National Cancer Institute 

CARCINOGENESIS 

Technical Report Series 

No. 15 

1977 

BIOASSAY OF 

CAPTAN 

FOR POSSIBLE CARCINOGENICITY 

CAS No. 133-06-2 

NCI-CG-TR-15 

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

Public Health Service 

National Institutes of Health 

BIOASSAY OF
 

CAPTAN
 

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
 

DHEW Publication No. (ΝΙΕ) 77-815
 

BIOASSAY OF
 

CAPTAN
 

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 captan 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 Tracor Jitco, Inc., prime contractor for the NCI 

carcinogenesis bioassay program. 

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

Weisburger*·»^ and R. R. Bates^»^; the doses were selected by
 

Drs. T. E. Shellenberger^»5, J. Η. Weisburger, and R. R. Bates.
 

Animal treatment and observation were supervised by Drs. Τ. Ε.
 

Shellenberger and H. P. Burchfield4

, 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.
 

Pathologists from NCI and Tracor Jitco have reviewed selected
 

slides and concur with the overall pathologic evaluation of the
 

study.
 

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?.
 

iii
 
This report was prepared at Tracor Jitco under the direction of
 

NCI. Those responsible for the report at Tracor Jitco were
 

Reichardt and Ms. L. A. Waitz bioscience writers; and Dr. E.
 

Dr. Marshall Steinberg?, Director of the Bioassay Program; Drs. 

J. F. Robens7

 and 0. G. Fitzhugh7 , toxicologists; Mr. W. D. 7 7 

,


W. Gunberg7, technical editor, assisted by Ms. Υ. Ε. Presley7.
 

The statistical analysis was reviewed by a member or members of
 

the Mathematical Statistics and Applied Mathematics Section of
 

NCI (Dr. John J. Gart, Mr. Jun-mo Nam, Dr. Hugh M. Pettigrew, and
 

Dr. Robert E. Tarone served as reviewers on an alternating
 

basis).
 

The following other 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. Or me
 

Dr. Robert A. Squire9
 

Dr. Jerrold M. Ward
 

^-Carcinogenesis Program, Divison of Cancer Cause and Prevention,
 

National Cancer Institute, National Institutes of Health,
 

Bethesda, Maryland.
 

^Now 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.
 

iv
 
5Now with the National Center for Toxicological Research,
 

Jefferson, Arkansas.
 

6EG&G Mason Research Institute, 1530 East Jefferson Street,
 

Rockville, Maryland.
 

^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.
 

^Now with the Division of Comparative Medicine, Johns Hopkins
 

University, School of Medicine, Traylor Building, Baltimore,
 

Maryland.
 

SUMMARY
 

A bioassay of technical-grade captan for possible carcinogenicity
 

was conducted by administering the test material in feed to
 

Osborne-Mendel rats and B6C3F1 mice.
 

Groups of 50 rats of each sex were administered one of two doses
 

of captan for 80 weeks, then observed for 33 or 34 weeks. The
 

time-weighted average doses for both sexes of rats were 2,525 or
 

6,050 ppm. Matched controls consisted of groups of 10 untreated
 

rats of each sex; pooled controls, used for statistical 

evaluation, consisted of the matched controls combined with 75 

untreated male and 75 untreated female rats from similar 

bioassays of six other test chemicals. All surviving rats were
 

killed at 113-114 weeks.
 

Groups of 50 mice of each sex were administered the test material
 

at one of two doses, either 8,000 or 16,000 ppm, for 80 weeks,
 

then observed for 11 weeks. Matched controls consisted of groups
 

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

statistical evaluation, consisted of the matched controls
 

combined with 80 untreated male and 80 untreated female mice from
 

similar bioassays of six other test chemicals. All surviving
 

mice were killed at 90-91 weeks.
 

The mean body weights of both low and high-dose rats and high-


dose mice were lower than those of the matched controls
 

throughout most of the study. Mortality rates did not show
 

statistically significant dose-related trends in either sex of
 

either species.
 

In rats, a positive dose-related trend and a difference between
 

incidences of tumors in high-dose and pooled-control groups were
 

found in females when the data for adrenal cortical adenoma were
 

combined with those for adrenal cortical carcinoma (pooled
 

controls 0/64, low-dose 2/50, high-dose 3/47, Ρ = 0.047). There
 

was also a positive dose-related trend for the incidence of
 

vii
 
C-cell adenoma of the thyroid in female rats (pooled controls
 

1/66, low-dose 1/49, high-dose 4/44, Ρ = 0.035). These
 

endocrine tumors in female rats are believed to have been
 

spontaneous, and not related to treatment.
 

In mice, the incidences of polypoid carcinoma (adenocarcinoma in
 

adenomatous polyp) of the duodenum were statistically significant
 

using tests for a positive dose-related trend both in male mice
 

(pooled controls 0/68, low-dose 1/43, high-dose 3/46, Ρ = 0.033)
 

and in female mice (pooled controls 0/68, low-dose 0/49, high-


dose 3/48, Ρ = 0.022). When the incidences of adenomatous polyp,
 

NOS (not otherwise specified), were combined with those of
 

polypoid carcinoma for statistical analysis, the tests for male
 

mice indicated a substantial increase in significance (pooled
 

controls 0/68, low-dose 3/43, high-dose 5/46, Ρ = 0.008).
 

It is concluded that under the conditions of this bioassay,
 

tumors in the duodenum of B6C3F1 mice were associated with
 

treatment with captan, but there was no convincing evidence that
 

the tumors observed in Osborne-Mendel rats were related to
 

treatment.
 

viii
 
TABLE OF CONTENTS
 

Page
 

I· Introduction · · 1
 

II. Materials and Methods 3
 

A. Chemical 3
 

Β. 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 12
 

H. Data Recording and Statistical Analyses 13
 

III. Results - Rats 19
 

A. Body Weights and Clinical Signs (Rats) 19
 

B. Survival (Rats) 19
 

C. Pathology (Rats) 22
 

D. Statistical Analyses of Results (Rats) 23
 

IV. Results - Mice 25
 

A. Body Weights and Clinical Signs (Mice) 25
 

. Survival (Mice) 27
 

C. Pathology (Mice) 27
 

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 Captan in the Diet 39
 

Table Al Summary of the Incidence of Neoplasms in
 

Male Rats Fed Captan in the Diet 41
 

Table A2 Summary of the Incidence of Neoplasms in
 

Female Rats Fed Captan in the Diet 45
 

ix
 
Page 

Appendix Β Summary of the Incidence of Neoplasms in 

Mice Fed Captan in the Diet 49 

Table Bl Summary of the Incidence of Neoplasms in 

Male Mice Fed Captan in the Diet 51 

Table B2 Summary of the Incidence of Neoplasms in 

Female Mice Fed Captan in the Diet 54 

Appendix C Summary of the Incidence of Nonneoplastic 

Lesions in Rats Fed Captan in the Diet 57 

Table CI Summary of the Incidence of Nonneoplastic 

Lesions in Male Rats Fed Captan in the Diet 59 

Table C2 Summary of the Incidence of Nonneoplastic 

Lesions in Female Rats Fed Captan in the Diet 63 

Appendix D Summary of the Incidence of Nonneoplastic 

Lesions in Mice Fed Captan in the Diet 67 

Table Dl Summary of the Incidence of Nonneoplastic 

Lesions in Male Mice Fed Captan in the Diet 69 

Table D2 Summary of the Incidence of Nonneoplastic 

Lesions in Female Mice Fed Captan in the Diet 72 

Appendix Ε Analyses of the Incidence of Primary Tumors 

in Rats Fed Captan in the Diet 75 

Table El Analyses of the Incidence of Primary Tumors 

in Male Rats Fed Captan in the Diet 77 

Table E2 Analyses of the Incidence of Primary Tumors 

in Female Rats Fed Captan in the Diet 80 

Appendix F Analyses of the Incidence of Primary Tumors 

in Mice Fed Captan in the Diet 87 

Table Fl Analyses of the Incidence of Primary Tumors 

in Male Mice Fed Captan in the Diet 89 

χ 
Page 

Table F2 Analyses of the Incidence of Primary Tumors 

in Female Mice Fed Captan in the Diet 94 

Appendix G Analysis of Formulated Diets for 

Concentrations of Captan 97 

TABLES 

Table 1

Table 2

 Design of Captan Chronic Feeding Studies 

in Rats

 Design of Captan Chronic Feeding Studies 

8 

in Mice 10 

FIGURES 

Figure 1

Figure 2

 Growth Curves for Rats Fed Captan 

in the Diet

 Survival Curves for Rats Fed Captan 

in the Diet

 20 

 21 

Figure 3 Growth Curves for Mice Fed Captan 

in the Diet 26 

Figure 4 Survival Curves for Mice Fed Captan 

in the Diet 28 

xi
 

I. INTRODUCTION
 

Captan (CAS 133-06-2; NCI C00077) is a broad-spectrum fungicide
 

which inhibits mycelial growth from germinating fungus spores
 

(EPA, 1974)· As a result, it has effective protective action,
 

although it will not eradicate a preexisting infection (Billings,
 

1974; EPA, 1974). Because captan is a nonpersistent fungicide
 

(EPA, 1974), directions for use indicate that it should be
 

reapplied every week as necessary to maintain control (Stauffer
 

Chemicals, 1975). It has been one of the most widely used
 

fungicides since its introduction in 1950 (EPA, 1974).
 

Captan is registered for use in foliar and soil applications for
 

growing vegetables, fruits, nut trees, and ornamental plants, and
 

for treatment of seeds and turf (EPA Compendium, 1975). It is
 

also used as an industrial fungicide in paints, plastics,
 

leather, and certain soaps and shampoos. Residue tolerances on
 

foods range from 2 to 100 ppm (EPA, 1975). The World Health
 

Organization has established an acceptable daily intake of 0-0.1
 

rag/kg (WHO, 1974).
 

Captan was selected for screening for carcinogenic activity
 

because there was a potential for long-term human exposure during
 

agricultural, industrial, or other applications, or from residues
 

in food products.
 

II . MATERIALS AND METHODS 

A. Chemical
 

Captan, the common name for N-((trichloromethyl)thio)-4-cyclohexene-l,2-dicarboximide, was obtained oin a single batch (Lot No.
 

5x-317) from the Chevron Chemical Company, Ortho Division, San
 

Francisco, California, for use in the chronic study. The
 

identity of the chemical was confirmed at Gulf South Research
 

Institute by infrared, nuclear magnetic resonance, and isobutane
 

chemical ionization mass spectra. Gas-liquid chromatography
 

(electron capture detector, 10% DC-200 column) showed a single
 

peak. No attempt was made to identify or quantitate impurities.
 

The chemical was stored at approximately 4°C.
 

B. Dietary Preparation
 

All diets were formulated using finely ground Wayne® Lab Blox
 

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

required amount of captan for each dietary concentration. A
 

given amount of the test chemical was first hand-mixed with an
 

approximately equal amount of feed. This mixture was then added
 

slowly with mechanical mixing to a larger quantity of feed to
 

give the desired concentration of the chemical. Acetone
 

(Mallinckrodt, Inc., St. Louis, Mo.) and corn oil (Louana®,
 

Opelousas Refinery Co., Opelousas, La.) were then added to the
 

3 
feed, each in an amount corresponding to 2% of the final weight
 

of feed. The diets were mixed mechanically for not less than 25
 

minutes to assure homogeniety of the mixture and evaporation of
 

the acetone. Formulated diets were stored at approximately 17°C
 

until used, but no longer than 1 week.
 

The stability of captan in feed was tested by determining the
 

concentration of the chemical in formulated diets at intervals
 

over a 7-day period. Diets containing 8,000 or 16,000 ppm captan
 

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 captan 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
 

samples tested was within 2.5% of the theoretical concentration,
 

and the coefficient of variation was never more than 5.9%. 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
 

4 
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. Upon
 

arrival at the laboratory, all animals were quarantined for an
 

acclimation period (rats for 6 or 7 days, mice for 15 days) and
 

then assigned to control and test 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 one time per week to
 

clean cages; later in the study, cages were changed every 2
 

weeks. Mice were transferred one time 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
 

5 
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 one time per week; at the same time each cage was
 

changed to a different position in the row within the same
 

column. Rats receiving captan, along with their matched
 

controls, were housed in a room by themselves. Mice receiving
 

captan were maintained in a room housing mice administered aldrin
 

(CAS 309-00-2) or photodieldrin (CAS 13366-73-9), together with
 

their respective matched controls.
 

