National Cancer Institute 

CARCINOGENESIS 

Technical Report Series 

No. 14 

1977 

BIOASSAY OF 

LINDANE 

FOR POSSIBLE CARCINOGENICITY 

CAS No. 58-89-9 

NCI-CG-TR-14 

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

Public Health Service 

National Institutes of Health 

BIOASSAY OF
 

LINDANE
 

FOR POSSIBLE CARCINOGENICITY
 

Carcinogen Bioassay and Program Resources Branch
 

Carcinogenesis Program
 

Division of Cancer Cause and Prevention
 

National Cancer Institute
 

National Institutes of Health
 

Bethesda, Maryland 20014
 

DREW Publication No. (NIH) 77-814
 

BIOASSAY OF
 

LINDANE
 

FOR POSSIBLE CARCINOGENICITY
 

Carcinogenesis Program
 

Division of Cancer Cause and Prevention
 

National Cancer Institute
 

National Institutes of Health
 

CONTRIBUTORS: This report presents the results of the bioassay
 

of lindane for possible carcinogenicity, conducted for the Carcinogen Bioassay and Program Resources Branch, Carcinogenesis
 

Program, Division of Cancer Cause and Prevention, National Cancer
 

Institute (NCI), Bethesda, Maryland. The bioassay was conducted
 

by Gulf South Research Institute, New Iberia, Louisiana, initially under direct contract to NCI and currently under a subcontract
 

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

Carcinogenesis bioassay program.
 

The experimental design was determined by Drs. J. H.
 

Weisburger^-'*• and R. R. Bates*'3; the doses were selected by Drs.
 

I. E. Shellenberger^'^, jt HP Weisburger, and R. R. Bates.
 

Animal treatment and observation were supervised by Drs. T. E.
 

Shellenberger and H. P. Burchfield^, with the technical
 

assistance of Ms. D. H. Monceaux^ and Mr. D. Broussard^.
 

Histopathology was performed by Drs. E. Bernal^ and B. Buratto^
 

at Gulf South Research Institute, and the diagnoses included in
 

this report represent the interpretation of these pathologists.
 

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

Mason Research Institute". Statistical analyses were performed
 

by Dr. J. R. Joiner^, using methods selected for the bioassay
 

program by Dr. J. J. Gart^. Chemicals used in this bioassay were
 

analyzed under the direction of Dr. H. P. Burchfield, and the
 

analytical results were reviewed by Dr. S. S. Olin^.
 

This report was prepared at Tracer Jitco under the direction of
 

NCI. Those responsible for the report at Tracor Jitco were Dr.
 

Marshall Steinberg^, Director of the Bioassay Program; Drs. J. F.
 

iii
 
Robens7

 and C. H. Williams?, toxicologists; Dr. R. L. Schueler7,
 

pathologist; Mr. W. D. Reichardt7

 and Ms. Y. E. Presley7,
 

technical writers; and Dr. E. W. Gunberg7, technical editor.
 

The following scientists at the National Cancer Institute were
 

responsible for evaluating the bioassay experiment, interpreting
 

the results, and reporting the findings:
 

Dr. Kenneth C. Chu
 

Dr. Cipriano Cueto, Jr.
 

Dr. J. Fielding Douglas
 

Dr. Dawn G. Goodman
 

Dr. Richard A. Griesemer
 

Dr. Thomas W. Orme
 

Mr. Harry A. Milman
 

Dr. Robert A. Squire^
 

Dr. Jerrold M. Ward
 

l-Carcinogenesis Program, Division of Cancer Cause Prevention,
 

National Cancer Institute, National Institutes of Health,
 

Bethesda, Maryland.
 

with the Naylor Dana Institute for Disease Prevention,
 

American Health Foundation, Hammond House Road, Valhalla,
 

New York.
 

with the Office of the Commissioner, Food and Drug
 

Administration, Rockville, Maryland.
 

^Gulf South Research Institute, Atchafalaya Basin Laboratories,
 

P. 0. Box 1177, New Iberia, Louisiana.
 

with the National Center for Toxicological Research,
 

Jefferson, Arkansas.
 

6

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

Rockville, Maryland.
 

iv
 
7

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

Maryland.
 

^Mathematical Statistics and Applied Mathematics Section,
 

Biometry Branch, Field Studies and Statistics, Division of
 

Cancer Cause and Prevention, National Cancer Institute, National
 

Institutes of Health, Bethesda, Maryland.
 

with the Division of Comparative Medicine, Johns Hopkins
 

University, School of Medicine, Traylor Building, Baltimore,
 

Maryland.
 

SUMMARY
 

A bioassay for possible carcinogenicity of lindane was conducted
 

by administering the test chemical in the diet to Osborne-Mendel
 

rats and B6C3F1 mice.
 

Groups of 50 rats of each sex were administered lindane at one of
 

two doses for 80 weeks, then observed for 29-30 weeks. Time-


weighted average doses for males were 236 or 472 ppm; those for
 

females were 135 or 270 ppm. Matched controls consisted of
 

groups of 10 untreated rats of each sex; pooled controls, used
 

for statistical evaluation, consisted of the matched-control
 

groups combined with 45 untreated male and 45 untreated female
 

rats from similar bioassays of four other test chemicals. All
 

surviving rats were killed at 108-110 weeks.
 

Groups of 50 mice of each sex were administered lindane at one of
 

two doses, either 80 or 160 ppm, for 80 weeks, then observed for
 

an additional 10-11 weeks. Matched controls consisted of groups
 

of 10 untreated mice of each sex; pooled controls, used for
 

statistical evaluation, consisted of the matched-control groups
 

combined with 40 untreated male and 40 untreated female mice from
 

similar bioassays of four other test chemicals. All surviving
 

mice were killed at 90-91 weeks.
 

Neither the mean body weights of rats nor those of mice showed
 

consistent effects from the administration of lindane. The
 

physical condition of the surviving treated mice deteriorated
 

during the final 6 weeks on study. Except for the female
 

matched-control group of rats, survival of all groups of rats and
 

mice was adequate for meaningful statistical analyses of the
 

incidence of tumors.
 

In rats, no tumor occurred at a statistically significant
 

incidence in the treated groups of either sex.
 

vii
 
In mice, the incidence of hepatocellular carcinoma in low-dose
 

males was significant when compared with that in the pooled
 

controls (controls 5/49, low-dose 19/49, P = 0.001). This
 

finding, by itself, is insufficient to establish the
 

carcinogenicity of lindane. The incidence of hepatocellular
 

carcinoma in high-dose male mice (9/46) was not significantly
 

different from that in the matched (2/10) or pooled controls.
 

It is concluded that under the conditions of this bioassay, 

lindane was not carcinogenic for Osborne-Mendel rats or B6C3F1 

mice. 

viii
 
TABLE OF CONTENTS
 

Page
 

I. Introduction 1
 

II. Materials and Methods 3
 

A. Chemical 3
 

B. Dietary Preparation 3
 

C. Animals 4
 

D. Animal Maintenance 5
 

E. Subchronic Studies 6
 

F. Designs of Chronic Studies 7
 

G. Clinical and Pathologic Examinations 10
 

H. Data Recording and Statistical Analyses 11
 

III. Results -Rats 17
 

A. Body Weights and Clinical Signs (Rats) 17
 

B. Survival (Rats) 17
 

C. Pathology (Rats) 20
 

D. Statistical Analyses of Results (Rats) 21
 

IV. Results - Mice 25
 

A. Body Weights and Clinical Signs (Mice) 25
 

B. Survival (Mice) 25
 

C. Pathology (Mice) 28
 

D. Statistical Analyses of Results (Mice) 30
 

V. Discussion 33
 

VI. Bibliography 37
 

APPENDIXES
 

Appendix A Summary of the Incidence of Neoplasms in
 

Rats Fed Lindane in the Diet 39
 

Table Al Summary of the Incidence of Neoplasms in
 

Male Rats Fed Lindane in the Diet 41
 

Table A2 Summary of the Incidence of Neoplasms in
 

Female Rats Fed Lindane in the Diet 44
 

ix
 
Page
 

Appendix B Summary of the Incidence of Neoplasms in 

Mice Fed Lindane in the Diet 49 

Table Bl Summary of the Incidence of Neoplasms in 

Male Mice Fed Lindane in the Diet 51 

Table B2 Summary of the Incidence of Neoplasms in 

Female Mice Fed Lindane in the Diet 54 

Appendix C Summary of the Incidence of Nonneoplastic 

Lesions in Rats Fed Lindane in the Diet 57 

Table Cl Summary of the Incidence of Nonneoplastic 

Lesions in Male Rats Fed Lindane 

in the Diet 59 

Table C2 Summary of the Incidence of Nonneoplastic 

Lesions in Female Rats Fed Lindane 

in the Diet 63 

Appendix D Summary of the Incidence of Nonneoplastic 

Lesions in the Mice Fed Lindane 

in the Diet 67 

Table Dl Summary of the Incidence of Nonneoplastic 

Lesions in the Male Mice Fed Lindane 

in the Diet 69 

Table D2 Summary of the Incidence of Nonneoplastic 

Lesions in Female Mice Fed Lindane 

in the Diet 71 

Appendix E Analyses of the Incidence of Primary Tumors 

in Rats Fed Lindane in the Diet 75 

Table El Analyses of the Incidence of Primary Tumors 

in Male Rats Fed Lindane in the Diet 77 

Table E2 Analyses of the Incidence of Primary Tumors 

in Female Rats Fed Lindane in the Diet 82 
Page 

Appendix F Analyses of the Incidence of Primary Tumors 

in Mice Fed Lindane in the Diet 89 

Table Fl Analyses of the Incidence of Primary Tumors 

in Male Mice Fed Lindane in the Diet 91 

Table F2 Analyses of the Incidence of Primary Tumors 

in Female Mice Fed Lindane in the Diet 94 

Appendix G Analysis of Formulated Diets for 

Concentrations of Lindane 97 

TABLES 

Table 1 Design of Lindane Chronic Feeding 

Studies in Rats 8 

Table 2 Design of Lindane Chronic Feeding 

Studies in Mice 9 

FIGURES 

Figure 1 Growth Curves for Rats Fed Lindane 

in the Diet 18 

Figure 2 Survival Curves for Rats Fed Lindane 

in the Diet 19 

Figure 3 Growth Curves for Mice Fed Lindane 

in the Diet 26 

Figure 4 Survival Curves for Mice Fed Lindane 

in the Diet 27 

xi
 

I. INTRODUCTION
 

Lindane (CAS 58-89-9; NCI C00204) is an organochlorine pesticide
 

that is registered for use in soil, foliar, and seed treatment
 

for a large variety of fruit and vegetable crops, and for use on
 

livestock, pets, and agricultural premises (EPA, 1971, 1973).
 

Residues of lindane may be persistent in soil and foods (Hayes,
 

1975). There may also be direct human exposure to lindane
 

through its use in pharmaceutical preparations or in public
 

health pest control.
 

Lindane was selected for testing because data regarding its
 

carcinogenicity were considered inadequate, and because there was
 

a potential for long-term human exposure due to its extensive use
 

and its persistence in the environment.
 

II. MATERIALS AND METHODS
 

A. Chemical
 

Lindane is the gamma isomer of 1,2,3,4,5,6-hexachlorocyclohexane.
 

It was obtained in two batches for the chronic study, the first
 

batch from City Chemical Co. , New York, N. Y. , and the second
 

batch from Diamond Shamrock Co., Agricultural Chemicals Division,
 

Cleveland, Ohio. Both suppliers reported the chemical to be
 

essentially 100% pure.
 

The identity and purity of these batches were confirmed at Gulf
 

South Research Institute using melting point; elemental analysis
 

(C, H, Cl) for CfcH^Cl^; infrared, nuclear magnetic resonance, and
 

mass spectroscopy; and thin-layer and gas-liquid chromatography.
 

The chemical was stored at approximately 4°C.
 

B. Dietary Preparation
 

All diets were formulated using finely ground Wayne® Lab Blox
 

meal (Allied Mills, Inc., Chicago, 111.) to which was added the
 

required amount of lindane for each dietary concentration. The
 

test chemical was first dissolved in a small amount of acetone
 

(Mallinckrodt, Inc., St. Louis, Mo.), which was then added to the
 

feed. Corn oil (Louana®, Opelousas Refinery Co., Opelousas, La.)
 

was also added to the feed, primarily as a dust suppressant, and
 

the diets were mixed mechanically to assure homogeneity of the
 

3
 
mixtures and evaporation of the acetone. Final diets, including
 

those for the control groups of animals, contained corn oil equal
 

to 2% of the final weight of feed. Formulated diets were stored
 

at approximately 17°C until used, but no longer than 1 week.
 

The stability of lindane in feed was tested by determining the
 

concentration of the chemical in formulated diets at intervals
 

over a 7-day period. Diets containing 80 or 320 ppm lindane
 

showed no change in concentration on standing at ambient
 

temperature for this period.
 

