National Cancer Institute 

CARCINOGENESIS 

Technical Report Series

No. 28 

1978 

BIOASSAY OF 

DIBROMOCHLOROPROPANE 

FOR POSSIBLE CARCINOGENICITY 

CAS No. 96-12-8 

NCI-CG-TR-28 

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

Public Health Service 

National Institutes of Health 

BIOASSAY OF
 

DIBROMOCHLOROPROPANE
 

FOR POSSIBLE CARCINOGEN1CITY
 

Carcinogenesis Testing Program
 

Division of Cancer Cause and Prevention
 

National Cancer Institute
 

National Institutes of Health
 

Bethesda, Maryland 20014
 

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

Public Health Service
 

National Institutes of Health
 

DHEW Publication No. (NIH) 78-828 

REPORT ON THE BIDASSAY OF DIBROMOCHLOROPROPANE
 

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
 

dibromochloropropane (DBCP) conducted for the Carcinogen Bioassay and
 

Program Resources Branch, Carcinogenesis Program, Division of Cancer
 

Cause and Prevention, National Cancer Institute (NCI), National Institutes of Health, Bethesda, Maryland. This bioassay was conducted
 

by Hazleton Laboratories America, Inc., Vienna, Virginia, initially
 

under direct contract to the NCI and currently under a subcontract to
 

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

Bioassay Program.
 

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

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

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

(3,5); the technical supervisor of animal treatment and observation
 

was Ms. K. J. Petrovics (3).
 

Histopathology was performed by Dr. R. H. Habermann (3) and
 

reviewed by Dr. R. W. Voelker (3) at the Hazleton Laboratories
 

America, Inc., and the diagnoses included in this report represent
 

the interpretation of these pathologists. Pathologists from NCI (1)
 

and Tracor Jitco (6) have reviewed selected slides and concur with
 

the overall pathologic evaluation of the study.
 

Compilation of individual animal survival, pathology, and
 

summary tables was performed by EG&G Mason Research Institute (7);
 

the statistical analysis was performed by Dr. J. R. Joiner (6) and
 

Mr. W. W. Belew (8), using methods selected for the Bioassay Program
 

by Dr. J. J. Gart (9).
 

This report was prepared at METREK, a Division of The MITRE
 

Corporation (8) under the direction of the NCI. Those responsible
 

for this report at METREK are the project coordinator, Dr. L. W.
 

Thomas (8), the task leader, Dr. M. R. Kornreich (8), and the senior
 

biologist, Ms. P. Walker (8). The final report was reviewed by
 

members of the participating organizations.
 

ill
 
The statistical analysis was reviewed by a member or members of
 

the Mathematical Statistics and Applied Mathematics Section of the
 

NCI: Dr. J. J. Gart (9), Mr. J. Nam (9), Dr. H. M. Pettigrew (9),
 

and Dr. R. E. Tarone (9).
 

The following other scientists at the National Cancer Institute
 

were responsible for evaluating the bioassay experiment, interpreting
 

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

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

R. A. Griesemer (1), Dr. T. W. Orme (1), Dr. R. A. Squire (10), and
 

Dr. J. M. Ward (1).
 

1.	 Carcinogenesis Program, Division of Cancer Cause and Prevention, National Cancer Institute, National Institutes of Heal; i,
 

Bethesda, Maryland.
 

2.	 Now with the Naylor Dana Institute for Disease Prevention,
 

American Health Foundation, Mammon House Road, Valhalla,
 

New York.
 

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

Vienna, Virginia.
 

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

Reston, Virginia.
 

5.	 No wit w h Rhodia, Inc., 23 Belmont Drive, Somerset, New Jersey.
 

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

Maryland.
 

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

Rockville, Maryland.
 

8.	 Th MITR e E Corporation, METREK Division, 1820 Dolley Madison
 

Boulevard, McLean, Virginia.
 

9.	 Mathematical Statistics and Applied Mathematics Section, Biometry
 

Branch, Field Studies and Statistics Program, Division of Cancer
 

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

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

University, School of Medicine, Traylor Building, Baltimore,
 

Maryland.
 

iv
 
SUMMARY
 

A bioassay for possible carcinogenicity of technical-grade dibromochloropropane (DBCP) was conducted using Osborne-Mendel rats and
 

B6C3F1 mice. DBCP in corn oil was administered by gavage 5 days a
 

week, at either of two dosages, to groups of 50 male and 50 female
 

animals of each species.
 

Initial dosage levels for the chronic bioassay were selected
 

on the basis of a preliminary subchronic toxicity test. Subsequent
 

dosage adjustments were made during the course of the chronic bioassay. The time-weighted average dosages of DBCP in the chronic study
 

were 29 mg/kg/day for the high dose rats of both sexes, and 15 mg/kg/
 

day for the low dose rats of both sexes. The time-weighted average
 

concentrations for the high dose male and female mice were 219 and
 

209 mg/kg/day, respectively. The time-weighted average concentrations for the low dose male and female mice were 114 and 110 mg/kg/
 

day, respectively.
 

For each species, 20 animals of each sex were placed on test as
 

vehicle controls. These animals were intubated with corn oil at the
 

same time that dosed animals were intubated with DBCP mixtures.
 

Twenty animals of each sex were placed on test as untreated controls
 

for each species. These animals received no gavage treatments.
 

DBCP was administered to the high dose male and high dose female
 

rats for 64 weeks prior to sacrifice, and to the low dose female rats
 

for 73 weeks prior to sacrifice. The low dose male rats were treated
 

for 78 weeks followed by an additional 5 weeks of observation. The
 

high dose male and female mice were treated for 47 weeks prior to
 

sacrifice; the low dose male mice were treated for 59 or 60 weeks
 

prior to sacrifice, and the low dose female mice were treated for 60
 

weeks prior to sacrifice.
 

In rats and mice of both sexes, statistically significant incidences of squamous-cell carcinomas of the forestomach occurred in
 

each dosed group and a significant positive association existed
 

between dosage level and tumor incidence. The incidences of adenocarcinomas of the mammary gland were statistically significant in
 

female rats when the treated groups were compared to the controls.
 

Toxic nephropathy was also observed at elevated incidences in all of
 

the treated rats and mice when compared to their respective untreated
 

or vehicle control groups.
 

Under the conditions of this study, DBCP is a stomach carcinogen
 

in rats and mice of both sexes and is carcinogenic to the mammary
 

gland in female rats.
 

v
 

TABLE OF CONTENTS
 

I. INTRODUCTION
 

II. MATERIALS AND METHODS
 

A. Chemicals
 

B. Dosage Preparation
 

C. Animals
 

D. Animal Maintenance
 

E. Gastric Intubation
 

F. Selection of Initial Dose Levels
 

G. Experimental Design
 

H. Clinical and Histopathologic Examinations
 

I. Data Recording and Statistical Analyses
 

III. CHRONIC TESTING RESULTS: RATS
 

A. Body Weights and Clinical Observations
 

B. Survival
 

C. Pathology
 

D. Statistical Analyses of Results
 

IV. CHRONIC TESTING RESULTS: MICE
 

A. Body Weights and Clinical Observations
 

B. Survival
 

C. Pathology
 

D. Statistical Analyses of Results
 

V. DISCUSSION
 

VI. BIBLIOGRAPHY
 

APPENDIX A SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

RATS TREATED WITH DBCP
 

APPENDIX B SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

MICE TREATED WITH DBCP
 

APPENDIX C SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN RATS TREATED WITH DBCP
 

APPENDIX D SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN MICE TREATED WITH DBCP
 

Page
 

1
 

4
 

4
 

4
 

4
 

5
 

6
 

7
 

8
 

12
 

13
 

18
 

18
 

20
 

23
 

25
 

33
 

33
 

33
 

37
 

38
 

41
 

44
 

A-l
 

B-l
 

C-l
 

vii
 

D-l 
Figure Number

 

1 

2 

3 

4 

Table Number
 

1
 

2
 

3
 

4
 

5
 

6
 

Al
 

LIST OF ILLUSTRATIONS 

GROWTH CURVES FOR DBCP CHRONIC STUDY RATS
 

SURVIVAL COMPARISONS OF DBCP CHRONIC STUDY
 

RATS
 

GROWTH CURVES FOR DBCP CHRONIC STUDY MICE
 

SURVIVAL COMPARISONS OF DBCP CHRONIC STUDY
 

MICE
 

LIST OF TABLES
 

DESIGN SUMMARY FOR OSBORNE-MENDEL RATS—DBCP
 

GAVAGE EXPERIMENT
 

DESIGN SUMMARY FOR B6C3F1 MICE—DBCP GAVAGE
 

EXPERIMENT
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS
 

AT SPECIFIC SITES IN MALE RATS TREATED WITH
 

DBCP
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS
 

AT SPECIFIC SITES IN FEMALE RATS TREATED
 

WITH DBCP
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS
 

AT SPECIFIC SITES IN MALE MICE TREATED WITH
 

DBCP
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS
 

AT SPECIFIC SITES IN FEMALE MICE TREATED
 

WITH DBCP
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

MALE RATS TREATED WITH DBCP
 

viii 

Page
 

19
 

21
 

34
 

35
 

Page
 

9
 

10
 

26
 

29
 

39
 

40
 

A-3
 
Table Number
 

A2
 

Bl
 

B2
 

Cl
 

C2
 

Dl
 

D2
 

LIST OF TABLES (Concluded) 

Page 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

FEMALE RATS TREATED WITH DBCP A-7 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

MALE MICE TREATED WITH DBCP B-3 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN 

FEMALE MICE TREATED WITH DBCP B-7 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN MALE RATS TREATED WITH DBCP C-3 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN FEMALE RATS TREATED WITH DBCP C-9 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN MALE MICE TREATED WITH DBCP 

D-3 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC 

LESIONS IN FEMALE MICE TREATED WITH DBCP D-7 

ix
 

I. INTRODUCTION
 

Dibromochloropropane (DBCP) (NCI No. C00500), a halogenated
 

aliphatic hydrocarbon and agricultural pesticide, was one of several
 

widely used pesticides selected for bioassay by the National Cancer
 

Institute. At that time there was a lack of adequate chronic toxicity
 

data on this compound.
 

The Chemical Abstracts Service (CAS) Ninth Collective Index
 

#
 

(1977) name for this compound is 1,2-dibromo-3-chloropropane. It is
 

also known in the agricultural trade as Nemagon , Fumazone and DBCP.
 

DBCP is used primarily as a soil fumigant for control of nematodes and is suitable for protection of a wide variety of field crops,
 

vegetables, fruits, nuts, and ornamental plants. In 1971, the most
 

recent year for which such data are available, approximately 255,000
 

acres of U.S. agricultural land were treated with 3.6 million pounds
 

of the nematocide, with more than 59 percent of total usage occurring
 

in the Pacific region (Andrilenas, 1974).
 

Recent production statistics for DBCP are considered proprietary
 

and are, therefore, not available; however, domestic production in
 

1969 amounted to more than 8.6 million pounds (Fowler and Mahan, 1975)
 

while domestic sales in 1972 exceeded 15 million pounds (U.S. International Trade Commission, 1977).
 

*
 

The CAS registry number is 96-12-8.
 

1 
Workers at pesticide formulating plants and agricultural workers
 

engaged in pesticide application may be exposed to DBCP either
 

through inhalation or skin contact. Since the compound is classified
 

as moderately volatile (evaporating at a rate similar to ethanol) and
 

undergoes biological degradation in moist soils (Shell Chemical
 

Company, 1972) exposure of the general population via ingestion of
 

contaminated food crops is probably negligible.
 

DBCP is harmful to humans following inhalation or ingestion; however, toxic amounts may also be absorbed through the skin. Inhalational intoxication results in nausea, irritation of the respiratory
 

tract membranes, narcosis, and possible pulmonary congestion following
 

prolonged or repeated exposure to high vapor concentrations. Symptoms
 

of acute ingestion include gastrointestinal distress and congestion
 

and edema of the lungs (Shell Chemical Company, 1972). DBCP is irritating to the skin, with repeated applications resulting in necrosis
 

of the dermis and mild erythema. Permanent eye injury may result
 

from chronic eye contact with vapors (Gleason et al., 1969). DBCP
 

was found to be mutagenic in bacteria, causing base-substitution
 

mutations in a histidine-requiring strain of Salmonella typhimurium
 

(The Ames Test using strain TA1530) (Rosenkranz, 1975).
 

In July and August, 1977, at least 60 male chemical plant workers
 

involved with the manufacture or formulation of DBCP pesticides were
 

found to be sterile (Peterson and Shinoff, 1977a; 1977b). Earlier
 

2
 
studies indicated sterility in laboratory animals (Torkelson et al.,
 

1961). Rats, guinea pigs, rabbits, and monkeys received at least
 

50 seven-hour exposures to 12 ppm DBCP in air. The most striking
 

observation at autopsy was severe atrophy and degeneration of the
 

testes of all species. In the rats this was characterized by degenerative changes in the seminiferous tubules, an increase in Sertoli
 

cells, reduction in the number of sperm cells, and development of
 

abnormal forms of sperm cells (Torkelson et al., 1961).
 

3 
II. MATERIALS AND METHODS
 

A. Chemicals
 

One batch of technical-grade l,2-dibromo-3-chloropropane (DBCP)
 

was purchased from Shell Chemical Company, Incorporated. Analysis
 

was performed by Hazleton Laboratories America, Inc., Vienna,
 

Virginia. Gas-liquid chromatography (GLC) suggested a purity greater
 

than 90 percent with 16 minor impurities. The infrared analysis was
 

not inconsistent with the structure of the compound.
 

A second GLC analysis, performed 24 months later, suggested a
 

similar degree of purity although some changes in the nature and
 

quantity of the impurities were noted in the chromatogram. Those
 

changes were reflected in a slightly altered infrared spectra.
 

Throughout this report the term DBCP is used to represent this
 

technical-grade material.
 

B. Dosage Preparation
 

Fresh mixtures of DBCP in Duke's corn oil (S. F. Sauer Company),
 

were prepared weekly, sealed, and stored in dark bottles at 34°F.
 

These DBCP solutions were considered generally stable for 10 days
 

under the indicated storage conditions. The concentrations of DBCP
 

in the corn oil were 1.0 to 1.5 percent for rats and 1.0 to 2.6 percent for mice.
 

C. Animals^
 

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

4
 
comparative study of the tumorigenic responsiveness to carbon tetrachloride of five different strains of rats (Reuber and Glover, 1970).
 

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

carcinogenesis bioassays and has proved satisfactory in this capacity.
 

Rats and mice of both sexes were obtained through contracts of
 

the Division of Cancer Treatment at the National Cancer Institute.
 

The Osborne-Mendel rats were procured from the Battelle Memorial
 

Institute, Columbus, Ohio, and the B6C3F1 mice were obtained from the
 

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

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

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

various treatment and control groups.
 

D. Animal Maintenance
 

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


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

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

conditioning system in the laboratory provided filtered air at a rate
 

of 12 complete changes of room air per hour. Fluorescent lighting
 

was provided on a 12-hour-daily cycle. The rats were individually
 

housed in suspended galvanized-steel wire-mesh cages with perforated
 

floors, while mice were housed by sex in groups of 10 in solid-bottom
 

polypropylene cages with filter tops. Sanitized cages with fresh
 

bedding (Sanichips , Shurfire) were provided once each week for mice.
 

Rats received sanitized cages with no bedding with the same frequency.
 

5
 
Food	 hoppers were changed and heat-sterilized once a week for the
 

first 10 weeks and once a month thereafter. Fresh heat-sterilized
 

glass water bottles were provided three times a week. Food (Wayne
 

Lab-Blox®, Allied Mills, Inc.) and water were available ad libitum.
 

DBCP-treated rats and their vehicle controls were housed in the
 

same room with rats treated with trichloroethylene (79-01-6), carbon
 

disulfide (75-15-0), 1,1-dichloroethane (75-34-3), and 1,2-dichloroethane (107-06-2). Untreated control rats for the DBCP bioassay were
 

housed in another room with rats treated with 1,1,2,2-tetrachloroethane (79-34-5), chloroform (67-66-3), 1,2-dibromoethane (106-93-4),
 

carbon tetrachloride (56-23-5), and allyl chloride (107-05-1). DBCP-


treated and both vehicle and untreated control mice were maintained
 

in the same room as mice receiving iodoform (75-47-8), 1,1,2-trichloroethane (79-00-5), 3-sulfolene (77-79-2), carbon disulfide (75-15-0),
 

1,1,2,2-tetrachloroethane (79-34-5), allyl chloride (107-05-1), 1,2­

dichloroethane (107-06-2), carbon tetrachloride (56-23-5), chloroform
 

(67-66-3), chloropicirin (76-06-2), tetrachloroethylene (127-18-4),
 

methylchloroform (71-55-6), hexachloroethane (67-72-1), 1,1-dichloroethane (75-34-3), trichloroethylene (79-01-6), trichlorofluoromethane
 

(75-69-4), and 1,2-dibromoethane (106-93-4).
 