E. Subchronic Studies
 

Subchronic studies were conducted to estimate the maximum
 

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

concentrations (hereinafter referred to as "low doses11

 and "high
 

doses") were determined for administration in the chronic
 

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

animal feed in twofold increasing concentrations, ranging from
 

500 to 32,000 ppm for both rats and mice. The treated and
 

control groups each consisted of five male and five female
 

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

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

weeks.
 

In both male and female rats, weight depression was apparent at
 

8,000 and 16,000 ppm during the first weeks. Later, these
 

animals appeared to adapt to the test chemical, and gains in
 

weight approached those of the controls. There were no deaths
 

in either male or female rats. The low and high doses for the
 

chronic studies in rats were set at 8,000 and 16,000 ppm.
 

In male and female mice, there was little, if any, adverse effect
 

on weight gain at dietary concentrations as high as 8,000 ppm.
 

At 16,000 ppm, captan caused a loss in weight among males during
 

the first 2 weeks and among females during the first week of the
 

feeding period; as the study progressed, treated animals of both
 

sexes recovered and their gains in weight were similar to those
 

of the controls. Weight losses were more marked at 32,000 ppm in
 

males and females during the initial weeks, and four males and
 

all females died during the study. The low and high doses for
 

the chronic study in mice were set at 8,000 and 16,000 ppm.
 

F. Designs of Chronic Studies
 

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

Initially, doses of 8,000 or 16,000 ppm were administered to
 

7 
Table 1. Design of Captan Chronic Feeding Studies in Rats
 

Sex and
 

Treatment
 

Group
 

MALE
 

Matched-Controla
 

Low-Dose
 

High-Dose
 

FEMALE
 

Ma tched-Controla
 

Low-Dose
 

High-Dose
 

Initial
 

No. of
 

Animalsk
 

10
 

50
 

50
 

10
 

50
 

50
 

Captan
 

in Diet
 

(ppm)
 

0
 

4,000
 

2,000
 

0
 

8,000
 

4,000
 

0
 

0
 

4,000
 

2,000
 

0
 

8,000
 

4,000
 

0
 

Time on Study
 

Treated Untreated
 

(weeks )ϋ (weeks )il
 

114
 

21
 

59
 

33
 

41
 

39
 

34
 

114
 

21
 

59
 

33
 

41
 

39
 

34
 

Time-Weighted
 

Average Dosee
 

(ppm)
 

2,525
 

6,050
 

2,525
 

6,050
 

a

The matched controls consisted of 5 animals of each sex, started
 

with the low-dose animals, and 5 animals of each sex, started with
 

the high-dose animals.
 

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

c

Doses of captan were lowered at week 21 during the study, since
 

it was believed that excessive mortality might occur before
 

termination of the study based on the mortality, weight changes,
 

and general condition of rats used in similar bioassays of
 

other chemicals at Gulf South Research Institute.
 

8
 
Table 1. Design of Captan Chronic Feeding Studies in Rats
 

(continued)
 

^When diets containing captan were discontinued, the high-dose rats
 

and their matched controls were fed the control diet without
 

corn oil for 6 weeks, then the control diet (2% corn oil added)
 

for an additional 28 weeks, while low-dose rats received only the
 

control diet (2% corn oil added) until termination of the study·
 

e

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

Σ(nο. of weeks receiving each dose)
 

9
 
Table 2. Design of Captan Chronic Feeding Studies in Mice
 

Sex and Initial Captan Time on Study 

Treatment No. of in Diet Treated Untreatedb 

Group Animalsa (ppm) (weeks) (weeks) 

MALE 

Matched-Control 10 0 91 

Low-Dose 50 8,000 80 

0 11 

High-Dose 50 16,000 80 

0 11 

FEMALE 

Matched-Control 10 0 90-91 

Low-Dose 50 8,000 80 

0 11 

High-Dose 50 16,000 80 

0 11 

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

bWhen diets containing captan were discontinued, all treated mice
 

and their matched controls were fed the control diet (2% corn oil
 

added) until termination of the study.
 

10
 
groups of rats of each sex. Because the chemical was highly
 

toxic at 16,000 ppm, tests at this dose were terminated after 18
 

weeks. Five matched controls of each sex were also terminated.
 

The groups receiving 8,000 ppm, originally designated "low-dose,11
 

were then redesignated lfhigh-dose,f! as indicated in table 1.
 

Seven males and one female of the groups fed at 8,000 ppm had
 

also died by week 18. These rats were replaced with healthy
 

animals selected from the groups fed at 16,000 ppm. Additional
 

groups of male and female rats, designated "low-dose11 as
 

indicated in table 1, were started at 4,000 ppm 20 weeks after
 

the beginning of the study. At the same time, five additional
 

matched controls of each sex were started. The time-weighted
 

average doses for the rats were 2,525 and 6,050 ppm.
 

For the mice, the initial doses of 8,000 and 16,000 ppm were
 

maintained throughout the study, as indicated in table 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 captan
 

were combined with matched controls from studies performed on
 

tetrachlorvinphos (CAS 961-11-5), malathion (CAS 121-75-5),
 

toxaphene (CAS 8001-35-2), endrin (CAS 72-20-8), lindane (CAS
 

58-89-9), and photodieldrin (CAS 13366-73-9). The pooled
 

11
 
controls for statistical tests using rats consisted of 75 males
 

and 75 females; using mice, 80 males and 80 females. The studies
 

on chemicals other than captan were also conducted at Gulf South
 

Research Institute and overlapped the captan study 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. Because additional
 

matched controls were started simultaneously with restarted
 

treatment groups for some of these chemicals, the number of
 

animals in the pooled-control groups varied.
 

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 icdes, heart,
 

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

small intestine, largo intestine, kidney, urinary bladder,
 

12
 
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.
 

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 experiment in
 

each group.
 

H. Data Recording and Statistical Analyses
 

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

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

clinical observations, survival, body weight, and individual
 

pathologic results, as recommended by the International Union
 

13
 
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 carcinogenicity are
 

discussed in the statistical narrative sections.
 

Probabilities of survival were estimated by the product-limit
 

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

report in the form of graphs. Animals were statistically
 

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

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

causes were not statistically censored. Statistical analyses for
 

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

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

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

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

departure from linearity test, which is only reported when its
 

two-tailed Ρ value is less than 0.05.
 

The incidence of neoplastic or nonneoplastic lesions has been
 

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

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

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

14
 
the denominators included only those animals for which that site
 

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

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

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

consist of the numbers of animals necropsied.
 

The purpose of the statistical analyses of tumor incidence is to
 

determine whether animals receiving the test chemical developed a
 

significantly higher proportion of tumors than did the control
 

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

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

a control group with that of a group of 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
 

 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 Ρ 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
 

15
 
of the dose-response curve is different from zero at the one-


tailed 0·05 level of significance. Unless otherwise noted, the
 

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

from linear trend.
 

A time-adjusted analysis was applied when numerous early deaths
 

resulted from causes that were not associated with the formation
 

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

first tumor was observed were excluded by basing the statistical
 

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

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

such an early tumor was found, comparisons were based exclusively
 

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

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

obtained, the standard procedures for analyses of the incidence
 

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

followed.
 

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;
 

16
 
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
 

17
 
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, Ρ < 0.050 when the control incidence is
 

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

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

the absence of a significant result while the upper limit
 

indicates that there is a theoretical possibility of the
 

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

detected under the conditions of this test.
 

18
 
III. RESULTS - RATS
 

A. Body Weights and Clinical Signs (Rats)
 

The mean body weights of both male and female low- and high-dose
 

rats were less than those of their corresponding controls
 

throughout the study (figure 1). During the first year of study,
 

the treated animals were generally comparable to the controls in
 

appearance and behavior.
 

Clinical signs including rough hair coats, alopecia, pale mucous
 

membranes, dermatitis, tachypnea, and hematuria were noted at a
 

low incidence in all treated groups of rats during the first half
 

of the second year, with a gradually increasing frequency during
 

the remainder of the study. A few treated females showed
 

evidence of vaginal bleeding.
 

B. Survival (Rats)
 

The Kaplan and Meier curves estimating the probabilities of
 

survival for male and female rats receiving captan at the doses
 

of this experiment, together with the controls, are shown in
 

figure 2.
 

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, 46% of the high-dose group, 58% of the
 

19
 
TIME ON STUDY (WEEKS) 

Figure 1. Growth Curves for Rats Fed Captan in the Diet
 

20
 
TIME ON STUDY (WEEKS) 

Figure 2. Survival Curves for Rats Fed Captan in the Diet
 

21
 
low-dose group, 80% of the low-dose controls, and 60% of the
 

high-dose controls survived to the end of the study. Survival in
 

female rats was higher, with over 75% of the treated animals, 60%
 

of the low-dose controls, and all of the high-dose controls
 

living to termination of the study. Survival was sufficient for
 

meaningful statistical analyses 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.
 

The types of tumors observed are not uncommon for this strain of
 

rat, and the distribution and frequency among animals of the
 

matched-control and captan-treated groups do not indicate any
 

trend of carcinogenic activity induced by the chemical.
 

Therefore, these lesions are considered to have occurred
 

spontaneously·
 

Likewise, a great variety of nonneoplastic lesions were observed
 

either sporadically or with approximately equal frequency among
 

animals of the control and treated groups. These lesions
 

frequently have been found in rats used in other experiments
 

independent of treatment.
 

22
 
The incidence and distribution of the neoplastic and non-


neoplastic lesions occurring in the rats in this study do not
 

implicate captan as the causative agent.
 

D. Statistical Analyses of Results (Rats)
 

Tables E1 and E2 in Appendix Ε 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, neither the results of the Cochran-Armitage test
 

for dose-related linear trend nor of the Fisher exact test for
 

the incidence of tumors at any specific site is significant at
 

the 0.05 level. Due to the early deaths in the high-dose males,
 

time-adjusted analyses, eliminating animals that died before 1
 

year on study, were performed on the incidence of chromophobe
 

adenoma of the pituitary; however, no significant result was
 

obtained.
 

When the incidences of cortical adenoma and cortical carcinoma of
 

the adrenal gland in female rats are combined, the Cochran-


Armitage test for positive linear dose-related trend has a
 

probability of 0.047, using the pooled controls. The Fisher
 

exact test results are not significant. The 95% confidence
 

interval for the relative risk in the high-dose group compared
 

23
 
with the pooled controls has a lower limit of 0,813, which is a
 

value less than one, indicating the possibility that no true
 

difference exists between these two groups. The incidence of
 

C-cell adenoma in the thyroid also has a positive linear trend
 

(P = 0.035), but the results of the Fisher exact test are not
 

significant. No other tumor or combination of tumors appears in
 

significantly larger numbers in the treated groups than in either
 

control group. The statistical conclusion is that there is
 

insufficient evidence for the dose association of the chemical
 

with the tumors in rats.
 

When groupings of types of tumors are made, as in cortical
 

adenoma or carcinoma of the adrenal gland in female rats, the
 

incidences of the individual components of the grouping are not
 

included in tables El and E2 unless the statistical tests are
 

significant. A list of the incidences of each type of tumor is
 

provided in Appendix A, tables Al and A2.
 

24
 
IV. RESULTS - MICE
 

A. Body Weights and Clinical Signs
 

Throughout the study, the mean body weights of the high-dose male
 

and female mice were notably lower than those of their controls,
 

whereas the mean body weights of the low-dose male and female
 

mice were only slightly lower (figure 3).
 

During the first 4 months of the study, the treated animals were
 

generally comparable to the controls in appearance and behavior,
 

except during the first 2 weeks, when the treated animals showed
 

loss of weight. At week 24, all of the high-dose females were
 

very excitable and a few had slight tremors (tremors were not
 

noted after this time). At the same time, a slight weight loss
 

occurred in high-dose females. At week 24, the high-dose males
 

showed a considerable weight loss but did not appear excitable.
 

During week 36, both treated and control animals showed weight
 

loss.
 

After 50 weeks, clinical signs including rough hair coats,
 

alopecia, and abdominal distention were observed in the treated
 

groups. At week 71 all of the low- and high-dose males appeared
 

to be very excitable. At week 80, all of the low- and high-dose
 

males evidenced abdominal distention. During the last 6 weeks of
 

the study, one low-dose male and one high-dose male had a
 

25
 
TIME ON STUDY (WEEKS)
 

Figure 3. Growth Curves for Mice Fed Captan in the Diet
 

26
 
purulent discharge from the penis. The perineal areas of a few
 

high-dose males were irritated and red in appearance.
 

B. Survival (Mice)
 

The Kaplan and Meier curves estimating the probabilities of
 

survival for male and female mice receiving captan at the doses
 

of this experiment, together with the controls, are shown in
 

figure 4.
 