As a quality control test on the accuracy of preparation of the
 

diets, the concentration of lindane was determined in different
 

batches of formulated diets during the chronic study. The
 

results are summarized in Appendix G. At each dietary
 

concentration, the mean of the analytical concentrations for the
 

checked samples was within 2.4% of the theoretical concentration,
 

and the coefficient of variation was never more than 5.1%. Thus,
 

the evidence indicates that the formulated diets were prepared
 

accurately.
 

C. Animals
 

Rats and mice of both sexes, obtained through contracts of the
 

Division of Cancer Treatment, National Cancer Institute, were
 

used in these bioassays. The rats were of the Osborne-Mendel
 
strain obtained from Battelle Memorial Institute, Columbus, Ohio,
 

and the mice were B6C3F1 hybrids obtained from Charles River
 

Breeding Laboratories, Inc., Wilmington, Massachusetts. On
 

arrival at the laboratory, all animals were quarantined for an
 

acclimation period (rats for 10 days, mice for 12 days) and were
 

then assigned to control and treated groups.
 

D. Animal Maintenance
 

All animals were housed in temperature- and humidity-controlled
 

rooms. The temperature range was 22-24°C, and the relative
 

humidity was maintained at 40-70%. The air in each room was
 

changed 10-12 times per hour. Fluorescent lighting provided
 

illumination 10 hours per day. Food and water were presented ad
 

libitum.
 

The rats were housed individually in hanging galvanized steel
 

mesh cages, and the mice were housed in plastic cages with filter
 

bonnets, five per cage for females, and two or three per cage for
 

males. Initially, rats were transferred once per week to clean
 

cages; later in the study, cages were changed every 2 weeks.
 

Mice were transferred once per week to clean cages with filter
 

bonnets; bedding used for the mice was Absorb-Dri® (Lab Products,
 

Inc., Garfield, N.J.). For rats, absorbent sheets under the
 

cages were changed three times per week. Feeder jars and water
 

bottles were changed and sterilized three times per week.
 
Cages for control and treated mice were placed on separate racks
 

in the same room. Animal racks for both species were rotated
 

laterally once per week; at the same time each cage was changed
 

to a different position in the row within the same column. Rats
 

receiving lindane, along with their matched controls, were housed
 

in a room by themselves. Mice receiving lindane were maintained
 

in a room housing mice administered safrole (CAS 94-59-7) or
 

N-2-fluorenylacetamide (CAS 53-96-3), together with their
 

respective matched controls.
 

E. Subchronic Studies
 

Subchronic studies were conducted to estimate the maximum
 

tolerated doses of lindane, on the basis of which low and high
 

concentrations (hereinafter referred to as "low doses" and "high
 

doses") were determined for administration in the chronic
 

studies. In these subchronic studies, lindane was added to the
 

animal feed in twofold increasing concentrations, ranging from
 

160 to 2,560 ppm for rats and from 40 to 1,280 ppm for mice.
 

Treated and control groups each consisted of five male and five
 

female animals. The chemical was provided in feed to the treated
 

groups for 6 weeks, followed by a period of observation for 2
 

weeks.
 

In male or female rats receiving 320 or 640 ppm, weight gains
 

decreased during the first 3 weeks. Later, weight gains of
 
treated male rats approached those of the controls. There were
 

no deaths at these doses; however, one male rat and two female
 

rats treated at 1,280 ppm died during week 2. The low and high
 

doses for the chronic studies using rats were set at 320 and 640
 

ppm.
 

In the mice, there was no effect on weight gains at 80 and 160
 

ppm, and no deaths occurred. At 320 ppm, one male died during
 

week 1 and two females died during week 5. By week 5, all mice
 

receiving 640 ppm died. The low and high doses for the chronic
 

studies using mice were set at 80 and 160 ppm.
 

F. Designs of Chronic Studies
 

The designs of the chronic studies are shown in tables 1 and 2.
 

Since the numbers of animals in the matched-control groups were
 

small, pooled-control groups also were used for statistical
 

comparisons. Matched controls from the current studies on
 

lindane were combined with matched controls from studies
 

performed on tetrachlorvinphos (CAS 961-11-5), toxaphene (CAS
 

8001-35-2), endrin (CAS 72-20-8), and malathion (CAS 121-75-5).
 

The pooled controls for statistical tests using rats consisted of
 

55 males and 55 females; using mice, 50 males and 50 females.
 

The studies on chemicals other than lindane were also conducted
 

at Gulf South Research Institute and overlapped the lindane study
 
Table 1. Design of Lindane Chronic Feeding Studies in Rats
 

Sex and 

Treatment 

Initial 

No. of 

Lindane 

in Dietb 

Time on Study 

Treated0 Untreatedd 

Time-Weighted 

Average Dose6 

Group Animals3 (ppm) (weeks ) (weeks) (ppm) 

Male 

Matched-Control 10 0 109 

Low-Dose 50 320 

160 

0 

38 

42 

30 

236 

High-Dose 50 640 

320 

0 

38 

42 

30 

472 

Female 

Matched-Control 10 0 108-109 

Low-Dose 50 320 

160 

80 

0 

2 

49 

29 

29-30 

135 

High-Dose 50 640 

320 

160 

0 

2 

49 

29 

30 

270 

a

All animals were 35 days of age when placed on study.
 

b

Doses of lindane were lowered during the study, as indicated, due to
 

deaths among the treated animals.
 

C

A11 animals were started on study on the same day.
 

diets containing lindane were discontinued, treated rats and their
 

matched controls were fed control diets without corn oil for 15 weeks,
 

then control diets (2% corn oil added) for an additional 15 weeks.
 

e

Time-weighted average dose = £ (dose in ppm x no. of weeks at that dose)
 

*• (no. of weeks receiving each dose)
 
Table 2. Design of Lindane Chronic Feeding Studies in Mice
 

Sex and Initial Lindane Time on Study 

Treatment No. of in Diet Treated5 Untreated0 

Group Animals3 (ppm) (weeks) (weeks ) 

MALE 

Matched-Control 10 0 90 

Low-Dose 50 80 80 

0 10 

High-Dose 50 160 80 

0 10 

FEMALE 

Matched-Control 10 0 90 

Low-Dose 50 80 80 

0 10 

High-Dose 50 160 80 

0 10-11 

a

All animals were 35 days of age when placed on study.
 

^All animals were started on study on the same day.
 

°When diets containing lindane were discontinued, treated mice and
 

their matched controls were fed control diets (2% corn oil added).
 
by at least 1 year. The matched-control groups for the different
 

test chemicals were of the same strain and from the same
 

supplier, and they were examined by the same pathologists.
 

G. Clinical and Pathologic Examinations
 

All animals were observed twice daily for signs of toxicity,
 

weighed at regular intervals, and palpated for masses at each
 

weighing. Animals that were moribund at the time of clinical
 

examination were killed and necropsied.
 

The pathologic evaluation consisted of gross and microscopic
 

examination of major tissues, major organs, and all gross lesions
 

from killed animals and from animals found dead. The following
 

tissues were examined microscopically: skin, lungs and bronchi,
 

trachea, bone and bone marrow, spleen, lymph nodes, heart,
 

salivary gland, liver, gallbladder (mice), pancreas, stomach,
 

small intestine, large intestine, kidney, urinary bladder,
 

pituitary, adrenal, thyroid, parathyroid, mammary gland, prostate
 

or uterus, testis or ovary, and brain. Occasionally, additional
 

tissues were also examined microscopically. The different
 

tissues were preserved in 10% buffered formalin, embedded in
 

paraffin, sectioned, and stained with hematoxylin and eosin.
 

Special staining techniques were utilized when indicated for more
 

definitive diagnosis.
 

10
 
A few tissues from some animals were not examined, particularly
 

from those animals that died early. Also, some animals were
 

missing, cannibalized, or judged to be in such an advanced state
 

of autolysis as to preclude histopathologic evaluation. Thus,
 

the number of animals from which particular organs or tissues
 

were examined microscopically varies, and does not necessarily
 

represent the number of animals that were placed on study in each
 

group.
 

H. Data Recording and Statistical Analyses
 

Pertinent data on this experiment have been recorded in an automatic data processing system, the Carcinogenesis Bioassay Data
 

System (Linhart et al., 1974). The data elements include descriptive information on the chemicals, animals, experimental design,
 

clinical observations, survival, body weight, and individual
 

pathologic results, as recommended by the International Union
 

Against Cancer (Berenblum, 1969). Data tables were generated for
 

verification of data transcription and for statistical review.
 

These data were analyzed using the statistical techniques
 

described in this section. Those analyses of the experimental
 

results that bear on the possibility of carcinogenic*ty are
 

discussed in the statistical narrative sections.
 

Probabilities of survival were estimated by the product-limit
 

11
 
procedure of Kaplan and Meier (1958) and are presented in this
 

report in the form of graphs. Animals were statistically
 

censored as of the time that they died of other than natural
 

causes or were found to be missing; animals dying from natural
 

causes were not statistically censored. Statistical analyses for
 

a possible dose-related effect on survival used the method of Cox
 

(1972) for testing two groups for equality and Tarone's (1975)
 

extensions of Cox's methods for testing for a dose-related trend.
 

One-tailed P values have been reported for all tests except the
 

departure from linearity test, which is only reported when its
 

two-tailed P value is less than 0.05.
 

The incidence of neoplastic or nonneoplastic lesions has been
 

given as the ratio of the number of animals bearing such lesions
 

ac a specific anatomic site (numerator) to the number of animals
 

in which that site is examined (denominator). In most instances,
 

the denominators included only those animals for which that site
 

was examined histologically. However, when macroscopic examination was required to detect lesions prior to histologic sampling
 

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

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

consist or the numbers of animals necropsied.
 

The purpose of the statistical analyses of tumor incidence is to
 

determine whether animal » receiving the test chemical developed a
 

12
 
significantly higher proportion of tumors than did the control
 

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

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

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

dose level. When results for a number of treated groups (k) are
 

compared simultaneously with those for a control group, a
 

correction to ensure an overall significance level of 0.05 may be
 

made. The Bonferroni inequality (Miller, 1966) requires that the
 

P value for any comparison be less than or equal to 0.05/k. In
 

cases where this correction was used, it is discussed in the
 

narrative section. It is not, however, presented in the tables,
 

where the Fisher exact P values are shown.
 

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

continuity correction (Armitage, 1971), was also used. Under the
 

assumption of a linear trend, this test determines if the slope
 

of the dose-response curve is different from zero at the one-


tailed 0.05 level of significance. Unless otherwise noted, the
 

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

from linear trend.
 

A time-adjusted analysis was applied when numerous early deaths
 

resulted from causes that were not associated with the formation
 

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

13
 
first tumor was observed were excluded by basing the statistical
 

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

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

such an early tumor was found, comparisons were based exclusively
 

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

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

obtained, the standard procedures for analyses of the incidence
 

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

followed.
 

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

incidence of tumors. Curves of the proportions surviving without
 

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

The week during which animals died naturally or were sacrificed
 

was entered as the time point of tumor observation. Cox's
 

methods of comparing these curves were used for two groups;
 

Tarone's extension to testing for linear trend was used for three
 

groups. The statistical tests for the incidence of tumors which
 

used life-table methods were one-tailed and, unless otherwise
 

noted, in the direction of a positive dose relationship.
 

Significant departures from linearity (P < 0.05, two-tailed test)
 

were also noted.
 

The approximate 95 percent confidence interval for the relative
 

risk of each treated group compared to its control was calculated
 
from the exact interval on the odds ratio (Gart, 1971). The
 

relative risk is defined as pt/pc where pt is the true binomial
 

probability of the incidence of a specific type of tumor in a
 

treated group of animals and pc is the true probability of the
 

spontaneous incidence of the same type of tumor in a control
 

group. The hypothesis of equality between the true proportion of
 

a specific tumor in a treated group and the proportion in a
 

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

excess of unity represent the condition of a larger proportion in
 

the treated group than in the control.
 

The lower and upper limits of the confidence interval of the
 

relative risk have been included in the tables of statistical
 

analyses. The interpretation of the limits is that in
 

approximately 95% of a large number of identical experiments, the
 

true ratio of the risk in a treated group of animals to that in a
 

control group would be within the interval calculated from the
 

experiment. When the lower limit of the confidence interval is
 

greater than one, it can be inferred that a statistically
 

significant result (a P < 0.025 one-tailed test when the control
 

incidence is not zero, P < 0.050 when the control incidence is
 

zero) has occurred. When the lower limit is less than unity, but
 

the upper limit is greater than unity, the lower limit indicates
 

the absence of a significant result while the upper limit
 

indicates that there is a theoretical possibility of the
 

15
 
induction of tumors by the test chemical, which could not be
 

detected under the conditions of this test.
 

16
 
III. RESULTS - RATS
 

A. Body Weights and Clinical Signs (Rats)
 

Mean body weights of the male rats were unaffected by lindane
 

(figure 1). The weights of the low-dose females were consistently higher than those of the matched-control and high-dose
 

females, and the weights of the high-dose females were higher
 

than those of the matched controls near the end of the study.
 

Except for occasional weight loss by individual animals, the
 

treated animals were generally comparable to the controls in
 

appearance and behavior during the first year of the study.
 

