC
 

CAPSULAR ABSCESS
 

INFLAMMATICN CHRONIC
 

ULCER FOCAL
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

POLYARTERITIS NODOSA
 POLYARTERITIS NODOSA
 

NEPHROSIS IOXIC
 

INFLAMMATION CHRONIC
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

ATROPHY
 

METAMORPHOSIS FATTY
 

NEPHROSIS TOXIC
 

INFLAMMATICN CHRONIC
 

Continued on next page
 
WEEKS ON DISP ANIMAL


STUDY CODE NUMBER
 

85 NATD 001
 

86 NATD 014
 

86 NATD 020
 

87 NATD 029
 

88 NATD 012
 

90 NATD 015.


94 NATD 025


Table XXXIa (continued). Individual Pathology - Trichloroethylene-Treated Male Rats
 

Low Dose Group
 

 TUMORS
 

TOPOGRAPHY MORPHOLOGY
 

 KIDNEY TUBULAR ADENOCARCINOMA,
 

UNILATERAL
 

 SUBCUT TISSUE/AXILLA FIBROMA
 

OTHER
 

TOPOGRAPHY
 

KIDNEY
 

LUNG
 

TRACHEA
 

KIDNEY
 

LUNG
 

PLEURA
 

TESTIS
 

TRACHEA
 

KIDNEY
 

LUNG
 

BONE MARROW
 

MYOCARDIUM
 

ATRIUM
 

AORTA
 

KIDNEY
 

LUNG
 

STOMACH
 

TESTIS
 

VENTRICLE
 

KIDNEY
 

LUNG
 

BONE MARROW
 

SKIN
 

SKIN
 

SKIN
 

SPLEEN
 

LIVER
 

KIDNEY
 

LUNG
 

MESENTERY
 

PANCREAS
 

RIGHT ADRENAL
 

MYOCARDIUM
 

MYOCARDIUM
 

LIVER
 

KIDNEY
 

LUNG
 

PLEURA
 

PATHOLOGY
 

MORPHOLOGY
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

INFLAMMATION
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

INFLAMMATION
 

ATROPHY
 

INFLAMMATION
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

METAMORPHOSIS FATTY
 

DEGENERATION
 

CALCIUM DEPOSITION
 

ARTERIOSCLEROSIS
 

NEPHROSIS IOXIC
 

INFLAMMATION CHRONIC
 

CALCIUM DEPOSITION
 

ATROPHY
 

CALCIUM DEPOSITION
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

METAMORPHOSIS FATTY
 

ACANTHOSIS
 

HYPERKERATOSIS
 

EPIDERMAL INCLUSION CYST
 

HEMATOPOIESIS £XTRAMED.
 

METAMORPHOSIS FATTY
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

POLYARTERITIS NODOSA
 

POLYARTERITIS NODOSA
 

CONa£NITAI MALFORMATION
 

DEGENERATICN
 

FIBROSIS
 

INFLAMMATION
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

INFLAMMATION
 

Continued on next page
 
Table XXXIa (continued). Individual Pathology - Trichloroethylene-Treated Male Rats
 

Low Dose Group 

WEEKS ON STUDY DISP CODE ANIMAL NUMBER 

TOPOGRAPHY 

TUMORS 

MORPHOLOGY 

OTHEE PATHOLOGY 

TOPOGRAPHY MORPHOLOGY 

94 NATD 049 KIDNEY LUNG 

SUBCUTANEOUS TISSUE OF HIND LEG 

EPIDIDYMIS 

NEPHROSIS TOXIC 

INFLAMMATION CHRONIC 

ABSCESS 

FAT NECROSIS WITH ENCAPSULATION 

96 NATD 021 KIDNEY 

LUNG 

TESTIS 

NEPHROSIS 1OXIC 

INFLAMMATION CHRONIC 

ATROPHY 

97 NATD 016 KIDNEY LUNG 

PANCREAS 

BONE MARROW 

NEPHROSIS TOXIC 

INFLAMMATION CHRONIC 

POLYARTERITIS NODOSA METAMORPHCSIS FATTY 

102 NATD 009 KIDNEY LUNG 

NEPHROSIS 1CXIC 

INFLAMMATION CHRONIC 

103 NATD 013 KIDNEY LUNG 

NEPHROSIS TOXIC 

INFLAMMATION CHRONIC 

103 NATD 027 ADRENAL 

ADRENAL KIDNEY LUNG 

PLEURA TESTIS 

ANGIECTASIS 

DEGENERATION NEPHROSIS TOXIC 

INFLAMMATION CHRONIC 

INFLAMMATION ATROPHY 

103 NATD 031 KIDNEY LUNG 

BONE MARROW 

NEPHROSIS TOXIC 

INFLAMMATION CHRONIC 

METAMORPHOSIS FATTY 

104 NATD 037 KIDNEY 

LUNG 

PANCREAS 

TESTIS 

PROSTATE 

NEPHROSIS TOXIC 

INFLAMMATION CHRONIC 

POLYARTERITIS NODOSA ATROPHY INFLAMMATION 

107 NATD 008 KIDNEY 

LUNG 

TRACHEA CERVICAL LYMPH NODE 

NEPHROSIS TOXIC 

INFLAMMATION CHRONIC 

INFLAMMATION INFLAMMATION 

107 NATD 045 KIDNEY LUNG 

PANCREAS 

PROSTATE 

NEPHROSIS TOXIC 

INFLAMMATION CHRONIC 

POLYARTERITIS NODOSA 

INFLAMMATION 

Continued on next page 
Table XXXIa (continued). Individual Pathology - Trichloroethylene-Treated Male Rats 

Low Dose Group 

WEEKS ON DISP ANIMAL TUMORS OTHER PATHOLOGY 

STUDY CODE NUMBER 

TOPOGRAPHY MORPHOLOGY TOPOGRAPHY MORPHOLOGY 

108 NATD 004 SUBCUT TISSUE/AXILLA FIBROSARCOMA KIDNEY NEPHROSIS TOXIC 

LUNG INFLAMMATION CHRONIC 

TESTIS ATROPHY 

TESTIS CALCIUM DEPOSITION 

110 TSAC 002 THYROID FOLLICULAR ADENOCARCINOMA LIVER METAMORPHCSIS FATTY 

KIDNEY HAMARTOMA, MEDULLA, UNILATERAL LIVER INFLAMMATION CYSTIC KIDNEY NEPHROSIS TOXIC 

LUNG INFLAMMATION CHRONIC 

TESTIS ATROPHY 

110 TSAC 011 SUBCUT TISSUE/AXILLA SQUAMOUS CELL CARCINOMA KIDNEY NEPHROSIS 1OXIC 

LUNG INFLAMMATION CHRONIC 

PROSTATE INFLAMMATION 

; no TSAC 017 SPLEEN HEMATOPOIESIS EXTRAMED. 

KIDNEY NEPHROSIS TOXIC 

LUNG INFLAMMATION CHRONIC 

PANCREAS POLYARTERITIS NODOSA MESENTERY POLYARTERITIS NODOSA TESTIS ATROPHY 

110 TSAC 022 KIDNEY NEPHROSIS TOXIC 

LUNG INFLAMMATION CHRONIC 

TESTIS ATROPHY 

110 TSAC 024 LIVER METAMCRPHCSIS FATTY KIDNEY NEPHROSIS TOXIC 

LUNG INFLAMMATION CHRONIC 

PANCREAS POLYARTERITIS NODOSA 

TESTIS ATROPHY BONE MARROW METAMORPHOSIS FATTY 

110 NATD 042 KIDNEY NEPHROSIS TOXIC 

TESTIS ATROPHY 

110 TSAC 048 KIDNEY NEPHROSIS TOXIC 

LUNG INFLAMMATION CHRONIC 

TESTIS ATROPHY BONE MARROW METAMORPHOSIS FATTY 

110 TSAC 050 MYOCARDIUM DEGENERATION 

KIDNEY NEPHROSIS TOXIC 

TESTIS ATROPHY BONE MARROW METAMORPHOSIS FATTY 

end of male rats—low dose 
Table XXXIa (continued). Individual Pathology - Trichloroethylene-Treated Male Rats 

WEEKS ON
 

STUDY
 

2
 

2
 

5
 

6
 

12
 

17
 

21
 

21
 

31
 

33
 

35
 

40
 

42
 

44
 

DISP
 

CODE
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

NAT.D
 

NATD
 

NATD
 

NATD
 

NATD
 

ANIMAL
 

NUMBER
 

TOPOGRAEHY
 

012
 

032
 

014
 

004
 

015
 

022
 

t>06
 

002
 

025
 

024
 

040
 

033
 

039
 

003
 

High Dose Group 

TUMORS OTHER PATHOLOGY
 

MORPHOLOGY TOPOGRAPHY
 

PERICARDIUM
 

LUNG
 

PLEURA
 

LUNG
 

PLEURA
 

BONE MARROW
 

PERICARDIUM
 

MYOCARDIUM
 

LUNG
 

PLEURA
 

KIDNEY
 

KIDNEY
 

LIVER/CENTRILOBULAR
 

LIVER
 

KIDNEY
 

LUNG
 

KIDNEY
 

KIDNEY
 

BONE MARROW
 

KIDNEY
 

PERICARDIUM
 

BONE MARROW
 

MYOCARDIUM
 

KIDNEY
 

KIDNEY
 

LIVER/CENTRILOBULAR
 

LIVER
 

KIDNEY
 

LIVER
 

LIVER
 

BONE MARROW
 

KIDNEY
 

MORPHOLOGY
 

INFLAMMATION
 

INFLAMMATION CHRONIC
 

INFLAMMATION
 

INFLAMMATION CHRONIC
 

INFLAMMATION
 

METAMORPHOSIS FATTY
 

INFLAMMATION
 

INFLAMMATION
 

INFLAMMATION CHRONIC
 

INFLAMMATION
 

NEPHROSIS TOXIC
 

NEPHROSIS TCXIC
 

METAMORPHOSIS FATTY
 

CONGESTION
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

NEPHROSIS TOXIC
 

NEPHROSIS TCXIC
 

METAMORPHOSIS FATTY
 

NEPHROSIS TOXIC
 

INFLAMMATION
 

METAMORPHOSIS FATTY
 

INFLAMMATION
 

NEPHROSIS TOXIC
 

NEPHROSIS TOXIC
 

METAMORPHOSIS FATTY
 

ANGIECTASIS
 

NEPHROSIS TCXIC
 

HEPATOCYTCKEGALY,
 

FOCAL
 

ANGIECTASIS
 

METAMORPHOSIS FATTY
 

NEPHROSIS TOXIC
 

Continued on next page 
WEEKS OK
 

STUDY
 

48
 

49
 

52
 

52
 

53
 

54
 

54
 

56
 

58
 

59
 

61
 

Table XXXIa (continued).

OISP ANIMAL TUMORS
 

CODE NUMBER
 

TOPOGRAEHY
 

NATD 018
 

NATO 030
 

NATD 005
 

NATD 041
 

NATD 037
 

NATD O1'3
 

NATD 017
 

NATD 027
 

NATD 047
 

NATD 009
 

NATD 029
 

 Individual Pathology - Trichloroethylene-Treated Male Rats 

High Dose Group 

OTHER PATHOLOGY
 

MORPHOLOGY TOPOGRAPHY"
 

BONE MARROW
 

KIDNEY
 

LIVER
 

KIDNEY
 

LIVER
 

BRONCHUS
 

KIDNEY
 

LIVER/CENTRILOBULAR
 

LIVER
 

LIVER
 

BRONCHUS
 

BONE MARROW
 

KIDNEY
 

EPIDIDYMIS
 

LUNG
 

PLEURA
 

KIDNEY
 

EPIDIDYMIS
 

BRONCHUS
 

LUNG
 

KIDNEY
 

LIVER
 

KIDNEY
 

LUNG
 

PLEURA
 

THYROID
 

KIDNEY
 

LIVER/CENTRILOBULAR
 

LIVER
 

BRONCHUS
 

KIDNEY
 

LUNG
 

KIDNEY
 

LUNG
 

PLEURA
 

KIDNEY
 

MORPHOLOGY
 

METAMORPHOSIS PATTY
 

NEPHROSIS TOXIC
 

METAMORPHOSIS FATTY
 

NEPHROSIS TOXIC
 

ΜΕΤΑMORPHCSIS FATTY
 

BRONCHIECTASIS
 

NEPHROSIS TOXIC
 

DEGENERATION
 

METAMORPHOSIS FATTY
 

ANGIECTASIS
 

BRONCHIECTASIS
 

METAMORPHOSIS FATTY
 

NEPHROSIS TCXIC
 

FAT NECROSIS WITH
 

ENCAPSULATION
 

INFLAMMATION CHRONIC
 

INFLAMMATION
 

NEPHROSIS TOXIC
 

FAT NECROSIS WITH
 

ENCAPSULATION
 

BRONCHIECTASIS
 

INFLAMMATION CHRONIC
 

NEPHROSIS TOXIC
 

ANGIECTASIS
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

INFLAMMATION
 

INFLAMMATION
 

NEPHROSIS ΤΟΠ Ο
 

DEGENERATION
 

METAMORPHOSIS FATTY
 

BRONCHIECTASIS
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

INFLAMMATION
 

NEPHROSIS TOXIC
 

Continued on next page 
Table XXXIa (continued). Individual Pathology - Trichloroethylene-Treated Male Rats
 