E.	 Gastric Intubation
 

Intubation was performed for five consecutive days per week on
 

a mg/kg body weight basis utilizing the most recently observed group
 

*'
 

CAS registry numbers are given in parentheses.
 

6
 
mean body weight as a guide for determining the dose. Mean body
 

weights for each group were recorded at weekly intervals for the
 

first 10 weeks and at monthly intervals thereafter. All animals of
 

one sex within a treatment group received the same dose. Animals
 

were gavaged with test solutions under a hood to minimize extraneous
 

exposure of other animals and laboratory personnel to the chemical.
 

F. Selection of Initial Dose Levels
 

In order to establish the maximum tolerated dosages of DBCP for
 

administration to treated animals in the chronic studies, subchronic
 

toxicity tests were conducted with both rats and mice. Six groups,
 

each consisting of five males and five females, were utilized for
 

each animal species. DBCP dissolved in corn oil was administered by
 

gavage to five of the six rat groups at concentrations of 25, 40, 63,
 

100, and 160 mg/kg/day and to five of the six mouse groups at concentrations of 100, 160, 251, 398 and 631 mg/kg/day. The sixth group of
 

each species served as a control group and was gavaged only with corn
 

oil. Intubation occurred 5 days per week for 6 weeks, followed by a
 

2-week observation period to detect any delayed toxicity.
 

A dosage inducing no mortality and resulting in a retardation
 

in body weight gain of approximately 20 percent was to be selected
 

as the initial high dose. When weight gain criteria were not applicable, mortality data alone were utilized.
 

All the male rats treated with 25 and 40 mg/kg/day and all the
 

female rats treated with 25 mg/kg/day survived the entire 6-week
 

7 
treatment period and the 2-week observation period, while deaths were
 

observed at higher doses. Body weight gain retardation, expressed as
 

a percentage of the weight gained by the controls, was 22 percent in
 

males and 33 percent in females receiving dosages of 25 mg/kg/day.
 

The initial high dose selected for use in the chronic bioassay was 24
 

mg/kg/day for both male and female rats.
 

All mice receiving dosages of 398 mg/kg/day or more died before
 

the subchronic study was completed. All other mice (with the exception of one male and one female) treated with 251 mg/kg/day or less
 

survived the entire 8-week period. No retardation in mean body
 

weight gain was observed in any of the male or female treated groups.
 

The initial high doses selected for the chronic bioassay were 160 and
 

120 mg/kg/day for males and females, respectively.
 

G. Experimental Design
 

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

untreated observation periods, and the time-weighted average dosages)
 

are summarized in Tables 1 and 2.
 

The low dose, high dose, and vehicle control rats were all
 

approximately 7 weeks old at the time the bioassay began. The untreated controls were approximately 16 weeks younger than the other
 

three groups, and were started on test a corresponding 16 weeks after
 

the other groups. They were observed for a total of 109 weeks. The
 

untreated controls were approximately 6 weeks old when they were
 

8
 
TABLE 1
 

DESIGN SUMMARY FOR OSBORNE-MENDEL RATS
 

DBCP GAVAGE EXPERIMENT
 

INITIAL OBSERVATION PERIOD TIME-WEIGHTED 

GROUP DBCP TREATED UNTREATED AVERAGE 

SIZE DOSAGE* (WEEKS) (WEEKS) DOSAGE 

MALE 

UNTREATED CONTROL 20 0 109 0
 

VEHICLE CONTROL0 20 0 78 5 0
 

LOW DOSEC 50 12 9 15
 

15 69
 

0 5
 

HIGH DOSEd 50 24 9 29
 

30 55
 

0
 

FEMALE
 

UNTREATED CONTROL 20 0 109 0
 

VEHICLE CONTROL0 20 0 78 5 0
 

LOW DOSE6 50 12 9
 

15 64 15
 

0
 

HIGH DOSEd 50 24 9 29
 

30 55
 

0
 

ja


Dosage, given in mg/kg body weight, was administered by gavage five
 

consecutive days per week.
 

These animals were sacrificed during week 83.
 

These animals were sacrificed during week 64.
 

These animals were sacrificed during week 73.
 

9
 

c
TABLE 2
 

DESIGN SUMMARY FOR B6C3F1 MICE
 

DBCP GAVAGE EXPERIMENT
 

INITIAL OBSERVATION PERIOD 

GROUP DBCP TREATED UNTREATED 

SIZE DOSAGE3 (WEEKS) (WEEKS) 

MALE 

UNTREATED CONTROL0 20 0 0 78 

VEHICLE CONTROL*1


LOW DOSE6


HIGH DOSE


FEMALE
 

UNTREATED CONTROL


VEHICLE CONTROL6


LOW DOSE6


HIGH DOSEf


 20 0 59 0 

 50 80 11 

100 14 

130 35 

0 

 50 160 11 

200 14 

260 22 

0 

 20 0 0 90 

 20 0 60 0 

 50 60 11 

100 14 

130 35 

0 

 50 120 11 

200 14 

260 22 

0 

TIME -WEIGHTED
 

AVERAGE
 

DOSAGE
 

0
 

0
 

114
 

219
 

0
 

0
 

110
 

209
 

Dosage, given in mg/kg body weight, was administered by gavage five
 

consecutive days per week.
 

£
 

These animals were sacrificed during week 78.
 

These animals were sacrificed during week 59.
 

These animals were sacrificed during week 60.
 

These animals were sacrificed during week 47.
 

10
 
started on test. The high and low dosages of DBCP initially utilized
 

for rats of both sexes were 24 and 12 mg/kg/day, respectively. Intubation was performed five consecutive days per week. In week 10,
 

dosages were raised to 30 and 15 mg/kg/day and maintained at these
 

levels until the end of the bioassay. High dose male and female rats
 

were sacrificed during week 64. Low dose female rats were sacrificed
 

in week 73. Low dose male rats and vehicle control rats of both sexes
 

were treated for 78 weeks, observed without treatment for 5 additional
 

weeks, and then sacrificed.
 

The low dose, high dose, and vehicle control mice were approximately 5 weeks old when the first gavage treatments were administered.
 

Male mice received initial high and low dosages of 160 and 80 mg/kg/
 

day, respectively, while female mice received high and low dosages of
 

120 and 60 mg/kg/day, respectively. In week 12, high and low dosages
 

administered to both male and female mice were increased to 200 and
 

100 mg/kg/day, respectively. Dosages were raised again in week 26,
 

to 260 and 130 mg/kg/day and these were the respective high and low
 

dosages utilized for the remainder of the bioassay.
 

Due to poor survival among the mice, all surviving high dose
 

males and females were sacrificed during week 47. All surviving male
 

vehicle control mice were sacrificed during week 59 and surviving
 

male and female low dose mice or female vehicle control mice were
 

sacrificed during weeks 59 and 60.
 

11
 
For both the rat and mouse control groups, the animals were
 

maintained and observed in the same manner as the dosed groups. The
 

untreated controls received no DBCP or corn oil, while the vehicle
 

controls received pure corn oil by gavage.
 

H. Clinical and Histopathologic Examinations
 

Animals were weighed immediately prior to initiation of the
 

experiment. From the first day, all animals were inspected daily
 

for mortality. Body weights, food consumption, and data concerning
 

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

and location of tissue masses were recorded at weekly intervals for
 

the first 10 weeks and at monthly intervals thereafter. The presence
 

of tissue masses was determined by observation and palpation of each
 

animal.
 

A necropsy was performed on each animal regardless of whether it
 

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

bioassay. The animals were euthanized by exsanguination under sodium
 

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

of major tissues, organs, or gross lesions taken from sacrificed
 

animals and, whenever possible, from animals found dead.
 

Slides were prepared from the following tissues: skin, subcutaneous tissue, lungs and bronchi, trachea, bone marrow, spleen,
 

lymph nodes, thymus, heart, salivary gland, liver, gallbladder (mice)
 

and bile duct, pancreas, esophagus, stomach, small intestine, large
 

12
 
intestine, kidney, urinary bladder, pituitary, adrenal, thyroid,
 

parathyroid, pancreatic islets, testis, prostate, seminal vesicle,
 

brain, eye, muscle, uterus, mammary gland, and ovary.
 

Tissues for which slides were prepared were preserved in 10 percent buffered formalin, embedded in paraffin, sectioned, and stained
 

with hematoxylin and eosin prior to microscopic examination. An
 

occasional section was subjected to special staining techniques for
 

more definitive diagnosis.
 

A few tissues were not examined for some animals, particularly
 

for those that died early. Also, some animals were missing, cannibalized, or judged to be in such an advanced state of autolysis as to
 

preclude histopathologic interpretation. Thus, the number of animals
 

for which particular organs, tissues, or lesions were examined microscopically varies and does not necessarily represent the number of
 

animals that were placed on experiment in each group.
 

I. Data Recording and Statistical Analyses
 

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

(Linhart et al., 1974). The data elements include descriptive information on the chemicals, animals, experimental design, clinical observations, survival, body weight, and individual pathologic results, as
 

recommended by the International Union Against Cancer (Berenblum,
 

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

13
 
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 used Tarone's (1975) extensions of Cox's methods for
 

testing a dose-related trend. One-tailed P-values have been reported
 

for all tests except the departure from linearity test, which is only
 

reported when its two-tailed P-value is less than 0.05.
 

The incidence of neoplastic or nonneoplastic lesions has been
 

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

specific anatomic site (numerator) to the number of animals in which
 

that site was examined (denominator). In most instances, the denominators included only those animals for which that site was examined
 

histologically. However, when macroscopic examination was required
 

to detect lesions prior to histologic sampling (e.g., skin or mammary
 

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

14
 
lymphomas), the denominators consist of the numbers of animals necropsied.
 

The purpose of the statistical analyses of tumor incidence is to
 

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

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

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

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

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

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

significance level of 0.05 may be made. The Bonferroni inequality
 

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

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

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

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

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

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

Under the assumption of a linear trend, this test determined if the
 

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


tailed 0.05 level of significance. Unless otherwise noted, the direction of the significant trend was a positive dose relationship.
 

This method also provides a two-tailed test of departure from linear
 

trend.
 

15
 
A time-adjusted analysis was applied when numerous early deaths
 

resulted from causes that were not associated with the formation of
 

tumors. In this analysis, deaths that occurred before the first tumor was observed were excluded by basing the statistical tests on
 

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

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

tumor was found, comparisons were based exclusively on animals that
 

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

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

Cochran-Armitage tests, etc.) were followed.
 

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

incidence of tumors. Curves of the proportions surviving without an
 

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

during which animals died naturally or were sacrificed was entered as
 

the time point of tumor observation. Cox's methods of comparing
 

these curves were used for two groups; Tarone's extension to testing
 

for linear trend was used for three groups. The statistical tests for
 

the incidence of tumors which used life-table methods were one-tailed
 

and, unless otherwise noted, in the direction of a positive dose
 

relationship. Significant departures from linearity (P < 0.05, two-


tailed test) were also noted.
 

The approximate 95 percent confidence interval for the relative
 

risk of each dosed group compared to its control was calculated from
 

16
 
the exact interval on the odds ratio (Gart, 1971). The relative risk
 

is defined as p /p where p is the true binomial probability of the
 

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

and p 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
 

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

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

would be within the interval calculated from the experiment. When
 

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

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

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

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

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

the lower limit indicates the absence of a significant result while
 

the upper limit indicates that there is a theoretical possibility
 

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

detected under the conditions of this test.
 

17
 
III.	 CHRONIC TESTING RESULTS: RATS
 

A.	 Body Weights and Clinical Observations
 

After the first 5 months of treatment, dose-related body weight
 

gain retardation was apparent in male and female rats (Figure 1).
 

All animals exhibited generally normal appearance and behavior
 

during the first 14 weeks of the study with the exception of occasional hunched appearance in a few treated rats. Beginning in week
 

18, an increasing number of treated males and females showed a
 

hunched appearance and by week 34, this observation was evident in
 

approximately 80 percent of the treated rats.
 

Urine staining of the abdominal area was the other predominant
 

clinical sign noted in this study. It was first observed in a few
 

high dose females in week 6, and in slightly increasing numbers of
 

treated males and females thereafter. By week 34, approximately 15
 

percent of the low dose and 50 percent of the high dose rats showed
 

abdominal urine stains. Urine stains were infrequently noted in the
 

vehicle controls during the first 15 months of the study, but more
 

frequently thereafter.
 

A declining survival rate, apparently from compound-related
 

toxicity, was observed beginning in week 30 in the treated females.
 

In week 36, 10 high dose females with large palpable tissue masses
 

were sacrificed. The majority of these masses were histopathologically diagnosed as mammary adenocarcinomas. Increasing mortality was
 

evident as the study progressed, particularly in the high dose rats.
 

18
 
TIME ON TEST (WEEKS) 

FIGURE 1 

GROWTH CURVES FOR DBCP CHRONIC STUDY RATS 

19 
For most of the DBCP-treated animals that died naturally or were
 

sacrificed, gross necropsy revealed tumors of the stomach, with metastases to adjacent organs in many cases*
 

Respiratory signs characterized by labored respiration, wheezing,
 

and/or nasal discharge were noted at a comparable and moderate incidence in all groups, including the vehicle controls, during the study.
 

The incidence increased as the animals aged. Other signs commonly
 

associated with aging that were noted at a similar rate in the vehicle
 

controls and treated animals included sores on the body and/or extremities, localized alopecia, reddish discharge or brown crust around
 

body orifices, and rough or stained fur. Isolated, apparently spontaneous clinical signs in one or two treated rats included head tilt,
 

circling, tremors, and unusual gait.
 

Palpable nodules or tissue masses were observed as early as
 

week 14 in a low dose female rat and were evident in a greater number
 

of treated animals than controls during the study. The increased
 

incidence of mammary adenocarcinomas in the DBCP-treated females was
 

confirmed by subsequent histopathology.
 

B.	 jSurviyal
 

The estimated probabilities of survival for male and female rats
 

in the control and DBCP-treated groups are shown in Figure 2.
 

In male rats the Tarone test indicated a significant (P < 0.001)
 

positive association between increased dosage and accelerated mortality.
 

The departure from linear trend was also significant (P < 0.001),
 

20
 
TIME ON TEST (WEEKS) 

FIGURE 2 

SURVIVAL COMPARISONS OF DBCP CHRONIC STUDY RATS 

21 
primarily because of the accelerated mortality noted in several of
 

the groups. Eighty percent of the high dose males had died by week
 

62; the other 10 were sacrificed in week 64. Ninety percent of the
 

low dose males were dead by week 83, at which point the remaining
 

five rats were sacrificed. Ten vehicle control rats were sacrificed
 

in week 64 and two in week 83; the others all died before week 83.
 

Despite the sacrifice of five rats in week 60, survival in the
 

untreated control group was adequate, with 55 percent of the male
 

rats surviving at least 88 weeks. Early deaths in the dosed males
 

may well have been tumor-related, since in both high and low dose
 

groups 47/50 (94 percent) had squamous-cell carcinomas of the stomach.
 

In female rats the Tarone test indicated a significant (P < 0.001)
 

positive association between dosage and mortality. The departure from
 

linear trend was also significant (P = 0.010), primarily because of
 

the accelerated mortality noted in treated rats. Ninety-four percent
 

of the high dose females were dead by week 61, the remaining three
 

were sacrificed in week 64. Eighty-four percent of the low dose
 

group were dead by week 73, at which point the remaining eight were
 

sacrificed. Seventy-five percent of the vehicle controls were dead
 

by week 77, including ten rats sacrificed in week 65; the remaining
 

five vehicle control rats were sacrificed in week 83. Survival in
 

the untreated controls was high; despite the sacrifice of five
 

animals in week 60, 65 percent of the rats survived until the end of
 

22
 
the study. Early deaths in the treated females may have been tumor-


related, since squamous-cell carcinomas of the stomach were discovered
 

in 38/50 (76 percent) of the low dose and 29/49 (59 percent) of the
 

high dose female rats.
 

C.	 Pathology
 

Histopathologic findings on neoplasms in rats are tabulated in
 

Appendix A (Tables Al and A2); findings on nonneoplastic lesions are
 

tabulated in Appendix C (Tables Cl and C2).
 