Clinical signs in all treated groups were noted at a low or
 

moderate incidence during the first half of the second year, and
 

with gradually increasing frequency during the remainder of the
 

study. These signs included rough and discolored hair coats
 

(primarily among the male animals), pale mucous membranes,
 

dermatitis, and vaginal bleeding.
 

B. Survival (Rats)
 

The Kaplan and Meier curves estimating the probabilities of
 

survival for male and female rats receiving lindane at the doses
 

used in this experiment, together with those of the matched
 

controls, are shown in figure 2.
 

17
 
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rva 

5 5 i S 

a 2 r m 8 i s a I m o > 

I! 

8 H z en 

r» 

i o"S 30 
In neither sex was the Tarone test result significant at the 0.05
 

level for positive dose-related trend in mortality over the
 

period. In male rats, 48% of the high-dose group, 50% of the
 

low-dose group, and 60% of the controls lived to the end of the
 

study. In females, only 40% of the controls survived to the end
 

of the study, while at least 60% of the low- and high-dose groups
 

survived. Early deaths of these rats were not tumor associated.
 

Over 80% of these animals lived at least as long as 52 weeks on
 

study, providing adequate numbers of rats of both sexes for
 

meaningful statistical analyses of the incidences of late-


developing tumors.
 

C. Pathology (Rats)
 

Histopathologic findings on neoplasms in rats are summarized in
 

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

are summarized in Appendix C, tables Cl and C2.
 

A variety of neoplastic, proliferative, degenerative, and
 

inflammatory lesions occurred in a random manner among rats of
 

the control and lindane-treated groups. The majority of these
 

lesions, commonly observed in this strain of rat, occurred with a
 

low incidence within a given group.
 

The incidences of most of these lesions in the treated groups
 

were comparable to those in the controls. Occasionally, however,
 

20
 
these lesions occurred only in animals of the treated groups.
 

This may reflect the small number of control animals rather than
 

any harmful effect caused by the chemical.
 

In the judgment of the pathologists, there was no evidence of
 

carcinogenicity induced in Osborne-Mendel rats by the administration of lindane under the conditions of this bioassay.
 

D. Statistical Analyses of Results (Rats)
 

Tables El and E2 in Appendix E contain the statistical analyses
 

of the incidences of those specific primary tumors that were
 

observed in at least 5% of one or more treated groups of either
 

sex.
 

In male rats, the Cochran-Armitage test for positive dose-related
 

trend in proportions for hemangioma of the spleen has a probability level of 0.030 using the pooled controls; however, this is
 

based on only a minimal incidence in the high-dose group (3/44,
 

7%), and under this condition, the analysis based on a hypothesis
 

of linear trend is questionable. Female rats do not show any
 

incidence of this tumor. There is inadequate statistical
 

evidence to conclude that the occurrence of this tumor is related
 

to treatment, since the results of the Fisher exact test are not
 

significant.
 

21
 
In the analyses of the incidence of chromophobe adenoma of the
 

pituitary in female rats, although the result of the Cochran-


Armitage test for positive dose-related trend in proportions is
 

not significant at the 0.05 level, the departure from linearity
 

has a probability level of 0.048 using the pooled controls, due
 

to the higher incidence in the low-dose group than in the
 

high-dose group. The Fisher exact test shows a P value of 0.033
 

when the incidence in the low-dose females is compared with that
 

in the pooled controls, but this probability level is above the
 

0.025 level required by the Bonferroni criterion for multiple
 

comparisons. In male rats, the statistical test results for the
 

incidence of this tumor are not significant in the positive direction. No direct relationship between the incidence of this tumor
 

and the administration of lindane can be concluded.
 

In female rats, the Fischer exact tests shows a P value of 0.049
 

when the incidence of C-cell adenoma of the thyroid in the low-


dose group is compared with that in the pooled-control group;
 

however, this probability level is above the 0.025 level required
 

by the Bonferroni criterion for multiple comparisons.
 

There are no other specific incidences of tumors for which the
 

Cochran-Armitage test or the Fisher exact test shows significance
 

in the positive direction. A negative Cochran-Armitage test
 

result is observed in the incidence of follicular-cell adenoma of
 

22
 
the thyroid in male rats when the incidence in the pooled
 

controls exceeds the incidences in the treated groups. In each
 

of the 95% confidence intervals for relative risk, shown in the
 

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

aspects of the results. It should also be noted that each of the
 

intervals has an upper limit greater than one, indicating the
 

theoretical possibility of the induction of tumors by this
 

chemical, which could not be detected under the conditions of
 

this test.
 

23
 

IV. RESULTS - MICE
 

A. Body Weights and Clinical Signs (Mice)
 

Mean body weights of the male mice were unaffected by lindane
 

(figure 3). The weights of the low-dose females were consistently lower than those of the matched-control and high-dose females.
 

During the first year of the study, the treated animals were
 

generally comparable to the controls in appearance and behavior.
 

A few treated animals had alopecia. During the second year, all
 

of the treated females appeared excitable when handled, and many
 

of the treated males were observed fighting. Clinical signs
 

including rough hair coats, alopecia, and abdominal distention
 

(predominantly in treated males) were noted with increasing
 

frequency during the remainder of the study. Treated mice were
 

generally in poor physical condition during the last 6 weeks of
 

the study.
 

B. Survival (Mice)
 

The Kaplan and Meier curves estimating the probabilities of
 

survival for male and female mice receiving lindane at the doses
 

used in this experiment, together with those of the matched
 

controls, are shown in figure 4.
 

25
 
40 

(£ 

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 MALE MICE 

 MATCHED CONTROL 

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 LOW DOSE 

 HIGH DOSE 

10 20 30 40 50

TIME ON STUDY (WEEKS) 

 60 70 80 90 

35 

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20- a

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10­ FEMALE MICE 

D MATCHED CONTROL 

O LOW DOSE 

5­ A HIGH DOSE 

10 20 30 40 50

TIME ON STUDY (WEEKS) 

 60 70 80 90 

Figure 3. Growth Curves For Mice Fed Lindane In The Diet 

26 
PROBABILITY OF SURVIVAL PROBABILITY OF SU

c> p p p p p p o o c3 _, cs o o o o o 

-* Vj uj A ui b> kj bo t S> 0 0 0 0 0 0 0 0 CS § i; S g g g § g 

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In neither sex was the Tarone test result significant for
 

positive dose-related trend in mortality over the period.
 

The survivals for treated and control groups within each sex are
 

comparable. At least 88% of the males and 80% of the females
 

lived to the end of the study, providing sufficient animals for
 

meaningful statistical analyses of the incidence of tumors.
 

C. Pathology (Mice)
 

Histopathologic findings on neoplasms in mice are summarized in
 

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

are summarized in Appendix D, tables Dl and D2.
 

A variety of neoplastic, proliferative, degenerative, and
 

inflammatory lesions occurred in a random manner among mice of
 

the control and lindane-treated groups. An exception was the
 

relatively high incidence of neoplastic lesions in the liver.
 

Hepatocellular carcinoma and neoplastic nodules of the liver, as
 

described by Squire and Levitt (1975), were the most numerous
 

lesions, occurring with approximately equal frequency among
 

control and high-dose mice. Hepatocellular carcinomas occurred
 

in 2/10 (20%) control males, 0/10 (0%) control females, 9/46
 

(20%) high-dose males, and 3/46 (7%) high-dose females. These
 

lesions were most frequent in the low-dose males (19/49 [39%]).
 

28
 
The hepatocellular carcinomas were quite variable in morphology.
 

Grossly, the lesions were large single masses (1 to 2 cm. in
 

diameter), or multiple discrete, confluent nodules varying in
 

size from microscopic to several millimeters in diameter. The
 

consistency also varied, but in most cases the tumors were soft,
 

particularly when there were extensive areas of necrosis. They
 

were well circumscribed and readily distinguishable from the
 

surrounding normal tissue. The major lobes were most frequently
 

affected with lesions within the parenchyma or bulging just
 

beneath the capsule. Microscopically, the most salient features
 

of the lesions were tinctorial and architectural. These features
 

varied not only from tumor to tumor, but also among different
 

areas of the same tumor. It was not uncommon to see nodular
 

structures within other nodules. The cytoplasm of the affected
 

cells was most commonly basophilic. The arrangement of the liver
 

plates was either nearly normal or so disorganized as to destroy
 

the trabecular pattern completely. Dilation of the sinuses also
 

contributed to the focal or diffuse distortion of the neoplastic
 

tissue. Because of these changes and the compression of the
 

adjacent parenchyma, the outline of the lesion was usually well
 

defined. The cytology was more or less that of well-


differentiated hepatocytes with some variation of size and shape.
 

Intracytoplasmic eosinophilic bodies were present in a large
 

number of cells. The nuclei were smaller, of the same size, or
 

29
 
larger than the normal ones. Abnormal mitoses were rare and
 

normal mitoses were relatively few. Occasionally, neoplastic
 

changes were associated with degenerative features such as lipid
 

deposition, focal or massive necrosis, hemorrhage, or with
 

inflammatory changes. Metastases to the lung (two animals),
 

diaphragm (one animal), or epididymis (one animal) were observed
 

in male mice only.
 

The nonneoplastic lesions were degenerative or inflammatory in
 

nature. The distribution of these nonneoplastic lesions among
 

the animals of the different groups suggests that they are not
 

attributable to the chemical tested; rather, they are
 

coincidental lesions.
 

In the judgment of the pathologists, there was insufficient
 

evidence to conclude that carcinogenicity in B6C3F1 mice was
 

induced by the administration of lindane under the conditions of
 

this bioassay.
 

D. Statistical Analyses of Results (Mice)
 

Tables Fl and F2 in Appendix F contain the statistical analyses
 

of the incidences of those specific primary tumors that were
 

observed in at least 5% of one or more treated groups of either
 

sex.
 

30
 
In male mice, although the Cochran-Armitage test result for
 

positive dose-related trend in proportions for hepatocellular
 

carcinoma is not significant at the 0.05 level, the departure
 

from linearity has a probability level of 0.002 using the pooled
 

controls, due to the higher incidence in the low-dose group than
 

in the high-dose group. The Fisher exact test shows a significantly higher proportion (P = 0.001) of this tumor in the low-


dose group than in the pooled controls, and the lower and upper
 

limits of the 95% confidence interval of the relative risk have
 

values greater than one. The incidence of hepatocellular
 

carcinoma in female mice is not statistically significant. When
 

hepatocellular carcinoma and neoplastic nodule of the liver in
 

females are considered together, the statistical test results of
 

the combined incidence are still not significant. In males,
 

after similar grouping, the result of the Cochran-Armitage test
 

of proportions remains not significant, with a probability level
 

of 0.011 for the departure from linearity, due to a higher
 

proportion in the low-dose group than in the high-dose group.
 

The Fisher exact test shows that the combined incidence in the
 

low-dose males is significantly higher than that in the pooled
 

controls (P = 0.010). The incidence in the high-dose group is
 

not significant, and this conclusion could not be attributed to
 

low survival, since the survival in the high-dose group was
 

comparable to that in the low-dose group. The results of the
 

31
 
historical controls compiled to date at this laboratory indicate
 

that hepatocellular carcinomas and neoplastic nodules of the
 

liver occurred in 75/360 (20.8%) of B6C3F1 male mice. Due to
 

this high spontaneous incidence, and because the incidence among
 

high-dose males in this study is not significant, these tumors
 

cannot be related conclusively to treatment.
 

There are no other specific incidences of tumors in mice for
 

which the statistical tests show significance. In each of the
 

95% confidence intervals of relative risk, shown in the tables,
 

the value of one is included; this indicates the negative aspects
 

of the results. It should also be noted that each of the
 

intervals has an upper limit greater than one, indicating the
 

theoretical possibility of the induction of tumors by lindane,
 

which could not be detected under the conditions of this test.
 

32
 
V. DISCUSSION
 

Lindane was toxic to both rats and mice, as shown by clinical
 

signs and effects on weight and/or mortality. During the first
 

year of the study, the treated rats were generally comparable to
 

the controls in appearance and behavior. Clinical signs,
 

including rough and discolored hair coats, pale mucous membranes,
 

dermatitis, and vaginal bleeding were noted with increasing
 

frequency in the treated animals during the second year of the
 

study.
 

Mean body weight gains in rats were not adversely affected by
 

lindane; however, individual treated animals lost weight at
 

various times during the study. Survival of the treated rats was
 

not statistically different from that of the matched controls;
 

however, there were several early deaths among the controls.
 

Survival to termination of the study was from 48% to 64% for each
 

of the treated groups.
 

In mice, the mean body weights were unaffected by treatment, with
 

the exception of the low-dose females. During the second year,
 

the female mice were excitable; among the males, increased
 

fighting, rough hair coats, alopecia, and abdominal distention
 

were noted. Treated mice were generally in poor physical
 

condition during the last 6 months on study; however, treatment
 

33
 
did not affect survival, and there were adequate numbers of
 

treated animals for meaningful statistical analyses of the
 

incidence of tumors.
 

In rats, a variety of neoplastic lesions occurred in a random
 

manner among both control and treated animals, but the incidences
 

in the treated groups were not significantly different from those
 

in either the matched- or pooled-control groups.
 