In neither sex was the Tarone test result significant for
 

positive dose-related trend in mortality over the period. More
 

than 90% of the animals survived to the end of the study,
 

providing sufficient numbers of animals for meaningful
 

statistical analyses of late-developing 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.
 

With the exception of the proliferative and/or neoplastic lesions
 

observed in the duodenum of both male and female treated mice,
 

the pathologic changes observed were not considered to be related
 

to the administration of captan.
 

27
 
TIME ON STUDY (WEEKS)
 

Figure 4. Survival Curves for Mice Fed Captan in the Diet
 

28
 
The duodenal lesions were located approximately 1 cm posterior to
 

the pylorus, usually in the antimesenteric portion of the
 

duodenal mucosa. Grossly, they were either single, well-circumscribed (3-5 mm across) and slightly elevated (1-2 mm) areas, or
 

single, thin mucosal projections up to 5 mm in height. The
 

lesions were inconspicuous on the serosal surface. Microscopically, the following three different lesions were classified:
 

(1) mucosal hyperplasia—a duplication of glands and
 

villi,
 

(2) adenomatous polyp—a more accentuated proliferative
 

process with glandular structures and villi aggregated
 

and branched around supporting stalks made up of
 

connective tissue (features of malignancy were not
 

observed), and
 

(3) adenocarcinoma in adenomatous polyp (polypoid
 

carcinoma)—the most advanced and aggressive-appearing
 

lesion, consisting of cellular polypoid structures with
 

numerous mitotic figures, disorganized microacini, and
 

areas where neoplastic infiltration was evident·
 

Tinctorial changes (basophilia) were also present.
 

The classification of these lesions was frequently difficult.
 

Nevertheless, the location and some common cellular character29
 
istics suggest that they are different developmental stages of
 

the same type of lesion. The distribution and incidence of the
 

duodenal alteration were as follows:
 

Males Females
 

Low High Low High 

Controls Dose Dose Controls Dose Dose 

Number examined (9) (43) (46) (9) (49) (48) 

Adenocarcinoma 0 1 3 0 0
 

Adenomatous polyp 0 2 2 1
 

Mucosal hyperplasia
 0
 0
 3
 0
 

1

0
 

3

0

0
 

The rarity of these lesions in the strain of mouse used suggests
 

that the lesions were caused by captan.
 

The nonneoplastic pathological changes were either inflammatory
 

or degenerative in nature, and their incidence and distribution
 

do not appear to be associated with captan treatment.
 

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.
 

The incidences of adenocarcinoma in adenomatous polyp of the
 

duodenum are in a significant linear trend for both male and
 

female mice, with Cochran-Armitage probability levels of 0.033
 

30
 
and 0.022, respectively, using the pooled controls. The Fisher
 

exact test results for both sexes are not significant. When the
 

incidences of adenocarcinoma in adenomatous polyp of the duodenum
 

are combined with those of adenomatous polyp, NOS (not otherwise
 

specified), for statistical analysis, the tests for male mice
 

show a substantial increase in significance when compared with
 

pooled controls. The test for positive linear trend is significant (P = 0.008), the Fisher exact test in the high-dose male
 

mice has a probability level of 0.009, and the 95% confidence
 

interval for relative risk has a lower limit of 1.849 using
 

pooled controls. The incidence of these combined tumors in
 

female mice is not significant. The overall consideration of
 

these various statistics suggests a dose association of the test
 

chemical with tumors in the duodenum in male mice.
 

Significant trends and results in the negative direction were
 

observed due to a lower incidence of liver tumors in the treated
 

groups of male mice than in either set of controls. When tumors
 

at the same site are grouped, as in neoplastic nodule and
 

hepatocellular carcinoma in male mice, the incidences of the
 

individual components of the grouping are not indicated in the
 

statistical analyses in the tables when they do not occur
 

individually at less than 5% incidence. No other incidences of
 

31
 
tumors were significant in the mice· A list of the incidences of
 

each type of tumor is provided in Appendix B, tables Bl and B2.
 

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

in the tables, with the exception of adenomatous polyp, NOS, or
 

adenocarcinoma in adenomatous polyp of the duodenum in high-dose
 

male mice, the lower limit is below the value of one; this
 

indicates the negative aspects of the results. It should also be
 

noted that each of the intervals, with the exception of liver
 

tumors in low-dose male mice, 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.
 

32
 
V. DISCUSSION
 

The doses of captan used in this bioassay adversely affected both
 

rats and mice. Mean body weights of both low- and high-dose rats
 

and high-dose mice were generally lower than controls throughout
 

most of the study. General clinical signs that were noted,
 

particularly during the second year of the study, included rough
 

hair coats, alopecia, pale mucous membranes, dermatitis,
 

tachypnea, and hematuria for treated rats, and rough hair coats,
 

alopecia, and abdominal distention for treated mice. The
 

high-dose female mice and both low- and high-dose males had
 

periods of excitability, and a few had slight tremors. Mortality
 

rates, however, did not show statistically significant dose-


related trends in either sex of either species.
 

In rats, there was a positive dose-related trend (P = 0.047) for
 

the combined incidence of cortical adenoma and cortical carcinoma
 

of the adrenal gland in high-dose females compared with the
 

incidence in the pooled controls. However, the spontaneous
 

incidence is variable in this strain of rat, and the incidence of
 

tumors was very low; one adrenal cortical adenoma and one
 

carcinoma were found in the low-dose animals and two adrenal
 

cortical adenomas and one carcinoma in the high-dose group.
 

There was also a positive dose-related trend for the incidence of
 

33
 
C-cell adenoma of the thyroid in female rats (pooled controls
 

1/66, low-dose 1/49, high-dose 4/44, Ρ = 0.035). The relationship of these tumors to treatment is not clearly established.
 

In mice, duodenal lesions, which are usually rare, occurred in
 

both sexes of treated animals. They were located approximately
 

1 cm posterior to the pylorus, usually in the antimesenteric
 

portion of the duodenal mucosa. Microscopically, the three
 

different lesions that were classified were mucosal hyperplasia,
 

adenomatous polyp, NOS, and adenocarcinoma in adenomatous polyp
 

(polypoid carcinoma). Among males, the mucosal hyperplasia was
 

found in three high-dose animals. Incidences of polypoid
 

carcinoma (adenocarcinoma in adenomatous polyp) of the duodenum
 

were statistically significant using tests for a positive dose-


related trend both in male mice (pooled controls 0/68, low-dose
 

1/43, high-dose 3/46, Ρ = 0.033) and in female mice (pooled
 

controls 0/68, low-dose 0/49, high-dose 3/48, Ρ = 0.022). When
 

the incidences of adenomatous polyp, NOS, were combined with
 

those of polypoid carcinoma for statistical analysis, the tests
 

for male mice indicated a substantial increase in significance
 

(pooled controls 0/68, low-dose 3/43, high-dose 5/46, Ρ = 0.008).
 

Only two females, a control and a low-dose animal, had
 

adenomatous polyp, and none had hyperplasia. Three high-dose
 

females had polypoid carcinoma; this incidence was significant,
 

34
 
using the test for positive dose-related trend, but was not
 

significant for direct comparison of the incidences in the high-


dose and control groups.
 

The absence of carcinogenicity of captan in rats in the present
 

bioassay agrees with findings of other studies reported in the
 

literature. In one study (Weir, 1956) groups of 10 rats of each
 

sex were fed diets containing 1,000 or 5,000 ppm of technical-


grade captan for 2 years. The incidence of tumors in each
 

treated group did not differ significantly from that in their
 

comparable control group. In a second study (Reyna et al.,
 

1974b), groups containing 50 male and 50 female rats were fed
 

diets containing 1,000 or 5,000 ppm of technical-grade captan for
 

2 years; the incidence of tumors was no higher in these treated
 

groups than in their comparable controls.
 

Previous studies using mice also reported negative findings.
 

Innes et al. (1969) treated groups of 18 mice of two hybrid
 

strains with captan for 18 months. Beginning with 7-day-old
 

mice, a dose of 215 mg/kg body weight of captan was given daily
 

by gavage for a period of 3 weeks; thereafter, a corresponding
 

dose of 560 ppm captan was added to the diet for the remainder of
 

the 18 months. There was no significant increase in the
 

incidence of tumors in the captan-treated groups compared with
 

35
 
that in the controls· In another 18-month study (Reyna et al. ,
 

1974a), three groups each of 50 male and 50 female Swiss mice
 

were fed diets containing 0, 3,700, or 7,500 ppm of technical-


grade captan. No difference in the incidence of tumors was found
 

in treated and control groups. These negative findings in mice
 

are not in agreement with the results of the present bioassay·
 

The differences may be related to the strains of mice used, since
 

C57BL/6 χ C3H/Anf and C57BL/6 χ AKR mice were used in the Innes
 

study, Swiss mice were used in the Reyna study, and B6C3F1
 

(C57BL/6 χ C3H/He) mice were used in the present study. It
 

should also be noted that the Innes study particularly considered
 

hepatomas, pulmonary tumors, and lymphomas, and the intestine may
 

not have been adequately examined.
 

It is concluded that under the conditions of this bioassay,
 

tumors in the duodenum in B6C3F1 mice were associated with
 

treatment with captan, but there was no convincing evidence that
 

the tumors observed in Osborne-Mendel rats were related to
 

treatment.
 

36
 
VI. BIBLIOGRAPHY
 

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

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

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

Panel on Carcinogenicity of the Cancer Research Commission
 

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

Geneva, 1969.
 

Billings, S. C , Pesticide Handbook - Entoma» The Entomalogical
 

Society of America, College Park, Md., 1974.
 

Cox, D. R. , Regression models and life tables. JN_ R^ Statist.
 

Soc. Β 34(2):187-220, 1972.
 

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

1970, pp. 48-52.
 

Environmental Protection Agency, Captan. EPA Compendium of
 

Registered Pesticides, Vol. II, Fungicides and Nematicides,
 

U.S. Government Printing Office, Washington, D.C., 1975,
 

C-10-00.06 - C-10-00.20.
 

Environmental Protection Agency, Initial Scientific and
 

Minieconomic Review of Captan, Office of Pesticide Programs,
 

Washington, D.C., 1975.
 

Environmental Protection Agency, Production, Distribution, Use
 

and Environmental Impact Potential of Selected Pesticides,
 

Office of Pesticide Programs, Washington, D. C , 1974.
 

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.
 

Innes, J. R. , Ulland, Β. Μ., Valerio, Μ. G. , Petrucelli, L. ,
 

Fishbein, L., Hart, E. R. , and Pallotta, A. J., Bioassay of
 

pesticides and industrial chemicals for tumorigenicity in
 

mice: A preliminary note. J^ Natl. Cancer Inst.
 

42,: 1101-1114, 1969.
 

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

incomplete observations. JN_ Am. Statist. Assoc. 53:457-481,
 

1958.
 

37
 
Linhart, Μ. S. , Cooper, J. Α., Martin, R. L. , Page, Ν. Ρ· , and
 

Peters, J. Α· , Carcinogenesis bioassay data system. Comp.
 

and Biomed. Res. ]j 230-248, 1974.
 

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

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

Reyna, M. S·, Kennedy, G. L. , and Keplinger, M. L., 18-month
 

carcinogenic study with captan technical in Swiss white
 

mice. Unpublished report of Industrial Bio-test Labs,
 

1973a, cited in 1973 Evaluations ^f Some Pesticide Residues
 

in Food; The Monographs, World Health Organization, Geneva,
 

1974a.
 

Reyna, M. S., Kennedy, G. L. , and Keplinger, M. L. , 2-year
 

carcinogenic study with captan technical in albino rats.
 

Unpublished report of Industrial Bio-test Labs, 1973b, cited
 i n

 1973 Evaluations of Some Pesticide Residues in Food: The
 

Monographs, World Health Organization, Geneva, 1974b.
 

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.
 

Stauffer Chemical Company, Captan (product label), Stauffer
 

Chemical Co., Westport, Conn., 1975.
 

Stauffer Chemical Company, Captan, Stauffer Chemical Co.,
 

Westport, Conn., 1972.
 

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

Biometrika 62(3):679-682, 1975.
 

Weir, R. J., Technical captan, recrystallized captan. Chronic
 

feeding - rats. Final report. Unpublished report of
 

Hazleton Laboratories, 1956, cited in World Health
 

Organization, 1969 Evaluations of Some Pesticide Residues in
 

Food: The Monographs, World Health Organization, Geneva,
 

1970.
 

World Health Organization, 1973 Evaluations og Some Pesticide
 

Residues in Food: The Monographs, World Health Organization, *
 

Geneva, 1974.
 