Squamous-cell carcinomas of the forestomach occurred in 47/50
 

(94 percent) low dose males, 47/50 (94 percent) high dose males,
 

38/50 (76 percent) low dose females, and 29/49 (59 percent) high dose
 

females. These tumors were not observed in control rats. Grossly,
 

the neoplasms appeared as white fibrous lesions in the forestomach.
 

Raised white and pale streaks were observed on the serosal surface.
 

Adhesions to adjacent abdominal organs and tissues were present in
 

some rats. Microscopically, the mucosal surface was acanthotic and
 

hyperkeratotic. The basal epithelial layer contained papillary cords
 

and nests of anaplastic squamous epithelium supported by dense bands
 

of fibrous connective tissue. The carcinoma extended through the
 

muscularis mucosa, submucosa, muscular layers, and serosa and metastasized (transcoelomic) to adjacent tissues. Pulmonary metastases were
 

observed in lung sections from 5/50 (10 percent) low dose males, 5/50
 

(10 percent) high dose males, and 3/44 (7 percent) high dose females.
 

23
 
Squamous-cell papillomas of the stomach occurred in 1/50 (2
 

percent) low dose males, 2/50 (4 percent) high dose males, 1/50 (2
 

percent) low dose females, and 9/49 (18 percent) high dose females.
 

These tumors were not observed in control rats.
 

In stomach sections from rats that did not have tumors of the
 

forestomach, both acanthosis and hyperkeratosis occurred in two high
 

dose males, two low dose males, nine high dose females, and four low
 

dose females. Acanthosis in the absence of hyperkeratosis occurred
 

in one low dose female.
 

Adenocarcinomas of the mammary gland were observed primarily
 

in treated females, 24/50 (48 percent) in the low dose group, 31/50
 

(62 percent) in the high dose group, and 2/20 (10 percent) in the
 

untreated controls. Mammary adenocarcinomas were also found in 1/50
 

(2 percent) low dose and 1/20 (5 percent) untreated control male rats.
 

Grossly, the neoplasms were light pink to red, subcutaneous, firm and
 

nodular, weighing 1 to 150 grams. Microscopically, these adenocarcinomas were characterized by irregular acini lined by anaplastic
 

epithelium and supported by a dense fibrous stroma. Acini were frequently lined by multiple layers of epithelium, and papillary infoldings or projections were present. Larger hyperchromatic cells were
 

present and mitoses were frequent in the more anaplastic adenocarcinomas .
 

Hemangiosarcomas occurred in the spleens of 9/50 (18 percent)
 

low dose males and 4/42 (10 percent) low dose females, but were not
 

observed in the high dose or control rats.
 

24
 
The other neoplasms that occurred in this study were comparable
 

in frequency and morphology to those tumors occurring naturally in
 

untreated Osborne-Mendel rats.
 

Toxic nephropathy occurred in 50/50 low dose males, 49/50 high
 

dose males, 42/42 low dose females, and 44/44 high dose females but
 

not in controls of either sex. Microscopically, toxic nephropathy
 

was characterized by degenerative changes in the proximal convoluted
 

tubules at the junction of the cortex and medulla, with cloudy
 

swelling, fatty degeneration, and necrosis of the tubular epithelium.
 

Some affected tubules were empty; others were filled with hyaline
 

casts. Occasionally the damaged tubules contained large basophilic
 

regenerative cells. At this stage, the kidneys often had infiltration of inflammatory cells, fibrosis, and calcium deposition.
 

Other inflammatory, proliferative, and degenerative lesions as
 

seen in the control and treated animals were similar in number and
 

kind to lesions occurring naturally in untreated rats.
 

In this study, DBCP treatment was associated with squamous-cell
 

carcinomas in the forestomach of male and female rats at both doses
 

and adenocarcinomas of the mammary gland in the female rats. This
 

chemical was also toxic to the kidneys, producing a toxic tubular
 

nephropathy in both male and female rats.
 

D. Statistical Analyses of Results
 

The results of the statistical analyses of tumor incidence in
 

rats are summarized in Tables 3 and 4. The analysis for every type
 

25
 
26

 

TABLE 3 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT 

SPECIFIC SITES IN MALE RATS TREATED WITH DBCPa 

TOPOGRAPHY '.MORPHOLOGY 

UNTREATED 

CONTROL 

VEHICLE 

CONTROL 

LOW DOSE HIGH 

DOSE 

Subcutaneous Tissue: Fibroma 

P Values0 >d 

Departure from Linear Trend 

Relative Risk (Untreated Control) 

Lower Limit 

Upper Limit 

Relative Risk (Vehicle Control)f 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

0/20(0.00) 

N.S. 

P = 0.047 

0/20(0.00) 

N.S. 

P = 0.047 

3/50(0.06) 

N.S. 

Infinite 

0.250 Infinite 

Infinite 

0.250 Infinite 

41 

0/50(0.00) 

N.S. 

Stomach: Squamous Cell Carcinoma^ 

P Valuesc'd 

Departure from Linear Trend6 

Relative Risk (Untr ated Control) 

Lowe Limit 

Uppe Limit 

Relative Risk (Vehi le Control)1 

Lowe Limit 

Uppe Limit 

Weeks to First Observed Tumor 

Stomach: Squamous Cell Papilloma 

P Values0 >d 

Relative Risk (Untreated Control) 

Lower Limit 

Upper Limit 

Relative Risk (Vehicle Control)1 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 

0/20(0.00) 

P­= 0.001 

P<O.001 

— 

— 

— 

— 

— 

— 

0/20(0.00) 

N.S. 

— 

— 

— 

— 

— 

0/19(0.00) 

P< 0.001 

P< 0.001 

— 

— 

— 

— 

— 

— 

0/19(0.00) 

N.S. 

47/50(0.94) 

P< 0.001* 

P< 0.001** 

Infinite 7.114 

Infinite 

Infinite 

6.779 Infinite 

52 

1/50(0.02) 

N.S. 

Infinite 

0.022 

Infinite 

Infinite 0.021 

Infinite 

58 

47/50(0.94) 

P^ 0.001* 

P< 0.001** 

Infinite 7.114 

Infinite 

Infinite 

6^775 Infinite 

34 

2/50(0.04) 

N.S. 

Infinite 0.123 

Infinite 

Infinite 0.117 

Infinite 

33 
27 

TOPOGRAPHY :MORPHOLOGY
 

All Sites: Hemangiosarcoma or Hemangioma
 

P Values0 >d
 

Departure from Linear Trend
 

Relative Risk (Untreated Control)^
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)1
 

Lower Limit
 

Upper Limit *•<­

Weeks to First Observed Tumor
 

Thyroid: Follicular-Cell Adenoma
 

P Values0 >d
 

c
 Relative Risk (Untreated Control)
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)1
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Thyroid: Follicular-Cell Adenoma or Carcinoma
 

P Values0 >d
 

Relative Risk (Untreated Control)
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 3 

(CONTINUED) 

UNTREATED VEHICLE LOW HIGH 

CONTROL CONTROL DOSE DOSE 

0/20(0.00) 0/20(0.00) 13/50(0.26) 2/50(0.04) 

N.S. N.S. P ­ 0.008* N.S. 

P ­ 0.008** 

P< 0.001 P' 0.001 

Infinite Infinite 

1.674 0.123 

Infinite Infinite 

Infinite Infinite 

1.674 0.123 

Infinite Infinite 

45 52 

0/19(0.00) 0/20(0.00) 3/48(0.06) 2/49(0.04) 

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

Infinite Infinite 

0.248 0.119 

Infinite Infinite 

Infinite Infinite 

0.261 0.125 

Infinite Infinite 

83 57 

1/19(0.05) 0/20(0.00) 5/48(0.10) 3/49(0.06) 

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

1.979 1.163 

0.246 0.1Q3 

91.529 59.809 

Infinite Infinite 

0.547 0.255 

Infinite Infinite 

90 81 57 
TABLE 3
 

(CONCLUDED)
 

treated groups received time-weighted average doses of 15 and 29 mg/kg by gavage.
 

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

Beneath the incidence of tumors in a control group is the probability level for the Cochran-Armitage test when P< 0.05; otherwise, not significant
 

(N.S.) is indicated. Beneath the incidence of tumors in a treated group is the probability level for the Fisher exact test for the comparison of
 

that treated group with the untreated control group (*) or with the vehicle control group (**) when P<0.05 for either control group; otherwise»
 

not significant (N.S.) is indicated.
 

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

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

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

28 
29 

TOPOGRAPHY : MORPHOLOGY
 

Stomach: Squamous Cell Carcinoma
 

P Valuesc>d
 

Departure from Linear Trend6
 

Relative Risk (Untreated Control)
 

Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Stomach: Squamous Cell Papilloma
 

P Values0 >d
 

Relative Risk (Untreated Control)
 

Lower Limit
 Upper Limit
 

Relative Risk (Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

All Sites: Hemangiosarcoma or Hemangioma
 

P Valuesc>d
 

Departure from Linear Trend*
 

Relative Risk (Untreated Control)
 Lower Limit
 

Upper Limit
 

Relative Risk (Vehicle Control)1
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 4
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN FEMALE RATS TREATED WITH DBCP3
 

UNTREATED	 VEHICLE


CONTROL	 CONTROL


0/20(0.00)	 0/20(0.00


P - 0.001	 P <= 0.001


P< 0.001	 P< 0.001
 

0/20(0.00) 0/20(0.00)


P = 0.003 P = 0.003


1/20(0.05)	 0/20(0.00)


N.S.	 N.S.


P - 0.015
 

109
 

 LOW
 

 DOSE
 

 38/50(0.76)
 

 P< 0.001*
 

P< 0.001**
 

Infinite
 

5 . 402
 

Infinite
 

Infinite
 

5.402
 

Infinite
 

47
 

 1/50(0.02)
 

 N.S.
 

Infinite
 

0.022
 

Infinite
 

Infinite
 

0.022
 Infinite
 

57
 

 6/50(0.12)
 

 N.S.
 

2.400
 

0.325
 

108.020
 

Infinite
 

0.667
 

Infinite
 

52
 

HIGH
 

DOSE
 

29/49(0.59)
 

P< 0.001*
 

P< 0.001**
 

Infinite
 

4.108
 

Infinite
 

Infinite
 

4.108
 

Infinite
 

33
 

9/49(0.18)
 

P =0.036*
 

P =0.036**
 

Infinite
 

1.119
 

Infinite
 

Infinite
 

1.119
 Infinite
 

36
 

1/50(0.02)
 

N.S.
 

0.400
 

0.005
 

30.802
 

Infinite
 

0.022
 

Infinite
 

45
 
30

 

TABLE 4
 

(CONCLUDED)
 

UNTREATED VEHICLE LOU HIGH
 

TOPOGRAPHY :MORPHOLOCT CONTROL CONTROL DOSE DOSE
 

Mammary Gland: Adenocarcinoma, NOSb 2/20(0.10) 0/20(0.00)	 24/50(0.48) 31/50(0.62)
 

P Valuesc>d P<0.001 P<0.001	 P< 0.001** P< 0.001**
 

P = 0.002* P< 0.001*
 

Relative Risk (Untreated Control)1 4.800 6.200
 

Lower Limit 1.390 1.860
 

Upper Limit 38.983 48.724
 

Relative Risk (Vehicle Control)f Infinite Infinite
 

Lower Limit
 3.281 4.322
 Upper Limit Infinite Infinite
 

Weeks to First Observed Tumor 109	 34 33
 

Mammary Gland: Adenocarcinoma, NOS or
 

Ftbroadenomab 4/20(0.20) 1/20(0.05) 24/50(0.48) 31/50(0.62)
 

P Values0 >d P - 0.002 P<0.001	 P •= 0.027* P - 0.002*
 

P < 0.001** P < 0.001**
 

Relative Risk (Untreated Control) 2.400 3.100
 

Lower Limit 0.989 I.i28
 

Upper Limit
 8.461 10.464
 

Relative Risk (Vehicle Cont)iol)£ 9.600 12.400
 

Lower Limit
 1.794 2.391
 

Upper Limit
 380.129 479.476
 

Weeks to First Observed Tumor 60	 83 34 33
 

treated groups received time-weighted average doses of 15 and 29 mg/kg by gavage.
 

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

C

Beneath the incidence of tumors in a control group is the probability level for the Cochran-Armitage test when P< 0.05; otherwise, not significant
 

(N.S.) is indicated. Beneath the incidence of tumors in a treated group is the probability level for the Fisher exact test for the comparison of that
 

treated group with the untreated control group(*) or with the vehicle control group (**) when P<0.05 for either control group; otherwise, not significant (N.S.) is indicated.
 

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

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

f

The 95% confidence interval of the relative risk between each treated group and the specified control group.
 
of tumor that was observed in more than 5 percent of any of the DBCP-


dosed groups of either sex is included.
 

Both male and female dosed rats developed a significant number
 

of squamous-cell carcinomas of the forestomach. For both sexes the
 

Cochran-Armitage test indicated a significant (P = 0.001) positive
 

association between increased dosage and elevated tumor incidence
 

when the dosed groups were compared to either the untreated controls
 

or the vehicle controls. However, the departure from linear trend
 

was significant (P < 0.001) due to the very high incidence levels
 

observed in not only the high dose but the low dose groups as well.
 

The Fisher exact tests comparing dosed groups to control groups were
 

all significant (P < 0.001); all confidence intervals on the relative
 

risk had a lower limit greater than the value one. The incidence
 

rates observed in the controls were not significantly different from
 

the historical spontaneous incidence rates observed to date for
 

Osborne-Mendel rats in the NCI Bioassay Program (male vehicle control
 

0/748; female vehicle control 0/741; male untreated control 0/196;
 

and female untreated control 1/198).
 

Squamous-cell papillomas of the stomach were also found in dosed
 

rats. For females the Cochran-Armitage test indicated a significant
 

(P = 0.003) positive dose^response association in females; the Fisher
 

exact tests comparing high dose group to either control had a probability level of P = 0.034, a marginal result which was not significant
 

31
 
under the Bonferroni criteria. For the male rats the incidence of
 

papillomas was not statistically significant.
 

Based upon these results, the statistical conclusion is that the
 

administration of DBCP was associated with a significantly increased
 

incidence of cancer of the forestomach in Osborne-Mendel rats under
 

the conditions of this experiment.
 

Among low dose males the incidence of either hemangioma or hemangiosarcoma was significantly higher (P = 0.008) than in either of the
 

control groups. This finding was not supported by significant results
 

in the high dose group. However, it is possible that some of the high
 

dose males were not at risk long enough to develop this tumor.
 

For females, when the dosed groups were compared to either control group, the Cochran-Armitage test indicated a significant (P <
 

0.001) association between dosage and an increased incidence of adenocarcinomas of the mammary gland. Fisher exact tests confirmed these
 

results (P < 0.003) when comparing either the high dose or the low
 

dose group to either control; for all comparisons the lower limit of
 

the confidence interval on the relative risk was greater than the
 

value one. When female rats with either an adenocarcinoma or a
 

fibroadenoma of the mammary gland were considered, all of the tests
 

remained significant.
 

Based upon these results, the statistical conclusion is that
 

the administration of DBCP at the dose levels of this experiment is
 

associated with the increased incidence of adenocarcinomas of the
 

mammary gland in female Osborne-Mendel rats.
 

32
 
IV. CHRONIC TESTING RESULTS: MICE
 

A. Body Weights and Clinical Observations
 

No appreciable differences in body weight gain patterns were
 

apparent among male or female mice (Figure 3).
 

During the first 34 weeks of the study, DBCP-treated and vehicle
 

control mice displayed essentially comparable appearance and behavior.
 

Clinical signs characterized by a hunched or thin appearance and
 

apparent compound-related deaths were observed in the high dose animals beginning in week 38 of the study. The incidence of clinical
 

signs and deaths in the high dose groups increased as the study
 

progressed. Gross necropsy of most of the DBCP-treated mice revealed
 

a high incidence of stomach tumors with metastases to adjacent organs
 

in many of the animals.
 

Signs often observed in group-housed laboratory mice were noted
 

at a comparable rate among control and treated mice. These signs
 

included sores on the body (more common in males due to fighting.) ,
 

penile/anal/vulvar irritation (sometimes with anal prolapse), hunched
 

posture, reddened or squinted eyes, soft feces, alopecia, and palpable
 

nodules. A bloated appearance and swelling along the abdominal area
 

were more frequently noted in the treated mice than in the vehicle
 

controls.
 

B. Survival
 

The estimated probabilities of survival for male and female mice
 

in the control and DBCP-treated groups are shown in Figure 4.
 