In mice, a relatively high incidence of neoplastic lesions in the
 

liver was observed in both control and treated animals. Hepatocellular carcinomas in the males occurred in 2/10 matched
 

controls, 5/49 pooled controls, 19/49 low-dose, and 9/46
 

high-dose mice. The incidence in the low-dose males was
 

significantly higher than that in the pooled controls (P =
 

0.001). However, neither the incidence in the high-dose males
 

nor the dose-related trend was significant, and the significance
 

of the findings was not increased when hepatocellular carcinomas
 

were combined with neoplastic nodules. No hepatic hyperplasia was
 

observed in treated mice of either sex. In control animals,
 

hepatocellular carcinomas and neoplastic nodules occurred in 23%
 

of all B6C3F1 male mice on study at the laboratory. Thus, the
 

incidence of hepatocellular carcinoma in male mice cannot clearly
 

be related to treatment. The incidence of hepatocellular
 

carcinoma among female mice was not significant.
 

34
 
The chronic toxicity of lindane has been studied previously.
 

Fitzhugh et al. (1950) fed lindane to groups of Wistar rats at
 

doses of 5 to 1,600 ppm for their life spans. At doses > 100
 

ppm, the chemical produced low incidences of liver enlargement
 

and small foci of necrosis, as seen by gross examination, and
 

hepatic-cell enlargement, hepatic-cell atrophy, fatty degeneration, and focal necrosis, as seen by microscopic examination.
 

The mortality was high in this study, and the mean age at death
 

for rats fed 100 ppm was 64 weeks.
 

Thorpe and Walker (1973) fed 400 ppm lindane (purity > 99.5%) to
 

groups of 30 male and 30 female CF1 mice for 2 years. Among
 

males, hyperplastic nodules of the liver occurred in 38% of the
 

treated animals and in 20% of the controls, while hepatic
 

neoplasms (types not delineated) occurred in 55% of the treated
 

animals and in only 4% of the controls. Among females, hyper-


plastic nodules of the liver occurred in 34% of the treated
 

animals and in 23% of the controls, while hepatic neoplasms
 

(types not delineated) occurred in 34% of the treated animals and
 

in none of the controls. Mortality was high in both sexes of
 

treated animals, and the calculations were based on the incidence
 

of tumors subsequent to appearance of the first liver tumor in
 

the group being analyzed. The concentration of lindane used in
 

35
 
this study was 2-1/2 times the high dose fed to the mice in the
 

present bioassay.
 

Herbst et al. (1975) fed NMRI mice diets containing 12.5, 25, and
 

50 ppm lindane for 80 weeks. No tumor-inducing effect was
 

observed in the livers of these animals. The highest dose used
 

in the Herbst study was less than the low dose used in the
 

present bioassay.
 

It is concluded that under the conditions of this bioassay,
 

lindane was not carcinogenic for Osborne-Mendel rats or B6C3F1
 

mice.
 

36
 
VI. BIBLIOGRAPHY
 

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

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

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

Panel of Carcinogenicity of the Cancer Research Commission
 

of UICC, Vol. 2^, International Union Against Cancer, Geneva,
 

1969.
 

Cox, D. R., Regression models and life tables. J. R. Statist.
 

Soc. B 34(2);187-220, 1972.
 

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

1970, pp. 48-52.
 

Environmental Protection Agency, EPA Compendium of Registered
 

Pesticides, U. S. Government Printing Office, Washington,
 

D.C., 1973, III-L-2.8.
 

Environmental Protection Agency, EPA Compendium of Registered
 

Pesticides, U. S. Government Printing Office, Washington,
 

D.C., 1971, III-L-2.6 - III-L-2.12.
 

Fitzhugh, 0. G., Nelson, A. A., and Frawley, J. P., The chronic
 

toxicities of technical benzene hexachloride and its alpha,
 

beta, and gamma isomers. J. Pharmacol. Exptl. Therap.
 

100(l):59-66.
 

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

significance tests, confidence limits and adjustments for
 

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

Hayes, W. J., Toxicology of Pesticides. Williams and Wilkins Co.,
 

Baltimore, Maryland, 1975, pp. 265-310.
 

Herbst, M. , Weisse, I., and Koellmer, H., A contribution to the
 

question of the possible heptocarcinogenic effects of
 

lindane. Toxicol. 4;91-96. 1975.
 

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

incomplete observations. J. Amer. Statist. Assoc.
 

13:457-481, 1958.
 

37
 
Linhart, M. S. , Cooper, J. A., Martin, R. L. , Page, N. P., and
 

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

and Biomed. Res. 7;230-248, 1974.
 

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

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

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

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

induced by different numbers of administrations of benzo(a)
 

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

Squire, R. A. and Levitt, M. H., Report on a workshop on
 

classification of specific hepatocellular lesions in rats.
 

Cancer Res. 35:3214-3223, 1975.
 

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

j32(3): 679-682, 1975.
 

Thorpe, E. and Walker, A. I. T., The toxicology of dieldrin
 

(HEOD). II. Comparative long-term oral toxicity studies in
 

mice with dieldrin, DDT, phenobarbitone, $-BHC and ^-BHC.
 

Fd. Cosmet. Toxicol. 2:433-442, 1973.
 

38
 
APPENDIX A
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS
 

IN RATS FED LINDANE IN THE DIET
 

39
 

TABLE A1.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS 

FED LINDANEINTHE DIET 

CONTROL LOW DOSE HIGH DOSE
 

ANIHALS INITIALLY IN STUDY 10 50 50
 

ANIMALS KEC6CESIED 10 48 49
 

ANIHALS EXAMINED HISICPATHOLOGICALLY 10 47 46
 

IKTIGUBEKTABY SYS1EB
 

*SKIH (10) (48) (49)
 

PJPILLOMA, NOS 1 (2X)
 

FIBROUS HISTIOCYTCMA, MALIGNANT 1 (2%)
 

*SOBCUT TISSUE (10) (48) (49)
 

FIEECBA 1 (2%) 3 (6»)
 

HESPIEA1C8Y SYSTEM
 

NCNE
 

HEMATCECIETIC SYSTEM
 

tSPLEEN (8) (44) (44)
 

HEHJNGICMJ 3 (7X)
 

CIECUL81CKY SYSTEM
 

NCNE
 

DIGESTIVE SYSTEM
 

ISALIVJEY GLAND (8) (39) (44)
 

SABCCMA. NOS 1 (3%)
 

tLIVEE (10) (45) (45)
 

NEOFLASIIC NODULE 3 (1%) 2 (4X)
 

UBINAEY SYSTEM
 

tKIDNEY (10) (46) (46)
 

1 (2X1
 

* NUHBEB OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUHEfE OF ANIHALS NECEOPSItC
 

41
 
TABLE A1. MALE RATS: NEOPLASMS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

HIXI E TUHOfi , MALIGNAN T 1 (2%) 

tHABJBTCM A 1 (2%) 

ENEOCBIKI SYSTEM
 

tPITOITAEY (10) (32) (35)
 

CAfiCINOHA,NOS 1 (3«)
 

ADJEKCMA, NCS 2 (6«)
 

CHECHOPHOBE ADENOMA 3 (9X) 1 (3X)
 

•ADRENAL	 (10) (37) (38)
 

COBTICAL AE£NOMA 1 (3X)
 

tTHYBOIC (6) (37) (37)
 

FOLLICULAB-CELL AEENOMA 1 (17S) 5 (14%)
 

FCLIICULAS-CELL CARCINOHA 1 <3X) U (11%)
 

C-CIIL ADENCMA 1 (17X) 3 <8X) 1 (3X)
 

tPABATHYBOIE (3) (23) (28)
 

AEENCflA, NOS 1 (H%)
 

*EANCBEATIC ISLETS (9) (41) (39)
 

ISLET-CELL ADENOMA 2 (22%) 1 (2«) 1 (3%)
 

BEFEOECC1IVE SYSTEM
 

*MA«MAEY GLANE (10) («8) (U9)
 

CABCINCHA,NOS 1 <2X)
 

ADENCMA, NOS 2 (4X)
 

NEBVOOS SYSTIH
 

NONE
 

SPECIAL EEBSE CRGANS
 

NCKE
 

BUSCUICSKiltTAL SYSTEt
 

* NUMBEB OF ANIMALS HUH 1ISSUI EXAMINED HICROSCOPICALLY
 

* NUMEEB OF ANIMALS UECBOFSIEE
 

'	 This is considered to be a benign form of the malignant mixed tumor
 

of the kidney and consists of lipocytes, tubular structures, and
 

fibroblasts in varying proportions.
 

42
 
TABLE A1. MALE RATS: NEOPLASMS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

BODY CAVITIES
 

*PLEUHA (10) (48) (49)
 

HESCTHELICKA, NOS 1 (2%)
 

ALL CTHEB SYSTEMS
 

NONE
 

ANIHAI EJSPCSITION SUEHABY
 

ANIHALS INITIALLY IK STUDY 10 50 50
 

Nil OPAL DEATHi 14 13
 

HORIEONE SACBIFICE 11 13
 

SCHICULED SACBIFICE
 

ACCIEEN1ALIY KILIEC
 

TEBHINAL SACBIFICE 25 24
 

ANIHAL HISSING
 

INCLUDES AUTCLIZED ANIHALS
 

TUMOB SOEHAGY
 

TOTAL ANIMALS WITH EBIMABY TUfiORS* 19 17
 

TOTAL PBIHABY TUBOBS 24 23
 

TOTAL ANIMALS WITH BEKIGN TUHCBS 13 13
 

TOTAL BEMIGN TOMOES 15 16
 

TOTAL ANIHALS HITH HALIGNANT TOHOBS 5 <t
 

TCTAI HALIGNANT TDMCBS 5 5
 

TOTAL ANIBALS NITU SECONDARY TUMORSt
 

TOTAL SICCNDABY TDDCHS
 

TOTAL ANIMALS HITH TOBOES UNCEBTAIB­ BBNIGN OB MALIGNANT 4 2
 

TOTAL UNCEBTAIN TOHOfiS 4 2
 

10TAL ANIHALS HITH TUHOES UNCERTAINPRIHAEY OB HETASTATIC
 

TOTAL UNCEBTAIN TUHOBS
 

* PBIHARY TUHOBS: ALL TOHOBS EXCEPT SECONDARY TOHOHS
 

t SECONIABY TOHOBS: HETASTATIC TUMORS OB TOHOBS INVASIVE INTO AN ADJACENT GBGAN
 

43
 
TABLE A2.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE RATS 

FED LINDANE INTHE DIET 

CONTROL LOW DOSE HIGH DOSE
 

ANIHAIS ISIT I ill! IN STOEY 10 50 50
 

ANIMALS SECBOESIED 10 50 50
 

ANIKMS E2AMISED HIS1CPATHOLOGICALLY 10 49 49
 

INTEGUaENTABY SYSTEM
 

fSUBCCI 1ISSOE (10) (50) (50)
 

SSEiCCHA, NOS 1 (10*) 1 (2%)
 

FIBRCHA 1 (2%)
 

LIECHA 1 (2X)
 

EESPIFATCBT SYS1EH
 

SLUNG (10) (US) (1*7)
 

SAflCCMA, NOS, HE1ASTATIC 1 (2»)
 

HEHATCECIITIC SYSIEH
 

KCNE
 

CIBCUIA1CBY SYS1EM
 

SCNE
 

DIGESTIVE SYSIEH
 

#LI¥EE (10) (48) (45)
 

Bill EOCT JDESOHA 1 (2X)
 

HEOPIAS1IC NODOIE U (8*) 2 <4X)
 

•BILE CUCT (10) (50) (50)
 

Bill COCT ADENOMA 2 (4X)
 

HAHAETCflA 1 (2*)
 

tSTCMACH (9) (<*8) (47)
 

LEICHYOMA 1 (2*)
 

OIISABY £YS1£H
 

# 80BBEB OF ? ,1I«ALS HITH 1ISSOE EXAHIBED MICROSCOPICALLY
 

OF ANIHALS NECEOPSIED
 

44
 
TABLE A2. FEMALE RATS: NEOPLASMS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

EBCCCBIKE SYS1JM
 

#PITUI1ABY (7) <»5> (41)
 

C4BCINOHA.NOS 1 (2%)
 

ADEKCMA, NCS 2 (5»)
 

CHBCMOPHCEE ADENOKA 3 («*3X) 14 (31X) 8 (2C«)
 

tADBENAL (9) (42) (44)
 

COE1ICAL JEENOMA 3 (7«) 2 (5*)
 

#THYBCIE (8) C*4) (42)
 

FOLIICULAB-CELL AEEHCMA 1 (2«) 1 (2X)
 

FCLIICUIAB-CELL CABCINOMA 1 <2X)
 

C-CIIl ADENOMA 4 (9X) 3 (7«)
 

fPANCSEATIC ISLETS (10) (48) (47)
 

ISLET-CELL ADENOMA 2 <4X)
 

BEPECECCIIVE SiSTJH
 

*BAH«AEY GLANE (10) (50) (50)
 

CAECINOBA, NCS 1 (10X) 1 (2X)
 

AtEKCBA , NCS 3 ((>%} 1 (2%)
 

ADENCCABCINOMA, NOS 1 (10%)
 