38
 
APPENDIX A
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

RATS FED CAPTAN IN THE DIET
 

39
 

TABLE ΑΙ. 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS 

FED CAPTAN IN THE DIET 

LOW DOSE HIGH DOSE
 

CONTROL CONTROL
 

ANIMALS INITIALLY IN STUDY 5 10 

ANIMALS NECROPSIED 5 5 

ANIMALS EXAMINED HISTOPATHOLOGICALLY 5 5 

INTEGUMENTARY SYSTEM 

•SKIN 

SQUAMOUS CELL CARCINOMA 

FIBROMA 

FIBROSARCOMA 

FIBROUS HISTIOCYTOMA, MALIGNANT 

(5) J5) 

• SUBCUT TISSUE 

FIBROMA 

LIPOMA 

(5) (5) 

RESPIRATORY SYSTEM 

#LUNG. (•») (5) 

TB.ANSITIONAL-CELL CARCINOMA, MET 

MIXED TUMOR, METASTATIC 

HEMATOPOIETIC SYSTEM 

•MULTIPLE	 ORGANS (5) (5) 

MALIG.LYMPHOMA, LYMPHOCYTIC TYPE 

#SPLEEN ( 4 ) (5) 

HEMANGIOMA 

HfiMANGIOSARCOMA 

tLIVjiR (5) (5) 

MALIG.LYMPHOMA, HISTIOCYTIC TYPE 

CIRCULATORY SYSTEM 

•HEART	 (4) (5) RHABDOMYOSARCOMA 

# NUM3ER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

• NUMBER OF ANIMALS NECROPSIED
 

LOW DOSE 

50 

50 

49 

(5 0) 

1 (2%) 

1 (2%) 

(50) 

1 (2%) 

(Uβ) 

1 (2%) 

(50) 

1 (2%) 

(49) 

1 {2%) 

1 (2%) 

(47) 

1 (2%) 

(48) 

1 (2%) 

HIGH DOSE 

50
 

49
 

49
 

(49) 

1 (2%) 

1 (2%) 

(19) 

1 (2%) 

1 [2%) 

(<*9) 

1 (2%) 

(49) 

(47) 

(*9) 

(49) 

41
 
TABLE A1. MALE RATS: NEOPLASMS (CONTINUED)

 

LOW DOSE HIGH DOSE LOW DOSE HIGH DOSE 

CONTROL CONTROL 

«MYOCARDIUM (4) (5) (48) (49) 

FIBROMA 1 (2%) 

DIGESTIVE SYSTEM 

•LIVER (5) (5) (47) (49) 

BILE DUCT CARCINOMA 1 (2%) 

NEOPLASTIC NODULE 1 (20%) 1 (2%) 2 (4%) 

•ESOPHAGUS ( 7 ) (5) 

FIBROUS HISTIOCYTOMA, MALIGNANT 1 (20%) 

•STOMACH (5) (5) (47) (44) 

SQUAMOUS CELL PAPILLOMA 1 (2%) 

HAMARTOMA 1 (2%) 

URINARY SYSTEM 

•KIDNEY 

TRANSITIONAL-CELL CARC

TUBULAR-CELL ADENOMA 

FIBROUS HISTIOCYTOMA,

MIXED TUMOR, MALIGNANT 

t HAMARTOMA 

INOMA 

 MALIGNANT 

(5) (5) 

1 (20%) 

(49 ) 

1

1

 (2%) 

 (2%) 

(49) 

1

1

1

1

 (2%) 

 (2%) 

 (2%) 

 (2%) 

•URINARY BLADDER 

FIBROUS HISTIOCYTOMA, MALIGNANT 

i5) (5) (46) (40) 

1 (3%) 

ENDOCRINE SYSTEM 

•PITUITARY (5) (5) (43) (45 ) 

CARCINOMA,NOS 1 (20%) 1 (2%) 

CHROMOPHOBE ADENOMA 1 (20%) 1 (20%) 9 (21% ) 5 (11%) 

•ADRENAL (5) (5) (47) (47) 

PHEOCHROMOCYTOMA 1 (2%) 

•THYROID (5) (5) (42) (47) 

C-CELL ADENOMA 1 (2%) 1 (2%) 

•PANCREATIC ISLETS (5) (45) (47) 

ISLET-CELL ADENOMA 1 (2%) 1 i2%) 

• NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

tThis 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) 

LOW DOSE 

CONTROL 

REPRODUCTIVE SYSTEM 

•MAMMARY GLAND 

ADENOCARCINOMA, NOS 

FIBROMA 

LIPOMA 

(5) 

•SEMINAL VESICLE 

ADENOCARCINOMA, NOS 

(5) 

tTESTI S 

INTERSTITIAL-CELL TUMOR 

(5) 

NERVOUS SYSTEM 

«BRAIN (5) 

GfiANULAR-CELL TUMOR, BENIGN 

SPECIAL SENSE ORGANS 

NONE 

MUSCULOSKELETAL SYSTEM 

NONE 

BODY CAVITIES 

NONE 

ALL OTHEE SYSTEMS 

•MULTIPLE ORGANS (5) 

FIBROUS HISTIOCYTOMA- MALIGNANT 

HIGH DOSE 

CONTROL 

(5) 

(5) 

(5) 

(5) 

(5) 

LOW DOSE HIGH DOSE 

(50) (49) 

1 (2%) 

1 (2%) 

1 (2%) 

i50) (49) 

1 (2%) 

(4 8) (48) 

2 (4%) 

(47) (45) 

2 (4%) 

(50) (49) 

1 (2%) 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

• NUMBER OF ANIMALS NE3ROPSIED
 

43 
TABLE A1. MALE RATS: NEOPLASMS (CONTINUED) 

LOW DOSE
 

CONTROL
 

ANIMAL DISPOSITION SUMMARY
 

ANIMALS INITIALLY IN STUDY
 

NATURAL DEATHo)
 

MORIBUND SACRIFICE
 

SCHEDULED SACRIFICE
 

ACCIDENTALLY KILLED
 

TERMINAL SACRIFICE
 

ANIMAL MISSING
 

@ INCLUDES AUTCLYZED ANIMALS
 

TUMOK SUMMARY
 

TOTAL ANIMALS WITH PRIMARY TUMORS*
 

TOTAL PRIMARY TUMORS
 

TOTAL ANIMALS WITH BENIGN TUMORS
 

TOTAL BENIGN TUMORS
 

TOT&L ANIMALS WITH MALIGNANT TUMORS
 

TOTAL MALIGNANT TUMORS
 

TOTAL ANIMALS WITH SECONDARY TUHORSt
 

TOTAL SECONDARY TUMORS
 

TOTAL ANIMALS WITH TUMORS UNCERTAINBENIGN OR MALIGNANT
 

TOTAL UNCERTAIN TUMORS
 

TOTAL ANIMALS WITH TUMORS UNCERTAINPRIMARY OR METASTATIC
 

TOTAL UNCERTAIN TUMORS
 

5
 

1
 

4
 

2
 

2
 

1
 

1
 

1
 

1
 

HIGH DOSE LOW DOSE 

CONTROL 

10 50 

1 6 

1 15 

3 29 

2 28 

3 32 

1 20 

1 21 

1 10 

2 10 

1 

1 

1 

1 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS
 

# SECONDARY TUMORS: METASTATIC TUMORS DR TUMORS INVASIVE INTO AN ADJACENT ORGAN
 

44
 

HIGH DOSE
 

50
 

12
 

15
 

23
 

20
 

24
 

14
 

14
 

5
 

8
 

1
 

1
 

2
 

2
 
TABLE A2.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE RATS 

FEDCAPTANINTHE DIET 

LOW DOSE HIGH DOSE
 CONTROL CONTROL
 

ANIMALS INITIALLY IN STUDY 5 10
 

ANIMALS NECROPSIED 4 5
 

ANIMALS EXAMINED HISTOPATHOLOGICALLY 4 5
 

INTEGUMENTARY SYSTEM
 

•SUBCUT TISSUE	 (4) (5)
 

FIBROUS HISTIOCYTOMA, MALIGNANT
 

AMELOBLASTOMA
 

RESPIRATORY SYSTEM
 

#LUNG (4) (5)
 

BILE DUCT CARCINOMA, METASTATIC
 

COPTICAL CARCINOMA, METASTATIC
 

HEMATOPOCETIC SYSTEM
 

•MULTIPLE ORGANS	 (4) (5)
 

MALI3.LYMPHOMA, LYMPHOCYTIC TYPE
 

#SPLi2EN (5)
 

BILE DUCT CARCINOMA, METASTATIC
 

CIRCULATORY SYSTEM
 

NONL·
 

DIGESTIVE SYSTEM
 

#LIVoR (*») (5)
 

NEOPLASTIC NODULE
 

HEPATOCELLULAR CARCINOMA
 

•BILr. DUCT (5)
 

RILE DUCT ADENOMA
 

# NUMbEB OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMdER DF ANIMALS NECROPSIED 

LOW DOSE
 

50
 

50
 

50
 

(50)
 

1 {2%)
 

1 (2%)
 

(50)
 

1 (2%)
 

(50)
 

(U9)
 

1 (2%)
 

(49)
 

4 (8%)
 

(50)
 

HIGH DOSE
 

50
 

50
 

50
 

(50)
 

1 [2%)
 

(50)
 

1 (2%)
 

(50)
 

(50)
 

1 (2%)
 

(50)
 

1 f 2*1
 

45
 
TABLE A2. FEMALE RATS: NEOPLASMS (CONTINUED) 

LOW DOSE HIGH DOSE LOW DOSE HIGH DOSE 

CONTROL CONTROL 

BILE DUCT

HAMARTOMA 

 CARCINOMA 

1 (20%) 

1 (2%) 

iPANCREAS 

BILE DUCT CARCINOMA, METASTATIC 

(45) 

1 (2%) 

(48) 

#SMALL INTESTINE 

BILE DUCT CARCINOMA, METASTATIC 

(4) (5) (47) 

1 (2%) 

(49) 

URINARY SYSTEM 

#KIDNEY ( 4) (5) (49) (47) 

TUBULAR-CELL ADENOMA 1 (2%) 

MIXED TUMOE, BENIGN 1 (2%) 

MIXED TUMOR, MALIGNANT 1 (2%) 

t HHMARTOMA 1 (2%) 1 (2%) 

#UPINARY BLADDER (5) (48 ) (43) 

BILE DUCT CARCINOMA, METASTATIC 1 (2%) 

ENDOCRINE SYSTEM 

#PITUITARY (4) (4) (48 ) (45) 

CARCINOMA,NOS 1 (2%) ADENOMA, NOS 1 (2%) 

CHROMOPHOBE ADENOMA 2 (50%) 1 (25%) 12 (25%) 4 (9%) 

#ADRENAL ( 4) (5) (50) (47) 

CORTICAL ADENOMA 1 (2%) 2 (4%) CORTICAL CARCINOMA 1 (2%) 1 (2%) 

PhEOCHROMOCYTOMA 1 (2%) 1 (2%) 

#THYROID (4) (5) (49) (44) 

FOLLICULAR-CELL ADENOMA 1 (20%) 

C-CELL ADENOMA 1 (2%) 4 (9%) 

C-CELL CARCINOMA 1 (2%) 

#PANCREATIC ISLETS (4) (45) (48) 

ISLET-CELL ADENOMA 1 (25%) 3 (6%) 

REPRODUCTIVE SYSTEM 

•MAMMARY GLAND (5) (50) (50) 

ADENOMA. NOS 3 16%) 1 (2%) 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBER OF ANIMALS NE3ROPSIED 

t 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.
 