WEEKS ON
 

STUDY
 

62
 

62
 

65
 

65
 

65
 

66
 

68
 

70
 

71
 

72
 

DISP
 

CODE
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

High Dose Group 

ANIMAL NUMBER 

TOPOGRAPHY 

 TUMORS 

MORPHOLOGY 

OTHER 

TOPOGRAPHY 

007 LUNG 

BONE MARROW KIDNEY 

038 ADRENAL ADRENAL LUNG 

KIDNEY LIVER 

011 THYROID FOLLICULAR ADENOCARCINOMA BRONCHUS 

BRONCHUS 

PLEURA BONE MARROW 

LIVER KIDNEY 

028 LUNG 

KIDNEY BONE MARROW 

045 LUNG 

KIDNEY TESTIS 

BONE MARROW 

049 LUNG 

KIDNEY 

BONE MARROW 

010 LUNG 

PLEURA KIDNEY 

020 KIDNEi 

EPIDIDYMIS 

016 KIDNEY 

034 SKIN PILOMATRIXOMA PERICARDIUM MYOCARDIUM LUNG 

PLEURA KIDNEY 

PATHOLOGY
 

MORPHOLOGY
 

INFLAMMATION CHRONIC
 

METAMORPHOSIS FATTY
 NEPHROSIS TOXIC
 

ANGIECTASIS
 

DEGENERATION
 

INFLAMMATION CHRONIC
 

NEPHROSIS 1OXIC
 

INFLAMMATION, DIFFUSE
 

ABSCESS
 

BRONCHIECTASIS
 

INFLAMMATION
 

METAMORPHOSIS FATTY
 

AN3IECTASIS
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

NEPHROSIS 1OXIC
 

METAMORPHOSIS FATTY
 

INFLAMMATION CHRONIC
 

NEPHROSIS IOXIC
 

ATROPHY
 METAMORPHOSIS FATTY
 

INFLAMMATION CHRONIC
 

NEPHROSIS TOXIC
 

METAMORPHOSIS FATTY
 

INFLAMMATION CHRONIC
 

INFLAMMATION
 

NEPHROSIS TOXIC
 

NEPHROSIS TOXIC
 

FAT NECROSIS WITH
 ENCAPSULATION
 

NEPHROSIS 1CXIC
 

INFLAMMATION
 

INFLAMMATION
 

INFLAMMATION CHRONIC
 

INFLAMMATION
 

NEPHROSIS TOXIC
 

Continued on next page
 
Table XXXIa (continued). Individual Pathology - Trichloroethylene-Treated Male Rats
 

High Dose Group
 

WEEKS ON DISP ANIMAL TUMORS OTHEU PATHOLOGY STUDY CODE NUMBER 

TOPOGRAPHY MORPHOLOGY TOPOGRAPHY MORPHOLOGY 

72 NATD 050 LUNG INFLAMMATION CHRONIC 

KIDNEY NEPHROSIS TOXIC 

ABDOMEN FAT NECROSIS WITH 

ENCAPSULATION 

75 NATD 044 LUNG INFLAMMATION CHRONIC 

KIDNEY NEPHROSIS TOXIC 

76 NATD 021 PERICARDIUM INFLAMMATION MYOCARDIUM INFLAMMATION LUNG INFLAMMATION CHRONIC 

PLEURA INFLAMMATION KIDNEY NEPHROSIS TOXIC 

82 NATD 019 HEART AORTIC BODY TUMOR LUNG INFLAMMATION CHRONIC KIDNEY NEPHROSIS TOXIC 

83 NATD 043 PITUITARY ANGIECTASIS 

LUNG INFLAMMATION CHRONIC KIDNEY NEPHROSIS TOXIC KIDNEY INFLAMMATION 

BONE MARROW METAMORPHOSIS FATTY 

88 NATD 031 HEART THROMBOSIS 

ENDOCARDIUM INFLAMMATION MYOCARDIUM INFLAMMATION LUNG INFLAMMATION CHRONIC 

LUNG ABSCESS KIDNEY NEPHROSIS TOXIC 

91 NATD 036 PANCREAS HEMANGIOSARCOMA PERICARDIUM INFLAMMATION MYOCARDIUM INFLAMMATION LUNG INFLAMMATION CHRONIC 

PLEURA INFLAMMATION KIDNEY NEPHROSIS TOXIC 

PROSTATE INFLAMMATION 

97 NATD 042 ADRENAL ANGIECTASIS 

LUNG INFLAMMATION CHRONIC KIDNEY NEPHROSIS TOXIC 

TESTIS ATROPHY 

99 NATD 008 LUNG INFLAMMATION CHRONIC 

KIDNEY NEPHROSIS TOXIC 

Continued on next page 
Table XXXIa (continued). Individual Pathology - Trichloroethylene-Treated Male Rats

 

High Dose Group 

WEEKS ON STODY DISP CODE ANIMAL NUMBER 

TOPOGRAPHY 

TUMORS 

MORPHOLOGY 

OTHER 

TOPOGRAPHY 

PATHOLOGY 

MORPHOLOGY 

102 NATD 035 LUNG 

BONE MARROW KIDNEY 

INFLAMMATION CHRONIC 

METAMORPHOSIS FATTY NEPHROSIS TOXIC 

103 NATD 001 LUNG 

KIDNEY INFLAMMATION CHRONIC NEPHROSIS TOXIC 

109 NATD 048 ADRENAL LUNG 

KIDNEY KIDNEY 

ANGIECTASIS 

INFLAMMATION CHRONIC 

NEPHROSIS TOXIC 

PYELONEPHRITIS 

110 TSAC 023 LUNG 

KIDNEY 

INFLAMMATION CHRONIC 

NEPHROSIS TOXIC 

110 TSAC 026 SPLEEN HEMANGIOSARCOMA LUNG KIDNEY INFLAMMATION CHRONIC 

NEPHROSIS TOXIC 

110 TSAC 046 LUNG KIDNEY TESTIS 

INFLAMMATION CHRONIC 

NEPHROSIS TOXIC 

ATROPHY 

end of male rats—high dose 
Table XXXIb. Individual Pathology - Trichloroethylene-Treated Female Rats
 

Control Group (Vehicle) 

WEEKS ON 

STUDY DISP CODE ANIMAL NUMBER 

TOPOGRAEHY 

TUMORS 

MORPHOLOGY TOPOGRAPHY 

OTHERR

MORPHOLOGY 

PATHOLOGY Y

25 NATD 008	 LUNG INFLAMMATION CHRONIC
 

BONE MARROW METAMORPHOSIS PATTY
 

47 NATD 016	 PERICARDIUM INFLAMMATION
 

MYOCARDIUM INFLAMMATION
 LUNG INFLAMMATION CHRONIC
 

PLEURA INFLAMMATION
 

U7 NATO 020	 LUNG INFLAMMATION CHRONIC
 

SPLEEN HEMATOPOIESIS EXTRAMSD»
 

6 6 NATD 006 PITUITARY CHROMOPHOBE ADENOMA	 LUNG INFLAMMATION CHRONIC
 

BONE MARROW METAMORPHOSIS FATTY
 

79 NATD 001	 LUNG INFLAMMATION CHRONIC
 

LUNG ABSCESS
 

87 NATD 007	 LUNG INFLAMMATION CHRONIC
 

PLEURA INFLAMMATION
 

97 NATD 011 MAMMARY GLAND FIBROADENOMA	 KIDNEY INFLAMMATION CHRONIC
 

LUNG INFLAMMATION CHRONIC
 

PLEURA INFLAMMATION
 STOMACH ULCER FOCAL
 

98 NATD 015	 LUNG INFLAMMATION CHRONIC
 

99 NATD 002	 LUNG INFLAMMATION CHRONIC
 

102 NATD 014	 PITUITARY CHROMOPHOBE ADENOMA ADRENAL ANGIECTASIS
 MAMMARY GLAND ADENOCARCINOMA LUNG INFLAMMATION CHRONIC
 

104 NATD 019 PITUITARY CHROMOPHOBE ADENOMA	 KIDNEY INFLAMMATION CHRONIC
 

LUNG INFLAMMATION CHRONIC
 

BONE MARROW METAMORPHOSIS FATTY
 

108 NATD 012	 KIDNEY INFLAMMATION CHRONIC
 

LUNG INFLAMMATION CHRONIC
 

BONE MARROW METAMORPHOSIS FATTY
 

110 TSAC 003 PITUITARY CHROMOPHOBE ADENOMA LUNG INFLAMMATION CHRONIC
 

MAMMARY GLAND FIBROADENOMA PLEURA INFLAMMATION
 D.ONE MARROW METAMORPHOSIS FATTY
 

Continued on next page
 
Table XXXIb (continued). Individual Pathology - Trichloroethylene-Treated Female Rats 

Control Group (Vehicle) 

WEEKS ON STUDY DISP CODE ANIMAL NUMBER 

TOPOGRAPHY 

TUHORS 

MORPHOLOGY 

OTHER 

TOPOGRAPHY 

PATHOLOGY 

MORPHOLOGY 

110 TSAC 004 SPLEEN 

OVARY RETICULUM CELL SARCOMA GRANULOSA CELL CARCINOMA LUNG INFLAMMATION CHRONIC 

110 TSAC 005 LUNG INFLAMMATION CHRONIC 

110 TSAC 009 LUNG INFLAMMATICN CHRONIC 

110 TSAC 010 MAMMARY GLAND FIBROADENOMA L£FT ADRENAL ADRENAL LUNG 

ANGIECTASIS 

DEGENERATION INFLAMMATION CHRONIC 

110 TSAC 013 KIDNEY 

LUNG 

INFLAMMATICN CHRONIC 

INFLAMMATION CHRONIC 

110 TSAC 017 LUNG 

BONE MARROW 

INFLAMMATICN CHRONIC 

METAMORPHOSIS FATTY 

110 TSAC 018 KIDNEY 

KIDNEY LUNG 

PLEURA LIVER 

LIVER PANCREAS 

PANCREAS 

INFLAMMATION CHRONIC 

INFLAMMATION CYSTIC 

INFLAMMATION CHRONIC 

INFLAMMATION METAMORPHOSIS FATTY 

INFLAMMATION ATROPHY INFLAMMATION CYSTIC 

end of female rats controls 
Table XXXIb (continued). Individual Pathology - Trichloroethylene-Treated Female Rats
 

WEEKS ON
 STUDY
 

2
 

2
 

2
 

3
 

5
 

7
 

15
 

16
 

16
 

19
 

21
 

21
 

21
 

22
 

22
 

25
 

26
 

DISP
 CODE
 

NATD
 

MISS
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

MISS
 

NATD
 

NATD
 

NATD
 

NATD
 

Low Dose Group
 

ANIMAL TUMORS
 

NUMBER
 

TOPOGRAPHY MORPHOLOGY
 

024
 

029
 

042
 

002 LUNG HEMANGIOSARCOMA,
 

HEART HEMANGIOSARCOMA,
 

020
 

047
 

C01
 

030
 

038
 

011
 

012
 

014
 

021
 

005
 

039
 

017
 

036
 

OTHER
 

TOPOGRAPHY
 

LUNG
 KIDNEY
 

LUNG
 

KIDNEY
 

KIDNEY
 

PERICARDIUM
 MYOCARDIUM
 

KIDNEY
 

PERICARDIUM
 

MYOCARDIUM
 LUNG
 

PLEURA
 

KIDNEY
 

KIDNEY
 

BONE MARROW
 KIDNEY
 

SUBCUT TISSUE
 

BONE MARROW
 

BONE MARROW
 

LUNG
 KIDNEY
 

BONE MARROW
 

KIDNEY
 

PATHOLOGY
 

MORPHOLOGY
 

INFLAMMATION CHRONIC
 

NEPHROSIS TOXIC
 

ANIMAL MISSING
 

INFLAMMATION CHRONIC
 

NEPHROSIS TOXIC
 

PYELONEPHRITIS
 

INFLAMMATION
 INFLAMMATION
 

NEPHROSIS TOXIC
 

INFLAMMATION
 

INFLAMMATION
 INFLAMMATION CHRONIC
 

INFLAMMATION
 

NEPHROSIS TOXIC
 

CALCIUM DEPOSITION
 METAMORPHOSIS FATTY
 

NEPHROSIS TOXIC
 

ABSCESS
 

METAMORPHOSIS FATTY
 

NO SIGNIFICANT DIAGNOSIS
 

ANIMAL MISSING
 

METAMORPHOSIS FATTY
 

INFLAMMATION CHRONIC
 

NEPHROSIS TOXIC
 

METAMORPHOSIS FATTY
 

NEPHROSIS TOXIC
 

Continued on next page
 
Table XXXIb (continued). Individual Pathology - Trichloroethylene-Treated Female Rats
 

WEEKS ON
 STUDY
 

23
 

33
 

34
 

40
 

42
 

57
 

60
 

61
 

Ο
 

68
 

69
 

75
 

75
 

77
 

86
 

96
 

96
 

DISP
 CODE
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

ANIMAL
 NUMBER
 

035
 

033
 

019
 

045
 

022
 

006
 

010
 

044
 

027
 

025
 

015
 

016
 

048
 

034
 

023
 

046
 

Low Dose Group 

TUMORS 

TOPOGRAPHY MORFHOLOGY 

LIVER RETICULUM CELL SARCOMA 

CERVICAL LYMPH NODE RETICULUM CELL SARCOMA 

ADRENAL ADRENAL CORTICAL CARCINOMA 

OTHER
 

TOPOGRAPHY
 

KIDNEY
 

KIDNEY
 

KIDNEY
 

BONE MARROW
 

ADRENAL
 

ADRENAL
 

KIDNEY
 

LUNG
 

KIDNEY
 

KIDNEY
 

BONE MARROW
 

KIDNEY
 

BRONCHUS
 

PLEURA
 

KIDNEY
 

LUNG
 

KIDNEY
 

LUNG
 

LUNG
 

KIDNEY
 

LUNG
 

KIDNEY
 

LUNG
 

KIDNEY
 

LUNG
 

PLEURA
 

KIDNEY
 

LUNG
 

KIDNEY
 

LUNG
 

PATHOLOGY
 

MORPHOLOGY
 

NEPHROSIS ICXIC
 

NEPHROSIS TCXIC
 

NEPHROSIS TCXIC
 

M£TAMORPHOSI3 FATTY
 

ANGIECTASIS
 

DEGENERATION
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

NEPHROSIS TOXIC
 

NEPHROSIS TOXIC
 

METAMORPHOSIS FATTY
 

NEPHROSIS TOXIC
 

ABSCESS
 

INFLAMMATION
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

INFLAMMATION CHRONIC
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

NEPHROSIS TCXIC
 

INFLAMMATION CHRONIC
 

INFLAMMATION
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

NEPHROSIS ICXIC
 

INFLAMMATION CHRONIC
 

Continued on next page
 
Table XXXIb (continued). Individual Pathology - Trichloroethylene-Treated Female Rats
 