FIEBCMA 2 (4X) 1 (2*)
 

FIBBCADENCBA 3 (30*) 12 (24X) 9 (18X)
 

tDTEBOS (9) (47) (44)
 

ADENCCABCINOHA, NOS 1 (2%)
 

LEICBTfOSAFCOMA 1 (2X)
 

ENCCHEIBIAI S180MAL PCLYP 1 (11S) 6 (13*) 7 (16S)
 

*tTEBO£/£NDCHETBIUM (9) (47) (44)
 

CABCINOHA.NOS 1 (2«)
 

iOVABY (8) (46) (44)
 

SEBTCLI-CE1L TOMOB 1 <2S)
 

SEBVODE SYSliB
 

#EHAIN (9) (49) (49)
 

OLIGCCENEEOGLIOMA 1 (2%)
 

SPECIAL SENS E CEGAN S 

* JiOMBEfi OF ANIMAL S HJ1H TISSUE EXAHINED MICROSCOPICALLY 

* SOflEEE OF ANIMAL S NECBCPSIE E 

45 
TABLE A2. FEMALE RATS: NEOPLASMS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE
 

HUSCUICSKELE-IAL SYS1EH
 

NONE
 

BODY CAVITIES
 

*AEDCHINJL CAVITY (10) (50) (50)
 

LIECFA 1 (2%)
 

ALL CTEIE SYSTEMS
 

*HULTIELE ORGANS (10) (50) (50)
 

SAECCBA, NCS 1 (2X)
 

FIBBCUS HISTIOCYTCMI, HALIGNANT 1 (2X)
 

SITE UtKNCH N
 

CAECINOHA.NOS 1
 

ANIBAL EISECSIIION SOKHAEY
 

ANIHilS ISITIAiLY IK STUDY 10 50 50
 

MitUEAL DEATHS 2 <t
 

HOEIEOND SACRIFICE 16 16
 

SCHitULED SACRIFICE
 

ACCIEENIALIY KILLED
 

TEBBINAL SACRIFICE 32 30
 

ANIBAL HISSING
 

* NUHBEE CF ANIMALS HITH TISSUE EXAMINED HICBOSCOPICALLY
 * NUMEE E OF JNIHALS NECROPSIEE
 

46
 
TABLE A2. FEMALE RATS: NEOPLASMS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE
 

TUHOB SGKMAE Y
 

TOTAL ANIHALS WITH EBIHABY ICHORS* 9 40 31
 

TOTAL PEIMARY TUMCES 10 65 42
 

TOTAL ANIHALS WITH EENIGN TUMORS 6 37 29
 

TCTAI BENIGN TUMOES 7 54 36
 

TOTAL INIflALS HITH MALIGNANT TUMORS 3 6 4
 

TCTAI MALIGNANT TOMC8S 3 7 4
 

TCTAL ANIHALS HITH SECONDARY TUMOBS* 1
 

TCIAI SECCNDABY TDHCHS 1
 

TOTAL AKIBALS WITH TUHCBS ONCEBTAIN-
 BENIGti OB HAIIGNANT 4 2
 

TCTAL UtlCHETAIN TOMOBS <t 2
 

TOTAL ANIHALS HITH IUHOES UNCERTAINfRIHAtY OE HETASTATIC
 

TOTAL UKCEBTAIN TUMCBS
 

* PBIKABY TUHCES: ALL TUHORS EXCEPT SECONDARY TUMOBS
 

# SECCNEARY TUHOBS-: HE1ASTATIC TUMORS OB TOBOBS INVASIVE INTO AN ADJACENT C8GAN
 

47
 

APPENDIX B
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS
 

IN MICE FED LINDANE IN THE DIET
 

49
 

TABLE B1. 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE 

FED LINDANEINTHE DIET 

CONTROL LOW DOSE HIGH DOSE 

ANIMA1S JMIIIAIIY Ili STCDY

ANIBAIS kECBOFSIED

ANIHA1S EXAHINiD HISTCPATHOLOGICA LtY

 10

 10

 10

 50

 50

 50

 50 

 47 

 47 

INTEGOHEKTA6Y SYSTEM 

*SKIN

FIB60US HISTIOCYTCMA

*SOBCUI TISSUE

HEUANGICHA

 (10)

 (10)

 (50)

 (50)

 (47) 

1 (2X) 

 (47) 

1 (2$) 

BESPIBA1CHY SYSTEM 

iLONG

HEPATOCILIOLAB CARCINOMA, BETAST

AIVICLAB/BBCNCHIOIAB ADtNOHA

 (10)

 2 (208)

 (50)

 2 (4X) 

2 (4X)

 (47) 

3 (6%) 

H£fllTC£CIETIC SYS1ZH 

*UULTI£IE C6GANS

HAIIGNANT IYHFHCBA, MOS

 (10) (50) (47) 

1 (2%) 

CIECOIA1C8Y SYSTEM 

NONE 

DICE SI Hi SISTER 

*LIVEB

HtOFLASTIC NODDLE

HEPA10CELIULAB CABCIBCHA

 (10)

 1 (10*)

 2 (20%)

 (49)

 19 (39X)

 (46) 

1 (2%) 

9 (20X) 

DEINABY SYSTEM 

t kOHBEE OF ANIMALS BITH IISSOE EXAMINED MICBOSCOPICALLY 

* NUBEEB OF ANIHALS NECBOPSIEE 

51
 
TABLE B1. MALE MICE: NEOPLASMS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

EHLCCBI*J SYSTEM 

NONE 

BEEBCCCC1IVE SYSTEM 

*EPICIEYMIS

HEEAIOCEIIULAB CABCINCBA, METAST

 (10) (50)

 1 (2%) 

 (47) 

NEBVOUS SYSTEM 

NONE 

SPECIAL SIKSE CBGANS 

NCKE 

HUSCOICSKELETAI SYSlEe 

NONE 

BOEX CAVI1IES 

NCKE 

ALL CTHEF SYS1EMS 

DIAPHtSGM 

HEEATCCELLDLAB CAECINCHA, HETAST 1 

ANIBAL CISECSITION SUHBABY 

SNIHA1S INITIALLY I* STUDY 

NA1UEAL EEATUd 

MOB1EUNE SACRIFICE 

SCHIIULED SACBIFICE 

ACCICENTAILY KILLED 

TEBBINAL SACBIEICE 

AHIKJ1L HISSING 

10 50 

144 

50 

* NUHEEF OF ANIMALS HITH 1ISSOE EXAMINED HICBOSCOPICALLY 

* NUHEEB OF ANIMALS NECBCESIEE 

52
 
TABLE B1. MALE MICE: NEOPLASMS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE
 

TUMOB SUKKAfiY
 

TOTAL JNIBALS HITS EBIBARY IUKOBS* 4 21 15
 

TOTAI PBIHABY TUHCBS 5 21 16
 

TOTAL INIBAiS HUB EENIGN TUBCBS 2 2 5
 

TOTAI BENIGN TUBOES 2 2 C
 

TOTAL ANIHALS WITH M4IIGNANT TUMOfiS 2 19 10
 

IOTJ1 MALIGNANT TUflCES 2 19 10
 

TOTAL ANIHAIS WITH SECCNDABY TUMORS* 3
 

TCTAI SECCNDAfiY TUBCfiS 4
 

TOTAI ANIHALS WITH TOHCES UNCERTAINBENIGN OR HALIGNAN1 1 1
 

TOIA1 UNCiBTAIN TUHCRS 1 1
 

TOTAI ANIflALS KITH 1UBOBS UNCERTAINPBIBABJ OB HETASTATIC
 

TOTAL UNCERTAIN 1DBCBS
 

* PBIMABY TUflOES: AIL TUHORS EXCEPT SECONDARY TUMOBS
 

# SECCNEABY TUMORS: ME1ASTATIC 1UBORS OR TUMORS INVASIVE INTO AN ADJACENT CBGAN
 

53
 
TABLE B2.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICE 

FED LINDANE INTHE DIET 

CONTROL LOW DOSE HIGH DOSE 

ANIMALS milAIIY IK STUDY 10 50 50
 

ANIHAIS KECEOESIEC 10 <»9 49
 

ANIBALS EXAMINED HIS10PATHOLOGICALLY 10 49 19
 

IHTEGOBEKTAEY SYSIEB
 

NC8E
 

BESEIEMCBY SYS1EB
 

*LUNG (10) (48) (48)
 

ALVECLAB/EEONCfclOLAB ADZNCBA 1 (2%) 2 («X)
 

AIVECLAfi/BECNCHIOLAB CAECINCMA 1 (10S)
 

OSTICS4ECCBA, HETSS1A1IC 1 (2*)
 

HEMA1CICIITIC SVS1EH
 

*SELEEK (8) (49) (48)
 

FIEEOUS HISTIOCYTCBi 1 (13S)
 

CIECUIAICBY SYS1EB
 

NONE
 

DIGES1IDI SYS1EB
 

tLIVEB (10) (47) (46)
 

NEOJLASTIC NOCULE 1 (10*) 2 (4X)
 

HEPA10CEILULAR CABCINOHA 2 (4X) 3 (7X)
 

tcEcua (1)
 

SA6CCBA, NCS 1 (100)1)
 

DBIMABY 5YS1EH
 

* NDBBEB CF ANIMALS HItH 1ISSUE EXAMINED MICROSCOPICALLY
 * NUMEEE OF ANIMALS NfiCEOESIEC
 

54
 
TABLE B2. FEMALE MICE: NEOPLASMS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

ENDCCBISE SYSIEB 

IADB£NAI

CORTICA L CARCINOM A

 (7) (46) (44) 

1 (2%) 

•THYKCJC

FOLLICULAE-CELL AEENCHA

 (7) (43)

 2

 (41) 

 (5X) 

H£PSCEUC1IV£ SSSTEH
 

*MAHHABY GLANE (10) (49) (49)
 

AC£KCHA, NOS 1 (10X)
 

tlTESUS (7) (44) (43)
 

SABCCHA, NOS 1 (2%)
 

NEE»00 £ SXS1EB 

NCN E 

SPECIAL £EilS£ CBGANS 

NCNE 

BUSCULC£KEL£1AI SISlfH
 

*SACHUH (10) (49) (49)
 

OSIICSAECCBA 1 (2%)
 

BOCK CAVI1IES
 

NCNE
 _ ^ «••_•• •> «M_v«_A^«o«K^_ •.•«_••*— —•.— ——•••*•—•«•——•*»••••»—•••— ««*««^*«»'V»»<- ———••-.••—— v^^^^l 

All 01EEB SIS1EHS 

* NOH££fi OF ANIMALS HUH IISSU6 EXAMINE D BICBOSCOPICALLY 

* HOflEEB OF ANIflAlS NECBOESIEE 

55 
TABLE B2. FEMALE MICE: NEOPLASMS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

ANIHAL USEOSITION SUHBABY
 

ANIflAIS INITIALLY IS STUDY 10 50 50
 

NA1CBAL DEATHS 1 2 2
 

B06IEUND SACBIFICI 1 3
 

SCHIIULED SACBIFICE
 

ACCIEENTALIY KIL1ED
 

TEBBINAI SACBIFICE 8 45 48
 

ANIHiL HISSING
 

a INCIOtIS AUTOLYZED ANIHALS
 

TUBOB SCKBABY
 

TOTAL ANIMALS WITH EBIMAB 4 Y TUKOBS* 7 8
 

TOTAL PBIMABY TUMOBS 4 7 9
 

TOTAL ANIMALS WITH 2 EENIGN TUHCBS 1 4
 

TCIAI BESIGN TUBOBS 2 1 4
 

TOTAL ANIMALS WITH BAIIGNAN 1 T TUMOBS 4 5
 

TCTAI MALIGNANT TUBOBS 1 4 5
 

TOTAL INIHALS KITH SECCNDABY TUMORS* 1
 

TCTAI SECCNDABY TDMCBS 1
 

T01AL ANIMALS WITH TUBOBS UNCEBTAINEENIGK OB MALIGNANT 1 2
 

TOliL ONCEBTAIN TUBOBS 1 2
 

TOTAL ANIHAIS WITH TOBOBS UNCEHTAINPSIHAEY OB HETASTATIC
 

TOTJI UNCEBTAIN TUBCBS
 

* PfilflABY TOBOBS: ALL TUBOBS EXCEPT SECONDARY TUBOBS
 « SECCNCABY TUBOfiS: HE1ASTATIC TUBOBS OB TUMOBS INVASIVE INTO AN ADJACENT CKGAN
 

56
 
APPENDIX C
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS
 

IN RATS FED LINDANE IN THE DIET
 

57
 

TABLE C1.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS 

FED LINDANEINTHEDIET 

CONTROL LOW DOSE HIGH DOSE
 

ANIHAIS milAlLY IS STUDY 10 50 50
 

ANIMALS MCFOESIED 10 46 49
 

ANIBALS EXAMINED HISTCFATHCLOGICALLY 10 47 46
 

INTEGCflESTABY SYSTEH
 

*SKIN (10) (48) (49)
 

HYEEBKEBATCSIS 1 (10X)
 

ACANIfcOSIS 1 (10X)
 