33
 
TIME ON TEST (WEEKS) 

FIGURES 

GROWTH CURVES FOR DBCP CHRONIC STUDY MICE 

34 
TIME ON TEST (WEEKS) 

FIGURE 4 

SURVIVAL COMPARISONS OF DBCP CHRONIC STUDY MICE 

35 
In both male and female mice the Tarone test indicated a significant (P < 0.001) positive association between increased dosage and
 

accelerated mortality. In males the departure from linear trend was
 

significant (P < 0.001) due to the accelerated mortality in dosed
 

groups.
 

Among the males, 80 percent of the high dose group died by the
 

end of week 47, at which point the remaining 10 mice were sacrificed.
 

Eighty-four percent of the low dose group died by week 59, after
 

which the 8 remaining mice were sacrificed. Ninety percent of the
 

vehicle control males were still alive by week 58, but these 18
 

mice were sacrificed in week 59. Eighty percent of the untreated
 

control mice died by the end of week 77, after which the remaining
 

4 mice were sacrificed.
 

Among the female mice, 61 percent of the high dose group died
 

by the end of week 47. The remaining 19 mice were sacrificed in week
 

47. Eighty-two percent of the low dose mice died by week 60, at
 

which point the remaining 9 mice were sacrificed. All 18 vehicle
 

control animals (90 percent) still alive in week 58 were sacrificed
 

in weeks 59 and 60. Ninety-five percent of the untreated controls
 

survived until the end of the study.
 

Early deaths in the treated mice may have been tumor-related as
 

over 90 percent of each dosed group was found to have squamous-cell
 

carcinomas of the stomach.
 

36
 
C. Pathology
 

Histopathologic findings on neoplasms in mice are tabulated in
 

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

tabulated in Appendix D (Tables Dl and D2).
 

Squamous-cell carcinomas of the stomach occurred in 43/46 (93
 

percent) low dose males, 47/49 (96 percent) high dose males, 50/50
 

(100 percent) low dose females, and 47/48 (98 percent) high dose
 

females but in no male or female controls. Microscopically, the
 

neoplasms of the forestomach were similar in appearance to the
 

squamous-cell carcinomas seen in the rats, with frequent metastases
 

to the abdominal viscera. Pulmonary metastases occurred in 4/46 (9
 

percent) low dose males, 3/48 (6 percent) high dose males, 10/50 (20
 

percent) low dose females, and 3/46 (7 percent) high dose females.
 

Other neoplasms that occurred in this study were observed at
 

similar incidences in control and treated mice.
 

Toxic nephropathy occurred in 11/46 (24 percent) low dose males,
 

45/48 (94 percent) high dose males, 14/50 (28 percent) low dose
 

females, and 43/46 (93 percent) high dose females but in no male or
 

female controls. Microscopically, toxic nephropathy in the mice
 

occurred primarily in the proximal tubules and was comparable in
 

appearance to renal lesions in the rats.
 

Other inflammatory, proliferative, and degenerative lesions seen
 

in the treated mice were similar to those seen in controls.
 

37
 
In this study DBCP treatment was associated with squamous-cell
 

carcinomas of the forestomach in male and female mice at both high
 

and low doses. This chemical was also toxic to the kidneys, producing a toxic tubular nephropathy in both male and female mice.
 

D. Statistical Analyses of Results
 

The results of the statistical analyses of tumor incidence in
 

mice are summarized in Tables 5 and 6. The analysis for every type
 

of tumor that was observed in more than 5 percent of any of the
 

DBCP-treated groups of either sex is included.
 

Both male and female dosed mice developed a significant number
 

of squamous-cell carcinomas of the stomach. For both sexes the
 

Cochran-Armitage test indicated a significant (P = 0.001) positive
 

association between increased dosage and elevated incidence when the
 

dosed groups were compared to either the untreated controls or the
 

vehicle controls. The Fisher exact tests comparing dosed groups to
 

control groups were all significant (P < 0.001); all confidence intervals on the relative risk had a lower limit greater than the value
 

one. Additionally, the incidence rates observed in the controls were
 

not significantly different from the historical spontaneous incidence
 

rates observed to date for B6C3F1 mice in the NCI Bioassay Program
 

(male vehicle control 1/887; female vehicle control 1/877; male untreated control 1/1544; and female untreated control 0/1503).
 

Based upon these results, the statistical conclusion is that
 

DBCP was a stomach carcinogen in B6C3F1 mice under the conditions of
 

this experiment.
 

38
 
39

 

TABLE 5
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN MALE MICE TREATED WITH DBCPa
 

UNTREATED VEHICLE LOW HIGH
 

TOPOGRAPHY :MORPHOLOGY CONTROL CONTROL DOSE DOSE
 

Stomach: Squamous Cell Carcinoma 0/20(0.00) 0/20(0.00)	 43/46(0.93) 47/49(0.96)
 

P Values0 >d P< 0.001 P < 0.001	 P< 0.001* P <0.001*
 

P< 0.001** P< 0.001**
 

Departure from Linear Trend	 P< 0.001 P< 0.001
 

Relative Risk (Untreated Control)£ Infinite Infinite
 

Lower Limit 7.052 7.416
 Upper Limit Infinite Infinite
 

Relative Risk (Vehicle Control)* Infinite Infinite
 

Lower Limit 7.052
 7.416
 Upper Limit Infinite Infinite
 

Weeks to First Observed Tumor	 39 27
 

treated groups received time-weighted average doses of 114 and 219 mg/kg.
 

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

°Beneath the incidence of tumors in a control group is the probability level for the Cochran-Armitage test when P<0,05; otherwise not significant
 

(N.S.) is indicated. Beneath the incidence of tumors in a treated group is the probability level for the Fisher exact test for the comparison of
 

that treated group with the untreated control group (*) or with the vehicle control group (**) when P<0.05 for either control group; otherwise,
 

not significant (N.S.) is indicated.
 

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

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

The 95% confidence interval of the relative risk between each treated group and the specified control group.
 
40 

TOPOGRAPHY :MORPHOLOGY
 

Stomach: Squamous Cell Carcinoma
 

P VatBesc>d
 

Departure from Linear Trend
 

Relative Risk (Untreated Control)£
 

Lower Limit
 Upper Limit
 

Relative Risk (Vehicle Control)f
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

TABLE 6
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS AT
 

SPECIFIC SITES IN FEMALE MICE TREATED WITH DBCP3
 

UNTREATED VEHICLE
 

CONTROL CONTROL
 

0/20(0.00) 0/20(0.00)
 

P< 0.001 P< 0.001
 

P< 0.001 P< 0.001
 

LOW
 

DOSE
 

50/50(0.100)
 

P < 0.001*
 

P <0.001**
 

Infinite
 

8.661
 Infinite
 

Infinite
 

8.661
 

Infinite
 

40
 

HIGH
 

DOSE
 

47/48(0.98)
 

P< 0.001*
 

P^0.001**
 

Infinite
 

7.845
 Infinite
 

Infinite
 

7.845
 Infinite
 

32
 

created groups received time-weighted average doses of 110 and 209 mg/kg.
 

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

°Beneath the incidence of tumors in a control group is the probability level for the Cochran-Armitage test when P<^ 0.05; otherwise, not significant
 

(N.S.) is indicated. Beneath the incidence of tumors in a treated group is the probability level for the Fisher exact test for the comparison of
 

that treated group with the untreated control group (*) or with the vehicle control group (**) when P< 0.05 for either control group; otherwise,
 

not significant (N.S.) is indicated.
 

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

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

The 95% confidence interval of the relative risk between each treated group and the specified control group.
 
V. DISCUSSION
 

Under the conditions of this bioassay, administration of
 

technical-grade DBCP was associated with a high incidence of squamouscell carcinomas of the forestomach and toxic nephropathy in male and
 

female rats and mice and with a high incidence of mammary adenocarcinoma in female rats.
 

The untreated controls for the rat bioassay were approximately
 

16 weeks younger than the dosed and vehicle control rats, were started
 

on test a corresponding 16 weeks later, and were housed in a separate
 

room. These differences in age and room assignments did not, however,
 

complicate statistical evaluation since vehicle controls are the preferred controls for a gavage study and statistical tests were applied
 

to the vehicle controls. In addition, there were no significant differences in tumor incidence between untreated and vehicle controls in
 

this bioassay.
 

Among the rats, squamous-cell carcinomas of the forestomach were
 

observed in 47/50 (94 percent) high dose males, 47/50 (94 percent)
 

low dose males, 29/49 (59 percent) high dose females, and 38/50 (76
 

percent) low dose females but in none of the untreated or vehicle controls. When the incidences of these neoplasms in the treated animals
 

were statistically compared to those in the control groups, there was
 

a significant positive association between dosage and incidence of
 

these neoplasms. Some of these carcinomas were accompanied by pulmonary metastases. Squamous-cell papillomas were also found in dosed
 

41
 
rats of both sexes, but not in vehicle or untreated controls. The
 

Cochran-Armitage test showed a significant association between increased dosage and increased incidence of squamous-cell papillomas
 

for females but not for males.
 

Squamous-cell carcinomas and papillomas of the forestomach are
 

uncommon lesions in control animals, and the high incidences of these
 

lesions in treated animals indicates that they are related to DBCP
 

treatment. It is possible, however, that the method of administration might have played a role in inducing these neoplasms, as DBCP is
 

a skin and mucous membrane irritant (Shell Chemical Company, 1972)
 

and proliterative lesions of the forestomach have been noted in other
 

bioassays where chemical irritants have been administered by gavage.
 

Adenocarcinomas of the mammary gland in females were an indication of the capacity of DBCP to induce tumors at other than the site
 

of application. These tumors occurred in 2/20 (10 percent), 24/50
 

(48 percent), and 31/50 (62 percent) of the untreated control, low
 

dose, and high dose groups, respectively. When the treated groups
 

were statistically compared to either control group, a significant
 

association between dosage and increased incidence of these tumors
 

was indicated.
 

Among mice, squamous-cell carcinomas of the forestomach were
 

observed in 47/48 (98 percent) high dose males, 43/46 (93 percent)
 

low dose males, 47/48 (98 percent) high dose females, and 50/50 (100
 

percent) low dose females but in none of the untreated or vehicle
 

42
 
control males or females. Statistically, there was a significant
 

association between dosage and incidence of these neoplasms when the
 

treated animals were compared to either of their respective control
 

groups. These carcinomas were frequently accompanied by metastases
 

to the abdominal viscera and occasionally by metastases to the lung.
 

The animals survived sufficiently long to develop the previously
 

mentioned tumors despite the frequency with which toxic nephropathy
 

occurred in the treated animals. This severely toxic lesion occurred
 

in 100 percent of the treated female and low dose male rats and 98
 

percent of the high dose male rats but in none of the untreated or
 

vehicle control rats. Additionally, toxic nephropathy in mice occurred in 45/48 (94 percent) high dose males, 11/46 (24 percent)
 

low dose males, 43/46 (93 percent) high dose females, and 14/50 (28
 

percent) low dose females, but in none of the untreated or vehicle
 

control groups.
 

Under the conditions of this study DBCP is a stomach carcinogen
 

in Osborne-Mendel rats and B6C3F1 mice of both sexes and is carcinogenic to the mammary glands of female rats. Chronic exposure to DBCP
 

also caused toxic nephropathy in both rats and mice.
 

43
 
VI. BIBLIOGRAPHY
 

Andrilenas, P.A., Farmers' Use of Pesticides in 1971—Quantities.
 

U.S. Department of Agriculture. Agricultural Economic Report
 

No. 252, 1974.
 

Armitage, P., Statistical Methods in Medical Research, Chapter 14.
 

J. Wiley & Sons, New York, 1971.
 

Berenblum, I., editor, Carcinogenicity Testing. International Union
 

Against Cancer, Technical Report Series, Vol. 2. International
 

Union Against Cancer, Geneva, 1969.
 

Chemical Abstracts Service, The Chemical Abstracts Service (CAS)
 

Ninth Collective Index, Volumes 76-85, 1972-1976. American
 

Chemical Society, Washington, B.C., 1977.
 

Cox, D.R., Analysis of Binary Data, Chapters 4 and 5. Methuen and
 

Co., Ltd., London, 1970.
 

Cox, D.R., "Regression Models and Life-Tables." Journal of the Royal
 

Statistical Society, Series "B" 34:187-220, 1972.
 

Fowler, D.L. and J.N. Mahan, The Pesticide Review 1975. U.S. Department of Agriculture, Washington, D.C., 1976.
 

Gart, J.J., "The Comparison of Proportion: A Review of Significance
 

Tests, Confidence Limits, and Adjustments for Stratification."
 

International Statistical Institute Review 39:148-169, 1971.
 

Gleason, M.N., R.E. Gosselin, H.C. Hodge, and R.P. Smith, Clinical
 

Toxicology of Commercial Products. Third edition. The .Williams
 

and Wilkins Company, Baltimore, Maryland, 1969.
 

Kaplan, E.L., and P. Meier, "Nonparametric Estimation from Incomplete
 

Observations." Journal of the American Statistical Association
 

^3:457-481, 1958.
 

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

"Carcinogenesis Bioassay Data System." Computers and Biomedical
 

Research 7:230-248, 1974.
 

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

Co., New York, 1966.
 

44
 
Peterson, B., and P. Shinoff, "Firms Had Sterility Data on Pesticide."
 

The Washington Post, p. Al, August 23, 1977a.
 

Peterson, B., and P. Shinoff, "Dow Recalling Pesticide DBCP." The
 

Washington Post, p. Al, August 26, 1977b.
 

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

in Male Rats Given Subcutaneous Carbon Tetrachloride." Journal
 

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

Rosenkranz, H.S., "Genetic Activity of l,2-Dibromo-3-Chloropropane, A
 

Widely Used Fumigant." Bulletin of Environmental Contamination
 

and Toxicology 14:8-12, 1975.
 

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

Kaufman, "Respiratory Tract Carcinogenesis in Hamsters Induced 

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

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

(R)
 Shell Chemical Company, Summary of Basic Data for Nemagon^ Soil
 

Fumigant. Shell Chemical Company/Agricultural Division,San
 

Ramon, California. Technical Data Bulletin SC:60-106.
 

September 1972.
 

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

62_:679-682, 1975.
 

Torkelson, T.R., S.E. Sadek, V.K. Rowe, J.K. Kodama, H.H. Anderson,
 

G.S. Loquvam, and C.H. Hine, "Toxicologic Investigations of
 

1,2-Dibromochloropropane." Journal of Toxicology and Applied
 

Pharmacology 3(5):545-559, 1961.
 

U.S.	 International Trade Commission, Synthetic Organic Chemicals.
 

United States Production and Sales, 1975. USITC Publication 804,
 

Washington, D.C., 1977.
 