46
 
TABLE A2. FEMALE RATS: NEOPLASMS (CONTINUED) 

_—__­_ —____ ___ . ___________ 

LOW DOSE 

CONTROL 

HIGH DOSE 

CONTROL 

LOW DOSE HIGH DOSE 

ADENOCARCINOMA, NOS 

SWEAT GLAND CARCINOMA 

INFILTRATING DUCT CAPCINOMA 

FIBROMA 

FIBROADENOMA 

2

4

 (4%) 

 (8%) 

1

1

1

3

5

 (2%) 

 (2%) 

 (2%) 

 (6%) 

 (10%) 

tUTERUS 

CARCINOMA,NOS 

SARCOMA, NOS 

EbDOMBTRIAL STROMAL POLYP 1 (25%) 

(5) 

1 (20%) 

(4 8) 

1

6

 (2%) 

 (13%) 

(45) 

1

7

 (2%) 

 (16%) 

#OVARY 

THECOMA 

GRANULOSA-CELL CAPCINOMA 

( 4 ) (5) (49) 

1

1

 (2%) 

 (2%) 

(46) 

NERVOUS SYSTEM 

ΝΟΝΞ 

SPECIAL SENSE ORGANS 

NONE 

MUSCULOSKELETAL SYSTEM 

NONE 

EODY CAVITIES 

•PERITONEUM 

BILE DUCT CARCINOMA,

SARCOMA, NOS 

 METASTATIC 

(5) (50) 

1 (2%) 

(50) 

1 (2%) 

ALL OTHER SYSTEMS 

NONE 

# NUMBER

* NUMBER

 OF

 OF

 ANIMALS

 ANIMALS

 WITH TISSUE

 NECROPSIED 

 EXAMINED MICROSCOPICALLY 

47
 
TABLE A2. FEMALE RATS: NEOPLASMS (CONTINUED) 

LOW DOSE 

CONTROL 

HIGH DOSE 

CONTROL 

LOW DOSE HIGH DOSE 

ANIMAL DISPOSITION SUMMARY 

ANIMALS INITIALLY IN STUDY 

NATURAL DEATHS 

MORIBUND SACRIFICE 

SCHEDULED SACRIFICE 

ACCIDENTALLY KILLED 

TERMINAL SACRIFICE 

ANIMAL MISSING 

5 

1 

1 

3 

10 

5 

50 

2 

10 

38 

50 

3 

9 

38 

 INCLUDES AUTCLYZED ANIMALS 

TUMOR SUMMARY 

TOTAL ANIMALS WITH PRIMARY TUMORS* 

TOTAL PRIMARY TUMORS 

3 

α 4 

4 

36 

47 

32 

41 

TOTAL ANIMALS WITH BENIGN

TOTAL BENIGN TUMORS 

 TUMORS 3 

4 

4 

4 

30 

36 

25 

32 

TOTAL ANIMALS WITH MALIGNANT

TOTAL MALIGNANT TUMORS 

 TUMORS 7 

7 

8 

9 

TOTAL ANIMALS WITH SECONDARY TUMORS» 

TOTAL SECONDARY TUMORS 

1 

6 

1 

1 

TOTAL ANIMALS WITH TUMORS

BENIGN OR MALIGNANT 

TOTAL UNCERTAIN TUMORS 

 UNCERTAIN4 

4 

TOTAL ANIMALS WITH TUMORS UNCERTAINPRIMARY OR METASTATIC 

TOTAL UNCERTAIN TUMORS 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS 

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

48
 
APPENDIX B
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

MICE FED CAPTAN IN THE DIET
 

49
 

TABLE B1.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE 

FED CAPTAN IN THE DIET 

HIGH DOSE
 

50
 

49
 

49
 

(49)
 

1 (2%)
 

(1)
 

1 (100%)
 

(49)
 

1 (2%)
 2 (4%)
 

(46)
 

2 (4%)
 

3 (7%)
 

ANIMALS INITIALLY IN STUDY
 

ANIMALS NECROPSIED
 

ANIMALS EXAMINED HISTOPATHOLOGICALLY
 

INTEGUMENTABY SYSTEM
 

NONE
 

RESPIRATORY SYSTEM
 

#LUNG
 

HEPATOCELLULAR CARCINOMA, METAST
 

ALVEOLAR/BBONCHIOLAR ADENOMA
 

ALVE0LAR/BR0NCHIOLAR CARCINOMA
 

HEMATOPOIETIC SYSTEM
 

#CECUM
 

MALIGNANT LYMPHOMA, NOS
 

CIRCULATORY SYSTEM
 

NONE
 

DIGESTIVE SYSTEM
 

#LIVER
 

NEOPLASTIC NODULE
 

HEPATOCELLULAR CARCINOMA
 

iDUODENUM
 

ADENOMATOUS POLYP, NOS
 

ADENOCA IN ADENOMATOUS POLYP
 

URINARY SYSTEM
 

NONE
 

CONTROL LOW DOSE
 

10
 

9
 

9
 

(9)
 

1 (11%)
 

2 (22%)
 

(9)
 

3 (33%)
 

(9)
 

50
 

48
 

47
 

(47)
 

2 (4%)
 

1 (2%)
 

(46)
 

1 (2%)
 

(43)
 

2 (5%)
 

1 (2%)
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

• NUMBER OF ANIMALS NECROPSIED
 

51
 
CONTROL LOW DOSE HIGH DOSE 

ENDOCRINE SYSTEM 

REPRODUCTIVE SYSTEM 

NONE 

NERVOUS SYSTEM 

NONE 

SPECIAL SENSE ORGANS 

NONE 

MUSCULOSKELETAL

NONE 

 SYSTEM 

BCDY CAVITIES 

NONiS 

ALL OTHER

NONE 

 SYSTEMS 

ANIMAL DISPOSITION SUMMARY 

ANIMALS INITIALLY IN STUDY 10 50

NATURAL DEATHS 1 3

MORIBUND SACRIFICE

SCHEDULED SACRIFICE 

ACCIDENTALLY KILLED 

TERMINAL SACRIFICE 9 47

ANIM\L MISSING 

3 INCLUDES AUTOLYZED ANIMALS 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBER OF ANIMALS NE3ROPSIED 

 50 

1 

1 

 48 

52
 
TABLE B1. MALE MICE: NEOPLASMS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE
 

TUMOR SUMMARY
 

TOTAL ANIMALS WITH PRIMARY TUMORS* 5 7 10
 

TOTAL PRIMARY TUMORS 5 7 10
 

TOTAL ANIMALS WITH BENIGN TUMORS 2 4 3
 

TOTAL BENIGN TUMORS 2 4 3
 

TOTAL ANIMALS WITH MALIGNANT TUMORS 3 3 6
 

TOTAL MALIGNANT TUMORS 3 3 6
 

TOTAL ANIMALS WITH SECONDARY TUMORS! 1
 

TOTAL SECONDARY TUMORS 1
 

TOTAL ANIMALS WITH TUMORS UNCERTAINBENIGN DR MALIGNANT 1
 

TOTAL UNCERTAIN TUMORS 1
 

TOTAL ANIMALS WITH TUMORS UNCERTAINPRIMARY OR METASTATIC
 

TOTAL UNCERTAIN TUMORS
 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS
 

# SECONDARY TUMORS: METASTATIC TUMORS DR TUMORS INVASIVE INTO AN ADJACENT ORGAN
 

53
 
TABLE B2.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICE 

FED CAPTAN IN THE DIET 

CONTROL LOW DOSE HIGH DOSE
 

ANIMALS INITIALLY IN STUDY 10 50 50
 ANIMALS NECROPSIED 10 50 49
 

ANIMALS EXAMINED HISTOPATHOLOGICALLY 10 50 48
 

INTEGUMENTARY SYSTEM
 

NONE
 

RESPIRATORY SYSTEM
 

NONJS
 

HEMATOPOIETIC SYSTEM
 

• MULTIPLE ORGANS (10) (50) (4 9)
 

MALIG.LYMPHOMA, HISTIOCYTIC TYPE 1 (10%) 1 (2%) 1 (2%)
 

LYMPHOCYTIC LEUKEMIA 1 (2%)
 

CIRCULATORY SYSTEM
 

NONE
 

DIGESTIVE SYSTEM
 

#LIVER (10) (4 9) (47) 

NEOPLASTIC NODULE 1 (10%) 1 (2%) 1 (2%) 

tSTOMACH (10) (49) (47)
 

SQUAMOUS CELL PAPILLOMA 1 (2%)
 

«DUODENUM (9) (49) (48) 

ADENOMATOUS POLYP, NOS 1 (11*) 1 (2%) 

ADENOCA IN ADENOMATOUS POLYP 3 (6%) 

URINARY SYSTEM
 

# NUMBER OP ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NE3ROPSIED
 

54
 
TABLE B2. FEMALE MICE: NEOPLASMS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

ENDOCRINE SYSTEM 

NONE 

REPRODUCTIVE SYSTEM 

• MAMMARY GLAND

ADENOMA, NOS

#OVARY

CYSTADENOMA, NOS

HEMANGIOMA

 (10 )

 (10)

 (50)

 1 (2%) 

 (46)

 (49) 

 (45) 

1 (2%) 

1 (2X) 

NERVOUS SYSTEM 

NONE 

SPECIAL SENSE ORGANS 

NONE 

MUSCULOSKELETAL SYSTEM 

NONE 

EODY CAVITIES 

NONE 

ALL OTHER SYSTEMS 

sau£ 

# NUMBEP OF ANIMALS

* NUMBED OP ANIMALS

 WITH TISSUE

 NECROPSIED 

 EXAMINED MICROSCOPICALLY 

55 
TABLE B2. FEMALE MICE: NEOPLASMS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE
 

ANIMAi, DISPOSITION SUMMAFY
 

ANIMALS INITIALLY IN STUDY 10 50 50
 

NATURAL DSATHo) 2 2
 

MORIBUND SACRIFICE 1 2
 

SCHEDULED SACRIFICE
 

ACCIDENTALLY KILLED
 

TERMINAL SACRIFICE 9 48 U6
 

ANIMAL MISSING
 

a) INCLUDES AUTOLYZED ANIMALS
 

TUMOR SUMMARY
 

TOTAL ANIMALS WITH PRIMARY TUMORS* 3 5 8
 

TOTAL PRIMARY TUMORS 3 5 8
 

TOTAL ANIMALS WITH BENIGN TUMORS 1 3 2
 

TOTAL BENIGN TUMORS 1 3 2
 

TOTAL ANIMALS WITH MALIGNANT TUMOPS 1 1 5
 

TOTAL MALIGNANT TUMORS 1 1 5
 

TOTAL ANIMALS WITH SECONDARY TUMORS*
 TOTAL SECONDARY TUHOFS
 

TOTAL ANIMALS WITH TUMORS UNCERTAIN­ BENIGN DR MALIGNANT 1 1 1
 TOTAL UNCERTAIN TUMORS 1 1 1
 

TOTAL ANIMALS WITH TUMORS UNCERTAINPRIMARY OR METASTATIC
 

TOTAL UNCERTAIN TUMORS
 

• PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMOPS
 

# SECONDARY TUMORS: METASTATIC TUMOPS DR TUMORS INVASIVE INTO AN ADJACENT ORGAN
 

56
 
APPENDIX C
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS
 

IN RATS FED CAPTAN IN THE DIET
 

57
 

TABLE C1. 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS 

FED CAPTAN IN THE DIET 

LOW DOSE HIGH DOSE LOW DOSE HIGH DOSE 

CONTROL CONTROL 

ANIMALS

ANIMALS

ANIMALS

 INITIALLY IN STUDY 

 NECROPSIED 

 EXAMINED HISTOPATHOLOGICALLY 

5 

5 

5 

10 

5 

5 

50 

50 

49 

50 

49 

49 

INTEGUMENTARY SYSTEM 

NONE 

RESPIRATORY SYSTEM 

#LUNG 

ATELECTASIS 

CONGESTION, NOS 

EDEMA, NOS 

BRONCHOPNEUMONIA SUPPURATIVE 

PNEUMONIA, CHRONIC MURINE 

BttONCHOPNEUMONIA, CHRONIC 

INFLAMMATION, CHRONIC FOCAL 

GRANULOMA, NOS 

INFLAMMATION, FOCAL GRANULOMATOU 

CALCIFICATION, METASTATIC 

(4) (5) (*8) 

1 (2%) 

1 (2%) 

1 (2%) 

1 (2%) 

1 (2%) 

(»9) 

1

1

1

1

1

1

 (2%) 

 (2%) 

 (2%) 

 (2%) 

 (2%) 

 (2%) 

#LUNG/ALVEOLI 

EMPHYSEMA, NOS 1 (25%) 

(5) 

1 (20%) 

(48) 

1 (2%) 

(49) 

1 (2%) 

HEMATOPOIETIC SYSTEM 

tSPLEEN 

FIBROSIS , FOCAL 

PERIARTERITIS 

HEMOSIDEROSIS 

HEMATOPOIESIS 

W 

1 (25%) 

(5) (4 9) 

1 (2%) 

1 (2%) 

1 (2%) 

1 (2%) 

(47) 

1 (2%) 

CIRCULATORY SYSTEM 

#MYOCfcRDIUM 

INFLAMMATION­ CHRONIC 

W (5) 

1 120%) 

(48) («9) 

I NUMBER OF ANIMALS

* NUMBER OF ANIMALS

 WITH TISSUE

 NECROPSIED 

 EXAMINED MICROSCOPICALLY 

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

LOW DOSE HIGH DOSE

CONTROL CONTROL 

INFLAMMATION, CHRONIC FOCAL

FIBROSIS, DIFFUSE

*AORIA	 (5) (5)