Low Dose Group
 

WEEKS ON DISP ANIMAL TUMORS OTHfiit PATHOLOGY STUDY CODE NUMBER 

TOPOGRAPHY MORPHOLOGY TOPOGRAPHY MORPHOLOGY 

98 NATD 032 SUBCUT TISSUE/CHEST LIPOSARCOMA	 KIDNEY NEPHROSIS 1CXIC
 

LUNG INFLAMMATION CHRONIC
 

100 NATD 013 SUBCOT TISSUE/NECK FIBROMA	 KIDNEY NcPHRCSIS TOXIC
 

LUNG INFLAMMATION CHRONIC
 

102 NATD 049	 KIDNEY NEPHROSIS TOXIC
 

LUNG INFLAMMATION CHRONIC
 

104 NATD 040 MAMMARY GLAND FIBROADENOMA	 SPLEEN HJSMATCPOIISI S EXTRAMED.
 

KIDNEY NEPHROSIS TOXIC
 

LUNG INFLAMMATION CHRONIC
 

110 TSAC 003	 TRACHEA INFLAMMATION
 

TRACHEAL LYMPH NODE INFLAMMATION
 

KIDNEY NEPHROSIS TOXIC
 

LUNG INFLAMMATION CHRONIC
 

110 TSAC 004	 KIDNEY NEPHROSIS TCXIC
 

LUNG INFLAMMATION CHRONIC
 

RIGHT OVARY CYST
 BONE MARROW METAMORPHOSIS FATTY
 

1 10 TSAC 007 PITUITARY CHROMOPHOBE ADENOMA	 ADRENAL ANGl£iCTASIS
 MAMMARY GLAND FIBROADENOMA	 MYOCARDIUM FIBROSIS
 

KIDNEY NEPHROSIS TOXIC
 

LUNG INFLAMMATION CHRONIC
 

110 TSAC 008	 KIDNEY NEPHROSIS TCXIC
 

LUNG INFLAMMATION CHRONIC
 

110 TSAC 009	 ADRENAL ANGIECTASIS
 

KIDNEY NEPHROSIS TOXIC
 

LUNG INFLAMMATION CHRONIC
 

RIGHT OVARY CYST
 

110 TSAC 018 MAMMARY GLAND FIBROADENOMA	 RIGHT ADRENAL ANGIECTASIS
 

KIDNEY NEPHROSIS TOXIC
 

LUNG INFLAMMATION CHRONIC
 

Continued on next page
 
Table XXXIb (continued). Individual Pathology - Trichloroethylene-Treated Female Rats 

Low Dose Group 

WEEKS ON DISP ANIMAL TUMORS 

STUDY CODE NUMBER 

TOPOGRAEHY MORPHOLOGY 

110 TSAC 026 ENDOMETRIUM SARCOMA 

110 TSAC 028 

110 T3AC 031 PITUITARY CHROMOPHOBE ADENOMA 

110 TSAC 037 

110 TSAC 041 

110 TSAC 043 MAMMARY GLAND FIBROADENOMA 

110 NATD 050 MAMMARY GLAND FIBROADENOMA, (MULTIPLE - 2) 

OTHER
 

TOPOGRAPHY
 

LEFT ADRENAL
 

ADRENAL
 

LEFT ADRENAL
 

KIDNEY
 

LUNG
 

OVARY
 

ADRENAL
 

ADRENAL
 

BILE DUCT
 

KIDNEY
 

LUNG
 

OVARY
 

KIDNEY
 

LUNG
 

ENDOMETRIUM
 

LEFT OVARY
 

RIGHT EYE
 

RETINA
 

KIDNEY
 

LUNG
 

BONE MARROW
 

PERICARDIUM
 

KIDNEY
 

LUNG
 

PLEURA
 

KIDNEY
 

LUNG
 

KIDNEY
 

LUNG
 

UTERUS
 

BILE DUCT
 

PATHOLOGY
 

MORPHOLOGY
 

AN3IECTASIS
 

DEGENERATION
 

INFLAMMATION CYSTIC
 

NEPHROSIS TCXIC
 

INFLAMMATION CHRONIC
 

CYST
 

ANGIECTASIS
 

DEGENERATION
 

INFLAMMATION PROLIFERATIVE
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

CYST
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

INFLAMMATION
 

CYST
 

CATARACT
 

DETACHMENT
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

METAMORPHOSIS FATTY
 

INFLAMMATION
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

INFLAMMATION
 

NEPHROSIS TCXIC
 

INFLAMMATION CHRONIC
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 INFLAMMATION
 

INFLAMMATION PROLIFERATIVE
 

end of female rats—low dose 
Table XXXIb (continued). Individual Pathology - Trichloroethylene-Treated Female Rats 

High Dose Group 

WEEKS ON STUDY DISP CODE ANIMAL NUMBER 

TOPOGRAPHY 

TUMORS 

MORPHOLOGY 

OTHER 

TOPOGRAPHY 

3 NATD 013 PERICARDIUM 

MYOCARDIUM LUNG 

PLEURA 

BONE MARROW KIDNEY 

5 NATD 002 PERICARDIUM 

MYOCARDIUM LUNG 

PLEURA BONE MARROW KIDNEY 

9 NATD 032 LUNG 

BONE MARROW 

KIDNEY 

21 NATD 036 KIDNEY BONE MARROW 

24 NATD 049 KIDNEY LUNG 

28 NATD 008 KIDNEY 

28 NATD 016 KIDNEY 

30 NATD 043 LUNG 

KIDNEY 

35 NATD 040 KIDNEY 

43 NATD 038 UTERUS 

46 NATD 045 LUNG 

KIDNEY 

50 NATD 004 LUNG 

PLEURA KIDNEY KIDNEY 

PATHOLOGY
 

MORPHOLOGY
 

INFLAMMATION
 INFLAMMATION
 INFLAMMATION CHRONIC
 

INFLAMMATION
 METAMORPHOSIS FATTY
 NEPHROSIS TOXIC
 

INFLAMMATION
 

INFLAMMATION
 INFLAMMATION CHRONIC
 

INFLAMMATION
 METAMORPHOSIS FATTY
 NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

METAMORPHOSIS FATTY
 

NEPHROSIS TOXIC
 

CALCIUM DEPOSITION
 METAMORPHOSIS FATTY
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

NEPHROSIS TOXIC
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

NEPHROSIS TOXIC
 

NEPHROSIS TOXIC
 

RETENTION FLUID
 

INFLAMMATION CHRONIC
 

NEPHROSIS IOXIC
 

INFLAMMATION CHRONIC
 

INFLAMMATION
 NEPHROSIS IOXIC
 

CALCIUM DEPOSITION
 

Continued on next page 
IEEKS ON DISP ANIMAL
 

STUDY CODE NUMBER
 

53 NATD 007
 

54 ATD 041
 

58 NATD 024
 

59 NATD 017
 

63 NATD 031
 

64 NATD 022
 

67 NATD 028
 

69 NATD 012
 

69 NATD 027
 

Table XXXIb (continued). Individual Pathology - Trichloroethylene-Treated Female Rats
 

High Dose Group 

TUMORS 

TOPOGBAFHY MORPHOLOGY 

OTHER
 

TOPOGRAPHY
 

LUNG
 

PLEURA
 

KIDNEY
 

LUNG
 

PLEURA
 

KIDNEY
 

LUNG
 

PLEURA
 

KIDNEY
 

BONE MARROW
 

LUNG
 

KIDNEY
 

BONE MARROW
 

PERICARDIUM
 

BRONCHUS
 

KIDNEY
 

LUNG
 

PLEURA
 

PERICARDIUM
 

MYOCARDIUM
 

KIDNEY
 

KIDNEY
 

BONE MARROW
 

KIDNEY
 

LUNG
 

PLEURA
 

I fJHG
 

PLEURA
 

KIDNEY
 

LUNG
 

KIDNEY
 

KIDNEY
 

UTERUS
 

BONE MARROW
 

PATHOLOGY
 

MORPHOLOGY
 

INFLAMMATION CHRONIC
 

INFLAMMATION
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

INFLAMMATION
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

INFLAMMATION
 

NEPHROSIS TOXIC
 

METAMORPHOSIS FATTY
 

INFLAMMATION CHRONIC
 

NEPHROSIS TOXIC
 

METAMORPHOSIS FATTY
 

INFLAMMATION
 

BRONCHIECTASIS
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

INFLAMMATION
 

INFLAMMATION
 

INFLAMMATION
 

NEPHROSIS TOXIC
 

CALCIUM DEPOSITION
 

METAMORPHOSIS FATTY
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

INFLAMMATION
 

INFLAMMATION CHRONIC
 

INFLAMMATION
 

NEPHROSIS TOXIC, MARKED
 

INFLAMMATION CHRONIC
 

NEPHROSIS TOXIC
 

CALCIUM DEPOSITION
 

RETENTION FLUID
 

METAMORPHOSIS FATTY
 

Continued on next page
 
Table XXXIb (continued). Individual Pathology - Trichloroethylene-Treated Female Rats
 

High Dose Group 

WEEKS ON DISP ANIMAL TUMORS OTHER PATH0L03Y STUDY CODE NUMBER 

TOPOGRAPHY MORPHOLOGY TOPOGRAPHY MORPHOLOGY 

70 NATD 018 LONG INFLAMMATION CHRONIC 

KIDNEY NEPHROSIS TOXIC 

SHALL INTESTINE ULC2R FOCAL 

70 NATD 020 LONG INFLAMMATION CHRONIC 

KIDNEY NEPHROSIS TOXIC ΒΟΙ MARROW METAMORPHOSIS FATTY 

72 NATD οία LONG INFLAMMATION CHRONIC KIDIEY NEPHROSIS TOXIC 

73 NATD 001 ADREHAL ANGIECTASIS 

LONG INFLAMMATION CHRONIC 

KIDNEY NEPHROSIS TOXIC ΒΟΙ MARROW METAMORPHOSIS FATTY 

73 NATD 0U2 BRONCHOS ABSCESS 

KIDNEY NEPHROSIS TOXIC KIDNEY CALCIUM DEPOSITION 

74 NATD 039 BRONCHOS BRONCHIECTASIS KIDNEY NEPHROSIS TOXIC 

82 NATD 026 LUNG INFLAMMATION CHRONIC 

KIDNEY NEPHROSIS TOXIC 

BONE MARROW METAMORPHOSIS FATTY 

86 NATD 04 8 LUNG INFLAMMATION CHRONIC 

PLEURA INFLAMMATION KIDNEY NEPHROSIS TOXIC 

MESENTERIC LYMPH NODE INFLAMMATION MESENTERIC LYMPH NODE INFLAMMATION CYSTIC 

89 NATD 047 ADRENAL ANGIECTASIS 

LUNG INFLAMMATION CHRONIC 

95 NATD 021 LUNG INFLAMMATION CHRONIC 

PLEURA INFLAMMATION KIDNEY NEPHROSIS TOXIC 

96 NATD 050 PITUITARY CHROMOPHOBE ADENOMA ADRENAL ANGIECTASIS 

LIVER METAMORPHOSIS FATTY LUNG INFLAMMATION CHRONIC KIDNEY NEPHROSIS TOXIC 

Continued on next page 
Table XXXIb (continued). Individual Pathology - Trichloroethylene-Treated Female Rats
 

High Dose Group 

WEEKS ON DISP ANIMAL TUMORS OTHER 

STUDY CODE NUMBER 

TOPOGRAPHY MORPHOLOGY TOPOGRAPHY 

101 NATD 029 MAMMARY GLAND FIBROADENOMA LUNG 

KIDNEY 

101 NATD 035 PITUITARY CHROMOPHOBE ADENOMA ADRENAL 

LUNG 

PLEURA 

KIDNEY 

102 NATD 019 LUNG 

KIDNEY 

103 NATD 033 MAMMARY GLAND FIBROADENOMA (MULTIPLE-3) LUNG 

KIDNEY 

104 NATD 034 BRAIN 

PERICARDIUM 

LUNG 

PLEURA 

KIDNEY 

110 TSAC 003 ADRENAL 

LUNG 

BILE DUCT 

KIDNEY 

BONE MARROW 

110 TSAC 005 PITUITARY CHROMOPHOBE ADENOMA ADRENAL 

LUNG 

KIDNEY 

BONE MARROW 

110 TSAC 006 THYMUS RETICULUM CELL SARCOMA LUNG 

KIDNEY 

OVARY 

110 TSAC 009 PITUITARY CHROMOPHOBE ADENOMA ADRENAL 

ΜΑΜΜΑΡ 3LAND FIBROADENOMA ADRENAL 

LUNG 

PLEURA 

KIDNEY 

PATHOLOGY
 

MORPHOLOGY
 

INFLAMMATION CHRONIC
 

NEPHROSIS TOXIC
 

ANGIECTASIS
 

INFLAMMATION CHRONIC
 

INFLAMMATION
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

NEPHROSIS IOXIC
 

HYDROCEPHALUS
 

INFLAMMATION
 

INFLAMMATION CHRONIC
 

INFLAMMATION
 

NEPHROSIS IOXIC
 

ANGIECTASIS
 

INFLAMMATION CHRONIC
 

DILATATION
 

NEPHROSIS TOXIC
 

METAMORPHOSIS FATTY
 

ANGIECTASIS
 

INFLAMMATION CHRONIC
 

NEPHROSIS TOXIC
 

METAMORPHOSIS FATTY
 

INFLAMMATION CHRONIC
 

NEPHROSIS TOXIC
 

INFLAMMATION
 

DEGENERATION
 

ANGIECTASIS
 

INFLAMMATION CHRONIC
 

INFLAMMATION
 

NEPHROSIS TOXIC
 

Continued on next page
 
Table XXXIb (continued) Individual Pathology - Trichloroethylene-Treated Female Rats
 

High Dose Group
 

end of female rats
 
Table XXXIIa. Individual Pathology - Trichloroethylene-Treated Male Mice
 

Control Group (Vehicle)
 