45
 

APPENDIX A
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS
 

IN RATS TREATED WITH DBCP
 

TABLE A1

 

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

HIGH DOSE 

01-053M 

50 

50 

50 

(50) 

1 (2X) 

1 (2X) 

(50) 

(50) 

5 (10X) 

(50) 

2 (4X) 

(50) 

1 (2X) 

(49) 

21 (H3X) 

(«8) 

1_I2S1 

CONTROL(UNTR) 

ANIMAL S INITIALLY IN STUDY 

ANIMAL S NECROPSIED 

ANIMAL S EXAMINE D HISTOPATHOLOGICA1LY ** 

INTEGUMENTAR Y SYSTEM 

*SKIN 

BASAL-CtLL CARCINOM A 

K rRATOACANTHOM A 

FIBROSARCOMA 

*SOBCDT TISSUE 

FIBROMA 

FIBROSARCONA 

HEHANGIOSARCOM A 

RESPIRATORY SYSTEM 

• LUHG 

SQUAMOUS CELL CARCINOMA, METASTA 

HFMANGIOSARCON A 

•HEMATOPOIETIC SYSTEM 

*MULTIPLB ORGAN S 

LYMPHOCYTIC LEUKEMI A 

SRANDLOCYTIC LEUKEMIA 

•ABDOMINAL CAVITY 

MALIG.LYMPHOMA, HISTIOCYTIC TYPE 

•SPLEEN 

SQUAMOUS CILL CARCINOMA, METASTA 

HEMANGIOM A 

HEMANGIOSARCOM A 

tCERVICAL LYMPH NODE 

__ HBEfiSiSGSfli _ . 

01-031M 

20 

20 

20 

(20) 

(20) 

1 (5X) 

(20) 

(20) 

1 <5X) 

(20) 

(20) 

(19) 

* NUMBER OF AHIMAL S WITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBER OF ANIMAL S NEC80PSIED 

"EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

A-3 

CONTROL (»EH)

01-051M 

20 

20 

20 

(20) 

(20) 

(20) 

(20) 

(20) 

(20) 

(20) 

 LOU DOSE 

01-052M 

50 

50 

50 

(50) 

1 (2X) 

2 («X) 

(50) 

3 <6X) 

1 (2X) 

(50) 

5 (10X) 

1 (2X) 

(50) 

(50) 

(50) 

17 (34X) 

2 <KX ) 

9 (18X) 

(tt) 
TABLE Al (CONTINUED) 

CONTROL (UNTR) CONTROL (VEH ) LOH DOSE ItlIGH DOSE 

01-031M 01-051M 01-052B 01-053B 

*M?SF»iT1?RIC L. NODE (19) (20) («4) (48) 

SQBAMOUS CELL CARCINOMA, METASTA 2 (5*) 4 (8X) HEMANGIOSAHCOMA , BETASTATIC 1 (2X) 

*THYBUS (13) (5) (8) (2)

SQUAKOU S CEL L CARCIHOMA. METASTA 1 (SOX) 

CIRCULATOR Y SYSTEM 

tENtlOCARDIUM (20) (20) (50) (50) 

SARCOMA, NOS 1 (2X) 

DIGESTIVE SYSTEM 

*SALIVAR Y GLAN D (14) (2) (5) 

CARCINOMA , NOS 1 (7X) 

*LIVER (20) (20) (50) (50) 

SQDAMOUS CELL CAHCIHOMA, METASTA 20 (40*) 27 (54X) 

HrNANGIOSARCOM A 1 (2«) 

tPANCREA S (20) (20) (50) (50) 

SQOAMOOS CELL CARCINOMA, BETASTA 16 (32X) 18 (36X) 

HEBAN6IOSARCOBA 2 (UX) 

H2HANGIOSARCOMA, METASTATIC 1 (2X) 

tSTOBACH (20) (19) (50) (50) 

SQUAMOOS CELL PAPILLOBA 1 (2*) 2 (<tX) 

SQOAHOUS CELL CARCINOMA 47 (9»X) 47 (94X) 

HEMANGIOSARCOMA 1 (2X) 

• SHALL INTFSTIHB (20) (20) (50) (50) 

SQUAMOUS CELL CARCINOMA, BETASTA 5 (10%) 5 (10X) PIBROSARCOMA 1 (5%) 

HTMANGIOSARCOMA, METASTATIC 1 <2X) 

tDOODENOM (20) (20) (50) (50) 

SQUAHOUS CELL CARCISOBA, METASTA 1 (2X) 

#LABG? INTESTINE (19) (20) (49) (49) 

SQOAMOOS CELL CARCINOMA, METASTA 3 (6X) 3 (6X) 

UPINAF Y SYSTEM 

*KIDNFY

SQUAMOOS CELL

BiaiEI2fli+ CARCINOMA, METASTA 

 (20) (20) (50) 

2 (4X) 

_ 1 I2SL 

(50) 

4 (8X) 

t NUMBE R OP ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* NONBFR OF ANIMAL S NECROPSIED + THIS IS CONSIDERED TO BE A BENIGN FORM OF THE MALIGNANT MIXED TUMOR OF THE KIDNEY AND CONSISTS OF PROLIFERATIVE 

LIPOCYTES, TUBULAR STRUCTURES, FIBROBLASTS, AND VASCULAR SPACES IN VARYING PROPORTIONS. 

A-4 
TABLE Al (CONTINUED) 

" CONTROL(UNTR) CONTROL (VEH ) 

01-031M 01-051M 

#UPINAP Y BLADDER (19) (19) 

SQUAHOOS CELL CARCINOMA, METASTA 

ENDOCEIN^ SYSTEM 

#PITUITARY (20) (20)

 CHROMOPHOBE ADENOMA 2 (10%)

 FIBROSARCOM A

 

f ADRFNA L (20) (20) 

SQUAMOUS CELL CARCINOMA, METASTA 

CORTICAL CARCINOMA 2 (10X)

 

#THYBOID (19) (20) FOLLICULAR-CELL ADENOM A

 

FOLLICULAR-CELL CARCINOM A 1 <5X)

 

REPEODOCTIVF SYSTEM 

*MAMMAR Y GLAN D (20) (20)

 CARCINOMA, HOS

 

ADENOCARCINOMA , NOS 1 (5X)

 FIBRO ADENOM A 1 (5X) 1 (5X)

 

fPPOSTATF. (20) (16)

 

SQUAMOUS CELL CARCINOMA, METASTA 

#TESTIS (20) (19)

 

SQUAMOHS CELL CARCINOMA, METASTA 

NEEVODS SYSTEM 

NON" 

SPECIAL SENSE ORGANS 

NONE 

BOSCOLOSKELETAL SYSTEM 

— NONE _ 

t NUMBER OF ANIMAL S KITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBER OF ANIMALS NECROPSIED 

LOW DOSE 

01-052M 

HIGH DOSE 

01-053M 

(49) 

3 (6X) 

(46) 

1 (2X) 

(49) 

1 (2*) 

1 <2X) 

(49) 

3 (6X) 

(48) 

1 (2X) 

(50) 

8 (16%) 

(«8) 

3 <6X) 2 (4X) 

(49) 

2 (4X) 

1 (2X) 

(50) 

1 (2X) 

(50) 

1 (2X) 

(39) 

1 (3X) 

(50) 

7 (14X) 

(12) 

(49) 

5 (10X) 

— _ 

A-5 
TABLE A-l (CONCLUDED) 

CONTROL (UNTR)

01-031M CONTROL (VEH) 01-051M LOU DOSE 01-052M HIGH DOSE  01-053H 

BODY CAVITIES 

•ABDOMINA L CAVIT Y

HEMANGIOSARCOM A

 (20) (20) (50)

 1 (2*)

 (50) 

1 (2%) 

• MESENTER Y

SQUAMOU S CELL CARCINOMA . BETASTA

 (20) (20) (50)

 2 (IX) 

 (50) 

A LL OTHER SYSTEMS 

NON E 

ANIMA L DISPOSITION SUMMAR Y 

ANIMAL S INITIALL Y IN STUDY

NATURA L DEATHS

MORIBUN D SACRIFICE

SCHEDULED SACRIFICE

ACCIDENT. LY KILLED 

TERMINA L SACRIFICE

ANIMA L MISSIN G 

 20

 11

 5

 <1

 20

 8

 10 

2

 50

 U2

 3

 5

 50 

 30 

 10 

 10 

9 INCLUDES AUTOLYZE D ANIMAL S 

TUMO R SUMMAR Y 

TOTA L ANIMAL S WIT H PRIMAR Y

TOTAL PRIMAR Y TUMOPS

 TUMORS * 6

 11

 1

 1

 H9

 81

 18 

 62 

TOTA L ANIMAL S WIT H BENIG N

TOTAL BFNIG N TUMORS

 2 TUMORS

3

1 

1

8 

 12

5 

5 

TOTA L ANIMAL S WIT H MALIGNAN T

TOTAL MALIGNAN T TUMORS

 TUMORS 6

 8

 U8

 69

 «7 

 57 

TOTA L ANIMAL S WIT H SECONDAR Y

TOTAL SECONDAR Y TUMOR S

 TUMORS * 2 8

 87

 3t 

 105 

TOTAL ANIMALS WITH TUMORS

BENIG N OR MALIGNAN T 

TOTAL UNCERTAI N TUMOR S 

 UNCERTAINTOTA L ANIMAL S WIT H TUMOR S

PRIMAR Y O R METASTATIC 

TOTAL UNCERTAI N TUMOR S 

 UNCERTAIN ­

* PRIMAR Y TUMORS : AL L TUMOR S EXCEPT SECONDAR Y TUMORS 

» SECONDAR Y TUMORS : METASTATIC TUMORS O R TUMOR S INVASIV E INTO A N ADJACEN T ORGA N 

A-6 
TABLE A2

 

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

CONTROL (UNTH) CONTROL (VEH) LOW DOSE HIGH DOSE
 

01-031F 01-051F 01-054F 01-055F
 

ANIMALS INITIALLY IN STUDY 20 20 50 50
 

ANIMALS NECROPSIED 20 20 50 50
 ANIMALS EXAMINED HISTOPATHOLOGICALLY ** 20 20 50 50
 

INTEGUMENTARY SYSTEM
 

*SOBCOT TISSUE (20) (20) (50) (50)
 

FIBROMA 1 (2*)
 HEMANGIOSARCOMA 2 (4*)
 

RESPIRATORY SYSTEM
 

*LUNG (20) (20) (42) (44)
 

SQUAMOOS CELL CARCINOMA, METASTA 3 (7%)
 

ADFNOCARCINOMA, NOS, METASTATIC 4 (10*) 3 (7X)
 

GRANDLOSA-CELL CARCINOMA, NETAST 1 (2X)
 

HEMANGIOSARCOMA 1 (2*)
 

HEMATOPOIETIC SYSTEM
 

tSPLFEN (20) (20) (42) (44)
 

SQUAMOUS CELL CARCINOMA, METASTA 5 (12X) 3 (7X)
 

HEMANGIOSARCOMA 4 (10X)
 

*MESENTERIC L. NODE (20) (18) (23) (43)
 

SQUAMOUS CELL CARCINOMA, METASTA 1 (4X) 1 (2X)
 GRANULOSA-CELL CARCINOMA, METAST 1 <4X)
 

CIRCULATORY SYSTEM
 

*HEART (20) (20) (42) (44)
 SARCOMA, NOS 1 (2X)
 MIXED TOMOR, METASTATIC 1 (5X)
 

DIGESTIVE SYSTEM
 

tLIVFR (20) (20) (42) (44)
 

SQUAMOUS CELL CARCINOMA, METASTA 6 (14X) 8 (18X)
 

HFPATOCELLOLAR CARCINOMA 1 (5X) 1 (2*)
 

HEMAN.gIOSARgO.BJ . __1 12SL
 

* NUMBE R OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBER OF ANIMAL S NECROPSIED 

**EXCLUBES PARTIALLY AUTOLYZED ANIMALS 

A-7 
TABLE A2 (CONTINUED) 

CONTROL (UNTR) CONTROL (VEH ) LOS DOSE HIGH DOSE 

01-031F 01-051F 01-054F 01-055F 

IPANCREA S (20) (19) C*2) (44) 

SQUANODS CELL CARCINOMA , METASTA 3 (7X| 7 (16X) HFMANGIOSARCOH A 1 (2X) 

#STOMAC H 

SQUAMOU S CELL PAPILLOMA 

(20) (20) (50) 

1 (2X) 

(49) 

9 (18%) 

SQUAMOOS CELL CARCINOMA 38 (76X) 29 (59X) 

iSMALL INTESTINE 

SQUAMOHS CELL CARCINOMA , METASTA 

(20) (20) (U2) 

1 (2X) 

<44) 

1 <2X) 

#LARG E INTESTINE (19) (19) (42) (44) 

SQUAMOU S CELL CARCINOMA , METASTA 1 <2X) 

UBINAR Y SYSTEM 

tKIDNE Y (20) (20) (D2) (44) 

SQUAHOUS CELL CARCINOMA , METASTA 1 (2X) 1 (2X) TRANSITIONAL-CELL CARCINOM A 1 (2X) 

ADFNOCARCINOMA , SOS, METASTATIC 1 <2X) TUBULAR-CELL ADENOMA 1 (2X) 

1IXED TUMOR , MALIGNAN T 1 (5*) HAMARTOMA + 2 (10X) 

ENDOCRIN E SYSTEM 

*PITUITARY (19) (20) (10) (41) 

CHROMOPHOBE ADENOM A 6 (32X) 1 (5X) 

#ADRENA L (20) (20) 0*0) (44) 

SQDAMOUS CELL CARCINOMA , METASTA 1 OX) 1 (2X) CORTICAL ADENOM A 1 <5X) 

CORTICAL CARCINOM A 1 (3X) 

#THYROID (20) (20) (39) (44) 

C-CELL ADENOM A 2 (10*) C-CELL CARCINOM A 2 (10X) 

*PA»ICREATIC ISLETS (20) (19) (42) (44) 

_ ISLS^CfLL.iPJNONA. . 1.15X1 

* NUMBER OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBE R OF ANIMAL S NECROPSIED +

 THIS IS CONSIDERED TO BE A BENIGN FORM OF THE MALIGNANT MIXED TUMOR OF THE KIDNEY AND CONSISTS OF PROLIFERATIVE 

LIPOCYTES, TUBULAR STRUCTURES, FIBROBLASTS, AND VASCULAR SPACES IN VARYING PROPORTIONS. 

A-8 
TABLE A2 (CONTINUED) 

COHTROL(UNTR ) CONTROL (VEH ) LOU DOSE 

01-931F 01-051F 01-054F 

prpRnnuCTIV F SYSTFJ1 

*MAMMAP Y GLAN D (20) (20) (50) 

ADENOCARCINOMA , NOS 2 (10*) 24 (USX ) 

"IBROAD^NOM A 2 (10X) 1 (5X) 

#(JTrP US (20) (20) (39) 

SQUAHOU S CrLL CARCINOMA , HETASTA 1 (3J) 

TNDOMFTRIA L STROMAL POLYP 1 (5X) 

H^HANGION A 1 (5%) 

*OVAP Y (20) (20) (39) 

SQOAMOU S CELL CARCINOMA , METASTA 2 (5X) 

CYSTAD^NOCARCINOMA , NOS 1 (5X) 

GP.ANBLOSA-CI:LL TOHOR 1 (3*) 

GRANULOSA-fLi CARCINOMA 1 (3J) 

NtPVOD S SYSTFB 

NON r 

SPECIAL SENSE ORGAN S 

NONr 

MUSCULOSKELFTA L SYSTEM 

NON r 

BODY CAVITIFS 

*MTS^NTER Y (20) (20) (50) 

SQUAMOU S CELL CARCINOMA , METASTA 1 (2X) 

A LL OTH^R SYSTEMS 

_ NONr _ _ 

* NUMBE R OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBPR OF ANIMAL S NECROPSIED 

HIGH DOSE 

01-055F 

(50) 

31 (62%) 

(43) 

1 (2%) 

(43) 

(50) 

1 <2X) 

A-9 
ANIMA L DISPOSITION SUMMARY
 

ANIMALS INITIALLY IN STUDY


NATURAL PSATH9


MORIBUND SACRIFICE
 

SCHEDULED SACEIFICE


ACCIDENTALLY KILLED
 

TERMINAL SACRIFICE


ANIMAL MISSING
 

59 INCLUDES AUTOLYZFD ANIMALS
 

TUMOR SUBMAPY
 

TOTAL ANIMALS WITH PRIMARY TUMOES*


TOTAL PRIMARY TUMORS


TOTAL ANIMALS WITH BENIGN TUMORS


TOTAL BENIGN TUMORS


•'"OTAL	 ANIMALS WITH MALIGNANT TUMOES


TOTAL MALIGNANT TUMORS


TOTAL ANIMALS WITH SECONDARY TUMORS*


TOTAL SECONDARY TUMORS


TOTAL ANIMALS WITH TUMORS UNCEETAINB^MIGN OP MALIGNANT
 

TOTAL UNCERTAIN TUMORS
 

TOTza ANIMAL S WITH TUMORS ONCERTAINFPTMAPY OF METASTATIC
 TOTAL UNCERTAIN TOMORS
 

TABLE A2 (CONCLUDED)
 

CONTROL (UHIE)


01-031F


 20


 2


 5


 13


 1 1t


 21


 12


 11


 6
 

7
 

1
 

1
 

 CONTROL (VEH)
 

 01-051F
 

 20
 

5
 

 10
 

5
 

 14
 

4
 

1
 

4
 

LOU DOSE
 

01-054F
 

50
 

31
 

11
 

8
 

44
 

78
 

2
 

2
 

44
 

75
 

12
 

29
 

1
 

1
 

* PHIMAB Y TUMOPS: ALL TUMORS EXCEPT SECONDAEY TUMORS
 

* S*CONDA"Y TUMORS: METASTATIC TUMORS OE TUMORS INVASIVE INTO AN ADJACENT ORGAN
 

A-IO 

HIGH DOSE
 

01-055F
 

50
 

25
 

22
 

3
 

43
 

72
 

11
 

11
 

42
 

61
 

13
 

30
 
APPENDIX B
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS
 

IN MICE TREATED WITH DBCP
 

TABLEB1

 