BESEIEATCBY SYSTEM
 

#LUNG (9) (46) (46)
 

AI£LECTASIS 1 (2X)
 

PNEUMONIA, CHRONIC BDBINE 1 (2%)
 

NECBCSIS, FOCAL 1 (2%)
 

HEMATOECIEIIC SYSTEM
 

#SELEES 

HEHCfifiHAGE 

INFLAMMATION, GBASUICBATOUS 

SCIEBOSIS 

HEHATCECIESIS 

HYPCEtASIA, LYHPHCID 

(8) (44) 

2 <5X) 

1 (2%) 

1 <2X) 

1 (2%) 

(44) 

1 (2X) 

#KESEBTEBIC L. NODE 

INFLAMMATION, NOS 

(10) 

1 (10%) 

(3D (34) 

CIBCUIATCBY SYSTEM 

tMYOCABEIUH INFLAMMATION , CHRONI C FOCAL 

(9) 

1 (11X) 

(46) (46) 

tEKDCCASDIUM 

SCLEBOSIS 

(9) (46) (46) 

1 (2X) 

t NUMBEB OF ANIMALS WITH TISSUE EXAHIMED MICROSCOPICALLY
 * NUHEEE OF ANIHAIS NECBOPSIEC
 

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

CONTROL LOW DOSE HIGH DOSE
 

DIGESIIlil SYS1EB
 

tilVEfl (10) (45) (45)
 

CIBEBOSIS, NOS 1 <2X)
 

DEGENEBATIGN, BALLOONING 1 (2X)
 

NECECSIS, FOCAL 1 (2X)
 

flETABOBEHOSIS FATIY 3 (7X) 5 (11X)
 

FOCAI CEL10LAR CHANGE 1 (10*) 1 <2X) 1 (2X)
 

ANGIECTASIS 1 (2X) 2 (4X)
 

*BILE EDCT (10) C*8) C«9)
 

DILATATION, NOS 1 (2X)
 

INFLAMMATION, CHECNIC 1 (2*)
 

INFIABBATICN, CHBCNIC FCCAL 1 (10%) 2 (UX)
 

HYEEEPLASIA, NOS 2 (20X) 3 <6X)
 

HYPEBPLASIA, FOCAL 2 (4X)
 

IEANCEEAS (9) (41) (39)
 

INFLAMMATION, CtBONIC 1 (11*)
 

•STCBACH	 (9) (43) (38)
 

EBOSICN 1 (11X) 1 (3X)
 

CALCIFICATION, DYSTBOPHIC 1 (3X)
 

IGASTEIC MUCOSA (9) (43) (38)
 

CALCIFICATION, DYSTBOPHIC 1 (2X)
 

UBINABl SYSIEH
 

tKIDMEY (10) (U6) («6)
 

INFLAMMATION, DIFFUSE 1 (2»)
 

INFLAMMATION, CHBCBIC 5 (SOX) 22 (4SX) 28 (61X)
 

PYEICNEPHEITIS, CHBOHIC 1 <2X)
 

tBENAL IAPILIA (10) (46) (46)
 

CALCIOM DEPOSIT 1 (2X)
 

*naiHABY BLADDEB (9) (34) (37)
 

INFLAMMATION, ACUTE 1 (11X)
 

*DBETHEi (10) (<»8) (49)
 

INFLAMMATION, SUPEUBATIVE 1 <10X)
 

ENDOCEINE SYS1EM
 

•PITUITABY	 (10) (32) (35)
 

qYSI. NOS 2 (2Q SI 2 J§?L 4 H1X)
 

* HOBBEB OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* SOBEEB OF ANIMALS NECBOPSIED
 

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

CONTROL LOW DOSE
 

DEGENERATION, CYS1IC 1 (3X)
 

ASGIECTASIS
 

tADRENAL MEEULLA (10) (37)
 

HYPBEPLASIA, NOS
 

tTHYECIt (6) (37)
 

HYPEEFLASIA, C-CEIL 3 <8X)
 

HYEEBPLASIA, FOLLICULAR-CELL 1 (17X) 4 (11X)
 

tPABAIHYECID (3) (23)
 

HYPEEPLASIA, NOS 1 (<**)
 

REEBOCUCTIVE S1STEB
 

fPBOSTME (9) (38)
 

INFLAMMATION, SUPPUHATIVE 1 (11X)
 

INFIABHATICN, ACUTE
 

INFIAMflATICN, CBECNIC 1 (3X)
 

tTESTIS (10) (43)
 

PEBIABTERITIS 1 (2X)
 

ATRCEHY, NOS 4 (40X) 1 (2X)
 

A1BOEHY, FCCAI 1 (2%)
 

ATBOfHY, DIFFUSE 1 (2%)
 

tTESTIS/TUEULE (10) (43)
 

DEGENERATION, NOS 1 (2%)
 

BEEVOCS SYSTEM
 

NONE
 

SPECIAL £ENSE CBGANS
 

NONE
 

BUSCULCSKELE1AI SYSIE8
 

NCNE
 

BCCY CAVITIES
 

*HESENTEBY (10) (48)
 

* NUMBEB OF ANIflAIS HITH TISSUE EXAHIMED MICBOSCOPICALLY
 * NUMEEE OF ANIMALS NECBOPSIED
 

HIGH DOSE
 

1 (3X)
 

1 (3X)
 

(38)
 

1 (3X)
 

(37)
 

1 (3X)
 

3 <8X)
 

(28)
 

2 (7X)
 

(37)
 

3 (8X)
 

(44)
 

1 (2X)
 

8 (18X)
 

(44)
 

(U9)
 

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

CONTROL LOW DOSE HIGH DOSE 

ALl CTHE6 SYSTEMS 

NCNE 

SPECIAL KCBEHCLCGY SUHHSRi 

NO IESICN REPORTED 1 10

AUTC/KECKCESY/HISIO PEBF 4

AU1C/NECRCESY/NO HISTO 1

ADlCI?SIS/JiO NECRCES? 2

* NUMBER GF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* NOHEEfi OF ANIMALS NICfiCESI£E 

6 

2 

3 

1 

62
 
TABLE C2.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS 

FED LINDANEINTHE DIET 

CONTROL LOW DOSE HIGH DOSE
 

ANIMALS INITIALLY IN STUEY 10 50 50
 

ANIMALS UCEOESIEC 10 50 50
 

ANIMALS EXAMINED HISICEATHOLOGICALLY 10 49 49
 

INTEGUMEKTABY SYSTEM
 

*SKIN (10) (50) (50)
 

GBANULOKA, NOS 1 (2*)
 

fiESEIEIICBY SYSTEM
 

tLUNG (10) (48) (47)
 

INFLAMMATION, NOS 1 (2%)
 

INFLAMMATION, FOCAL 1 (2%)
 

ABSCISS, NOS 1 (2%)
 

INFLAMMATION, CHBCNIC 2 (IX)
 

HEMAIOECIETIC SYSTEM
 

*ABBCMINAL LYMPH NODE (9) (37) (39)
 

INFLAMMATION, CHBCNIC 1 (3S)
 

tMESENJEEIC L. NOEE (9) (37) (39)
 

INFLAMMATION, NOS 1 (11S)
 

CIBCULA1C8Y SYSTEM
 

NCNE
 

DIGESTIVE SYSTEM
 

*SALIV«BY GLAND (9) (45) (43)
 

DIGESEEATION, CYSTIC 1 (2X)
 

*LIV£B (10) (<*8) (45)
 

2 (4X) 2 (4«1
 

t NUMBEB OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUHEEB OF ANIMALS NECECFSIED
 

63
 
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

EASCEHILIC CYTO CHANGE 2 (4X)
 

FOCAL CELLULAR CHANGE 5 (10X) 4 (9X)
 

ANGIECTASIS 5 (10X) 2 <4X)
 

*BILE DUCT (10) (50) (50)
 

INFLAMMATION, NOS 1 (2%)
 

HYEEEELASIA, NOS 1 (2X)
 

•ESCEHAGUS	 (1) (15) (12)
 

HYEEBKEBATCSIS 1 (7X) 1 (8X)
 

UEINARY SYSTEM
 

•KICNEY	 (10) (46) (46)
 

CYST, NOS 1 (2%)
 

INFLAMMATION, CHBCNIC 3 (7X) 5 (10%)
 

•KICKEY/EELVIS	 (10) (46) (48)
 

BEIAELASIA, NOS 1
 

ENDOCBINE SYSTEM
 

•PITUITARY	 (7) ("5) (41)
 

CYST, NOS 1 (2%)
 

•ADRENAL	 (9) (42) (44)
 

HEMCBBHAGE 3 (7X)
 

DEGENERATION, CYSTIC 1 (2*) 2 <5X)
 

ANGIECTASIS 2 (22%) 3 (7X)
 

•ADRENAL	 MECULLA (9) (42) (44)
 

HYPfBPLASIA, NOS 1 (11X)
 

•THYBCIE	 (8) (44) (42)
 

HYEESPLASIA, C-CE1L 1 (13)1)
 

HYEEBELASIA, FOLLICULAR-CELL 2 (5X) 3 (7X)
 

BEEEOCCC1IVE SYSTEM
 

*KAMflAEY GLAKt (10) (50) (50)
 

HYEEEPLASIA, NOS 1 (10*)
 

ADEKCSIS 1 (10X)
 

•UTEROS	 (9) (47) (44)
 

HYCBCHEIBA 1 (2*1
 

t NUHBEB OF ANIHAIS HUH TISSUE EXAMINED MICROSCOPICALLY
 * NONEEfi OF ANIMALS NICBOPSIEE
 

64 
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

NEfiVCCS SYSTEM
 

NONE
 

SPECIAI EESSE CEGAKS
 

HCNE
 

HUSCULCSKELETAL SYSTEM
 

NCNE
 

BODY CAVJ1IIS
 

KCNI
 

ALI C1HIJ SXS1IHS
 

NCNE
 

SFHCIAI eCEEHCLCGY SOBHABY
 

NO IESICN EEPCBTEt 5 7
 

AUTC/KECEOESY/HIS10 PEBF 2
 

A01C/NECBOESY/NO HISTO 1 1
 

* NUHEES OF ANIMALS HUH TISSUE BXAHISBD MICROSCOPICALLY
 * NUHEEB OF ANIMALS NECBOESIEE
 

65
 

APPENDIX D
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS
 

IN MICE FED LINDANE IN THE DIET
 

67
 

TABLE 01.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE MICE 

FED LINDANEINTHE DIET 

CONTROL LOW DOSE HIGH DOSE
 

ANIMALS INITIALLY IN STGDY 10 50 50
 

ANIMALS KECSOFSIED 10 50 <J7
 

ANIHAIS EXAMINED HISTCPAIHOLOGICALLY 10 50 47
 

IKIIGOCEKTAEY SYS1EH
 

NCNE
 

EESPIEAICEY SYSTEM
 

tiCKE
 

HEMATCICIETIC SYSTEM
 

«SPLEE» (10) (50) (47)
 

CONGESTION, PASSIVE 1 (2JS)
 

HYPEEPLASIA, LYMPHOID 1 (2%)
 

tCEBVICAL LYMPH NODE (9) (42) (37)
 

INFLAMMATION, FOCAL 1 (3%)
 

CIHCOIA1CBY SYSTEM
 

KCNE
 

DIGESIIVE SYSlEfl
 

tllVEB (10) (49) (46)
 

INfLAMHATICN, CKECNIC 1 (2X)
 

CIOtEY SHELLING 3 (7«)
 

FOCAI CELLULAB CHANGE 1 (2%)
 

*BILE COCT (10) (50) (47)
 

DILATATION, NOS 1 (10X)
 

UEINABY £YSTEH
 

tKIDNEY (10) (50) (47)
 1
 * NUflEEB OF ANIMALS WITH 1ISSDE EXAMINED MICfiOSCOPICALLY
 * NOMEEE OF ANIHALS NSCBOPSIEC
 

69
 
TABLE D1. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE
 

IHFLABHATICN, CHRONIC FCCAL 1 (2X)
 

INEOCB1KI SYS1EB
 

NCNE
 

EEPEODCCIIVE SYSTEK
 

SCNE
 

KEBVCUS SYSIEB
 

NCNE
 

SPECIAL SENSE CEGANS
 

NCKE
 

MUSCULCSKELETA1 SYSTEK
 

NONE
 

EOCY CAVI1IEE
 

NCNE
 

ALL CTHEF SYS1EHS
 

NCNE
 

SPECIAL eCBEHCIOGY SUCMSR1
 

SO IESICN BEPCBTEE 6 26 25
 

ADTC/KECECESY/HIS1C PEKF 1 1
 

A01CLYSIS/NO NECROPSY 3
 

» NUHEEE OF ANIMALS HITH TISSUt EXAMINED MICROSCOPICALLY
 * NUMBER OF ANIHALS NECBOPSIEE
 

70
 
TABLE D2.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE 

FED LINDANE INTHE DIET 

CONTROL LOW DOSE HIGH DOSE
 

ANIMALS mTIAIIY IK STUDY 10 50 50
 

ANIMALS KECBCESIEC 10 49 49
 

ANIMALS EXAMINED HISIOPATHOLOGICALLI 10 49 49
 

INTEGUMiKTABY SYSTEM
 

NCNE
 

BESEIFAICBY SYSTEM
 

NCNE
 

HEMATCICIETIC SYSTEM
 

tSELEEN (8) (49) (48)
 