WEEKS ON STUDY DISP CODE ANIMAL NUMBER 

TOPOGRAPHY 

 TUMORS 

MORPHOLOGY 

OTHER PATHOLOGY 

TOPOGRAPHY MORPHOLOGY 

32 NATD 010 NO SIGNIFICANT DIAGNOSIS 

39 NATD 009 SPLEEN 

KIDNEY AMYLOIDOSIS PYELONEPHRITIS 

60 NATD 008 KIDNEY 

KIDNEY KIDNEY 

INFLAMMATION CHRONIC 

HYDRONEPHROSIS 

AMYLOIDOSIS 

61 NATD 007 KIDNEY KIDNEY INFLAMMATION CHRONIC 

AMYLOIDOSIS 

64 NATD 006 KIDNEY KIDNEY SPLEEN 

INFLAMMATION CHRONIC 

HYDRONEPHROSIS 

AMYLOIDOSIS 

66 NATD 020 MESENTERIC LYMPH 

SUBCUT TISSUE 

NODE ANGIECTASIS 

ABSCESS 

68 NATD 005 KIDNEY KIDNEY KIDNEY SPLEEN LIVER ENDOCARDIUM 

IKFLAMMATICN CHRONIC 

AMYLOIDOSIS 

HYDRONEPHROSIS 

AMYLOIDOSIS 

HYPERPLASIA HYPERPLASIA 

72 NATD 019 LIVER HEPATOCELLULAR CARCINOMA SPLEEN AMYLOIDOSIS 

76 NATD 004 KIDNEY 

KIDNEY 

INFLAMMATION CHRONIC 

HYDRONEPHROSIS 

76 «SAC 018 LIVER 

SPLEEN MESENTERIC LYMPHNODE PROSTATf 

SEMINAL VESICLES 

RETICULUM CELL SARCOMA RETICULUM CELL SARCOMA RETICULUM CELL SARCOMA RETICULUM CELL SARCOMA RETICULUM CELL SARCOMA 

77 NATD 017 SUBCOT TISSUE/BACK FIBROSARCOMA SPLEEN 

KIDNEY KIDNEY 

AMYLOIDOSIS 

INFLAMMATION CHRONIC 

AMYLOIDOSIS 

Continued on next page 
Table XXXIIa (continued). Individual Pathology - Trichloroethylene-Treated Male Mice
 

Control Group (Vehicle) 

WEEKS ON DISP ANIMAL TOHORS OTHSR STUDY CODE NUMBER 

TOPOGRAPHY HORPHOLOGY TOPOGRAPHY 

78 NATD 003 

90 TSAC 001 SKIN OP CHEST FIBROSARCOMA KIDNEY 

KIDNEY 

90 TSAC 002 KIDNEY 

KIDNEY 

90 TSAC 011 SOBCUT TISSUE/BACK FIBROSARCOMA 

90 TSAC 012 

90 TSAC 013 SKIN 

SKIN 

90 TSAC 014 

90 TSAC 015 KIDNEY KIDNEY KIDNEY 

90 TSAC 016 

PATHOLOGY
 

MORPHOLOGY
 

NO SIGNIFICANT DIAGNOSIS
 

INFLAMMATION CHRONIC
 

HYDRONEPHROSIS
 

INFLAMMATICN CHRONIC
 

HYDRONEPH5OSIS
 

NO SIGNIFICANT DIAGNOSIS
 

ACANTHOSIS
 

INFLAMMATION
 

NO SIGNIFICANT DIAGNOSIS
 

HYDRONEPHROSIS
 

INFLAMMATION CHRONIC
 

AMYLOIDOSIS
 

NO SIGNIFICANT DIAGNOSIS
 

end of male mice controls
 
Table XXXIIa (continued). Individual Pathology - Trichloroethylene-Treated Male Mice
 

WEEKS ON
 

STUDY
 

16
 

18
 

31
 

51
 

53
 

58
 

63
 

65
 

65
 

77
 

81
 

DISP
 

CODE
 

NATD
 

KATD
 

ACCK
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

ANIMAL
 NUMBER
 

010
 

050
 

009
 

030
 

040
 

008
 

039
 

020
 

038
 

019
 

028
 

 TUMORS
 

TOPOGRAPHY
 

THYMUS
 

BRONCHIAL LYMPHNODE
 

LUNG
 

KIDNEY
 PROSTATE
 BONE MARROW
 

CERVICAL LYMPHNODE
 SPLEEN
 

LIVER
 

SPLEEN
 LIVER
 

Low Dose Group
 

MORPHOLOGY
 

LYMPHOSARCOMA
 LYMPHOSARCOMA
 LYMPHOSARCOMA
 LYMPHOSARCOMA
 LYMPHOSARCOMA
 LYMPHOSARCOMA
 

LYMPHOSARCOMA
 LYMPHOSARCOMA
 

LYMPHOSARCOMA
 

LYMPHOSARCOMA
 LYMPHOSARCOMA
 

OTHER PATHOLOGY
 

TOPOGRAPHY MORPHOLOGY
 

NO SIGNIFICANT DIAGNOSIS
 

KIDNEY NEPHROSIS TOXIC
 

NO SIGNIFICANT DIAGNOSIS
 

KIDNEY NEPHROSIS TOXIC
 

KIDNEY NEPHROSIS 1OXIC
 

HEART ORGANIZED THHCMBUS
 

ENDOCARDIUM INFLAMMATION
 

MYOCARDIUM INFLAMMATION
 

KIDNEY PYELONEPHRITIS
 

KIDNEY HYDRONEPHROSIS
 

SPLEEN AMYLOIDOSIS
 

URINARY BLADDER INFLAMMATION
 

PROSTATE INFLAMMATION
 

PANCREAS ATROPHY
 

PANCREAS INFLAMMATIOH
 BONE INFLAMMATION
 

KIDNEY NEPHROSIS IOXIC
 

KIDNEY NEPHROSIS TOXIC
 

LIVER HYPERPLASIA
 KIDNEY NEPHROSIS TOXIC
 

KIDNEY NEPHROSIS TOXIC
 

BRONCHUS ABSCESS
 

CERVICAL LYMPH NODE INFLAMMATION
 

KIDNEY NEPHROSIS TOXIC
 

KIDNEY NEPHROSIS TOXIC
 

Continued on next page
 
Table XXXIIa (continued).


WEEKS ON DISP ANIBAL TUHORS
 

STUDY CODE NUMBER
 

TOPOGRAPHY
 

81 NATD 029 LONG 

LIVER 

66 NATD 049 LIVER 

88 NATD 007	 LIVER
 

LUNG
 

LONG
 

88 NATD 018
 

90 TSAC 001	 LIVER
 

LONG
 

90 TSAC 002	 S0BC0T TISSUE/BACK
 

90 TSAC 003	 LIVER
 

90 TSAC 004	 LIVER
 

LONG
 

HESENTERIC LYHPHNODE
 ILEUM
 

RENAL LYHPHNODE
 

90 TSAC 005	 LIVER
 

90 TSAC 006	 LIVER
 

90 TSAC 011 LIVER
 

LUNG
 

90 TSAC 012 LIVER
 

90 TSAC 013 LIVER
 

90 TSAC 014	 HARDERIAN GLAND
 

 Individual Pathology - Trichloroethylene-Treated Male Mice
 

Low Dose Group
 

OTHER PATHOLOGY
 

MORPHOLOGY	 TOPOGRAPHY MORPHOLOGY
 

ADENOMA
 KIDNEY NEPHROSIS TOXIC
 HEPATOCELLULAR CARCINOMA
 

HEPATOCELLULAR CARCINOMA KIDNEY
 NEPHROSIS ICXIC
 STOMACH HYPERKERATOSIS
 

STOMACH ACANTHOSIS
 

HEPATOCELLULAR CARCINOMA KIDNEY
 NEPHROSIS TCXIC
 HEPATOCELLULAR CARCINOMA METASTA
 

ADENOMA
 

MESENTERIC LYMPH NODE ANGIECTASIS
 KIDNEY NEPHROSIS TOXIC
 

HEPATOCELLULAR CARCINOMA KIDNEY	 NEPHROSIS
 ICXIC
 ADENOMA
 

FIBEOMA
 KIDNEY NEPHROSIS TCXIC
 

HEPATOCELLULAR CARCINOMA KIDNEY
 NEPHROSIS TCXIC
 

HEPATOCELLULAR CARCINOMA
 SPLEEN AMYLOIDOSIS
 ADENOMA	 KIDNEY NEPHROSIS
 TOXIC
 RETICULUM CELL SARCOMA
 

RETICULUM CELL SARCOMA
 

RETICULUM CELL SARCOMA
 

HEPATOCELLULAR CARCINOMA LIVER
 THROMBOSIS
 SPLEEN HEMATOPOIESIS EXIRAMED.
 KIDNEY NEPHROSIS TOXIC
 

HEPATOCELLULAR CARCINOMA
 MESENTERIC LYMPH NODi: ANGIECTASIS
 KIDNEY INFLAMMATION CHRONIC
 KIDNEY NEPHROSIS TOXIC
 

HEPATOCELLULAR CARCINOMA KIDNEY
 NEPHROSIS TCXIC
 HEPATOCELLULAR CARCINOMA METASTA
 

HEPATOCELLULAR CARCINOMA KIDNEY NEPHROSIS
 TCXIC
 

HEPATOCELLULAR CARCINOMA
 MESENTERIC LYMPH NODE INFLAMMATION
 KIDNEY NEPHROSIS TOXIC
 

ADENOMA
 KIDNEY NEPHROSIS ICXIC
 

Continued on next, page
 
Table XXXIIa (continued) . Individual Pathology - Trichloroethylene-Treated Male Mice
 

WEEKS ON
 STUDY
 

90
 

90
 

90
 

90
 

90
 

90
 

90
 

90
 

90
 

90
 

90
 

90
 

90
 

90
 

90
 

90
 

90
 

DISP
 CODE
 

TSAC
 

TSAC
 

TSAC
 

TSAC
 

TSAC
 

TSAC
 

TSAC
 

TSAC
 

TSAC
 

TSAC
 

TSAC
 

TSAC
 

TSAC
 

TSAC
 

TSAC
 

TSAC
 

TSAC
 

ANIMAL
 NUMBER
 

TOPOGRAPHY
 

015 EPIDIDYMIS 

PROSTATE SPLEEN 

LIVER LUNG 

016 

017 LIVER 

021 LIVER 

022 

023 

024 LIVER 

025 

026 LIVER 

027 

031 LIVER 

032 LIVER 

033 LIVER 

034 LIVER 

035 LIVER 

036	 LIVER
 

037	 LUNG
 

LIVER
 

TUMORS
 

Low Dose Group
 

MORPHOLOGY
 

RETICULUM CELL SAHCOHA
 

RETICULUM CELL SARCOHA
 RETICULUM CELL SAfiCOHA
 

HEPATOCELLULAR CABCIIOHA
 HEPATOCELLULAR CARCI1OHA BETAS A
 

HEPATOCELLULAR CAECIBOHA
 

HEPATOCELLULAR CA ECHO HA
 

HEPATOCELLULAR C ARCI HO HA
 

HEPATOCELLULAR CABCIIOHA
 

HEPATOCELLULAR CA EC 110 HA
 

HEPATOCELLULAR CABCIIOHA
 

HEPATOCELLULAR CAECIIOHA
 

HEPATOCELLULAR CABCIIOHA
 

HEPATOCELLULAR CAHCIBGHA
 

HEPATOCELLULAR CABCI1CHA
 

ADENOMA
 

HEPATOCELLULAR CABCIIOHA
 

OTHER
 

TOPOGRAPHY
 

KIDNEY
 

KIDNEY
 

KIDNEY
 

KIDNEY
 

KIDNEY
 

KIDNEY
 

KIDNEY
 

KIDNEY
 

KIDNEY
 

KIDNEY
 

KIDNEY
 

KIDNEY
 

KIDNEY
 

KIDNEY
 

KIDNEY
 KIDNEY
 

KIDNEY
 

KIDNEY
 

PATHOLOGY
 

MORPHOLOGY
 

NEPHROSIS TOXIC
 

NEPHROSIS TOXIC
 

NEPHROSIS TOXIC
 

NEPHROSIS TOXIC
 

NEPHROSIS TOXIC
 

NEPHROSIS TOXIC
 

NEPHROSIS TOXIC
 

NEPHROSIS TOXIC
 

NEPHROSIS TOXIC
 

NEPHROSIS TOXIC
 

NEPHROSIS TOXIC
 

NEPHROSIS TOXIC
 

NEPHROSIS TOXIC
 

NEPHROSIS TOXIC
 

INFLAMMATION CHRONIC
 

NEPHROSIS TOXIC
 

NEPHROSIS TOXIC
 

NEPHROSIS TOXIC
 

Continued on next page
 
Table XXXIIa (continued). Individual Pathology - Trichloroethylene-Treated Male Mice 

Low Dose Group 

WEEKS ON DISP ANIMAL TUMORS OTHER PATHOLOGY
 STUDY CODE NUMBER
 

TOPOGRAPHY MORPHOLOGY TOPOGRAPHY MORPHOLOGY
 

90 TSAC 041 LIVER HEPATOCELLULAR CARCINOMA KIDNEY STOMACH STOMACH MESENTERIC LYMPH RIGHT EYE RIGHT EYE HARDERIAN GLAND 

NEPHROSIS TOXIC 

ACANTHOSIS 

HYPERKERATOSIS 

NODE INFLAMMATION CONGENITAL MALFORMATION PHTHISIS EULEI INFLAMMATION 

90 TSAC 042 KIDNEY NEPHROSIS TOXIC 

90 TSAC 043 LIVER LUNG 

HEPATOCELLULAR 

HEPATOCELLULAR 

CARCINOMA 

CARCINOMA METASTA 

KIDNEY 

STOMACH STOMACH STOMACH MESENTERIC LYMPH 

NEPHROSIS TCXIC 

ACANTHOSIS 

HYPERKERATOSIS 

INFLAMMATION EXUDATIVE NODE INFLAMMATION 

90 TSAC 044 KIDNEY NEPHROSIS TOXIC 

90 TSAC 045 KIDNEY 

LIVER 

NEPHROSIS TOXIC 

HYPERPLASIA 

90 TSAC 046 KIDNEY NEPHROSIS TOXIC 

90 TSAC 047 KIDNEY LIVER 

NEPHROSIS TCXIC 

INFLAMMATION 

90 TSAC 048 LIVEB HEPATOCELLULAR CARCINOMA KIDNEY NEPHROSIS TOXIC 

end of male mice—low dose 
{continued)^ Individual Pathology - Trichloroethylene-Treated Male Mice
 