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

'—-——— »——- «i rifi-— I — I IU- «•—-.—_ __» H-M-— .-..-_•­

CONTROL(ONTR) CONTROL (VEH) LOR DOSE HIGH DOSE 

02-M061 02-M051 02-M052 02-M053 

ANIMALS INITIALLY IH STUDY 20 20 50 50 ANIMALS NECROPSIED 19 20 H7 50 ANIMALS EXAMINED HISTOPATHOLOGICALLY *» 19 20 l»5 50 

INTEGUMENTARY SYSTEM 

*SKIN (19) (20) («7) (50) 

SQUAHOUS CELL CARCINOMA. METASTA 8 (17X) 5 (10%) 

•SDBCUT TISSUE (19) (20) (47) (50)

 

FIBROSARCOMA 1 (5%)

 

RESPIRATORY SYSTEM 

• LON G (18) (20) (46) (48) 

SQDAMOOS CELL CARCINOMA, BETASTA 4 (9X) 3 (6*) AL7EOLAR/BROHCHIOLAR ADEHOMA 1 (2X) 

HEBATOPOITTIC SYSTEM 

*MOLTIPLE ORGAN S (19) (20) (47) (50) MALIG.LYMPHOMA . DNDIFPEH-TYPE 1 (5%)

 MALIG.LYMPHOHA, LYMPHOCYTIC TYPE 1 (5X)

 GRANOLOCYTIC LEDKEHIA 1 (5%)

 

*SPLEE» (19) (20) (45) (47) 

SQUAMODS CELL CARCINOHA, METASTA 8 (18%) 14 (30*) 

*LYMPH NOD? (18) (18) (39) (42) 

SQUAMODS CELL CARCINOMA , METASTA 1 (3»)

 

tCERTICAL LYMP H NODE (18) (18) (39) (42) 

SQDA800S CELL CARCINOMA, METASTA 3 <8X)

 

CIRCOLATORY SYSTEM 

_ flosi _ _ 

* SOMBER OF ANIMAL S KITH TISSOE EXAMINED MICROSCOPICALLY *

 NUMBER OF ANIMAL S NECROPSIED 

**EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

B-3 
TABLE Bl (CONTINUED) 

CONTROL (UNTR) CONTROL (VEH) 

02-M061 02-M051 

DIGESTIVE SYSTEM 

*LTVR (19) (20) 

SQUAMOU S C'LL CARCINOMA , HETASTA 

NrOPIASTIC NODDLE 1 (5S)

 

H'-PATOC-LLULAR CARCINOM A 1 (5)1)

 

*GALLBLADBT:R (19) (20) 

SQUAMOU S CELL CARCINOMA , METASTA 

0PANCRFA S (19) (19) 

SQUAMOU S CELL CARCINOMA , METASTA 

ESOPHAGUS (18) 

SQUAKOU S CELL CARCINOMA, METASTA 

*STOKAC H (20) (20) 

SQUABOU S CFLL CARCINOM A 

tSMAL L INTESTINE (18) (19)

 

SQUAMOU S CELL CARCINOMA , METASTA 

• DUODENU M (18) (19)

 

SQUAMOUS CFLL CARCINOMA, HETASTA 

•COLON (19) (19)

 

SQUABOUS C"LL CARCINOBA, BETASTA 

UPINAR Y SYSTEM 

tKIDNE Y (19) (20)

 

SQUAMOD S CELL CARCINOMA , METASTA 

*UPINAR Y BLADDE R (17) (19) 

SQUAMOU S C"LL CARCINOMA , BETASTA

 

ENDOCRIN E SYSTEM 

#PDRENA L (19) (20) 

SQUABOUS CFLL CARCINOMA, BETASTA 

REPRODUCTIV E SYSTEM 

#PPOSTAT? (18) (20) 

S2UiMOUS_CELL_CARQINfiMii _JSE.TAST4_

 

t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

LOH DOSE 

02-B052 

(46) 

20 (43X) 

1 (2%) 

(47) 

2 (4X) 

(45) 

19 (42X) 

(3)

1 (33X) 

(46) 

43 (93X) 

(44) 

2 (5X) 

(44) 

1 <2X) 

(46) 

2 (4X) 

(46) 

4 (9*) 

(46) 

2 (<tX) 

(42) 

21 (SOX) 

(33) 

. __i_12SL__ 

HIGH DOSE 

02-M053 

(48) 

17 (35X) 

(50) 

2 (4X) 

(48) 

14 (29X) 

(7)

2 (29%) 

(49) 

47 (96X) 

(47) 

(47) 

(47) 

(48) 

4 (8X) 

(47) 

(49) 

13 (27X) 

(38) 

i_I2SL 

B-4
 
TABLE Bl (CONTINUED) 

CORTBOL(DITB) CONTROL (? EH) LOU DOSE HIGH DOSE 

02-H061 02-M051 02-H052 02-H053 

»SFMINAL VF.SICLE 

SQ.UABOOS CELL CARCINOMA, HETASTA 

(19) (20) (U7) (50) 

1 (2%) 

tTESTIS 

SQDAMOIIS CELL CARCINOMA, HBTASTA 

(19) (20) (15) 

1 (2*) 

(18) 

3 (6X) 

*?PIDIDYHIS 

SQUANOOS CELL CARCINOMA, HBTASTA 

(19) (20) <U7) 

8 (17X) 

(50)

5 (1C*) 

HERVOOS SYSTEM 

NONE 

SPECIAL SENSE ORGANS 

*EY'J 

MALIGNANT HE LA SOBA 

(19) 

1 (5*) 

(20) (U7) (50) 

HOSCULOSKELETAL SISTEB 

*NDSCLE OF TRUNK 

FIBROSARCOHA 

(19) (20) (»7) 

1 (2») 

(50) 

•HDSCL! RIP/THIGH 

SQOAHODS CELL CABCIiOBA, HETASTA 

(19) (20) (17) 

1 (2*) 

(50) 

1 (2X) 

BODY CAVITIES 

*ABDOBIIIAL CAfITT 

SQOAHOUS CELL CABCIIOBA. HBTASTA 

(19) (20) (47) 

1 (2X) 

(50) 

1 (2X) 

*SESEHTHRY (19) (20) (17) (50) 

SQUAHOOS CELL CABCIIOBA, MBTASTA 1 (2X) 

ALL OTHER SYSTE8S 

DIAPHRAGM 

52]lifl225 C£U_£AB£UUHUUJUXA5XA_ 1 

* HOBBFR OF ANIMALS KTH TISSUE EXAHIHED BICROSCOPICALLI 

* NUMBER OF AHIBALS IBCBOPSIZO 

B-5 
TABLE Bl (CONCLUDED)
 

CONTROL (UNTR) CONTROL (VEH) LOW DOSE HIGH DOSE
 

02-M061 02-M051 02-H052 02-M053
 

ANIMAL DISPOSITION SUMMARY
 

ANIMALS INITIALLY III STUDY 20 20 50 50
 

NATURAL DEATHS 16 2 41 39
 MORIBUND SACRIFICE 1 1
 

SCHEDULED SACRIFICE
 

ACCIDENTALLY KILLED
 TERMINAL SACRIFICE 1* 1 8 8 10
 ANIMAL MISSING
 

S INCLUDES AUTOLYZED ANIMALS
 

TUMOR SUMMARY
 

TOTAL ANIMALS WITH PRIMARY TUMORS* 5 2 13 17
 

TOTAL PRIMARY TUMORS 5 2 l»5 «8
 

TOTAL ANIMALS WITH BENIGN TUMORS 1
 

TOTAL BENIGN TUMORS 1
 

TOTAL ANIMALS WITH MALIGNANT TUMORS 5 1 43 «7
 

TOTAL MALIGNANT TUMORS 5 1 15 17
 

TOTAL ANIMALS WITH SECONDARY TUMORS* 36 30
 

TOTAL SECONDARY TUMORS 110 88
 

TOTAL ANIMALS WITH TUMORS UNCERTAINBENIGN OH MALIGNANT 1
 

TOTAL UNCERTAIN TUMORS 1
 

TOTAL ANIMALS WITH TUMORS UNCERTAINPRIMARY OR METASTATIC
 

TOTAL UNCFRTAIN TUMORS
 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDARY TUMORS
 

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

B-6
 
SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICE TREATED WITH DBCP 

CONTROL (UNTR) CONTROL (VEH ) LOW DOSE HIGH DOSE 

02-F061 02-F051 02-F054 02-F055 

ANIMALS INITIALLY IN STUDY 20 20 50 50 

ANIMAL S HISSING 1 

ANIMALS NECROPSIED 20 20 50 U7 

ANIMAL S EXAMINE D HISTOPATHOLOGICALLY ** 20 20 50 17 

INTEGUMENTARY SYSTEM 

*SKIN (20) (20) (50) (t7) 

SQUAMOUS CELL CARCINOMA , METASTA 13 (26X) 8 (17%) 

*SD3COT TISSUE (20) (20) (50) C*7)

SQUAMODS CELL CARCINOMA, METASTA 6 (13X) 

PFSPIRATOR Y SYSTEM 

*LUNG (20) (20) (50) (t6)

 

CARCINOMA, NOS, METASTATIC 1 (5S) 

SQUAMOU S CELL CARCINOMA , METASTA 10 (20X) 3 (7X)

 

AD"NOCARCINOMA, NOS, METASTATIC 1 (2X)

 

ALVEOLAR/BRONCHIOLA R ADENOMA 2 C4X) 

ALVFOLAR/BRONCHIOLAR CARCINOMA 1 (2%)

 

HEMATOPOI'TIC SYSTEM 

*MULTIPLr ORGAN S (20) (20) (50) («7)

 

MALIG.LYMPHOMA , UNDIFFER-TYPE 1 (2X)

 

1ALIG.LYMPHOMA , LYMPHOCYTIC TYPE U (20X) 

1ALIGNANT LYMPHOMA, MIXED TYPE 1 (5X) 

*SPL^TN (20) (20) (50) (H6) 

SQUAMOU S CELL CARCINOMA , METASTA 10 (20X) 11 (2UX) 

*LYMP H NODE (20) (19) («6) (39) 

SQUAMOUS CELL CARCINOMA, METASTA 1 (3X)

 

#CF°VICAL LYMPH NODE (20) (19) (46) (39) 

SQUAMOU S CELL CARCINOMA , BETASTA 15 (33X) 5 (13X) 

tMESFNTERIC L. NODE (20) (19) (16) (39) 

-__ SO.UAMOUS_CELL_CARCINOMA.._J1ETASTA -2 iSSL 2_i5.SL _. 

* MUMB^ P OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* !'UHPr'P OF ANIMAL S NECROPSIED 

"EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

B-7 
TABLE B2 (CONTINUED) 

~ — -—— CONTROL (ONTR) 

02-P061 

CONTROL (TEH) 

02-P051 

LOW DOSE 

02-F05K 

HIGH DOSE 

02-F055 

CIRCULATORY

NONE 

 SYSTEM 

DIGESTIVE SYSTEH 

fLIVEF 

SQUAHOOS CELL CARCIHOHA, HETASTA 

HCPATOCELLULAR CARCINOMA 

•GALLBLADDER 

SQUAMOUS CELL CARCINOMA, HETASTA 

•PANCHPAS 

CARCIHOHA, NOS, HBTASTATIC 

SQUAHOnS CELL CARCINOMA, HETASTA 

•ESOPHAGUS 

SQOAHODS CELL CARCINOMA, HETASTA 

tSTOHACH 

SQDAHOUS CELL CARCINOHA 

«SHALL INTESTINE 

SQUAMOOS CELL CARCINOHA, HETASTA 

(20) 

(20) 

(19) 

1

(20) 

(20) 

(20) 

 (5X) 

(20) 

(20) 

(20) 

(20) 

(20) 

(50) 

21 (K8%) 

(50) 

1 (8X) 

(50) 

27 (5»%) 

(1) 

1 (100%) 

(50) 

50 (100X) 

<U2) 

1 (2X) 

<«6) 

13 (28*) 

1 (2X) 

(47) 

1 (2X) 

(«6) 

14 (30X) 

(48) 

47 (98X) 

(46) 

OHINAFI STSTEH 

*KIDNri 

SQUAMOUS CELL CARCINOHA,

•URINAR Y BLADDER 

SQUAMOUS CELL CARCINOHA,

 HETASTA 

 METASTA 

(20) 

(18) 

(20) 

(19) 

(50) 

1 (2*) 

(47) 

2 («X) 

(46) 

1 (2X) 

(46) 

2 (4X) 

ENDOCRINE SYSTEM 

tADRTNA L 

SQUAHOUS CELL CARCINOHA. HETASTA 

*THYBOID 

F<?I.LICOLA.R-C?LL C^^^NPHA 

(19) 

(20) 

!_JSSU . 

(19) 

(20) 

* NUMBER OP ANIHALS KITH TISSUE EXAHINED MICROSCOPICALLY 

* NOHB?R OP ANIHALS NECROPSIED 

(U7) 

12 (26X) 

(»9) 

(42) 

9 (21X) 

(45) 

B-8 
TABLE B2 (CONTINUED) 

CONTROL (OHTR) CONTROL (TEH) LOW DOSE HIGH DOSE 

02-F061 02-F051 02-F051 02-F055 

REPRODUCTIVE SYSTEM 

*MABMAR Y GLAND (20) (20) (50) («7) 

SQOAHOUS CELL CARCIHOHA, HETASTA "» (8X) 

ADENOCARCINOBA, NOS 2 (IX) 

*OTERUS (20) (20) (45) (»5) 

SQUAMOOS CELL CARCINOMA, METASTA 1 (2%) 2 (»%) 

BNDOMETRIAL STROBAL POLYP 1 (5X) 

tOVARY/OVIDOCT (20) (20) (45) (15) 

SQOAMODS CELL CARCINOMA, BETASTA 3 (7X) 5 (11X) 

tOTAR Y (20) (20) (U3) (<tl() 

CARCIHOBA.NOS 1 (5X) 

SQOAMODS CELL CASCINOBA, BETASTA 10 (23X) 5 (11X) 

NERVOU S SYSTEB 

NON B 

SPECIAL SENSE ORGANS 

NONE 

BDSCULOSKELETAL SYSTEH 

*BUSCLE RIP/THIGH (20) (20) (50) (H7) 

SQDAMODS CELL CARCINOBA, BETASTA 1 (2X) 

BODY CAVITIES 

NONF 

ALL OTHFR SYSTEBS 

NONF . _ _ . . . , _ J . _ . . _ .___ i r _ J , _ . .___ ,. 