HYEE6ELASIA, NODULAfi 1 (2*)
 

HYPEfiPLASIA, LYMEHOID 2 (4%) 3 (6%)
 

CIBCOLATCEY SYSTEM
 

NCNE
 

DIGESTIVE SYSTEM
 

#IIVEE (10) (47) (46)

 

METAMCHfHOSIS FATTY 1 (10X) 

*BILE CCCT (10) (49) (49) 

CYST, NOS 1 (2X) 

fFANCFEAS (9) (46) (48) 

DILATATICJi/DUCTS 1 (2%) 

tEEYEES EA1CH (9) (48) (48)

 

2 

* NUMEEf i OF ANIMAL S HUB TISSUE EXAMINE D MICBOSCOPICALLY 

* NUMEh E OF ANIMAL S lifCEOPSIEC 

71 
TABLE D2. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

UHINAEY SYSTEM
 

#KICNEY
 

INFLAMMATION, INIEBSTITIAL
 

INEIAHMATICli, CHBCHIC
 

ENECCBIKE SYSTEM
 

NCN E
 

REEBCEGCTIVE SYSTEM
 

#UTEBUE
 

HYIECHEIEA
 

*C1ERU£/JNCCM£TEIUM
 

HYfEFELASIA, HOS
 

HYEEBELASIS, FOCAL
 

HiPEBPLASIA, CYSTIC
 

ICVAEY
 

FCIIICULAR CYST, NOS
 

INE1AHMATICN, NOS
 

INFLAHMATICN, SUFEUBA1IVE
 

IN5IABMATICN, CHBCNIC
 

KEBVOUS SYSTEM
 

NCNE
 

SPECIA L SENS E CEGAN S 

NCN E 

HUSCULCSKELETAL SYSTEK
 

NONE
 

BCCY CA1/ITIES 

JJIJB1 

CONTROL LOW DOSE HIGH DOSE 

(10) (49) (47) 

2 (20S) 

1 (10SS) 1 (2X) 

(7) (44) (43) 

1 (2X) 

(7) (44) (43) 

1 (25!) 

1 (2*) 

3 (7X) 2 <5X) 

(7) (42) (46) 

2 (5*) 2 (4S) 

2 (29X) 2 (58) 5 (11X) 

1 (14X) 10 (24X) 2 (4S) 

1 (2«) 

* NUHEE6 OF ANIMALS HUH TISSUE EXAMINED MICBOSCOPICALLY
 * NUMEEfi Of ANlflALS KECBOPSIEt
 

72
 
TABLE D2. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

A LL CTHi F SYSTEM S 

NCN E 

SPECIAL KCBPHCLOGY SUBMABY 

NO IHSICN EEPCRTEE 3 26 27 

AUTC/KECECPSY/HISTO PEBF 1 

AUTCIYSIS/NO NECBCPSY 1 1 

* NOHBEB OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 * NUHE£E OF ANIMALS NICECESIEE
 

73
 

APPENDIX E
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS
 

IN RATS FED LINDANE IN THE DIET
 

75
 

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

Fed Lindane in the Dieta
 

Topography: Morphology


Subcutaneous Tissue: Fibromab


P Valuesc»d


Relative Risk (Matched Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Pooled Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Liver: Neoplastic Noduleb


P Valuesc»d


Relative Risk (Matched Control) f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Pooled Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Matched


 Control


 0/10 (0.00)


 N.S.


 —
 

 0/10 (0.00)


 N.S.


 —
 

 Pooled
 

 Control
 

 0/49 (0.00)
 

 N.S.
 

 0/49 (0.00)
 

 N.S.
 

Low
 

Dose
 

1/48 (0.02)
 

N.S.
 

Infinite
 

0.012
 

Infinite
 

Infinite
 

0.055
 

Infinite
 

108
 

3/45 (0.07)
 

N.S.
 

Infinite
 

0.149
 

Infinite
 

Infinite
 

0.656
 

Infinite
 

110
 

High
 

Dose
 

3/49 (0.06)
 

N.S.
 

Infinite
 

0.136
 

Infinite
 

Infinite
 

0.602
 

Infinite
 

69
 

2/45 (0.04)
 

N.S.
 

Infinite
 

0.073
 

Infinite
 

Infinite
 

0.322
 

Infinite
 

110
 
00 

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

(continued)
 

Topography: Morphology


Pituitary: Chromophobe
 

Adenomab


P Valuesc'd


Relative Risk (Matched Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Pooled Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Thyroid: Follicular-cell
 

Adenomab


P Values0 »d


Departure from Linear Trend6


Relative Risk (Matched Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Pooled Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Fed Lindane in the Diet3
 

Matched Pooled
 

 Control Control
 

 0/10 (0.00) 6/47 (0.13)
 

 N.S. N.S.
 

 1/6 (0.17) 3/42 (0.0,7)
 

P = 0.028 (N) N.S.
 

P = 0.048
 

 109
 

Low
 

Dose
 

3/32 (0.09)
 

N.S.
 

Infinite
 

0.209
 

Infinite
 

0.734
 

0.125
 

3.148
 

110
 

5/37 (0.14)
 

N.S.
 

0.811
 

0.131
 

37.268
 

1.892
 

0.397
 

11.410
 

110
 

High
 

Dose
 

1/35 (0.03)
 

N.S.
 

Infinite
 

0.017
 

Infinite
 

0.224
 

0.005
 

1.720
 

110
 

0/37 (0.00)
 

N.S.
 

0.000
 

0.000
 

3.028
 

0.000
 

0.000
 

1.869
 

—
 
Table El. Analyses of the Incidence of Primary Tumors in Male Rats
 

Fed Lindane in the Diet3
 

(continued) 

Topography: Morphology

Matched 

 Control 

Pooled

Control

 Low 

 Dose 

High 

Dose 

Thyroid: Follicular-cell 

Carcinomab 0/6 (0.00) 1/42 (0.02) 1/37 (0.03) 4/37 (0.11) 

P Valuesc'd N.S. N.S. N.S. N.S. 

Relative Risk (Matched Control)f 

Lower Limit 

Upper Limit 

Infinite 

0.010 

Infinite 

Infinite 

0.185 

Infinite 

Relative Risk (Pooled Control)f 

Lower Limit 

Upper Limit 

1.135 

0.015 

86.730 

4.541 

0.479 

217.008 

Weeks to First Observed Tumor 85 93 

Thyroid: Follicular-cell 

Adenoma or Carcinoma^ 1/6 (0.17) 3/42 (0.07) 6/37 (0.16) 4/37 (0.11) 

P Valuesc'd , N.S. N.S. N.S. N.S. 

Relative Risk (Matched Control)f 

Lower Limit 

Upper Limit 

0.973 

0.171 

43.453 

0.649 

0.093 

31.077 

Relative Risk (Pooled Control)f 

Lower Limit 

Upper Limit 

0.270 

0.524 

13.112 

1.514 

0.274 

9.698 

Weeks to First Observed Tumor 109 85 93 
Table El. Analyses of the Incidence of Primary Tumors in Male Rats
 

Fed Lindane in the Diet3
 

(continued)
 

Matched Pooled Low
 

Topography: Morphology Control Control Dose
 

Thyroid: C-cell Adenomab

 1/6 (0.17) 2/42 (0.05) 3/37 (0.08)
 

P Valuesc>d N.S. N.S. N.S.
 

Relative Risk (Matched Control) ^ 0.486
 

Lower Limit 0.056
 

Upper Limit 24.871
 

Relative Risk (Pooled Control)f 1.703
 

Lower Limit 0.206
 

Upper Limit 19.427
 

oo
 

o
 

Weeks to First Observed Tumor 109 110
 

Adrenal: Cortical Adenomab 0/10 (0.00) 2/54 (0.04) 0/37 (0.00)
 

P Valuesc»d N.S. N.S. N.S.
 

Relative Risk (Matched Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Pooled Control)f 0.000
 

Lower Limit 0.000
 

Upper Limit 4.894
 

Weeks to First Observed Tumor —
 —
 

High
 

Dose
 

1/37 (0.03)
 

N.S.
 

0.162
 

0.003
 

12.423
 

0.568
 

0.010
 

10.429
 

110
 

1/38 (0.03)
 

N.S.
 

Infinite
 

0.015
 

Infinite
 

0.711
 

0.012
 

13.091
 

103
 
Table El. Analyses of the Incidence of Primary Tumors in Male Rats
 

Fed Lindane in the Diet3
 

(continued)
 

Matched Pooled Low High
 

Topography: Morphology Control Control Dose Dose
 

Spleen: Hemangiomab 0/8 (0.00) 0/52 (0.00) 0/44 (0.00) 3/44 (0.07)
 

P Valuesc»d N.S. P = 0.030 N.S. N.S.
 

Relative Risk (Matched Control) ^ Infinite
 

Lower Limit 0.126
 

Upper Limit Infinite
 

Relative Risk (Pooled Control)f Infinite
 

Lower Limit 0.711
 

Upper Limit Infinite
 

oo
 

Weeks to First Observed Tumor — 97
 

a

Treated groups received time-weighted average doses of 236 or 472 ppm in feed.
 

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

c

Beneath the incidence of tumors in a control group is the probability level for the Cochran-


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

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

the comparison of that treated group with the matched-control group (*) or with the pooled-


control group (**) when P < 0.05 for either control group; otherwise, not significant (N.S.)
 

is indicated.
 

<*A negative trend (N) indicates a lower incidence in a treated group than in a control group.
 

e

The probability level for departure from linear trend is given when P < 0.05 for any comparison.
 

*The 95% confidence interval of the relative risk between each treated group and the specified
 

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

Fed Lindane in the Dieta
 

Matched Pooled Low High
 

Topography: Morphology Control Control Dose Dose
 

Subcutaneous Tissue: Fibroma*5 0/10 (0.00) 0/54 (0.00) 1/50 (0.02) 0/50 (0.00)
 

P Values0 »d N.S. N.S. N.S. N.S.
 

Relative Risk (Matched Control)f Infinite
 

Lower Limit 0.012
 

Upper Limit Infinite
 

Relative Risk (Pooled Control)f Infinite
 

Lower Limit 0.058
 

Upper Limit Infinite
 00
 

KJ
 

Weeks to First Observed Tumor — 110
 _.—.


Liver: Neoplastic Noduleb 0/10 (0.00) 1/49 (0.02) 4/48 (0.09) 2/45 (0.04)
 

P Valuesc»d N.S. N.S.' N.S. N.S.
 

Relative Risk (Matched Control)f Infinite Infinite
 

Lower Limit 0.215 0.073
 

Upper Limit Infinite Infinite
 

Relative Risk (Pooled Control)f 4.083 2.178
 

Lower Limit 0.424 0.117
 

Upper Limit 196.654 125.581
 

Weeks to First Observed Tumor — 104 110
 
00 

Table E2. Analyses of the Incidence of Primary Tumors in Female Rats
 

Fed Lindane in the Diet3
 

(continued)
 

Topography: Morphology


Pituitary: Chromophobe
 

Adenomab


P Valuesc»d


Departure from Linear Trend6
 

Relative Risk (Matched Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Pooled Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Thyroid: Follicular-cell
 

Adenomab


P Valuesc»d


Relative Risk (Matched Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Pooled Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Matched
 

 Control
 

 3/7 (0.43)
 

 N.S.
 

 109
 

 0/8 (0.00)
 

 N.S.
 

Pooled


Control


6/46 (0.13)


N.S.


P = 0.048
 

0/48 (0.00)


N.S.


 —
 

 Low
 

 Dose
 

 14/45 (0.31)
 

P = 0.033**
 

0.726
 

0.321
 

3.371
 

2.385
 

0.954
 

6.877
 

87
 

 1/44 (0.02)
 

 N.S.
 

Infinite
 

0.011
 

Infinite
 

Infinite
 

0.059
 

Infinite
 

110
 

High
 

Dose
 

8/41 (0.20)
 

N.S.
 

0.455
 

0.175
 

2.294
 

1.496
 

0.498
 

4.786
 

95
 

1/42 (0.02)
 

N.S.
 

Infinite
 

0.011
 

Infinite
 

Infinite
 

0.061
 

Infinite
 

110
 
__ 

Table E2. Analyses of the Incidence of Primary Tumors in Female Rats
 

Fed Lindane in the Dieta
 

(continued)
 

Matched Pooled Low
 

Topography: Morphology Control Control Dose
 

Thyroid: Follicular-cell
 

Carcinomab 0/8 (0.00) 0/48 (0.00) 1/44 (0.02)
 

P Valuesc»d N.S. N.S. N.S.
 

Relative Risk (Matched Control)f Infinite
 

Lower Limit 0.011
 

Upper Limit Infinite
 

Relative Risk (Pooled Control)f Infinite
 

Lower Limit 0.059
 

oo
 Upper Limit Infinite
 

Weeks to First Observed Tumor — 110
 

Thyroid: Follicular-cell
 

Adenoma or Carcinomab

 0/8 (0.00) 0/48 (0.00) 2/44 (0.05)
 

P Valuesc»d N.S. N.S. N.S.
 