MUDIA L CALCIFICATION


CALCIFICATION, METASTATIC


DIGESTIVE SYSTEM 

#SALIVARY GLAND

INFLAMMATION, CHRONIC

 (5) (5)

#LIV£R

PERIARTERITIS

DEGENERATION, BALLOONING

DEGENERATION PARENCHYMATOUS

METAMORPHOSIS FATTY

FOCAL CELLULAR CHANGE

ANGIECTASIS

 (5)

 1 (20%) 

 1 (20%)

 (5)

 2 (40%)

 1 (20%)

• BILi. DUCT

INFLAMMATION, CHRONIC

HYPERPLASIA, NOS

 (5)

 1 (20%) 

 (5)

 *

#PANCREAS

THROMBOSIS, NOS

INFLAMMATION, CHRONIC FOCAL

PERIARTERITIS

 (4)

 1 (25%)

 (5)

 1 (20%)

#STOMACH

EROSION

N&CROSIS, FOCAL

CALCIFICATION, NOS

 (5) (5)

#GASTRIC MUCOSA

HEMORRHAGE

CALCIFICATION, METASTATIC

 (5) (5)

URINARY SYSTEM
 

#KIDNEY (5) (5)


INFLAMMATION, CHRONIC 3 (60%) 5 (100%)


INFLAMMATION, CHRONIC FOCAL


GLOMERULOSCLEROSIS. NOS


# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NE3ROPSIED
 

 LOW DOSE

 HIGH DOSE 

1 (2%) 

1 (2%) 

 (50)	 (49)
 

 1 (2%)
 

 1 (2%)
 

 (47)	 (43)
 

 1 (2%)
 

 (47) (49)
 

 1 (2%)
 

 5 (11%) 10 (20%)
 

 9 (19%) 14 (29%)
 

 1 (2%) 5 (10%)
 

 4 (9%) 3 (6%)
 

 (50)	 (49)
 

 1 (2%)
 

 (45) (47)
 

 1 (2%)
 

 1 (2%)
 

 1 (2%) 4 (9%)
 

 (47) (44)
 

 1 (2%)
 

 1 (2%)
 

 1 (2%)
 

 (47) (44)
 

 1 (2%)
 

 1 (2%)
 

 (49) (49)
 

 25 (51%)	 33 (67%)
 

 1 (2%)
 

 1 12%)
 

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

LOW DOSE 

CONTROL 

HIGH DOSE 

CONTROL 

LOW DOSE HIGH DOSE 

ENDOCRINE SYSTEM 

tPITUITARY 

MULTIPLE CYSTS 

CONGESTION, NOS 

HEMORRHAGE 

DEGENERATION, CYSTIC 

HYPERPLASIA, NOS 

ANGEECTASIS 

(5) 

1 (20%) 

(5) 

1 

1 

(20%) 

(20%) 

(*3) 

2 (5%) 

2 15%) 

2 (5%) 

6 (14%) 

(*5) 

1 (2%) 

#ADR£NAL 

DEGENERATION,

ANGEECTASIS 

 CYSTIC 

( 5 ) (5) (47) 

1 (2%) 

(47) 

1 (2%) 

#ADRUN& L CORTEX 

HEMORRHAGE 

NECROSIS, FOCAL 

METAMORPHOSIS FATTY 

( 5) 15) (47) 

1

1

1

 (2%) 

 (2%) 

 (2%) 

(47) 

3

3

 (6%) 

 (6%) 

tTHYROED 

FOLLICULAR CYST, NOS 

HEMORRHAGE 

HYPERPLASIA, C-CELL 

HYPERPLASIA, FOLLICULAR-CELL 

(5) 

1 (20%) 

(5) 

1 

2 

(20%) 

(40%) 

(42) 

2

1

1

1

 (5%) 

 i2%) 

 (2%) 

 (2%) 

(47) 

1

1

6

 (2%) 

 ί 2%) 

 (13%) 

tPARATHYROID 

HYPERPLASIA,

HYPERPLASIA,

 NOS 

 SECONDARY 

(3) (2) (21) 

1 (5%) 

(33) 

2

1

 (6%) 

 13%) 

REPRODUCTIVE SYSTEM 

#PROSTATE 

INFLAMMATION,

INFLAMMATION,

 ACUTE 

 ACUTE SUPPURATIVE 

(*) (5) (44) (45) 

1

1

 (2%) 

 (2%) 

#TESIIS 

EDEMA, NOS 

PERIARTERITIS 

DEGENERATION, NOS 

ATROPHY, NOS 

ATROPHY, FOCAL 

(5) 

3 (60%) 

(5) 

1 (20%) 

(4 8) 

3

2

14

 (6%) 

 (4%) 

 (29%) 

(48) 

1

4

2

6

2

 (2%) 

 (8%) 

 (4%) 

 (13%) 

 (4%) 

•EPIDIDYMIS 

HEMORRHAGE 

(5) J5) (50) (49) 

1 12%) 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED

* NUMBER OF ANIMALS NECROPSIED 

 MICROSCOPICALLY 

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

INFLAMMATION, CHRONIC
 

FIBROSIS
 

NERVOUS SYSTEM
 

NONE
 

SPECIAL SENSE ORGANS
 

NONE
 

MUSCULOSKELETAL SYSTEM
 

•FEMUR
 

OSTEOPOROSIS
 

EODY CAVITIES
 

•MEShNTERY
 

PERIARTERITIS
 

ALL OTHER SYSTEMS
 

NONE
 

SPECIAL MORPHOLOGY SUMMARY
 

NO LESION REPORTED
 

AUTO/NECROPSY/NO HISTO
 

AUTOLYSIS/NO NECROPSY
 

# NUMBER OF ANIMALS WITH TISSUE

* NUMBER OF ANIMALS NECROPSIED 

LOW DOSE HIGH DOSE LOW DOSE
 

CONTROL CONTROL
 

(5) (5) (50) 

(5) 

1 (20%) 

(5) 

1 J 20%) 

(50) 

1 (2%) 

 EXAMINED MICROSCOPICALLY 

3 

1 

62
 

HIGH DOSE
 

1 {2%)
 

1 (2%)
 

(<*9)
 

1 (2%)
 

(49)
 

 (16%)
 

1
 
TABLE C2. 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS
 

FED CAPTAN IN THE DIET
 

LOW DOSE HIGH DOSE LOW DOSE HIGH DOSE
 

CONTROL CONTROL
 

ANIMALS INITIALLY IN STUDY 5 10 50 50
 ANIMALS NECROPSIED 4 5 50 50
 ANIMALS EXAMINED HISTOPATHOLOGICALLY 4 5 50 50
 

INTEGUMENTARY SYSTEM
 

•SKIN	 (5) (50) (50)
 

DEGENERATION, CYSTIC 1 (2%)
 

RESPIRATORY SYSTEM
 

#LUNG (5) (50) (49)
 

EMPHYSEMA, NOS 1 (2%)
 

ATELECTASIS 1 (2%)
 

INFLAMMATION, FOCAL 1 (2%) 1 (2%)
 

PNEUMONIA, ASPIRATION 1 (25%)
 

PNEUMONIA, CHRONIC MURINE 1 (2%)
 

INFLAMMATION, FOCAL GPANULOMATOU 1 [2%)
 

NECROSIS, NOS 1 (2%)
 

HEMATOPOIETIC SYSTEM
 

#SPLEEN (Ό (5) (49) (50)
 

CONGESTION, CHRONIC PASSIVE 1 (2%)
 

INFLAMMATION, CHRONIC 1 [2%)
 

FIBROSIS 1 (2%)
 

HYPERPLASIA, NOS 1 (2%)
 

HYPERPLASIA, LYMPHOID 1 (2%)
 

HEMATOPOIESIS 1 (2%)
 

#LYMPH NODE t5) (44) (41)
 

INFLAMMATION, NOS 1 (2%)
 

tCERVICAL LYMPH NODE (U) (5) (44) (U1)
 

INFLAMMATION ACUTE AND CHRONIC 1 (20%)
 

CIFCULVTOR Y SYSTEM
 

#MYOCARDIUM (") (50) (SO)
 

INFLAMMATION. INTERSTITIAL 1 i2%)
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBER OF \NI^ALS NECROPSIED 

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

LOW DOSE HIGH DOSE LOW DOSE HIGH DOSE 

CONTROL CONTROL 

# ENDOCARDIUM (4) (50) (50 ) 

FIBROSIS, FOCAL 1 (2%) 

DIGESiIVE SYSTEM 

#LIVLR (5) (49 ) (50 ) 

DEGENERATION PARENCHYMATOUS 2 (50%) 3 (60%) 1 (2% ) 6 (12%) 

NLCROSIS , FOCAL 1 (2%) 

METAMOPPHOSIS FATTY 1 (25%) 4 (8%) 5 (10%) 

FuCAL CELLULAR CHANGE 1 (2%) 2 (4%) 

ANGIECTASIS 1 (20%) 3 (6%) 8 (16%) 

•BILL·	 DUCT (5) (50 ) (50 )
 

IWFLAMMATICN, CHRONIC 1 (20%)
 

FIBROSIS 1 i2% )
 

HYPERPLASIA, NOS 2 (U%)
 

[*) 

#GASTRIC MUCOSA (5) (4 9) (49 )
 

EtOSION 1 (2% )
 

URINAL·/ SYS^M 

#KIDNEY (5) (49 ) (4*7) 

CYST, N3S 1 (2%) 

INFLAMMATION, CHRONIC 2 (U0%) 7 (14%) 4 (9%) 

ENDOChINE SYSTEM 

#PITJITAPΥ ( 4)	 CM (43 ) (45 ) 

CYST, NDS 1 (2%) 

CONGESTION, NOS 1 [2%) 

HEMORRHAGIC CYST 1 (2%) 

DEGENERATION, CYSTIC 1 (2%) 

HYPERPLASIA, NOS 1 (25%) 2 (4% ) 1 (2%) 

HYPERPLASIA, FOCAL 1 (2%) 

fNGIECTASIS 8 (17% ) 2 (4%) 

#ADRi.NAL	 (5) (50 ) (47 ) {*) 

CuNGSSTION, NOS 1 (2%) 1 (2%) 

HEMORRHAGE 4 13%) 2 (4%) 

DrGENEPATICN, CYSTIC 1 (25%) 

NLCRQSIS, NOS 1 i2%) 1 i2%L 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMDER OF ANIMALS NECROPSIED 

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

LOW DOSE HIGH DOSE LOW DOSE HIGH DOSE 

CONTROL CONTROL 

METAMORPHOSIS FATTY 1 (2%) 1 (2%) 

HEMOSIDEROSIS 1 (2%) 

HYPERPLASIA,, NOS 1 (2%) 

ANGIECTASIS 1 (2%) 

#ADRhNAL CORTEX (5) (50) (47) 

HEMORRHAGE 2 (4%) 1 (2%) 

D&GENERATICN, CYSTIC 1 (25%) 1 (2%) 

NECROSIS, NOS 1 (2%) 

NECROSIS, HEMORRHAGIC 1 (2%) 

McTAMDRPHOSIS FATTY 1 (2%) 

ATROPHY, NCS 1 (2%) 

HYPERPLASIA,, FOCAL 1 (25%) 

ANGIECTASIS 2 (4%) 

#THYROID i5) (44) 

FOLLICULAR CYST, NOS 1 (25%) 

HYPERPLASIA, C-CELL 3 (6%) 2 i5%) 

HYPERPLASIA, FOLLICULAR-CELL 1 (20%) 1 (2%) 2 (5%) 

REPRODUCTIVE SYSTEM 

•MAMMARY GLAND (<O (5) (50) (50) 

HYPERPLASIA, NOS 2 |tt%) 

•MAMMARY LOBULE (5) {50) (50) 

HYPERPLASIA, NOS 1 (20%) 3 (6%) 

#OVARY i<0 (5) (4 9) (46) 

FOLLICULAR CYST, NOS 1 (2%) 

NERVOUS SiSTEM 

NONE 

SPECIAL SENSE CFGANS 

NONE 

MUSCULOSKELETAL SYSTEM 

•BONE W (5) (50) (50) 

OSTEOPOROSIS I i2%L 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

• NUMBER OF ANIMALS NECROPSIED 

65 
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED)
 