WEEKS ON DISP ANIMAL TUMORS 

STUDY CODE NUMBER 

TOPOGRAPHY 

13 NATD 010 

14 NATD 040 

15 NATD 049 

15 ACCK 050 

24 NATD 019 

24 NATD 020 

25 NATD 047 

25 NATD 048 

27 NATD 046 LIVER 

28 NATD 018 LIVER 

28 NATD 045 

29 NATD 030 

30 NATD 009 

30 NATD 017 LIVER 

36 NATD 008 LIVER 

42 NATD 007 

53 NATD 039 LIVER 

60 NATD 005 

60 NATD 006 

61 . NATD 016 LIVER 

70 NATD 038 LIVER 

71 NATD 015 LIVER 

High Dose Group
 

MORPHOLOGY 

HEPATOCELLULAR CARCINOMA
 

HEPATOCELLULAR CARCINOMA
 

HEPATOCELLULAR CABCINCMA
 

HEFATOCELLULAR CARCINOMA
 

HEPATOCELLULAR CAFCINCMA
 

HEPATOCELLULAR CARCINOMA
 

HEPATOCELLULAR CARCINOMA
 

HEPATOCELLULAR CARCINOMA
 

OTHER

TOPOGRAPHY

KIDNEY
 

KIDNEY
 

KIDNEY
 

KIDNEY
 

LIVER
 

KIDNEY
 

KIDNEY
 

KIDNEY
 

KIDNEY
 

KIDNEY
 

KIDNEY
 

KIDNEY
 

KIDNEY
 

KIDNEY
 

KIDNEY
 

KIDNEY
 

KIDNEY
 

KIDNEY
 

KIDNEY
 

KIDNEY
 

KIDNEY
 

KIDNEY
 

 PATHOLOGY 

 MORPHOLOGY 

NEPHROSIS TOXIC
 

NEPHROSIS TOXIC
 

NEPHROSIS ICXIC
 

NO SIGNIFICANT DIAGNOSIS
 

NEPHROSIS TOXIC
 

HYPfiRPLASIA
 

NEPHROSIS TOXIC
 

NEPHROSIS TOXIC
 

NEPHRCSIS TOXIC
 

NEPHROSIS TOXIC
 

NiiPHROSIS TOXIC
 

N£PHROSIS ICXIC
 

INFLAMMATION CHRONIC
 

NEPHRCSIS TCXIC
 

INFLAMMATION
 

NEPHRCSIS TCXIC
 

NEPHRCSIS TOXIC
 

NEPHROSIS TOXIC
 

NEPHROSIS TCXIC
 

NEPHROSIS TOXIC
 

NSPHROSIS TOXIC
 

NEPHROSIS TCXIC
 

NEPHROSIS TOXIC
 

Continued on next page 
Table XXXIIa (continued). Individual Pathology - Trichloroethylene-Treated Male Mice
 

High Dose Group _____^ _
 

WEEKS ON 

STUDY 

DISP 

CODE 

ANIMAL

NUMBER 

TOPOGRAPHY 

 TUMORS 

MORPHOLOGY 

OTHER

TOPOGRAPHY 

 PATHOLOGY 

MORPHOLOGY 

72 NATD 037 AUTOLYSIS 

74 NATD 029 LIVER HEPATOCELLULAR CARCINOMA KIDNEY 

LIVER/CENTRILOBULAR 

NEPHROSIS

NECROSIS 

 TOXIC 

75 NATD 004 KIDNEY 

LIVER/CENTRILOBULAR 

SALIVARY GLAND 

STOMACH 

STOMACH 

NEPHROSIS TOXIC 

NECROSIS 

INFLAMMATION CYSTIC 

ACANTHOSIS 

HYPERKERATOSIS 

75 NATD 036 AUTOLYSIS 

78 MSAC 044 IIVER 

TISSUE MUSCLE OF 

BACK 

HEPATOCELLULAR

NEUROFIBROMA 

 CARCINOMA KIDNEY NEPHROSIS TCXIC 

83 NATD 035 LIVER 

LUNG 

LUNG 

BRONCHIAL LYMPH NODE 

SKIN OF CHEST 

AORTA 

HEPATOCELLULAR CARCINOMA 

HEPATOCELLULAR CARCINOMA

ALVEOLAR ADENOCARCINOMA 

CARCINOMA METASTATIC 

CARCINOMA METASTATIC 

CARCINOMA METASTATIC 

 METASTA 

KIDNEY NEPHROSIS TOXIC 

88 MSAC 003 LIVER 

SPLEEN 

MESENTERIC

KIDNEY 

 LYMPHNODE 

HEPATOCELLULAR

LYMPHOSARCOMA 

LYMPHOSARCOMA 

LYMPHOSARCOMA 

 CARCINOMA TESTIS 

HARDERIAN GLAND 

KIDNEY 

ATROPHY 

INFLAMMATION 

NEPHROSIS TOXIC: 

90 TSAC 001 LIVER HEPATOCELLULAR CARCINOMA KIDNEY NEPHROSIS TOXIC 

90 SAC 002 LIVER HEPATOCELLULAR CARCINOMA THYROID 

KIDNEY 

KIDNEY 

HYPERPLASIA CYSTIC 

INFLAMMATION CHRONIC 

HYDRONEPHROSIS 

MESENTERIC LYMPH NODS INFLAMMATION 

KIDNEY NEPHROSIS TOXIC 

90 TSAC 011 LIVER HEPATOCELLULAR CARCINOMA KIDNEY NEPHROSIS TOXIC 

90 TSAC 012 LIVER HEPATOCELLULAR CARCINOMA KIDNEY NEPHROSIS TCXIC 

90 TSAd 013 KIDNEY 

LIVER 

ADENOMA 

HEPATOCELLULAR CARCINOMA 

LIVER 

KIDNEY 

HYPERPLASIA 

NEPHROSIS TOXIC 

Continued on next page 
Table XXXIIa (continued). Individual Pathology - Trichloroethylene-Treated Male Mice
 

High Dose Group 

WEEKS ON STUDY DISP CODE ANIMAL NUMBER 

TOPOGRAPHY 

TUMORS 

MORPHOLOGY 

OTHER PATHOLOGY 

TOPOGRAPHY MORPHOLOGY 

90 TSAC 014 LUNG 

LIVER HEPATOCELLULAR CARCINOMA METASTA HEPATOCELLULAR CARCINOMA KIDNEY KIDNEY KIDNEY 

HYDRONEPHfCSIS 

INFLAMMATION CHRONIC 

NEPHROSIS TOXIC 

90 TSAC 021 LIVER HEPATOCELLULAR CARCINOMA KIDNEY KIDNEY HYDRONEPHROSIS 

NEPHROSIS TOXIC 

90 TSAC 022 LIVER HEPATOCELLULAR CARCINOMA MESENTERIC LYMPH 

KIDNEY 

NODE INFLAMMATION 

NEPHROSIS TOXIC 

90 TSAC 023 LIVER HEPATOCELLULAR CARCINOMA KIDNEY NEPHROSIS TOXIC 

9C TSAC 024 MESENTERIC LYMPHNODE 

SPLEEN PANCREAS KIDNEY LIVER 

RETICULUM CELL SARCOMA 

RETICULUM CELL SARCOMA RETICULUM CELL SARCOMA RETICULUM CELL SARCOMA RETICULUM CELL SARCOMA 

KIDNEY NEPHROSIS TOXIC 

90 TSAC 025 LIVER HEPATOCELLULAR CARCINOMA KIDNEY NEPHROSIS TOXIC 

90 TSAC 026 LIVER HEPATOCELLULAR CARCINOMA KIDNEY NEPHROSIS TOXIC 

90 TSAC 027 LIVER HEPATOCELLULAR CARCINOMA KIDNEY NEPHROSIS TOXIC 

90 TSAC 028 LIVER HEPATOCELLULAR CARCINOMA KIDNEY NEPHROSIS TOXIC 

90 TSAC 031 LUNG ADENOMA KIDNEY NEPHROSIS TOXIC 

90 TSAC 032 LIVER LUNG 

HEPATOCELLULAR CARCINOMA HEPATOCELLULAR CARCINOMA METASTA KIDNEY MESENTERIC LYMPH 

NEPHROSIS TOXIC 

NODE ANGIECTASIS 

90 TSAC 033 LIVER STOMACH HEPATOCELLULAR CARCINOMA 

PAPILLOMA 

STOMACH 

KIDNEY 

HYPERKERATOSIS 

NEPHROSIS TOXIC 

90 NATD 034 KIDNEY LIVER ABDOMENAL CAVITY ADRENAL PANCREAS 

MESENTERIC LYMPHNODE STOMACH 

FIBROSARCOMA, METASTATIC 

HEPATOCELLULAR CARCINOMA FIBROSARCOMA, PRIMARY FIBROSARCOMA, METASTATIC 

FIBROSARCOMA, METASTATIC 

FIBROSARCOMA, METASTATIC 

FIBROSARCOMA, METASTATIC 

KIDNEY NEPHROSIS TOXIC 

Continued on next page 
_ Table XXXIIa (continued). Individual Pathology - Trichloroethylene-Treated Male Mice 

High Dose Group 

WEEKS ON DISP ANIMAL TUMORS OTHER PATHOLOGY 

STUDY CODE NUMBER 

TOPOGRAPHY MOPPHOLOGY TOPOGRAPHY MORPHOLOGY 

90 TSAC 041 LIVER HEPATOCELLULAR CARCINOMA KIDNEY NEPHROSIS TOXIC
 

90 TSAC 042 LIVER HEPATOCELLULAR CARCINOMA KIDNEY NEPHROSIS ICXIC
 

90 TSAC 043 LIVER HEPATOCELLULAR CARCINOMA KIDNEY NEPHROSIS TOXIC
 

LUNG HEMANGIOSARCOMA
 

end of male mice—high dose 
Table XXXIIb. Individual Pathology - Trichloroethylene-Treated Female Mice
 

Control Group (Vehicle) 

WEEKS ON DISP ANIMAL STUDY CODE NUMBER 

TOPOGRAPHY 

TUMORS 

MORPHOLOGY 

OTH^R

TOPOGRAPHY

 PATHOLOGY 

 MORPHOLOGY 

20 ACCK 010
 NO SIGNIFICANT DIAGNOSIS
 

37 ACCK 009
 ENDOMETRIUM HYPERPLASIA CYSTIC
 

83 ACCK 008 SOFT TISSUES OF OSTEOSARCOMA
 BACK
 

90 TSAC 001
 OVARY	 CYST
 

90 TSAC 002
 LUNG INFLAMMATION CHRONIC
 

UTERUS INFLAMMATION
 OVARY CYST
 OVARY INFLAMMATION
 

90 TSAC 003 ENDOMETRIUM ADENOCARCINOMA
 

90 TSAC 004
 NO SIGNIFICANT DIAGNOSIS
 

90 TSAC
 005	 ENDOMETRIUM HYPERPLASIA CYSTIC
 

OVARY CYST
 

90 TSAC
 006	 ENDOMETRIUM HYPERPLASIA CYSTIC
 

90 TSAC 007 LUNG	 ADENOMA
 

90 TSAC 011	 OVARY CYST
 ENDOMETRIUM HYPERPLASIA CYSTIC
 

90 TSAC 012
 NO SIGNIFICANT DIAGNOSIS
 

90 TSAC 013
 ENDOMETRIUM HYPERPLASIA CYSTIC
 

90 TSAC 014 MESENTERIC LYMPH NODE RETICULUM CELL SARCOMA	 LUNG INFLAMMATION CHRONIC
 

ENDOMETRIUM HYPERPLASIA CYSTIC
 

90 TSAC 015	 OVARY CYST
 

90 TSAC 016
 ENDOMETRIUM HYPERPLASIA CYSTIC
 

90 TSAC 017
 ENDOMETRIUM HYPERPLASIA CYSTIC
 

90 TSAC 018	 OVARY CYST
 ENDOMETRIUM HYPERPLASIA CYSTIC
 

90 TSAC 019
 NO SIGNIFICANT DIAGNOSIS
 

90 TSAC 020
 ENDOMETRIUM HYPERPLASIA CYSTIC
 

end of female mice controls 
Table XXXIIb (continued). Individual Pathology - Trichloroethylene-Treated Female Mice
 

Low Dose Group
 

WEEKS ON DISP ANIMAL TUMORS OTHER PATHOLOGY STUDY CODE NUMBER 

TOPOGRAPHY MORPHOLOGY TOPOGRAPHY MORPHOLOGY 

10 NATD 020 NO SIGNIFICANT DIAGNOSIS 

32 ACCK 010 KIDNEY NEPHROSIS TOXIC 

37 SAC 019 KIDNEY NEPHROSIS TOXIC 

38 NATD 018 NO SIGNIFICANT DIAGNOSIS 

4 1 ACCK 009 KIDNEY NEPHROSIS TOXIC 

55 NATD 030 UTERUS INFLAMMATION 

63 NATD 029 NO SIGNIFICANT DIAGNOSIS 

66 NATD 040 KIDNEY NEPHROSIS TOXIC 

76 NATD 017 LIVER RETICULUM CELL SARCOMA KIDNEY NEPHROSIS TOXIC 

UTERUS RETICULUM CELL SARCOMA CERVICAL LYMPHNODE RETICULUM CELL SARCOMA 

81 NATD 016 SPLEEN LYMPHOSARCOMA KIDNEY NEPHROSIS 1CXIC 

MESENTERIC LYMPHNODE LYMPHOSARCOMA ILEUM LYMPHOSARCOMA 

ADRENAL LYMPHOSARCOMA CERVICAL LYMPHNODE LYMPHOSARCOMA 

90 TSAC 00 1 LIVER RETICULUM CELL SARCOMA KIDNEY NEPHROSIS 1CXIC 

VA3INA RETICULUM CELL SARCOMA SPLEEN RETICULUM CELL SARCOMA 

90 TSAC 002 UTERUS INFLAMMATION KIDNEY NEPHROSIS TOXIC 

90 TSAC 003 LIVER HEPATOCELLULAR CARCINOMA KIDNEY NEPHROSIS TOXIC 

UTERUS FIBROSARCOMA 

90 TSAC 004 ENDOMETRIUM HYPERPLASIA CYSTIC 

KIDNEY NEPHROSIS TOXIC 

90 TSAC 005 KIDNEY NEPHROSIS TCXIC 

90 TSAC 006 ENDOMETRIUM HYPERPLASIA CYSTIC 

KIDNEY NEPHROSIS TOXIC 

Continued on next page 
Table XXXIIb (continued). Individual Pathology - Trichloroethylene-Treated Female Mice
 