Relative Risk (Matched Control)f Infinite
 

Lower Limit 0.061
 

Upper Limit Infinite
 

Relative Risk (Pooled Control)f Infinite
 

Lower Limit 0.323
 

Upper Limit Infinite
 

Weeks to First Observed Tumor — 110
 

High
 

Dose
 

0/42 (0.00)
 

N.S.
 

1/42 (0.02)
 

N.S.
 

Infinite
 

0.011
 

Infinite
 

Infinite
 

0.061
 

Infinite
 

110
 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats
 

Fed Lindane in the Dieta
 

(continued) 

Topography: Morphology

Matched 

 Control 

Thyroid: C-cell Adenomab 0/8 (0.00) 

P Values0 >d N.S. 

Relative Risk (Matched Control)f 

Lower Limit 

Upper Limit 

CO 

Ul 

Relative Risk (Pooled Control)f 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor — 

Adrenal: Cortical Adenomab 0/9 (0.00) 

P Valuesc>d N.S. 

Relative Risk (Matched Control)f 

Lower Limit 

Upper Limit 

Relative Risk (Pooled Control)f 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor — 

Pooled


Control


0/48 (0.00)


N.S.


0/51 (0.00)


N.S.


 Low
 

 Dose
 

 4/44 (0.09)
 

P = 0.049**
 

Infinite
 

0.194
 

Infinite
 

Infinite
 

1.013
 

Infinite
 

101
 

 3/42 (0.07)
 

 N.S.
 

Infinite
 

0.145
 

Infinite
 

Infinite
 

0.731
 

Infinite
 

105
 

High
 

Dose
 

3/42 (0.07)
 

N.S.
 

Infinite
 

0.132
 

Infinite
 

Infinite
 

0.689
 

Infinite
 

103
 

2/44 (0.05)
 

N.S.
 

Infinite
 

0.068
 

Infinite
 

Infinite
 

0.343
 

Infinite
 

103
 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats 

(continued) 

Topography: Morphology


Mammary Gland: Adenoma, NOSb


P Values0 »d


Relative Risk (Matched Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Pooled Control)f
 

Lower Limit
 

Upper Limit
 oo

o\ 

Weeks to First Observed Tumor
 

Mammary Gland: Adenoma or
 

Carcinoma, NOSb


P Valuesc'd


Relative Risk (Matched Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Pooled Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Fed Lindane in

Matched
 

 Control
 

 0/10 (0.00)
 

 N.S.
 

 1/10 (0.10)
 

 N.S.
 

 the Diet3 

Pooled


Control


0/47 (0.00)


N.S.


1/47 (0.02)


N.S.


 100
 

 Low
 

 Dose
 

 3/50 (0.06)
 

 N.S.
 

Infinite
 

0.134
 

Infinite
 

Infinite
 

0.566
 

Infinite
 

103
 

 4/50 (U.08)
 

 N.S.
 

0.800
 

0.097
 

38.616
 

3.760
 

0.390
 

181.269
 

103
 

High
 

Dose
 

1/50 (0.02)
 

N.S.
 

Infinite
 

0.012
 

Infinite
 

Infinite
 

0.050
 

Infinite
 

101
 

1/50 (0.02)
 

N.S.
 

0.200
 

0.003
 

15.415
 

0.940
 

0.012
 

72.331
 

101
 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats 

(continued) 

Tomography: Morphology


Mammary Gland: Fibroadenomab


P Valuesc»d


Relative Risk (Matched Control)f
 

Lower Limit 

Upper Limit 

Relative Risk (Pooled Control)£ 

Lower Limit 

oo Upper Limit 

Weeks to First Observed Tumor

Uterus: Endometrial 

Stromal Polypb

P Valuesc>d

Relative Risk (Matched Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Pooled Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Fed Lindane

Matched
 

 Control
 

 3/10 (0.30)
 

 N.S.
 

 50
 

 1/9 (0.11)
 

 N.S.
 

 109
 

 in the Diet3 

Pooled


Control


8/42 (0.19)


N.S.


4/52 (0.08)


N.S.


 Low
 

 Dose
 

 12/50 (0.24)
 

 N.S.
 

0.800
 

0.293
 

3.948
 

1.260
 

0.528
 

3.227
 

44
 

 6/4'/ (0.13)
 

 N.S.
 

1.149
 

0.177
 

51.671
 

1.660
 

0.421
 

7.527
 

87
 

High
 

Dose
 

9/50 (0.18)
 

N.S.
 

0.600
 

0.202
 

3.093
 

0.945
 

0.358
 

2.573
 

61
 

7/44 (0.16)
 

N.S.
 

1.432
 

0.235
 

62.922
 

2.068
 

0.566
 

9.021
 

93
 
Table E2. Analyses of the Incidence of Primary Tcmors in Female Rats
 

Fed Lindane in the Dieta
 

(continued)
 

a

Treated groups received time-weighted avarage doses of 135 or 270 ppm in feed.
 

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

c

Beneath the incidence of tumors in a control group is the probability level for the Cochran-


Armitage test when P < 0.05; otherwise, not significant (N.S.) is indicated. Beneath the incidence of tumors in a treated group is the probability level for the Fisher exact test for the
 

comparison of that treated group with the matched-control group (*) or with the pooled-control
 

group (**) when P < 0.05 for either control group; otherwise, not significant (N.S.) is indicated.
 

'JA negative trend (N) indicates a lower incidence in a treated group than in a control group.
 

oo
 e oo
 The probability level for departure from linear trend is given when P < 0.05 for any comparison.
 

*Ihe 95% confidence interval of the relative risk between each treated group and the specified
 

control group.
 
APPENDIX F
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS
 

IN MICE FED LINDANE IN THE DIET
 

89
 

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

Topography: Morphology


Lung: Alveolar/Bronchiolar
 

Adenomab


P Valuesc'd


Relative Risk (Matched Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Pooled Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Liver: Hepatocellular
 

Carcinomab


P Valuesc»d


Departure from Linear Trend6


Relative Risk (Matched Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Pooled Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Fed Lindane in the Diet3 

Matched Pooled 

 Control Control 

 2/10 (0.20) 3/48 (0.06) 

 N.S. N.S. 

 77 

 2/10 (0.20) 5/49 (0.10) 

 N.S. N.S. 

P = 0.002 

 77
 

Low 

Dose 

High 

Dose 

2/50 (0.04) 3/47 (0.06) 

N.S. N.S. 

0.200 

0.018 

2.592 

0.319 

0.046 

3.576 

0.640 

0.056 

5.345 

1.021 

0.143 

7.264 

90 90 

19/49 (0.39) 9/46 (0.20) 

P = 0.001** N.S. 

1.939 

0.615 

15.778 

0.978 

0.263 

8.635 

3.800 

1.516 

11.891 

1.917 

0.626 

6.755 

60 79 
Table Fl. Analyses of the Incidence of Primary Tumors in Male Mice
 

Fed Lindane in the Dieta
 

(continued) 

Matched Pooled Low High 

Topography: Morphology Control Control Dose Dose 

Liver: Hepatocellular Carcinoma
 

or Neoplastic Noduleb 3/10 (0.30) 8/49 (0.16) 19/49 (0.39) 10/46 (0.22)
 

P Valuesc>d N.S. N.S. P = 0.010** N.S.
 

Departure from Linear Trend6 P = 0.011
 

Relative Risk (Matched Control) f 1.293 0.725
 

Lower Limit 0.518 0.253
 

Upper Limit 6.021 3.657
 

Relative Risk (Pooled Control)f 2.375 1.332
 

Lower Limit 1.109 0.519
 

Upper Limit 5.616 3.536
 

Weeks to First Observed Tumor 77 60 79
 

a

Treated groups received doses of 80 or 160 ppm in feed.
 

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

c

Beneath the incidence of tumors in a control group is the probability level for the Cochran-


Armitage test when P < 0.05; otherwise, not significant (N.S.) is indicated. Beneath the incidence of tumors in a treated group is the probability level for the Fisher exact test for the
 

comparison of that treated group with the matched-control group (*) or with the pooled-control
 

group (**) when P < 0.05 for either control group; otherwise, not significant (N.S.) is indicated.
 
Table Fl. Analyses of the Incidence of Primary Tumors in Male Mice
 

Fed Lindane in the Diet3
 

(continued)
 

^A negative trend (N) indicates a lower incidence in a treated group than in a control group.
 

e

The probability level for departure from linear trend is given when P < 0.05 for any comparison.
 

95% confidence interval of the relative risk between each treated group and the specified
 

control group.
 

VD
 

OJ
 
Table F2. Analyses of the Incidence of Primary Tumors in Female Mice
 

Topography: Morphology


Lung: Alveolar/Bronchiolar
 

Adenomab


P Valuesc>d


Relative Risk (Matched Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Pooled Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Lung: Alveolar/Bronchiolar
 

Adenoma or Carcinoma

b


P Valuesc»d


Relative Risk (Matched Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Pooled Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Fed Lindane in the Dieta 

Matched 

 Control 

Pooled

Control

 Low 

 Dose 

High 

Dose 

 0/10 (0.00) 1/48 (0.02) 1/48 (0.02) 2/48 (0.04) 

 N.S. N.S. N.S. N.S. 

Infinite 

0.012 

Infinite 

Infinite 

0.068 

Infinite 

1.000 

0.013 

76.886 

2.000 

0.108 

115.535 

— 90 91 

 1/10 (0.10) 2/48 (0.04) 1/48 (0.02) 2/48 (0.04) 

 N.S. N.S. N.S. N.S. 

0.208 

0.003 

16.043 

0.417 

0.026 

24.099 

0.500 

0.009 

9.277 

1.000 

0.075 

13.306 
Table F2. Analyses of the Incidence of Primary Tumors in Female Mice
 

Fed Lindane in the Diet3
 

(continued)
 

Topography: Morphology

Liver: Hepatocellular 

Carcinoma13

P Valuesc'

d

Relative Risk (Matched Control)

f 

Lower Limit 

Upper Limit 

Relative Risk (Pooled Control)f 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor

Liver: Hepatocellular Carcinoma 

or Neoplastic Noduleb

P Valuesc»d

Relative Risk (Matched Control)

f 

Lower Limit 

Upper Limit 

Relative Risk (Pooled Control) 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor
 

Matched 

 Control 

 0/10 (0.00) 

 N.S. 

— 

 1/10 (0.10) 

 N.S. 

90
 

Pooled Low High 

Control Dose Dose 

2/47 (0.04) 2/47 (0.04) 3/46 (0.07) 

N.S. N.S. N.S. 

Infinite Infinite 

0.069 0.145 

Infinite Infinite 

1.000 1.533 

0.075 0.184 

13.295 17.650 

90 91 

3/47 (0.06) 4/47 (0.09) 3/46 (0.07) 

N.S. N.S. N.S. 

0.851 0.652 

0.104 0.064 

41.020 33.512 

1.333 1.022 

0.237 0.143 

8.665 7.260 

90 91 
Table F2. Analyses of the Incidence of Primary Tumors in Female Mice
 

Fed Lindane in the Diet3
 

(continued)
 

a

Treated groups receiving doses of 80 or 160 ppm in feed.
 

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

c

Beneath the incidence of tumors in a control group is the probability level for the Cochran-


Armitage test when P < 0.05; otherwise, not significant (N.S.) is indicated. Beneath the incidence of tumors in a treated group is the probability level for the Fisher exact test for the
 

comparison of that treated group with the matched-control group (*) or with the pooled-control
 

group (**) when P < 0.05 for either control group; otherwise, not significant (N.S.) is indicated.
 

"A negative trend (N) indicates a lower incidence in a treated group than in a control group.
 

vo eThe probability level for departure from linear trend is given when P < 0.05 for any comparison,
 o**
 

*The 95% confidence interval of the relative risk between each treated group and the specified
 

control group.
 
APPENDIX
 

ANALYSIS OF FORMULATED DIETS FOR
 

CONCENTRATIONS OF LINDANE
 

97
 

APPENDIX G
 

Analysis of Formulated Diets for
 

Concentrations of Lindane
 

A 100-g sample of the diet was shaken with 125 ml hexane for 16
 

hrs. , then filtered through Celite with hexane washes and reduced
 

in volume to 10 ml. After appropriate dilutions, the solution
 

was quantitatively analyzed for lindane by gas-liquid
 

chromatography (electron-capture detector, 10% DC-200 on
 

Gas-Chrom Q column). Recoveries were checked with spiked
 

samples, and external standards were used for calibration.
 

Theoretical No. of Sample Coefficient of Range (ppm) 

Concentration Samples Analytical Variation (%) 

in Diet (ppm) Mean (ppm) 

80 26 79.2 4.0% 74.2-85.6
 

160 33 160.2 4.0% 144-174
 

320 16 317.8 3.2% 296-337
 

640 11 655.5 5.1% 600-708
 

•ft U.S. GOVERNMENT PRINTING OFFICE: 1977- 260-899:3136 

99 

DHEW Publication No. (NIH) 77-814 
DHEW Publication No. (NIH) 77-814