LOW DOSE HIGH DOSE LOW DOSE HIGH DOSE 

CONTROL CONTROL 

BODY CAVITIES 

NONt, 

ALL O1HER SYSTEMS 

NONt 

SPECIAL MORPHOLOGY SUMMARY 

NO LESICN EEPORTED 6 4 AUTOLYSIS/NO NECROPSY 1 

# NUMbEF OF ANIMALS WITH TISSUE EXAMINED MICEOSCOPICALLΥ 

* NUMBER OF ANIMALS NECROPSIED 

66
 
APPENDIX D
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS
 

IN MICE FED CAPTAN IN THE DIET
 

67
 

TABLE D1.
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE MICE 

FED CAPTAN IN THE DIET 

CONTROL LOW DOSE HIGH DOSE 

ANIMALS INITIALLY IN STUDY 10 50 50 

ANIMALS NECROPSIED 9 48 49 

ANIMALS EXAMINED HISTOPATHOLOGICALLY 9 47 49 

INTEGUMENTARY SYSTEM 

NONL· 

RESPIRATORY SYSTEM 

tLUNG (9) (47) (4 9) 

HYPERPLASIA, ALVEOLAR EPITHELIUM 2 (4%) 

HEMATOPOIETIC SYSTEM 

• MESENTERIC L. NODE (9) (37) ί*1> 

CONGESTION, CHRONIC PASSIVE 1 (3X) 

INFLAMMATION, NOS 1 (2%) 

CIRCULATORY SYSTEM 

#MYOCARDIUM (9) (47) (49) 

DEGENERATION, NOS 1 (2%) 

DIGESTIVE SYSTEM 

tLEVER 

DEGENERATION PARENCHYMATOUS 

NECROSIS, FOCAL 

(9) (46) 

2

1

 (4%) 

 (2%) 

(49) 

#PANCREAS 

DILATATION/DUCTS 

INFLAMMATION, NOS 

(9) (47) (49) 

1

1

 (2%) 

 (2%) 

tDUODENUM 

HYPERPLASIA­ FOCAL 

(9) (43) (4 6) 

1 f 2%) 

# NUMBER

* NUMBER

 OF

 OF

 ANIMALS

 ANIMALS

 WITH TISSUE

 NECROPSIED 

EXAMINED MICROSCOPICALLY 

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

CONTROL LOW DOSE HIGH DOSE 

#DUODENAL MUCOSA (9) (43) (46)
 

HYPERPLASIA, NOS 1 (2%)
 

HYPERPLASIA, FOCAL 2 (4%)
 

URINAHY SYSTEM
 

NONE
 

ENDOCRINE SYSTEM
 

NONE
 

REPRODUCTIVE SYSTEM
 

NONE
 

NERVOUS SYSTEM
 

NONE
 

SPECIAL SENSE ORGANS
 

NONE
 

MUSCULOSKELETAL SYSTEM
 

NONE
 

EODY CAVITIES
 

NONE
 

ALL OIHER SYSTEMS
 

NONE
 

SPECIAL MORPHOLOGY SUMMARY
 

NO LESION BEPORTED 4 35 34
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

70 
TABLE D1. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED)

 

CONTROL LOW DOSE HIGH DOSE 

AUTO/NECRCESY/NO HISTO 

AUTOLYSIS/NO NECROPSY 1 

1 

2 1 

#

*

 NUMBER

 NUMBER

 OF

 OF

 ANIMALS

 ANIMALS

 HITH TISSU E

 NECROPSIED 

 EXAMINED MICROSCOPICALLY 

71 
TABLE D2.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE 

FED CAPTAN IN THE DIET 

HIGH DOSE
 

50
 

49
 

48
 

(48)
 

(48)
 

(36)
 

(47)
 

1 (2%)
 

(47)
 

ANIMALS INITIALLY IN STUDY
 

ANIMALS NECROPSIED
 

ANIMALS EXAMINED HISTOPATHOLOGICALLY
 

INTEGUMENTARY SYSTEM
 

NONE
 

RESPIRATORY SYSTEM
 

#LUNG
 

HYPERPLASIA, ALVEOLAF EPITHELIUM
 

HEMATOPOIETIC SYSTEM
 

#SPLLEN
 

CONGESTION, ACUTE
 

HYPERPLASIA, LYMPHOID
 

HEMATOPOIESIS
 

«MESENTERIC L. NODE
 

INFLAMMATION, NOS
 

CIRCULATORY SYSTEM
 

NONE
 

DIGESTIVE SYSTEM
 

•LIVER
 

INFLAMMATION, NOS
 

DEGENERATION PARENCHYMATOUS
 

METAMORPHOSIS FATTY
 

•PANCREAS
 

HAfUI3PQII£IS
 

CONTROL LOW DOSE
 

10 50
 

10 50
 

10 50
 

(10)	 (49)
 

1 (2%)
 

(10) (49)
 

1 (2%)
 

1 (2%)
 

1 (2%)
 

(10)	 (42)
 

1 (2%)
 

(10)	 (49)
 

1 (10%)
 

7 {14%)
 

1 (10%)
 

(10) (48)
 

1 llftft.-.
 

# NUMuER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER DF ANIMALS NECROPSIED
 

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

 

CONTROL LOW DOSE
 

URINAH* SYSTEM
 

#KIDNEY (8) (50)
 

INFLAMMATION, CHRONIC 1 (2%)
 

ENDOCRINE SYSTEM
 

NONE
 

REPRODUCTIVE SYSTEM 

#UTEKUS (10) (45) 

HEMORRHAGE 1 (2%) 

#UTERUS/ENDOMETRIUM MO) (45) 

HYPERPLASIA, CYSTIC 1 (10%) 3 (7%) 

#OVARY (10) (46) ' 

INFLAMMATION, NOS 3 (30%) 

INPLAMMATION, SUPPURATIVE 1 (10%) 3 (7%) 

NERVOUS SYSTEM
 

NONE
 

SPECIAL SENSE ORGANS
 

NONE
 

MUSCULOSKELETAL SYSTEM
 

NONE
 

BODY CAVITIES
 

NONE
 

ALL OTHER SYSTEMS
 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NE3ROPSIED
 

HIGH DOSE
 

(48)
 

(43)
 

(43)
 

3 (7%)
 

(45)
 

73 
TABLE D2. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

SPECIAL MORPHOLOGY SUMMARY 

NO LESION REPORTED

AUTO/NECROPSY/NO HISTO

AUTOLYSIS/NO NECROPSY

 2 29 38 

1 

1 

# NUMBER OF ANIMALS

* NUMBER OF ANIMALS

 WITH TISSUE

 NECROPSIED 

 EXAMINED MICROSCOPICALLY 

74 
APPENDIX E
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS
 

IN RATS FED CAPTAN IN THE DIET
 

75
 

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

Fed Captan in the Dieta
 

Topography: Morphology 

Pooled 

Control 

Matched 

Control 

Low 

Dose 

High 

Dose 

Thyroid: C-cell Adenomab 2/65 (3) 0/10 (0) 1/42 (2) 1/47 (2) 

 Valuesc,d N.S. N.S. N.S. N.S. 

Relative Risk (Pooled Control)f 

Lower Limit 

Upper Limit 

0.774 

0.013 

14.321 

0.691 

0.012 

12.844 

Relative Risk (Matched Control)f 

Lower Limit 

Upper Limit 

Infinite 

0.014 

Infinite 

Infinite 

0.012 

Infinite 

Weeks to First Observed Tumor 113 75 

Pancreatic Islets:
 

Islet-cell Adenoma*5 3/72 (4) 0/9 (0) 1/45 (2) 1/47 (2)
 

 Valuesc , d N.S. N.S. N.S. N.S.
 

Relative Risk (Pooled Control)f 0.533 0.511
 

Lower Limit 0.010 0.010
 

Upper Limit 6.370 6.108
 

Relative Risk (Matched Control)f Infinite Infinite
 

Lower Limit 0.012 0.011
 

Upper Limit Infinite Infinite
 

Weeks to First Observed Tumor — —
 114 
Table El. Analyses of the Incidence of Primary Tumors in Male Rats
 

Fed Captan in the Diet3
 

(continued)
 

Topography: Morphology
 

Pituitary: Chromophobe
 

Adenoma**
 

 Values0

>d
 

Relative Risk (Pooled Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Liver: Neoplastic
 

Noduleb
 

 Values0»d
 

Relative Risk (Pooled Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pooled
 

Control
 

8/62 (13)
 

N.S.
 

2/73 (3)
 

N.S.
 

——
 

Matched
 

Control
 

2/10 (20)
 

M.S.
 

113
 

1/10 (10)
 

N.S.
 

113
 

Low
 

Dose
 

9/43 (21)
 

N.S.
 

1.622
 

0.603
 

4.414
 

1.047
 

0.283
 

9.208
 

60
 

1/47 (2)
 

N.S.
 

0.777
 

0.013
 

14.431
 

0.213
 

0.003
 

16.378
 

....
 

High
 

Dose
 

5/45 (11)
 

N.S.
 

0.861
 

0.235
 

2.768
 

0.556
 

0.116
 

5.436
 

86
 

2/49 (4)
 

N.S.
 

1.490
 

0.111
 

19.881
 

0.408
 

0.025
 

23.619
 

114
 
Table El. Analyses of the Incidence of Primary Tumors in Male Rats
 

Fed Captan in the Dieta
 

(continued)
 

a

Treated groups received time-weighted average doses of 2,525 or 6,050 ppm.
 

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

c

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


Armitage test when Ρ < 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 Ρ < 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 Ρ < 0.05 for any comparison.
 

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

control group.
 
Table Ε2· Analyses of the Incidence of Primary Tumors in Female Rats 

Fed Captan in the Diet a 

Pooled Matched Low
 

Topography: Morphology Control Control Dose
 

Pituitary: Chromophobe
 

Adenomab 12/62 (19) 3/8 (38) 12/48 (25)
 

 Valuesc,d M.S. = 0.014(N) N.S.
 

Relative Risk (Pooled Control)f 1.292
 

Lower Limit 0.531
 

Upper Limit 2.842
 

Relative Risk (Matched Control)f 0.667
 

Lower Limit 0.266
 

Upper Limit 3.220
 

Weeks to First Observed Tumor — 113 97
 

Liver: Neoplastic
 

Noduleb 1/71 (1) 0/9 (0) 4/49 (8)
 

 Values0»d N.S. N.S. N.S.
 

Departure from Linear Trende - 0.010
 

Relative Risk (Pooled Control)f 5.796
 

Lower Limit 0.594
 

Upper Limit 279.251
 

Relative Risk (Matched Control)^ Infinite
 

Lower Limit 0.192
 

Upper Limit Infinite
 

Weeks to First Observed Tumor _—
 — 113
 

High
 

Dose
 

4/45 (9)
 

N.S.
 

0.459
 

0.115
 

1.398
 

0.237
 

0.059
 

1.433
 

0/50 (0)
 

N.S.
 

0.000
 

0.000
 

26.485
 

——
 

—
 

97 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats
 

(continued)
 

Topography: Morphology
 

Liver: Neoplastic Nodule
 

or Hepatocellular
 

Carcinoma*5
 

 Values0»d
 

Departure from Linear Trende
 

Relative Risk (Pooled Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Adrenal: Cortical Adenoma
 

or Cortical Carcinoma**
 

 Values0

»d
 

Relative Risk (Pooled Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Fed Captan in the Dieta
 

Pooled Matched
 

Control Control
 

1/71 (1) 0/9 (0)
 

N.S. N.S.
 

 - 0.039
 

0/64 (0) 0/9 (0)
 

 - 0.047 N.S.
 

Low
 

Dose
 

4/49 (8)
 

N.S.
 

5.796
 

0.594
 

279.251
 

Infinite
 

0.192
 

Infinite
 

113
 

2/50 (4)
 

N.S.
 

Infinite
 

0.377
 

Infinite
 

Infinite
 

0.060
 

Infinite
 

1 1 1
 

High
 

Dose
 

1/50 (2)
 

N.S.
 

1.420
 

0.018
 

109.277
 

Infinite
 

0.011
 

Infinite
 

3/47 (6)
 

N.S.
 

Infinite
 

0.813
 

Infinite
 

Infinite
 

0.130
 

Infinite
 

«7
 

114 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats
 

Fed Captan in the Dieta
 

(continued)
 

Topography: Morphology
 

Pancreatic Islets: Islet-cell
 

Adenoma
 

 Values0>d
 

Relative Risk (Pooled Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Mammary Gland: Adenocarcinoma, NOS,
 

Adenoma, NOS, or
 

Infiltrating
 

Duct Carcinoma**
 

 Values0>d
 

Relative Risk (Pooled Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 —
 

Low
 

Dose
 

0/45 (0)
 

N.S.
 

0.000
 

0.000
 

28.540
 

0.000
 

0.000
 

3.329
 

—
 

3/50 (6)
 

N.S.
 