Low Dose Group 

WEEKS ON

STUDY

 DISP ANIMAL 

 CODE NUMBER 

TOPOGRAFHY 

TUMORS 

MORPHOLOGY 

OTHER

TOPOGRAPHY

 PATHOLOGY 

 MORPHOLOGY 

90 TSAC 007 LIVER HEPATOCELLULAR CARCINOMA ENDOMETRIUM 

HARDERIAN SLAND 

KIDNEY 

HYPEEFLASIA CYSTIC 

HYPERPLASIA 

NEPHROSIS TOXIC 

90 TSAC 008 STOMACH 

STOMACH 

KIDNEY 

ACANTHOSIS 

HYPERKERA1OSIS 

NEPHROSIS TOXIC 

90 TSAC 011 UTERUS 

KIDNEY 

RETSNTION

NEPHROSIS

 FLUID 

 1OXIC 

90 TSAC 012 LIVER HEPATOCELLULAR CARCINOMA KIDNEY NEPHROSIS TOXIC 

90 TSAC 013 OVARY 

UTERUS 

KIDNEY 

CYST 

RETENTION

NEPHROSIS

 FLUID 

 TOXIC 

90 TSAC 014 UTERUS 

UTERUS 

KIDNEY 

POLYP 

INFLAMMATION 

NEPHROSIS TOXIC 

90 TSAC 015 UTERUS 

KIDNEY 

RETENTION FLUID 

NEPHROSIS TOXIC 

90 TSAC 021 KIDNEY NEPHHOSIS TOXIC 

90 TSAC 022 KIDNEY NEPHROSIS TOXIC 

90 TSAC 023 ENDOMETRIUM 

KIDNEY 

HYPERPLASIA CYSTIC 

NEPHROSIS TOXIC 

90 TSAC 024 £NDOMETRIUM 

KIDNEY 

HYPERPLASIA CYSTIC 

NJSPHROSIS TOXIC 

90 TSAC 025 ENDOMETRIUM 

KIDNEY 

HYPERPLASIA CYSTIC 

NfiPHROSIS TOXIC 

90 TSAC 026 KIDNEY NEPHROSIS TCXIC 

90 TSAC 027 LUNG ALVEOLAR ADENOCARCINOMA KIDNEY 

KIDNEY 

PANCREAS 

KIDNEY 

INFLAMMATICN INTERSTITIAL 

INFLAMMATION PROLIFERATIVfi 

ATROPHY 

NEPHROSIS TOXIC 

Continued on next page 
Table XXXIIb (continued). Individual Pathology - Trichloroethylene-Treated Female Mice
 

Low Dose Group
 

WEEKS ON DISP ANIMAL TUMORS OTHER PATHOLOGY 

STUDY CODE NUMBER 

TOPOGRAPHY MORPHOLOGY TOPOGRAPHY MORPHOLOGY 

90 TSAC 028 ENDOMETRIUM HYPERFLASIA CYSTIC 

KIDNEY NEPHROSIS TOXIC 

90 TSAC 031 KIDNEY N£PHfiOSIS TCXIC 

90 TSAC 032 LIVER HEPATOCELLULAR CARCINOMA UTERUS RETENTION J1UIC 

KIDNEY NEPHROSIS TOXIC 

90 TSAC 033 OVARY GRANULOSA CELL CARCINOMA UTERUS INFLAMMATION 

KIDNEY NEPHfiOSIS TOXIC 

90 TSAC 034 ENDOMETRIUM HYPSRPLASIA CYSTIC 

OVARY CYST 

KIDNEY NEPHROSIS TOXIC 

90 TSAC 035 SUBCUT TISSU/ABDOMEN PIBROSARCOMA ENDOMETRIUM HYPERPLASIA CYSTIC 

KIDNEY NEPHROSIS TOXIC 

90 TSAC 036 LUNG ADENOMA KIDNEY NEPHfiOSIS TOXIC 

SPLEEN LYMPHOSARCOMA 

90 TSAC 037 SNDOMETRIUM HYPSRPLASIA CYSTIC 

KIDNEY NEPHROSIS TOXIC 

90 TSAC 038 MAMMARY GLAND ADENOCARCINOMA OVARY CYST 

KIDNEY NSPHROSIS TOXIC 

90 TSAC 039 KIDNEY NEPHROSIS TCXIC 

90 TSAC 041 KIDNEY NEPHROSIS TCXIC 

90 TSAC 042 LUNG ADENOMA OVARY RETENTION ILUID 

THYMUS LYMPHOSAECOMA KIDNEY NEPHROSIS TOXIC 

90 TSAC 043 OVARY CYSTADENOMA THYROID INFLAMMATION CYSTIC 

LUNG ALVEOLAE ADENOCARCINOMA KIDNEY NEPHROSIS TOXIC 

HARDERIAN GLAND ADENOMA 

90 TSAC 044 iSNDOMETRIUM HYPERPLASIA CYSTIC 

THYROID INFLAMMATION CYSTIC 

KIDNEY NEPHROSIS TOXIC 

90 TSAC 045 UTERUS RETENTION FLUID 

RIGHT OVARY CYST 

KIDNEY NEPHROSIS TOXIC 

Continued on next page 
Table XXXIIb (continued). Individual Pathology - Trichloroethylene-Treated Female Mice 

Low Dose Group 

WEEKS ON STUDY DISP ANIMAL CODE NUMBER 

TOPOGRAPHY

 TUMORS

 MORPHOLOGY

 OTHER PATHOLOGY 

 TOPOGRAPHY MORPHOLOGY 

TSAC 046 KIDNEY NEPHROSIS TCXIC 

91 TSAC 047 ENDOMETRIUM KIDNEY 

HYPERPLASIA CYSTIC 

NEPHROSIS TOXIC 

91 TSAC 048 UTERUS KIDNEY RETENTION ELUIC 

NEPHROSIS TOXIC 

91 TSAC 049 ENDOMETRIUM KIDNEY 

HYPERPLASIA CYSTIC 

NEPHROSIS TOXIC 

91 TSAC 050 UTERUS KIDNEY RETENTION fLUID NEPHROSIS TOXIC 

end of female mice—lew dose 
Table XXXIIb (continued). Individual Pathology - Trichloroethylene-Treated Female Mice
 

WEEKS ON 

STUDY 

14
 

22
 

26
 

32
 

32
 

33
 

38
 

39
 

UO
 

69
 

88
 

91
 

91
 

91
 

91
 

91
 

91
 

91
 

91
 

DISP ANIMAL
 

CODE NUMBER
 

MISS
 

MISS
 

MISS
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

NATD
 

TSAC
 

TSAC
 

TSAC
 

TSAC
 

TSAC
 

TSAC
 

TSAC
 

TSAC
 

010
 

040
 

050
 

030
 

049
 

039
 

038
 

020
 

037
 

009
 

036
 

001
 

002
 

003
 

004
 

005
 

006
 

007
 

008
 

High Dose Group
 

TUMORS 

TOPOGRAPHY MORPHOLOGY 

SPLEEN LYMPHOSARCOMA
 

SPLEEN RETICULUM CELL SARCOMA
 KIDNEY RETICULUM CELL SARCOMA
 LUNG RETICULUM CELL SARCOMA
 STOMACH RETICULUM CELL SARCOMA
 

MESENTERIC LYMPH NODE RETICULUM CELL SARCOMA
 ILEUM RETICULUM CELL SARCOMA
 

LIVER HEPATOCELLULAR CARCINOMA
 

LUNG ALVEOLAR ADENOCARCINOMA
 LIVER HEPATOCELLULAR CARCINOMA
 

OTHER

TOPOGRAPHY 

KIDNEY
 

KIDNEY
 

KIDNEY
 

KIDNEY
 

KIDNEY
 

KIDNEY
 

KIDNEY
 

OVARY
 

KIDNEY
 

KIDNEY
 

LIVER
 OVARY
 

KIDNEY
 

KIDNEY
 UTERUS
 

KIDNEY
 

ENDOMETRIUM
 

KIDNEY
 

KIDNEY
 

KIDNEY
 

 PATHOLOGY 

MORPHOLOGY 

ANIMAL MISSING
 

ANIMAL MISSING
 

ANIMAL MISSING
 

NEPHROSIS 1CXIC
 

NEPdRCSIS
 

NEPHROSIS 1OXIC
 

NEPHROSIS TOXIC
 

NEPHROSIS TOXIC
 

N£PHROSIS TOXIC
 

NEPHROSIS 1OXIC
 

CYST
 

NEPHROSIS TOXIC
 

NEPHROSIS IOXIC
 

HYPERPLASIA
 CYST
 

NEPHROSIS TOXIC
 

NEPHROSIS TOXIC
 

RETENTION FLUID
 

NEPHROSIS TOXIC
 

HYPERPLASIA CYSTIC
 

NEPHROSIS TOXIC
 

NEPHROSIS TCXIC
 

NEPHROSIS TOXIC
 

Continued on next page 
Table XXXIIb (continued). Individual Pathology - Trichloroethylene-Treated Female Mice 

WEEKS ON
 

STUDY
 

91
 

91
 

91
 

91
 

91
 

91
 

91
 

91
 

91
 

91
 

91
 

91
 

91
 

91
 

91
 

91
 

DISP ANIMAL
 

CODE NUMBER
 

TSAC
 

TSA.C
 

TSAC
 

TSAC
 

TSAC
 

TSAC
 

TSAC
 

TSAC
 

TSAC
 

TSAC
 

TSAC
 

TSAC
 

TSAC
 

TSAC
 

TSAC
 

TSAC
 

011
 

012
 

013
 

014
 

015
 

016
 

017
 

018
 

019
 

021
 

022
 

023
 

024
 

025
 

026
 

027
 

TUMORS 

TOPOGRAPHY 

LIVER
 

LIVER
 

LUNG
 

CERVICAL LYMPH NODE
 

LIVER
 

KIDNEY
 

SPLEEN
 

LUNG
 

LIVER
 

LUNG
 

LIVER
 

LIVER
 

High Dose Group 

MORPHOLOGY 

HEFATOCELLULAR CARCINOMA
 

HEPATOCELLULAR CARCINOMA
 

ADENOMA
 

MALIGNANT LYMPHOMA
 

LYMPHOSARCOMA
 

LYMPHOSARCOMA
 

LYMPHOSARCOMA
 

ADENOMA
 

HEPATOCELLULAR CARCINOMA
 

ALVEOLAR ADENOCARCINOMA
 

HEPATOCELLULAR CARCINOMA
 

HEPATOCELLULAR CARCINOMA
 

OTHER PATHOLOGY 

TOPOGRAPHY MORPHOLOGY 

KIDNEY NEPHROSIS 1CXIC
 

KIDNEY NEPHROSIS TOXIC
 

UTERUS RETENTION FLUID
 

KIDNEY NEPHRCSIS TOXIC
 

OVARY CYST
 

KIDNEY NEPHROSIS TCXIC
 

ENDOMETRIUM INFLAMMATION
 

KIDNEY NEPHROSIS TOXIC
 

KIDNEY NEPHRCSIS TOXIC
 

MESENTERIC LYMPH NODE ANGIECTASIS
 

UTERUS RETENTION FLUID
 

KIDNEY NEPHROSIS TOXIC
 

KIDNEY NEPHRCSIS TOXIC
 

KIDNEY NEPHROSIS TCXIC
 

KIDNEY NEPHROSIS TCXIC
 

UTERUS RETENTION FLUID
 

OVARY CYST
 

UTERUS RETENTION FLUID
 

KIDNEY NEPHROSIS TOXIC
 

KIDNEY NEPHROSIS TCXIC
 

LIVER HYPERPLASIA
 

BRONCHUS BRONCHIECTASIS
 

KIDNEY NEPHROSIS TOXIC
 

KIDNEY NEPHROSIS TOXIC
 

OVARY CYST
 

KIDNEY NEPHROSIS TOXIC
 

KIDNEY NEPHROSIS TOXIC
 

Continued on next page 
Table XXXIIb (continued). Individual Pathology - Trichloroethylene-Treated Female Mice
 