* NUMBFR OF ANIBALS KITH TISSDE EXABINED MICROSCOPICALLY 

* SOMBER OF ANIBALS NECROPSIED 

B-9 
ANIHHL DISPOSITION SUMMARY
 

ANIMALS INITIALLY IN STUDY


"JATUBAL DEATH3


MORIBUND SACRIFICE
 SCHEDULED SACRIFICE
 

ACCIDENTALLY KILLED
 TERMINAL SACRIFICE


ANIMAL HISSING
 

9 INCLUDTS AUTOLYZED ANIMALS
 

TUMOR SUMMARY
 

TOTAL ANIMALS HITH PRIMARY TUMORS*


TOTAL PRIMARY TOMORS


TOTAL ANIMALS »ITH BENIGN TUMORS


TOTAL BENIGN TDMORS


CONTROL (ONTR)


02-F061


 20


 1


 19


 6


 8


 1
 

1
 

TOTAL ANIMALS HITH MALIGNANT TOMORS 6


TOTAL MALIGNANT TOMORS 7


TOTAL ANIMALS KITH SECONDARY TOMORS* 1


TOTAL SECONDARY TBMOES 2


TOTAL ANIMALS WITH TOMORS ONCERTAINBFNIGN OR MALIGNANT
 

TOTAL UNCERTAIN TOMOHS
 

TOTAL ANIMALS KITH TOMORS ONCERTAINPRIMARY OR METASTATIC
 

TOTAL UNCERTAIN TOMORS
 

TABLE B2 (CONCLUDED) 

 CONTROL (VEH)


 02-F051


 20


 2


 18


 LOW DOSE HIGH DOSE
 

 02-F054 02-F055
 

 50 50
 

 11 28
 

2
 

9 19
 

1
 

 50 47
 

 51 53
 

2
 

2
 

 50 47
 

 51 51
 

 »1 30
 

 141 89
 

* PRIMARY TOMORS: ALL TOMORS EXCEPT SECONDARY TDBOBS
 

* SECONDARY TUHOBS: METASTATIC TOMORS OB TUMORS INVASIVE INTO AN ADJACENT ORGAN
 

B-10
 
APPENDIX C
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN RATS TREATED WITH DBCP
 

TABLE Cl

 

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

CONTROL (UNTR) CONTROL (VEH) 

01-031M 01-051M 

ANIHALS INITIALLY IN STUDY ANIMALS NECROPSIBD ANIHALS EXAMINED HISTOPATHOLOGICALLY ** 20

20

20

 20 

 20 

 20 

INTEGUMENTARY SYSTEM
 

*SKIN (20) (20)
 INFLAMMATION, NOS 1 <5X)
 

RESPIRATORY SYSTEM
 

*LONG (20) (20)
 EDEMA, NOS
 INFLAMMATION, NOS
 

INFLAMMATION, FOCAL
 

ABSCESS, NOS
 

PNEUMONIA, CHRONIC HORINE 16 (SOX) 20 (100%)
 

CALCIUM DEPOSIT 1 (5*)
 

HEMATOPOIETIC SYSTEM
 

ISPLEEN (20) (20)
 THROMBOSIS, NOS
 

HEMORRHAGIC CYST
 

INFLAMMATION, NOS
 NECROSIS, NOS
 

NECROSIS, FOCAL
 HYPOPLASIA, NOS
 

ATROPHY, NOS
 

ANGIECTASIS
 

LEUKEHOID REACTION
 HE8ATOPOIESIS 1 (5«) 1 <5X)
 

ICERVICAL LYMPH NODE (19) (20)
 INFLAMMATION, NOS 1 <5X)
 

*THYMUS (13) (5)
 

£m*_sa§
 * NUMBER OF ANIMAL S WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBER OF ANIHALS NECROPSIED 

"EXCLUDES PARTIALLY AUTOLYEED ANIMALS 

LOB DOSE
 

01-052H
 

50
 

50
 

50
 

(50)
 

(50)
 

1 (2X)
 

2 («*)
 

1 <2X)
 

35 (70X)
 

(50)
 

1 (2X)
 

1 (2X)
 

2 (<•*)
 

1 <2X)
 

1 (2X)
 

1 (2X)
 t (8X)
 

(««)
 

(8)
 

1 11ISL
 

HIGH DOSE
 

01-053M
 

50
 

50
 

50
 

(50)
 

(50)
 

3 <6X)
 

5 (10X)
 

1 (2X)
 

21 (12X)
 

(19)
 

1 (2X)
 

1 (2X)
 

1 (2X)
 

1 (2*)
 

(K8)
 

(2)
 

C-3 
TABLE CJ (CONTINUED) 

CONTROL (ONTR)

01-031H

 CONTROL (VEH)

 01-051M

 LOH DOSE

 01-052H

 HIGH DOSE 

 01-053B 

CIRCULATOR Y SYSTEH 

tHEAH T

MINERALIZATIO N

CALCIUM DEPOSIT

 (20)

 1 (5X) 

 (20) (50)

 1 <2X )

 (50) 

1 (2%) 

#MYOCARDIU M

INFLAMMATION ,

DEGENERATION,

 NOS

 NOS

 (20)

 2 (10X)

 1 (5X) 

 (20)

 2 (10*)

 (50)

 2 («X)

 (50) 

3 (6X) 

IENDOCARDIO M

INFLAMMATION , NOS

INFLAMMATIO N PROLIFERATIVE

FIBROSIS

HYPEHPLASIA , NOS

 (20)

 1 (5%) 

 (20) (50)

 1 (2%) 

1 (2X) 

 (50) 

1 (2X) 

*AORTA

MEDIA L CALCIFICATION

CALCIFICATION, NOS

 (20)

 2 (10%) 

 (20)

 1 (5X)

 (50) (50) 

1 (2X) 

*MESENTEBIC ARTER Y

MEDIA L CALCIFICATION

 (20)

 1 (5X) 

 (20) (50) (50) 

DIGESTIVE SYSTEM 

*LIVER

CONGESTION, NOS

INFLAMMATION , NOS

DEGENERATION , NOS

DEGENERATION, HYDROPIC

NECROSIS, NOS

NECROSIS, FOCAL

METAMORPHOSI S FATTY

HYPERTROPHY , NOS

HYPERPLASIA , NOS

HYPERPLASIA , FOCAL

ANGIECTASIS

HEMATOPOIESIS

 (20)

 1 (5X)

 2 (10*)

 3 (15X) 

 (20)

 1 (5X)

 2 (10*)

 (50)

 1 (2%) 

2 (4X) 

 10 (20X)

 6 (12X)

 3 (6X) 

1 (8X)

 1 (2X) 

1 (2X) 

 (50) 

1 (2%) 

1 (2X) 

2 (UX) 

1 (2%) 

2 (IX) 

1 (2X) 

#LIVER/CENTHILOBBLA R

DEGENERATION, NOS

NECROSIS, NOS

 (20) (20) (50)

 8 (16X) 

1 (2X)

 (50) 

1 (2X) 

*BILE DUCT

INFLAMMATION , NOS

INFLAMMATION , CHRONIC

HI£SBELASiit,Nos

 (20)

 !t.J2CXL

 (20)

 i_i5SL

 (50)

 1 (2X) 

 2.i<»SL

 (50) 

1 (2X) 

2_i<m__ 

* NUMBER OF ANIMALS HITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBER OF ANIMALS NECROPSIED 

C-4
 
TABLE Cl (CONTINUED) 

CONTROL (UNTH) CONTROL (VEH)
 

01-031M 01-051M
 

* PANCREAS (20) (20)
 PERIARTBRITIS 4 (20X) 1 (5%)
 ATROPHY, FOCAL 1 <5X)
 

tPANCREATIC ACINUS (20) (20)
 ATROPHY, NOS
 

#STOHACH (20) (19)
 MINERALIZATION
 

ULCER, FOCAL 1 <5X)
 CALCIUM DEPOSIT 2 (10X)
 CALCIFICATION, SOS 1 <5X)
 HYPERKEHATOSIS
 

ACANTHOSIS
 

ILARGE INTESTINE (19) (20)
 PARASITISM 1 (5X)
 

fCOLON (19) (20)
 NEMATODIASIS 1 (5X)
 PARASITISM
 

URINARY SYSTEM 

tKIDNEY 

MINERALIZATION 

(20) (20) 

HYDRONEPHROSIS 

CONGESTION, NOS 

HEMORRHAGE 

1 (5%) 

PYELONEPHRITIS, NOS 

INFLAMMATION, CHRONIC 

NEPHROPATHY, TOXIC NEPHROSIS, CHOLEHIC 

CALCIUM DEPOSIT 

CALCIFICATION, NOS 

1

15

1

 (5X) 

 (75%) 

 (5X) 

15 (75X) 

tURINARY BLADDER 

INFLAMMATION, NOS 

INFLAMMATION, CHRONIC 

(19) 

1 (5X) 

(19) 

ENDOCRINE SYSTEM
 

*PITOITAHY (20) (20)
 CYST, NOS 1 (5X)
 

4.HSIKI&5IS __ l.ISJi- „
 

I NUMBER OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBER OF ANIMALS NECROPSIED 

LOW DOSE
 

01-052M
 

(50)
 

1 (2X)
 

(50)
 

2 <4X)
 

(50)
 

2 (IX)
 

2 (IX)
 

2 (KX)
 

2 (4X)
 

(U9)
 

(K9)
 

1 (2X)
 

(50)
 

5 (10X)
 

3 (6X)
 

1 (2X)
 

1 (2X)
 

26 (52X)
 50 (100X)
 

1 (2X)
 

1 (2%)
 

(«9)
 

1 (2X)
 

(49)
 

1 (2X)
 

HIGH DOSE
 

01-053M
 

(50)
 

(50)
 

(50)
 

M (8%)
 

3 (6X)
 

3 (6X)
 

(49)
 

(49)
 

(50)
 

1 (2X)
 

17 (34%)
 

49 (98S)
 

(46)
 

1 (2X)
 1 (2X)
 

(48)
 

C-5 
TABLE Cl (CONTINUED) 

•ADRENAL
 

DEGENERATION, NOS
 

CYTOLOGIC DEGENERATION
 

ATROPHY. NOS
 HYPERTROPHY, NOS
 

•ADRENAL CORTEX
 

DEGENERATION, NOS
 

HYPERTROPHY, NOS
 

HYPERTROPHY, FOCAL
 

ANGIECTASIS
 

*THY80ID
 

ULTIMOBRANCHIAL CYST
 CYST, NOS
 

HYPERPLASIA, C-CELL
 

HYPERPLASIA, POLLICOLAR-CELL
 

IPARATHYROID
 HYPERPLASIA, NOS
 

REPRODUCTIVE SYSTEM
 

•PROSTATE
 

INFLAMMATION, NOS
 

INFLAMMATION, SOPPURATIYE
 

ATROPHY, NOS
 

*SEMINAL VESICLE
 

INFLAMMATION, NOS
 

ATROPHY, NOS
 

tTESTIS
 

GRANULONA, SPERMATIC
 ATROPHY, NOS
 

*EPIDIDYBIS
 

NECROSIS, FOCAL
 NECROSIS, FAT
 ATROPHY, NOS
 

NERVOUS SYSTEM
 

•BRAIH/MENINGES
 

I»EtiafliIIS£x-fla5
 

CONTROL (ONTR)
 

01-031H
 

(20)
 

(20)
 

1 (5*)
 

(19)
 

2 (11*)
 

1 (5*)
 

1 (5*)
 

(3)
 

2 (67%)
 

(20)
 

5 (25*)
 

(20)
 

1 (5*)
 

(20)
 

1 (5»)
 11 <55*>
 

(20)
 

1 (5«)
 

3 (15*)
 

(20)
 

CONTROL (TEH)
 

01-0511
 

(20)
 

1 (5*)
 

1 (5«)
 

(20)
 

2 (10»)
 

1 (5«)
 

1 (5*)
 

(20)
 

1 (5*)
 

2 (10*)
 

(D


1 (100*)
 

(16)
 

1 (6*>
 

(20)
 

(19)
 

« (21*)
 

(20)
 

1 (5«)
 

(20)
 

LOW DOS!
 

01-052H
 

(19)
 

12	 (2«*>
 

« (BS)
 

(«9)
 

19 (39*)
 

1 (2«)
 

(«8)
 

1 (2*)
 

1 (2«)
 

(39)
 

1 (3*)
 

1 (3«)
 

1 O*)
 

(50)
 

1 (2X)
 

(50)
 

38 (76*)
 

(50)
 

(50)
 

BIGR DOSE
 

01-0538
 

(50)
 

9 (18*)
 

1 (2*)
 

1 (2*>
 

(50)
 

35 (70*)
 

7 (1««)
 

(»9)
 

3 (6*)
 

1 (2*)
 

2 («S)
 

(12)
 

1 (8*)
 

1 (8*)
 

(50)
 

2 («*)
 

<U9)
 

«7 (96X)
 

(50)
 

1 (2*)
 

(50)
 

_l.i2JL
 

I NUHBE H OF ANIHAL S WITH TISSDE ElAMIMED HICROSCOPICALLT * NUMBE R OF ANIBALS NECROPSIBO 

C-6 
TABLE Cl (CONTINUED)
 

CONPROL(UNTR) CONTROL (VEH) LOW DOSE
 

01-031M 01-051M 01-052M
 

IB8AIN (20) (20) (50)
 

HYDROCEPHALUS, INTERNAL 1 (2X)
 INFLAMMATION, NOS
 

SPECIAL SENSE ORGANS
 

*EYE/LACRIMAL GLAND (20) (20) (50)
 INFLAMMATION, NOS 1 (5X)
 

*HARDEHIAN GLAND (20) (20) (50)
 INFLAMMATION, NOS 1 (5X>
 

MUSCULOSKELETAL SYSTEM
 

*SKELETAL MUSCLE (20) (20) (50)
 MINERALIZATION 1 (2*)
 DEGENERATION, NOS 1 (2X)
 

BODY CAVITIES
 

*ABDOMINAL CAVITY (20) (20) (50)
 THEOMBOSIS, NOS 1 (2%)
 

*PERITONEUM (20) (20) (50)
 INFLAMMATION, NOS 1 (5X)
 

*PLEDRA (20) (20) (50)
 INFLAMMATION, NOS
 

•PERICARDIUM	 (20) (20) (50)
 INFLAMMATION, NOS 2 (10X)
 INFLAMMATION, SUPPURATIVE 1 <2X)
 

*EPICARDIOM (20) (20) (50)
 INFLABMATION, NOS 3 (15*) 1 (2%)
 

*MESENTERY (20) (20) (50)
 PEHIARTERITIS 1 (20X)
 

ALL OTHER SYSTEMS
 

__8QSS
 

t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 * NUMBER OF ANIMALS NECBOPSIED
 

HIGH DOSE
 01-053M
 

(50)
 

1 (2X)
 

(50)
 

(50)
 

(50)
 

(50)
 

(50)
 

(50)
 

1 (2«)
 

(50)
 

2 <UX)
 

(50)
 

(50)
 

C-7
 
TABLEC1 (CONCLUDED) 

CONTROL (UNTB)

01-031H

 CONTBOL(VEH)

 01-051B

 LOU DOSE

 01-052M

 HIGH DOSE 

 01-053H 

SPECIAL MORPHOLOGY SUMMARY 

NONE 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICBOSCOPICALLY * NUMBER OF ANIMALS NECHOPSIED 

C-8 
TABLE C2

 

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

TM"—*~— iTM_-i « ..-mil- rr TM—_..«_ -—'

J

 L«TM— 

CONTROL (UNTR) CONTROL (VEH) LOU DOSE HIGH DOSE 

01-031F 01-051F 01-054F 01-055P 

ANIMAL S INITIALLY IN STODY 20 20 50 50 

ANIMALS NECROPSIED 20 20 50 50 

ANIMALS EXAMINED HISTOPATHOLOGICALLY** 20 20 50 50 

INTEGUMENTARY SYSTEM 

*SUBCUT TISSOE (20) (20) (50) (50) 

ABSCESS, SOS 1 (2X) 

RESPIRATORY SYSTEM 

tLUNG (20) (20) (12) <«»)

INFLAMMATION, NOS U (9X) 

INFLAMMATION, FOCAL 1 (2») 1 (2X) 

ABSCESS, NOS 3 (15X) 1 (2X) 

PNEUMONIA, CHRONIC MDRINE 18 (90S) 16 (BOX) 34 (81X) 30 (68X) 

FIBROSIS 1 (2X) 

METAPLASIA, SQOAHOOS 1 (2%) 

HEMATOPOIETIC SYSTEM 

tSPLEEN (20) (20) C»2) <*«) 

THROMBUS, ORGANIZED '1 <2«) 

HEMATOPOIESIS 15 (36%) 13 (SOX) 

#MES ENTERIC L. NODE (20) (18) (23) (43) 

ATROPHY, NOS 1 (2X) 

CIRCULATORY SYSTEM 

tHEAST (20) (20) (U2) •e«t) MINERALIZATION 1 (2X) 

tMYOCARDIUM (20) (20) (i»2) (t|«) 

INFLAMMATION, NOS 1 (2X) 

FIBROSIS 1 (5%) 

f TNDOCARDIUM (20) (20) <U2 ) (»*> ISFLAMflATION. Nps ^ ,,r ^ ^_ - 1_I2SL__ 

t NUMBE R OF ANIMAL S KITH TISSUE EXABIDED MICROSCOPICALLY 

* NUHBFP OF ANIMAL S NECROPSIED 

"EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

C-9 
TABLE C2 (CONTINUED) 

CONTROL (UNTR) CONTROL (»EH) 

01-031F 01-051F 

*AORT A (20) (20) "IFDIAL CALCIFICATION 1 (5X)

 

DIG"STIVF SYSTFM 

*LIVR (20) (20)

 INFLAMMATION , NOS

 

INFLAMMATION , NECROTIZING 

NECROSIS, NOS 

tl^TAMORPHOSIS FATTY 1 <5X)

 HYPERTROPHY , NOS

 

HYPERPLASIA , NOS 1 <5X)

 ANGIECTASIS 1 (5X)

 HEMATOPOIBSIS

 

*LIVFR/CFNTRILOBULAB (20) (20) FIBROSIS

 

NFCROSIS, NOS 1 <5X)

 M^TAMOFPHOSIS FATTY

 

fLIVFR/HFPATOCYTES (20) (20) INFLAMMATION , NOS

 

*BTLF DUCT (20) (20)

 HYPERPIASIA, NOS

 

*PANCR*AS (20) (19) FIBROSIS

 

NECROSIS, FAT 

ATROPHY, NOS 

#STOMACH (20) (20)

 INFLAMMATION , NOS 1 <5X)

 ULCER, NOS

 

ULCER, FOCAL 1 (5X)

 HYPERKERATOSI S 1 (5X)