Infinite
 

0.800
 

Infinite
 

Infinite
 

0.122
 

Infinite
 

97
 

High
 

Dose
 

3/48 (6)
 

N.S.
 

4.313
 

0.359
 

221.533
 

0.500
 

0.051
 

25.730
 

114
 

3/50 (6)
 

N.S.
 

Infinite
 

0.800
 

Infinite
 

Infinite
 

0.122
 

Infinite
 

114
 

Pooled
 

Control
 

1/69 (1)
 

N.S.
 

—
 

0/67 (0)
 

N.S.
 

Matched
 

Control
 

1/8 (13)
 

N.S.
 

98
 

0/9 (0)'
 

N.S.
 

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

Fed Captan in the Diet a 

(continued) 

Topography: Morphology

Pooled 

 Control 

Matched 

Control 

Mammary Gland: 

Fibromab

 1/72 (1) 0/9 (0) 

 Valuesc » d N.S. N.S. 

Relative Risk (Pooled Control)f 

Lower Limit 

Upper Limit 

oo 

u> 

Relative Risk (Matched Control)f 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

Mammary Gland: 

Fibroadenoma15

 8/72 (11) 0/9 (0) 

 Valuesc » d N.S· N.S· 

Relative Risk (Pooled Control)f 

Lower Limit 

Upper Limit 

Relative Risk (Matched Control)f 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor — 

Low
 

Dose
 

2/50 (4)
 

N.S.
 

2.880
 

0.155
 

166.479
 

Infinite
 

0.060
 

Infinite
 

102
 

4/50 (8)
 

N.S.
 

0.720
 

0.166
 

2.523
 

Infinite
 

0.188
 

Infinite
 

75
 

High
 

Dose
 

3/50 (6)
 

N.S.
 

4.320
 

0.358
 

222.074
 

Infinite
 

0.122
 

Infinite
 

114
 

5/50 (10
 

N.S.
 

0.900
 

0.244
 

2.920
 

Infinite
 

0.257
 

Infinite
 

38
 
Table Ε2· Analyses of the Incidence of Primary Tumors in Female Rats
 

Fed Captan in the Dieta
 

(continued)
 

Topography: Morphology
 

Mammary Gland: Adenoma, NOS,
 

or Fibroadenoma*5
 

 Values0»d
 

Relative Risk (Pooled Control)^
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Uterus: Endometrial
 

Stromal Polypb
 

 Valuesc»d
 

Relative Risk (Pooled Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Pooled
 

Control
 

3/72 (11)
 

N.S.
 

mm mm
 ,


7/67 (10)
 

N.S.
 

—
 

Matched
 

Control
 

0/9 (0)
 

N.S.
 

2/9 (22)
 

N.S.
 

113
 

Low
 

Dose
 

7/50 (14)
 

N.S.
 

1.260
 

0.413
 

3.701
 

Infinite
 

0.396
 

Infinite
 

75
 

6/48 (13)
 

N.S.
 

1.196
 

0.354
 

3.876
 

0.563
 

0.134
 

5.293
 

66
 

High
 

Dose
 

6/50 (12)
 

N.S.
 

1.080
 

0.328
 

3.311
 

Infinite
 

0.326
 

Infinite
 

38
 

7/45 (16)
 

N.S.
 

1.489
 

0.476
 

4.608
 

0.700
 

0.178
 

6.391
 

114
 
Table E2. Analyses of the Incidence of Primary

Fed Captan in the Diet a 

(continued) 

Pooled Matched
 

Topography: Morphology Control Control
 

Thyroid: C-cell
 

Adenoma*5 1/66 (2) 0/9 (0)
 

 Valuesc>d	 « 0.035 N.S.
 

Relative Risk (Pooled Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Control)f
 

Lower Limit
 

Upper Limit
 00 

Weeks to First Observed Tumor
 «...	 «...


Thyroid: C-cell Adenoma
 

or Carcinoma*5 2/66 (3) 0/9 (0)
 

 Values0>d	 N.S. N.S.
 

Relative Risk (Pooled Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor ——
 ——
 

 Tumors in Female Rats 

Low	 High
 

Dose	 Dose
 

1/49 (2) 4/44 (9)
 

N.S.	 N.S.
 

1.347 6.000
 

0.017 0.619
 

103.614	 288.319
 

Infinite Infinite
 

0.011 0.214
 

Infinite Infinite
 

113	 114
 

2/49 (4) 4/44 (9)
 

N.S.	 N.S.
 

1.347 3.000
 

0.101 0.451
 

17.968	 31.881
 

Infinite Infinite
 

0.061 0.214
 

Infinite Infinite
 

113	 107
 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats
 

Fed Captan in the Dieta
 

(continued)
 

a

Treated groups received time-weighted average doses of 2,525 or 6,050 ppm.
 

^Number of tumor-bearing animals/number of animals examined at site (percent)·
 

c

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


Armitage test when Ρ < 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 Ρ < 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 Ρ < 0.05 for any comparison.
 

^The 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 CAPTAN IN THE DIET
 

87
 

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

Fed Captan in the Dieta 

Pooled Matched Low High 

Topography: Morphology Control Control Dose Dose 

Lung: Alveolar/Bronchiolar 

Adenoma or Carcinoma*5 5/66 (8) 2/9 (22) 3/47 (6) 1/49 (2) 

 Valuesc > d 

N.S. = 0.034(N) N.S. N.S. 

Relative Risk (Pooled Control)f 0.843 0.269 

Lower Limit 0.136 0.006 

Upper Limit 4.093 2.296 

Relative Risk (Matched Control)f 0.287 0.092 

Lower Limit 0.043 0.002 

Upper Limit 3.205 1.657 

Weeks to First Observed Tumor __ 91 91 91 
Table Fl.


(continued)
 

Topography: Morphology
 

Liver: Neoplastic Nodule
 

or Heptatocellular
 

Carcinoma
 

 Values0>d
 

Departure from Linear Trende
 

Relative Risk (Pooled Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

 Analyses of the Incidence of Primary Tumors in Male Mice
 

Fed Captan in the Dieta
 

Pooled Matched
 

Control Control
 

14/76 (18) 3/9 (33)
 

 = 0.012(N) N.S.
 

P^= 0·003
 

91
 

Low
 

Dose
 

1/46 (2)
 

 = 0.012*(N)
 

 = 0.006**(N)
 

0.118
 

0.003
 

0.729
 

0.065
 

0.001
 

0.738
 

91
 

High
 

Dose
 

3/49 (6)
 

 = 0.042*(N)
 

 = 0.041**(N)
 

0.332
 

0.064
 

1.109
 

0.184
 

0.033
 

1.233
 

84
 
Table Fl. Analyses of the Incidence of Primary Tumors in Male Mice
 

(continued)
 

Topography: Morphology
 

Duodenum; Adenomatous
 

Polyp, NOSb
 

 Values0>d
 

Relative Risk (Pooled Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Duodenum: Adenocarcinoma
 

in Adenomatous Polypi
 

 Values0>d
 

Relative Risk (Pooled Control)f
 

Lower Limit
 

Upper Limit
 

Relative Risk (Matched Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Fed Captan in the Dieta 

Pooled 

Control 

Matched 

Control 

Low 

Dose 

0/68 (0) 0/9 (0) 2/43 (5) 

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

Infinite 

0.463 

Infinite 

Infinite 

0.069 

Infinite 

91 

0/68 (0) 0/9 (Ο)­ 1/43 (2) 

 = 0.033 N.S. N.S. 

Infinite 

0.085 

Infinite 

Infinite 

0.012 

Infinite 

— 91 

High
 

Dose
 

2/46 (4)
 

N.S.
 

Infinite
 

0.433
 

Infinite
 

Infinite
 

0.065
 

Infinite
 

91
 

3/46 (7)
 

N.S.
 

Infinite
 

0.882
 

Infinite
 

Infinite
 

0.132
 

Infinite
 

91
 
Table Fl· Analyses of the Incidence of Primary Tumors in Male Mice
 

Fed Captan in the Dieta
 

(continued) 

Pooled Matched Low High 

Topography: Morphology Control Control Dose Dose 

Duodenum: Adenomatous Polyp,
 

NOS, or Adenocarcinoma in
 

Adenomatous Polypi 0/68 (0) 0/9 (0) 3/43 (7) 5/46 (11)
 

 Values0>d 

 = 0.008 N.S. N.S. = 0.009**
 

Relative Risk (Pooled Control)f Infinite Infinite
 

Lower Limit 0.943 1.849
 

Upper Limit Infinite Infinite
 

Relative Risk (Matched Control)f Infinite Infinite
 

Lower Limit 0.142 0.280
 

Upper Limit Infinite Infinite
 

Weeks to First Observed Tumor — — 91 91
 

a

Treated groups received doses of 8,000 or 16,000 ppm.
 

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

c

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


Armitage test when Ρ < 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 Ρ < 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 Captan in the Dieta
 

(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 Ρ < 0.05 for any comparison.
 

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

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

Fed Captan in the Dieta 

Pooled Matched Low High 

Topography: Morphology Control Control Dose Dose 

Liver: Neoplastic 

Noduleb 2/67 (3) 1/10 (10) 1/49 (2) 1/47 (2) 

 Values0 > d N.S. N.S. N.S. N.S. 

Relative Risk (Pooled Control)^ 0.684 0.713 

Lower Limit 0.012 0.012 

Upper Limit 12.716 13.241 

Relative Risk (Matched Control)f 0.204 0.213 

Lower Limit 0.003 0.003 

Upper Limit 15.723 16.378 

Weeks to First Observed Tumor 91 91 91 

Duodenum: Adenomatous 

Polyp, NOSb 1/68 (1) 1/9 (11) 1/49 (2) 0/48 (0) 

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

Relative Risk (Pooled Control)^ 1.388 0.000 

Lower Limit 0.018 0.000 

Upper Limit 106.757 26.404 

Relative Risk (Matched Control)f 0.184 0.000 

Lower Limit 0.003 0.000 

Upper Limit 14.153 3.512 

Weeks to First Observed Tumor — — 91 — 
__ 

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

Fed Captan in the Diet a 

(continued) 

Pooled Matched Low High
 

Topography: Morphology Control Control Dose Dose
 

Duodenum: Adenocarcinoma
 

in Adenomatous Polypi 0/68 (0) 0/9 (0) 0/49 (0) 3/48 (6)
 

 Valuesc>d  = 0.022 N.S. N.S. N.S.
 

Relative Risk (Pooled Control)f Infinite
 

Lower Limit — 0.844
 

Upper Limit — Infinite
 

Relative Risk (Matched Control)f Infinite
 

Lower Limit — 0.127
 

Upper Limit — Infinite
 

Weeks to First Observed Tumor _—
 91
 

Duodenum: Adenomatous Polyp,
 

NOS, or Adenocarcinoma in
 

Adenomatous Polyp" 1/68 (1) 1/9 (11) 1/49 (2) 3/48 (6)
 

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

Relative Risk (Pooled Control)f 1.388 4.250
 

Lower Limit 0.018 0.351
 

Upper Limit 106.757 218.317
 

Relative Risk (Matched Control)f 0.184 0.563
 

Lower Limit 0.003 0.056
 

Upper Limit 14.153 28.937
 

Weeks to First Observed Tumor — —
 91 91
 
Table F2. Analyses of the Incidence of Primary Tumors in Female Mice
 

Fed Captan in the Dieta
 

(continued)
 

a

Treated groups received doses of 8,000 or 16,000 ppm.
 

^Number of tumor-bearing animals/number of animals examined at site (percent),
 

c

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


Armitage test when Ρ < 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 Ρ < 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 Ρ < 0.05 for any comparison.
 

f

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

control group.
 
APPENDIX G
 

ANALYSIS OF FORMULATED DIETS FOR CONCENTRATIONS
 

OF CAPTAN
 

97
 

APPENDIX G
 

Analysis of Formulated Diets for
 

Concentrations of Captan
 

A 10-g sample of the diet mixture was shaken with 125 ml of
 

benzene at room temperature for 16 hours, then filtered through
 

Celite with benzene washes· The extracts were evaporated almost
 

to dryness under dry nitrogen· After appropriate dilutions, the
 

solution was quantitatively analyzed for captan 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) 

Concentrations Samples Analytical Variation (%) 

in Diet (ppm) Mean (ppm) 

2,000 12 2,014 3.6% 1,870-2,130
 

4,000 18 4,012 5.9% 3,550-4,340
 

8,000 31 8,186 5.0% 7,250-9,230
 

16,000 26 15,748 4.9% 14,100-17,350
 

ftU.S.GOVERNMENTPRINTINGOFFICE: 1977 - 260-899.3134 

99 


DHEW Publication No. (NIH) 77-815