High Dose Group
 

WEEKS ON DISP ANIMAL TUMORS OTHER PATHOLOGY STUDY CODE NUMBER 

TOPOGRAPHY MORPHOLOGY TOPOGRAPHY MORPHOLOGY 

91 TSAC 028 KIDNEY NEPHROSIS TOXIC 

ENDOMETRIUM HYPERPLASIA CYSTIC 

OVARY CYST 

91 TSAC 029 KIDNEY NEPHROSIS TOXIC 

OVARY CYST 

91 TSAC 031 KIDNEY NEPHROSIS TOXIC 

UTERUS RETENTION fLUID 

91 TSAC 032 KIDNEY NEPHROSIS TOXIC 

91 TSAC 033 KIDNEY NEPHROSIS TOXIC 

91 TSAC 03a LUNG ADENOMA KIDNEY NEPHROSIS TOXIC 

91 TSAC 035 KIDNEY NEPHROSIS TOXIC 

91 TSAC 041 LUNG ADENOMA KIDNEY NEPHROSIS TOXIC 

MESENTERIC LYMPHNODE LYMPHOSARCOMA SPLEEN LYMPHOSARCOMA 

CERVICAL LYMPHNODE LYMPHOSARCOMA LIVER HEPATOCELLULAR CARCINOMA 

91 TSAC 042 LIVER HEPATOCELLULAR CARCINOMA KIDNEY NEPHRCSIS TOXIC 

91 TSAC 043 KIDNEY NEPHROSIS TOXIC 

91 TSAC 044 LIVER HEPATOCELLULAR CARCINOMA KIDNEY NEPHROSIS TOXIC 

UTERUS RETENTION FLUID 

91 TSAC 045 KIDNEY NEPHROSIS TOXIC 

UTERUS RETENTION FLUID 

91 TSAC 046 LIVER HEPATOCELLULAR CARCINOMA KIDNEY NEPHRCSIS TCXIC 

LUNG ADENOMA 

91 TSAC 047 KIDNEY NEPHROSIS TOXIC 

MESENTERIC LYMPH NODE ANGIECTASIS 

STOMACH ACANTHOSIS 

STOMACH HYPERKERATOSIS 

OVARY CYST 

91 TSAC 048 KIDNEY NEPHROSIS 

OVARY CYST 

end of female mice—high dose 

APPENDIX E: POSITIVE CONTROLS
 

177
 
Table XXXIIIa. Liver Histopathology for Positive Controls
 

(Carbon Tetrachloride) - Low Dose Male Rats
 

Individual
 

Number
 

001
 

002
 

003
 

004
 

005
 

006
 

007
 

008
 

009
 

010
 

011
 

012
 

013
 

014
 

015
 

016
 

017
 

Week of
 

Death
 

82
 

110
 

103
 

82
 

110
 

110
 

13
 

106
 

65
 

95
 

102
 

62
 

73
 

44
 

99
 

107
 

66
 

Diagnosis
 

Portal cirrhosis
 

Cholangietasis
 

Fatty
 

Bile duct proliferation
 

Advanced autolysis
 

Hepatocellular carcinoma
 

Portal cirrhosis
 

Bile duct proliferation
 

Fatty
 

Pigment deposition
 

Peliosis
 

Bile duct proliferation
 

Fibrosis (around bile ducts)
 

Centrilobular necrosis
 

Fatty
 

Portal cirrhosis
 

Cholangiectasis
 

Fatty
 

Bile duct proliferation
 

Portal cirrhosis
 

Fatty
 

Bile duct proliferation
 

Cholangiectasis
 

Fatty
 

Portal cirrhosis
 

Fatty
 

Bile duct proliferation
 

Portal cirrhosis
 

Fatty
 

Bile duct proliferation
 

Fatty (diffuse)
 

Periportal necrosis
 

Degeneration
 

Fatty
 

Fibrosis
 

Periportal degeneration
 

Fibrosis
 

Bile duct proliferation
 

Portal cirrhosis
 

Pigment deposition
 

Portal cirrhosis
 

Bile duct proliferation
 

Fatty
 

Neoplastic nodule
 

178
 
Table XXXIIIa. Liver Histopathology for Positive Controls
 

(Carbon Tetrachloride) - Low Dose Male Rats
 

(continued)
 

018
 

019
 

020
 

021
 

022
 

023
 

024
 

025
 

026
 

027
 

028
 

029
 

030
 

031
 

032
 

033
 

110
 

94
 

110
 

105
 

110
 

106
 

110
 

107
 

110
 

63
 

75
 

110
 

90
 

110
 

60
 

73
 

Portal cirrhosis
 

Bile duct proliferation
 

Fatty
 

Portal cirrhosis
 

Bile duct proliferation
 

Fatty
 

Bile duct proliferation
 

Angiectasis
 

Fatty
 

Regenerative nodules
 

Portal cirrhosis
 

Bile duct proliferation
 

Fatty
 

Angiectasis
 

Bile duct proliferation
 

Fatty
 

Fibrosis
 

Advanced autolysis
 

Portal cirrhosis
 

Fatty
 

None
 

Portal cirrhosis
 

Bile duct proliferation
 

Regenerative nodules
 

Regenerative nodules
 

Fatty
 

Angiectasis
 

Portal cirrhosis
 

Bile duct proliferation
 

Fibrosis
 

Cholangiectasis
 

Bile duct proliferation
 

Fatty
 

Fibrosis
 

Myelogenous leukemia
 

Portal cirrhosis
 

Bile duct proliferation
 

Regenerative nodules
 

Portal cirrhosis
 

Bile duct proliferation
 

Fatty
 

Organizing thrombus
 

Hepatocellular carcinoma
 

Regenerative nodules
 

Portal cirrhosis
 

Fatty
 

Bile duct proliferation
 

179
 
Table XXXIlla. Liver Histopathology for Positive Controls
 

(continued)
 

034
 

035
 

036
 

037
 

038
 

039
 

040
 

041
 

042
 

043
 

044
 

045
 

046
 

047
 

048
 

049
 

050
 

(Carbon Tetrachloride) - Low Dose Male Rats
 

71 Portal cirrhosis 

Bile duct proliferation 

Fatty 

Regenerative nodules 

71 Portal cirrhosis 

Bile duct proliferation 

Regenerative nodules 

82 Portal cirrhosis 

Bile duct proliferation 

110 Fatty 

101 Portal cirrhosis 

Hepatic abscess 

108 Portal cirrhosis 

Bile duct proliferation 

109 Bile duct proliferation 

Fibrosis 

Fatty 

Regenerative nodules 

58 Portal cirrhosis 

Bile duct proliferation 

30 Portal cirrhosis 

Bile duct proliferation 

Fatty 

110 Portal cirrhosis 

Bile duct proliferation 

Fatty 

Regenerative nodules 

69 Portal cirrhosis 

Bile duct proliferation 

Fatty 

110 Neoplastic nodule 

110 Portal cirrhosis 

Bile duct proliferation 

Regenerative nodules 

109 Portal cirrhosis 

Bile duct proliferation 

104 Portal cirrhosis 

Bile duct proliferation 

36 Reticulum cell sarcoma (multicentric) 

110 Portal cirrhosis 

Bile duct proliferation 

Foci of altered cells 

Regenerative nodules 

180
 
Table XXXIIIb. Liver Histopathology for Positive Controls
 

(Carbon Tetrachloride) - High Dose Male Rats
 

Individual
 

Number
 

001
 

002
 

003
 

004
 

005
 

006
 

007
 

008
 

009
 

010
 

011
 

012
 

013
 

014
 

015
 

016
 

017
 

018
 

Week of
 

Death
 

107
 

110
 

100
 

97
 

80
 

96
 

96
 

109
 

91
 

77
 

55
 

58
 

64
 

81
 

78
 

61
 

110
 

68
 

Diagnosis
 

Portal cirrhosis
 

Bile duct proliferation
 

Fatty
 

Portal cirrhosis
 

Bile duct proliferation
 

Advanced autolysis
 

Portal cirrhosis
 

Bile duct proliferation
 

Hepatocellular carcinoma
 

Regenerative nodules
 

Advanced autolysis
 

Bile duct proliferation
 

Portal cirrhosis
 

Portal cirrhosis
 

Bile duct proliferation
 

Regenerative nodules
 

Portal cirrhosis
 

Bile duct proliferation
 

Hepatitis
 

Portal cirrhosis
 

Bile duct proliferation
 

Fatty
 

Portal cirrhosis
 

Fatty
 

Portal cirrhosis
 

Bile duct proliferation
 

Advanced autolysis
 

Regenerative nodules
 

Portal cirrhosis
 

Bile duct proliferation
 

Fatty
 

Portal cirrhosis
 

Bile duct proliferation
 

Fatty
 

Portal cirrhosis
 

Bile duct proliferation
 

Portal cirrhosis
 

Bile duct proliferation
 

Fatty
 

Neoplastic nodule
 

181
 
Table XXXlIlb. Liver Histopathology for Positive Controls
 

(Carbon Tetrachloride) - High Dose Male Rats
 

(continued)
 

019
 

020
 

021
 

022
 

023
 

024
 

025
 

026
 

027
 

028
 

029
 

030
 

031
 

032
 

033
 

034
 

035
 

036
 

037
 

40
 

97
 

110
 

110
 

77
 

55
 

104
 

79
 

90
 

100
 

97
 

65
 

43
 

109
 

98
 

93
 

88
 

110
 

90
 

Portal cirrhosis
 

Fatty
 

Portal cirrhosis
 

Bile duct proliferation
 

Cholangiectasis
 

Portal cirrhosis
 

Bile duct proliferation
 

Regenerative nodules
 

Portal cirrhosis
 

Bile duct proliferation
 

Portal cirrhosis
 

Bile duct proliferation
 

Portal cirrhosis
 

Fatty
 

Bile duct proliferation
 

Lymphocytic leukemia
 

Hepatocellular carcinoma
 

Portal cirrhosis
 

Bile duct proliferation
 

Portal cirrhosis
 

Bile duct proliferation
 

Portal cirrhosis
 

Bile duct proliferation
 

Organizing thrombus
 

Portal cirrhosis
 

Bile duct proliferation
 

Fatty
 

Portal cirrhosis
 

Fatty
 

Bile duct proliferation
 

Advanced autolysis
 

Portal cirrhosis
 

Bile duct proliferation
 

Portal cirrhosis
 

Bile duct proliferation
 

Portal cirrhosis
 

Bile duct proliferation
 

Portal cirrhosis
 

Fatty
 

Bile duct proliferation
 

Foci of altered cells
 

Regenerative nodules
 

Portal cirrhosis
 

Bile duct proliferation
 

Portal cirrhosis
 

Bile duct proliferation
 

182
 
Table XXXIIIb. Liver Histopathology for Positive Controls
 

(Carbon Tetrachloride) - High Dose Male Rats
 

(continued)
 

038
 

039
 

040
 

041
 

042
 

043
 

044
 

045
 

046
 

047
 

048
 

049
 

050
 

14
 

100
 

83
 

109
 

73
 

77
 

110
 

107
 

96
 

98
 

110
 

54
 

85
 

Fat deposition, diffuse
 

Portal cirrhosis
 

Bile duct proliferation
 

Portal cirrhosis
 

Bile duct proliferation
 

Advanced autolysis
 

Portal cirrhosis
 

Regenerative nodules
 

Portal cirrhosis
 

Bile duct proliferation
 

Fatty
 

Portal cirrhosis
 

Bile duct proliferation
 

Fatty
 

Portal cirrhosis
 

Bile duct proliferation
 

Portal cirrhosis
 

Bile duct proliferation
 

Portal cirrhosis
 

Bile duct proliferation
 

Portal cirrhosis
 

Bile duct proliferation
 

Portal cirrhosis
 

Bile duct proliferation
 

Regenerative nodules
 

Portal cirrhosis
 

Fatty
 

Portal cirrhosis
 

Bile duct proliferation
 

183
 
Table XXXIIIc. Liver Histopathology for Positive Controls
 

(Carbon Tetrachloride) - Low Dose Female Rats
 

Individual
 

Number
 

001
 

002
 

003
 

004
 

005
 

006
 

007
 

008
 

009
 

010
 

011
 

012
 

013
 

014
 

015
 

016
 

017
 

018
 

019
 

020
 

Week of
 

Death
 

100
 

110
 

88
 

110
 

71
 

96
 

105
 

61
 

110
 

110
 

101
 

110
 

104
 

110
 

86
 

107
 

110
 

110
 

57
 

110
 

Diagnosis
 

Portal cirrhosis
 

Bile duct proliferation
 

Neoplastic nodule
 

Regenerative nodules
 

Bile duct proliferation
 

Foci of altered cells
 

Regenerative nodules
 

Fatty
 

Portal cirrhosis
 

Bile duct proliferation
 

Portal cirrhosis
 

Bile duct proliferation
 

Fatty
 

Portal cirrhosis
 

Bile duct proliferation
 

Portal cirrhosis
 

Bile duct proliferation
 

Fatty
 

Foci of altered cells
 

Regenerative nodules
 

Portal cirrhosis
 

Regenerative nodules
 

Reticulum cell sarcoma
 

Portal cirrhosis
 

Fatty
 

Portal cirrhosis
 

Bile duct proliferation
 

Fatty
 

Neoplastic nodule
 

Hepatocellular carcinoma
 

Portal cirrhosis
 

Fatty
 

Portal cirrhosis
 

Bile duct proliferation
 

Portal cirrhosis
 

Bile duct proliferation
 

Regenerative nodules
 

Portal cirrhosis
 

Fatty
 

Bile duct proliferation
 

Portal cirrhosis
 

Fatty
 

Portal cirrhosis
 

Regenerative nodules
 

184
 
Table XXXIIIc. Liver Histopathology for Positive Controls
 

(Carbon Tetrachloride) - Low Dose Female Rats
 

(continued)
 

021
 

022
 

023
 

024
 

025
 

026
 

027
 

028
 

029
 

030
 

031
 

032
 

033
 

034
 

035
 

036
 

94
 

110
 

100
 

88
 

110
 

110
 

76
 

110
 

88
 

110
 

41
 

93
 

13
 

94
 

73
 

91
 

Portal cirrhosis
 

Bile duct proliferation
 

Fatty
 

Portal cirrhosis
 

Bile duct proliferation
 

Regenerative nodules
 

Portal cirrhosis
 

Organizing thrombus
 

Pigment deposition
 

Bile duct proliferation
 

Portal cirrhosis
 

Regenerative nodules
 

Portal cirrhosis
 

Angiectasis
 

Hepatocellular carcinoma
 

Bile duct proliferation
 

Portal cirrhosis
 

Foci of altered cells
 

Regenerative nodules
 

Portal cirrhosis
 

Fatty
 

Bile duct proliferation
 

Hepatitis
 

Regenerative nodules
 

Fatty
 

Bile duct proliferation
 

Regenerative nodules
 

Portal cirrhosis
 

Bile duct proliferation
 

Fatty
 

Portal cirrhosis
 

Bile duct proliferation
 

Regenerative nodules
 

Portal cirrhosis
 

Bile duct proliferation
 

Advanced autolysis
 

Portal cirrhosis
 

Bile duct proliferation
 

Advanced autolysis
 

Portal cirrhosis
 

Fatty
 

Portal cirrhosis
 

Bile duct proliferation
 

Fatty
 

Portal cirrhosis
 

Bile duct proliferation
 

Angiectasis
 

185
 
Table XXXIIIc. Liver Histopathology for Positive Controls
 

(Carbon Tetrachloride) - Low Dose Female Rats
 

(continued)
 