 ACANTHOSI S 1 (5X)

 

•LARGE INTESTINE (19) (19) 

ElIASISStt __ 2-I11SL 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECBOPSIED
 

c-io 

LOU DOSE 

01-054F 

(50) 

(U2) 

2 (5X) 

1 (2X) 

8 (19X) 

1 (2X) 

1 (2X) 

(t2) 

4 (10X) 

(12) 

(50) 

1 (2X) 

(it 2) 

1 (2X) 

1 (2X) 

(50) 

1 <2X) 

1 (2X) t (8X) 

5 (10X) 

(12) 

_-l_12SL 

HIGH DOSE 

01-055F 

(50) 

(11)

3 (7X) 

1 (2X) 

II (9X) 

2 (5X) 

2 (5X) 

1 (2X) 

(11)

1 (2X) 

2 (5X) 

1 (2X) 

(11) 

1 (2X) 

(50) 

5 (10X) 

(11) 

1 <2X) 

(19) 

3 (6X) 

10 (20X) 

10 (20X) 

(11) __«»«__ «» 
TABLE C2 (CONTINUED)
 

CONTROL (ONI8) CONTROL (VEH ) 

01-031F 01-051F 

URINAR Y SYSTEM 

tKIDNEY (20) (20) 

MINERALIZATION 

INFLAMMATION , FOCAL 

INFLAMMATION, CHRONIC 9 (45%) 9 (151) 

NFPHROPATHY, TOXIC 

CALCIUM DEPOSIT 1 (5%) 

ENDOCRINE SYSTEM 

tPITOITARY (19) (20)

 

CYST, BOS 1 (5%)

 

HYPEHPLASIA, CHROHOPHOBE-CELL 1 (5*)

 

*ADRENAl (20) (20)

 

DEGENERATION, NOS 1 (5%) 

ATROPHY, NOS 

HYPERTROPHY, NOS 

ANGIECTASIS 1 (5%) 

HEMATOPOI2SIS 

iADR?NAL CORTEX (20) (20)

 

DEGENERATION, NOS 1 (5%)

 

HYPERTFOPHY, NOS 1 (5%) 

ANGircTASIS 3 (15*)

 

#THYROID (20) (20)

 

HYPERPLASIA, C-CELL 4 (20%) 1 (5%)

 

IPARATHYROID d)

HYPERPLASIA, NOS 1 (100%)

 

REPRODUCTIVE SISTEH 

tUTERUS (20) (20) 

HYDROMETRA 4 (20%) 2 (10%) 

fUTERUS/ENDOMFTRIOM (20) (20)

 

INFLAMMATION, NOS 1 (5%) 1 (5%)

 

HYPERPLASIA, CYSTIC 1 (5%)

 

tOVARY (20) (20)

 

r.OillCIiifi.B-SIs.lt.BOS. 

» NUMBER OF ANIMALS HITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

C-ll
 

LOU DOSE 

01-051P 

(«2) 

1 (2%) 

9 (21%) 

12 (10PX) 

(HO) 

(10) 

1 (3%) 

1 (3%) 

3 (8%) 

(«0) 

33 (83%) 

(39) 

(39) 

3 (8%) 

(39) 

(39) 

HIGH DOSE 

01-055F 

(tt) 

1 (2%) 

9 (20%) 

44 (100%) 

(«1) 

(44) 

9 (20%) 

1 (2%) 

(44) 

31 (70%) 

(It «) 

1 (2S) 

(43) 

3 (7%) 

(43) 

(43) 

1_i2Sl 
TABLE C2 (CONCLUDED) 

CONTROL (ONTR) CONTROL (VEH) LOU DOSE HIGH DOSE 

01-031F 01-051P 01-054F 01-055P 

NF.RVOUS SYSTEM 

NOSF 

SPECIAL SENSE ORGANS 

*FY1; (20) (20) (50) (50) CATARACT 1 (5X)

 

MDSCULOSKELETA L SYSTEM 

NONF 

BODY CAVITIES 

*PL2URA (20) (20) (50) (50) 

INFLAMMATION , NOS 1 (5X) 

•PERICARDIUM (20) (20) (50) (50) 

INFLAMMATION , NOS 3 (15*) 3 (6%) 5 (10%) 

ALL OTHER SYSTFMS 

NON^ 

SPECIAL BOKPHOLOGY SUMMAR Y 

NO LESION REPORTED 4 3 

* NUIB^R OF ANIMAL S WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBER OF ANIMAL S HECFOPSIED 

C-12 
APPENDIX D
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC
 

LESIONS IN MICE TREATED WITH DBCP
 

TABLE Dl

 

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

~" L" TM•"•""•—~TMTM'- — u__««____»-•-•- HT •••"•"——
 

CONTROL (ONTR) CONTROL (VEH)
 

ANIMALS INITIALLY IH STDDI
 

AHIflALS NECROPSIED
 

ANIMALS EXABINED HlSTOPATHOLOGICALLY **
 

INTEGUMENTARY SYSTEM
 

*SKIN
 

INFLAHBATION, NOS
 

CALCIUM DEPOSIT
 

*SUBCOT TISSUE
 

ABSCESS, NOS
 

RESPIRATORY SYSTEM
 

*LUN6
 

EDEMA, NOS
 

INFLAMMATION, NOS
 

INFLAMMATION, SUPPURATITE
 

PNEUMONIA, CHRONIC BDRINE
 

HEBATOPOIETIC SYSTEM
 

*BONE MARROW
 

NECROSIS, NOS
 

#SPLEE»
 

INFLAMMATION, NOS
 

AMYLOIDOSIS
 

ATROPHY, NOS
 

HEMATOPOIESIS
 

CIRCULATORY SYSTEM
 

* HEART
 

ABSCESS, NOS
 

_Qil£IU£ATl(JiI_]i2S__
 

02-H061 02-H051
 

20 20
 

19 20
 

19 20
 

(19) (20)
 

(19)	 (20)
 

2 (11%)
 

(18)	 (20)
 

1 (6%)
 

1 (6%)
 

1 (6%)
 

1 (6%)
 

(17)	 (2)
 

1 (6%)
 

(19) (20)
 

12 (63%)
 

2 (11%)
 

(19)	 (20)
 

1 (5%)
 

3.116SL
 

t NUMBER OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBER OF ANIMALS NECROPSIED 

**EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

D-3 

LOR DOSE HIGH DOSE
 

02-H052 02-M053
 

50 50
 «7 50
 »5 50
 

(47) (50)
 

1 (2%)
 

1 (2%)
 

(17)	 (50)
 

1 (2X)
 

(46) (48)
 

3 (7%) 3 (6*)
 

(t5) (48)
 

(45)	 (47)
 

1 (2%)
 « (9%)
 

15 (33%) 4 (9%)
 

(46) (48)
 

.—•.»»«._«»_«
 
TABLE Dl (CONTINUED)
 

CONTROL (UNTR) CONTROL (VEH )

02-M061 02-M051 

* MYOCARDIU M (19) (20)

 

INFLAMMATION , SUPPOEATIVE 1 (5%)

 DEGENERATION , (JOS 1 (5*)

 

DIGTSTIV" SYSTEM 

*LIVP (19) (20)

 

THROMBUS , ORGANIZE D 1 (5%)

 

INFLAMMATION , NOS 

NECROSIS, FOCAL 1 (5X)

 

AMYLOIDOSIS 12 (63%)

 

METAHORPHOSIS FATTY 

CALCIFICATION, NOS 1 <5X) 

HYPF.RPLASIA, NODULAR 

LT-UKEMOID REACTION 

HEHATOPOIESIS 

*LIVER/CENTRILOBULAR (19) (20) 

DEGENERATION, NOS 3 (16%)

 

NECROSIS, NOS 1 <5X)

 

*BILF DUCT (19) (20)

 

DILATATION, NOS 

tPANCREA S (19) (19) 

AMYLOIDOSIS 6 (32X)

 

*FSOPHAGUS (18) 

INFLAMMATION, NOS 

#STOMACH (20) (20) 

INFLAMMATION , FOCAL 1 (5%)

 

CALCIFICATION, NOS 3 (15X)

 

*SMALL INTESTINE (18) (19) 

NEMATODIASIS 1 <6X)

 

tLARGE INTESTINE (19) (19)

 

NEMATODIASIS 1 <5X)

 

*RECTUM (19) (20) 

PROLAPSE 

* NUMBER OF ANIMALS IIITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

D-4
 

 LOW DOSE 

02-M052 

(46) 

1 (2*) 

(<t6) 

3 <7X) 

1 (2X) 

2 (<»X) 

1 (2X) 

1 <2X) 

(46) 

(47) 

(45) 

(3) 

(46) 

(44) 

(46) 

(47) 

HIGH DOSE 

02-B053 

(48) 

(48) 

5 (10X) 

1 (2X) 

2 (4X) 

(48) 

(50) 

1 (2*) 

(48) 

(7) 

2 (29X) 

(49) 

(47) 

(47) 

(50) 

1_12S1_ 
TABLE Dl (CONTINUED) 

CONTROL (ONTR)

02-M061

 CONTROL (VEH)

 02-M051

 LOB DOSE

 02-M052

 HIGH DOSE 

 02-H053 

URINAR Y SYSTEH 

tKIDNEY 

PYELONEPHRITIS SUPPURATIVE 

INFLAMMATION, CHROMIC 

NEPHROPATHY, TOXIC 

AMYLOIDOSIS 

CALCIFICATION, HOS 

LEOKEMOID REACTION 

(19)

1 (5%) 

15 (79X)

6 (32X)

1 <5X) 

 (20)

 2 (10X) 

 (16)

11 (24*)

 1 (2X) 

1 (2X) 

 (18) 

 45 (94X) 

IDRINARY BLADDER 

INFLAMMATION, FOCAL 

(17)

1 (6X) 

 (19) (46) (47) 

ENDOCRINE SYSTEM 

* ADRENA L 

AHYLOIDOSIS 

ANGIECTASIS 

(19)

2 (11X) 

1 <5X) 

 (20) (42) («9) 

REPSODDCTIVE SYSTEM 

*PBEPOTIAL GLAN D 

ABSCESS, NOS 

(19) (20)

1 (5X) 

 (47) (50) 

ITESTIS 

GRANOLOMA, SPERMATIC 

ATROPHY, NOS 

(19)

1 (5X) 

 (20) (45) (48) 

1 <2X) 

NE8V00S SYSTEM 

tBRAIN 

CALCIFICATION, NOS 

(18)

1 <6X) 

 (20) (46) (48) 

SPECIAL SENSE ORGANS 

NONE 

MOSCDLOSKELETAL SYSTEH 

t NUMBER OF ANIMALS BITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBER OF ANIMALS NECROPSIED 

*MOSCLE HIP/THIGH 

£Al£i2fl_2Sf2§Il — 

(19) (20) (47)

1 12SL 

 (50) 

D-5
 
TABLE Dl (CONCLUDED) 

CONTROL (ONTR)

02-B061

 CONTROL (VEH)

 02-M051

 LOR DOSE

 02-H052

 HIGH DOSE 

 02-M053 

BODY CAVITIES 

*PERITON50S

INFLAMMATION , NOS

 (19) (20) (U7) (50) 

1 (2X) 

ALL OTHER SYSTEMS 

NOME 

SPECIAL MORPHOLOGY SOMfiARY 

HO LESION REPORTED 1« AUTO/NECROPSY/HISTO PERF 1 

AOTO/HECHOPSY/MO HISTO

AOTOLYSIS/HO NFCROPSY 1

* NOBBER OF ANIMALS BITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBER OF ANIMALS NECROPSIED 

2 

2 

3 

D-6
 
TABLE D2

 

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

c:ONTROL(ONTR) CONTROL(TEH ) LOH DOSE HIGH DOSE 

02-F061 02-F051 02-F054 02-F055 

ANIMAL S INITIALLY IN STODY 20 20 50 50 

ANIMAL S MISSING 1 

ANIMAL S NECROPSIED 20 20 50 47 

ANIMAL S EXAMINE D HISTOPATHOLOGICALLY** 20 20 50 47 

INTEGUMENTARY SYSTEM 

*SKIN (20) (20) (50) (47) 

INFLAMMATION , CHRONIC 1 (5%) 

RESPIRATORY SYSTEM 

fLUN G (20) (20) (50) (»6) 

PNFUMONIA, CHRONIC HUBINE 11 (55*) 2 (4(1 

HYPERPLASIA, FOCAL 1 (2S) 

HYPERPLASIA, LYMPHOID 1 (5*) 

HF.MATOPOIETIC SYSTEM 

#SPLEEN (20) (20) (50) («6) 

HEMATOPOIESIS 3 <6X) 3 (7») 

tMESFNTERIC L. NODE (20) (19) (46) (39) 

ANGIECTASIS 1 <5X) 

#THYMOS (20) (20) (50) (46) 

HYPERPLASIA, LYMPHOID 1 (5*) 

CIRCULATORY SYSTEM 

#BYOCARDIUM (20) (20) (50) (16) 

INFLAMMATION , NOS 1 (5X( 

DIGFSTIVE SYSTEM 

iLIVR (20) (20) (50) (46) 

IflFJAJU&U&JU-ISS _. 2_1SSL_ —J-iZSi­

* NUMBE R OF ANIMAL S KITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBER OF ANIMAL S NECROPSIED 

"EXCLUDES PARTIALLY AUTOLYZED ANIMALS 

D-7 
TABLE D2 (CONTINUED)
 

CONTROL (UNTR) CONTROL (VEH) 

02-FC61 02-F051 

#LIVFP/CFNTRILOBULA R (20) (20)

 

DEGENERATION, UOS 2 (10X)

 

fEcpoSIS, NOS 1 (5X)

 

*BILF DOCT (20) (20)

 

INFLAMMATION , NOS 1 (5»)

 

INELAMNATION , FOCAL 1 (5X)

 

HKINAP Y SYSTEM 

tKIWY (20) (20) 

PYELONEPHRITIS , NOS 

INFLAMMATION , CHRONIC 13 (65X)

 

VEPHROPATHY , TOXIC 

OUPINAR Y BLADDER (18) (19)

 

INFLAMMATION , FOCAL 1 (22%) 

rNTOCRIN E SYSTEM 

tADRENA L (19) (19) 

INFLAMMATION , NOS 

PEPRODUCTIV E SYSTEM 

#OTERa S (20) (20) 

HYDBOMETR A 4 (20%} 

*aTEPUS/ENDOMETRIUM (20) (20) 

HYPERPLASIA , CYSTIC 17 (85X) 3 (15*) 

*OVAR Y (20) (20) 

CYST, NOS 3 (15») 

PAROVARIA N CYST 4 (20%) 

INFLAMMATION , NOS 1 (5X) 

ANGITCTASIS 1 <5X) 

NFEVOD S SYSTEM 

*BEAIN/MENINGES (20) (20) 

IBZlifllJillONx.rogai, .. _ i_.im_ _ 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY
 

* NUMBER OF ANIMALS NECROPSIED
 

LOB DOSE 

02-F054 

(50) 

(50) 

(50) 

3 <6X) 

14 (28X) 

(47) 

(47) 

1 <2X) 

(45) 

3 (7X) 

(45) 

2 (4X) 

(43) 

(50) 

HIGH DOSE 

02-F055 

(46) 

(47) 

(46) 

43 (93X) 

(46) 

(42) 

(45) 

2 (4*) 

(45) 

(44) 

(46) 

D-8
 
TABLE D2 (CONCLUDED) 

CONTROL (ONTE)

02-P061

 CONTROL (TEH)

 02-F051

 LOB DOSE

 02-F05U

 HIGH DOSE 

 02-F055 

•SPINAL COED

CYST, NOS

 (20)

 1 (5*) 

 (20) (50) (47) 

SPECIAL SENSS ORGANS 

NONF 

MUSCULOSKELETAL SYSTEM 

• SKELETAL MUSCLE

INFLAMMATION, FOCAL

 (20)

 1 (5») 

 (20) (50) (47) 

BODY CAVITIFS 

NOKF 

ALL OTHER SYSTEMS 

NONF 

SPECIAL MORPHOLOGY SUMMARY 

HO LESION REPORTED ANIMAL MISSING/NO NECROPSY

AUTOLYSIS/NO NECROPSY

* NUMB'R OF ANIMALS KITH TISSUE EXAMINED * NOHBER OF AMIHALS NBCBOPSIED 

 12 

 MICROSCOPICALLY 

1 

2 

ft US GOVERNMENT PRINTING OFFICE 1978-260-8993205 

D-9 


DHEW Publication No. (NIH) 78-828