037
 

038
 

039
 

040
 

041
 

042
 

043
 

044
 

045
 

046
 

047
 

048
 

049
 

050
 

110
 

79
 

104
 

74
 

12
 

104
 

110
 

110
 

110
 

58
 

39
 

48
 

110
 

110
 

Portal cirrhosis
 

Bile duct proliferation
 

Advanced autolysis
 

Portal cirrhosis
 

Fatty
 

Advanced autolysis
 

Portal cirrhosis
 

Bile duct proliferation
 

Fatty
 

Portal cirrhosis
 

Fatty
 

Bile duct proliferation
 

Portal cirrhosis
 

Fatty
 

Portal cirrhosis
 

Bile duct proliferation
 

Hepatocellular carcinoma
 

Portal cirrhosis
 

Fatty
 

Bile duct proliferation
 

Regenerative nodules
 

Portal cirrhosis
 

Fatty
 

Portal cirrhosis
 

Fatty
 

Bile duct proliferation
 

Portal cirrhosis
 

Fatty
 

Portal cirrhosis
 

Bile duct proliferation
 

Portal cirrhosis
 

Fatty
 

Neoplastic nodule
 

Hepatocellular carcinoma
 

Bile duct proliferation
 

Fatty
 

Portal cirrhosis
 

Bile duct proliferation
 

186
 
Table XXXIIId. Liver Histopathology for Positive Controls
 

(Carbon Tetrachloride) - High Dose Female Rats
 

Individual
 

Number
 

001
 

002
 

003
 

004
 

005
 

006
 

007
 

008
 

009
 

010
 

011
 

012
 

013
 

014
 

015
 

016
 

017
 

018
 

Week of
 

Death
 

110
 

110
 

16
 

71
 

66
 

110
 

38
 

103
 

110
 

110
 

110
 

82
 

110
 

110
 

110
 

12
 

1
 

48
 

Diagnosis
 

Portal cirrhosis
 

Bile duct proliferation
 

Fatty
 

Portal cirrhosis
 

Fatty
 

Regenerative nodules
 

Portal cirrhosis
 

Portal cirrhosis
 

Fatty
 

Portal cirrhosis
 

Fatty
 

Bile duct proliferation
 

Portal cirrhosis
 

Bile duct proliferation
 

Fatty
 

Regenerative nodules
 

Portal cirrhosis
 

Fatty
 

Portal cirrhosis
 

Bile duct proliferation
 

Portal cirrhosis
 

Fatty
 

Portal cirrhosis
 

Bile duct proliferation
 

Fatty
 

Regenerative nodules
 

Portal cirrhosis
 

Bile duct proliferation
 

Regenerative nodules
 

Advanced autolysis
 

Portal cirrhosis
 

Bile duct proliferation
 

Portal cirrhosis
 

Bile duct proliferation
 

Portal cirrhosis
 

Bile duct proliferation
 

Neoplastic nodule
 

Neoplastic nodule
 

Portal cirrhosis
 

Fatty
 

Bile duct proliferation
 

Portal cirrhosis
 

Angiectasis
 

Portal cirrhosis
 

Bile duct proliferation
 

187
 
Table XXXIIId. Liver Histopathology for Positive Controls
 

(Carbon Tetrachloride) - High Dose Female Rats
 

(continued)
 

019
 

020
 

021
 

022
 

023
 

024
 

025
 

026
 

027
 

028
 

029
 

030
 

031
 

032
 

033
 

034
 

035
 

036
 

037
 

038
 

039
 

80
 

62
 

52
 

13
 

4
 

30
 

19
 

69
 

110
 

34
 

76
 

34
 

80
 

70
 

4
 

110
 

110
 

33
 

110
 

99
 

12
 

Portal cirrhosis
 

Bile duct proliferation
 

Portal cirrhosis
 

Bile duct proliferation
 

Portal cirrhosis
 

Fatty
 

Bile duct proliferation
 

Regenerative nodules
 

Portal cirrhosis
 

Fatty
 

Portal cirrhosis
 

Fatty
 

Portal cirrhosis
 

Fatty
 

Portal cirrhosis
 

Fatty
 

Portal cirrhosis
 

Bile duct proliferation
 

Hepatitis
 

Portal cirrhosis
 

Bile duct proliferation
 

Neoplastic nodule
 

Portal cirrhosis
 

Bile duct proliferation
 

Fatty
 

Portal cirrhosis
 

Foci of altered cells
 

Regenerative nodules
 

Lost
 

Portal cirrhosis
 

Bile duct proliferation
 

Regenerative nodules
 

Portal cirrhosis
 

Bile duct proliferation
 

Fatty
 

Regenerative nodules
 

Portal cirrhosis
 

Fatty
 

Regenerative nodules
 

Regenerative nodules
 

Portal cirrhosis
 

Bile duct proliferation
 

Regenerative nodules
 

Advanced autolysis
 

Portal cirrhosis
 

Bile duct proliferation
 

Portal cirrhosis (early)
 

Fatty
 

188
 
Table XXXIIId. Liver Histopathology for Positive Controls
 

(Carbon Tetrachloride) - High Dose Female Rats
 

(continued)
 

040
 

041
 

042
 

043
 

044
 

045
 

046
 

047
 

048
 

049
 

050
 

95
 

104
 

9
 

53
 

29
 

15
 

44
 

1
 

50
 

3
 

110
 

Portal cirrhosis
 

Bile duct proliferation
 

Hepatocellular carcinoma
 

Portal cirrhosis
 

Fatty
 

Bile duct proliferation
 

Fatty
 

Sinusoidal ectasia
 

Portal cirrhosis
 

Bile duct proliferation
 

Portal cirrhosis
 

Bile duct proliferation
 

Portal cirrhosis
 

Bile duct proliferation
 

Fatty
 

Regenerative nodules
 

Fatty
 

Portal cirrhosis
 

Bile duct proliferation
 

Fatty
 

Portal cirrhosis
 

Bile duct proliferation
 

Regenerative nodules
 

189
 
Table XXXIVa. Liver Histopathology for Positive Controls
 

(Carbon Tetrachloride) - Low Dose Male Mice
 

Individual
 

Number
 

001
 

002
 

003
 

004
 

005
 

006
 

007
 

008
 

009
 

010
 

011
 

012
 

013
 

014
 

015
 

016
 

017
 

018
 

019
 

020
 

021
 

022
 

023
 

024
 

025
 

026
 

027
 

028
 

029
 

030
 

031
 

032
 

033
 

034
 

035
 

036
 

037
 

038
 

039
 

040
 

Week of
 

Death
 

80
 

74
 

72
 

72
 

72
 

66
 

65
 

63
 

50
 

48
 

80
 

75
 

74
 

74
 

73
 

67
 

67
 

65
 

63
 

55
 

82
 

81
 

79
 

76
 

76
 

72
 

70
 

64
 

61
 

60
 

86
 

82
 

81
 

80
 

75
 

74
 

71
 

64
 

54
 

42
 

Diagnosis
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Autolysis
 

190
 
Table XXXIVa. Liver Histopathology for Positive Controls
 

(Carbon Tetrachloride) - Low Dose Male Mice
 

(continued)
 

041
 

042
 

043
 

044
 

045
 

046
 

047
 

048
 

049
 

050
 

84


80


77


77


75


72


72


69


64


50


 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

191
 
Table XXXIVb. Liver Histopathology for Positive Controls
 

(Carbon Tetrachloride) - High Dose Male Mice
 

Individual
 

Number
 

001
 

002
 

003
 

004
 

005
 

006
 

007
 

008
 

009
 

010
 

on
 

012
 

013
 

014
 

015
 

016
 

017
 

018
 

019
 

020
 

021
 

022
 

023
 

024
 

025
 

026
 

027
 

028
 

0 29
 

030
 

031
 

032
 

033
 

034
 

035
 

036
 

037
 

038
 

039
 

040
 

Week of
 

Death
 

75
 

75
 

75
 

66
 

63
 

63
 

60
 

60
 

55
 

52
 

90
 

79
 

77
 

74
 

69
 

64
 

56
 

53
 

42
 

30
 

77
 

74
 

66
 

65
 

62
 

56
 

48
 

48
 

48
 

26
 

74
 

74
 

70
 

65
 

63
 

60
 

51
 

51
 

47
 

47
 

Diagnosis
 

Hepatocellular


Hepatocellular


Hepatocellular


Hepatocellular


Hepatocellular


Hepatocellular


Hepatocellular


Hepatocellular


Hepatocellular


Hepatocellular


Hepatocellular


Hepatocellular


Hepatocellular


Hepatocellular


Hepatocellular


Hepatocellular


Hepatocellular


Hepatocellular


Hepatocellular


Hepatocellular


Hepatocellular


Hepatocellular


Hepatocellular


Hepatocellular


Hepatocellular


Cannibalized
 

Hepatocellular


Autolysis
 

Hepatocellular


Hepatocellular


Hepatocellular


Hepatocellular


Hepatocellular


Hepatocellular


Hepatocellular


Hepatocellular


Hepatocellular


Hepatocellular


Hepatocellular


Hepatocellular


 carcinoma
 

 carcinoma
 

 carcinoma
 

 carcinoma
 

 carcinoma
 

 carcinoma
 

 carcinoma
 

 carcinoma
 

 carcinoma
 

 carcinoma
 

 carcinoma
 

 carcinoma
 

 carcinoma
 

 carcinoma
 

 carcinoma
 

 carcinoma
 

 carcinoma
 

 carcinoma
 

 carcinoma
 

 carcinoma
 

 carcinoma
 

 carcinoma
 

 carcinoma
 

 carcinoma
 

 carcinoma
 

 carcinoma
 

 carcinoma
 

 carcinoma
 

 carcinoma
 

 carcinoma
 

 carcinoma
 

 carcinoma
 

 carcinoma
 

 carcinoma
 

 carcinoma
 

 carcinoma
 

 carcinoma
 

 carcinoma
 

192
 
Table XXXIVb. Liver Histopathology for Positive Controls
 

(Carbon Tetrachloride) - High Dose Male Mice
 

(continued)
 

041
 

042
 

043
 

044
 

045
 

046
 

047
 

048
 

049
 

050
 

73


73


71


70


70


58


56


35


26


16


 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Portal cirrhosis
 

Bile duct proliferation
 

193
 
Table XXXIVc. Liver Histopathology for Positive Controls
 

(Carbon Tetrachloride) - Low Dose Female Mice
 

Individual
 

Number
 

001
 

002
 

003
 

004
 

005
 

006
 

007
 

008
 

009
 

010
 

on
 

012
 

013
 

014
 

015
 

016
 

017
 

018
 

019
 

020
 

021
 

022
 

023
 

024
 

025
 

026
 

027
 

028
 

029
 

030
 

031
 

032
 

033
 

034
 

035
 

036
 

037
 

038
 

039
 

040
 

Week of
 

Death
 

68
 

77
 

74
 

74
 

70
 

68
 

62
 

62
 

58
 

45
 

80
 

80
 

79
 

75
 

70
 

68
 

65
 

64
 

36
 

29
 

81
 

80
 

75
 

70
 

66
 

55
 

48
 

46
 

33
 

11
 

86
 

84
 

79
 

78
 

75
 

66
 

66
 

61
 

54
 

30
 

Diagnosis
 

Hepatocellular carcinoma
 

Organizing thrombus
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Cannibalized
 

Hepatocellular carcinoma
 

Cannibalized
 

Autolysis
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Organizing thrombus
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Autolysis
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Autolysis
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Autolysis
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

Hepatocellular carcinoma
 

194
 
Table XXXIVc. Liver Histopathology for Positive Controls
 

(Carbon Tetrachloride) - Low Dose Female Mice
 

(continued)
 

041
 

042
 

043
 

044
 

045
 

046
 

047
 

048
 

049
 

050
 

81


79


78


72


71


71


69


64


55


16


 Hepatocellular carcinoma
 

 Cannibalized
 

 Cannibalized
 

 Cannibalized
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Cannibalized
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

195
 
Table XXXIVd. Liver Histopathology for Positive Controls
 

(Carbon Tetrachloride) - High Dose Female Mice
 

Individual
 

Number
 

001
 

002
 

003
 

004
 

005
 

006
 

007
 

008
 

009
 

010
 

011
 

012
 

013
 

014
 

015
 

016
 

017
 

018
 

019
 

020
 

021
 

022
 

023
 

024
 

025
 

026
 

027
 

028
 

0 29
 

030
 

031
 

032
 

033
 

034
 

035
 

036
 

037
 

038
 

039
 

040
 

Week of
 

Death


92


68


61


61


58


51


34


19


19


14


74


66


63


60


57


52


51


47


43


42


78


73


71


70


67


67


67


58


54


13


80


74


68


59


57


57


56


41


38


35


 Diagnosis
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Lost
 

 Hepatocellular carcinoma
 

 Cannibalized
 

 Toxic hepatitis
 

Cirrhosis
 

Bile duct proliferation
 

Fatty
 

 Hepatocellular carcinoma
 

r carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Cannibalized
 

 Autolysis
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Portal cirrhosis
 

Bile duct proliferation
 

 Hepatocellular carcinoma
 

r a
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

196
 
Table XXXIVd. Liver Histopathology for Positive Controls
 

(Carbon Tetrachloride) - High Dose Female Mice
 

(continued)
 

041
 

042
 

043
 

044
 

045
 

046
 

047
 

048
 

049
 

050
 

80


78


77


73


67


64


46


43


42


36


 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Cannibalized
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

 Hepatocellular carcinoma
 

197
 
198
 


DHEW Publication No. (NIH) 76-802