1978 

National Cancer Institute 

CARCINOGENESIS 

Technical Report Series 

No. 88
 

BIOASSAY OF 

1H-BENZOTRIAZOLE 

FOR POSSIBLE CARCINOGENICITY 

CAS No. 95-14-7 

NCI-CG-TR-88 

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

Public Health Service 

National Institutes of Health 

BIOASSAY OF
 

IH-BENZOTRIAZOLE
 

FOR POSSIBLE CARCINOGENICITY
 

Carcinogenesis Testing Program
 

Division of Cancer Cause and Prevention
 

National Cancer Institute
 

National Institutes of Health
 

Bethesda, Maryland 20014
 

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

Public Health Service
 

National Institutes of Health
 

DHEW Publication No. (NIH) 78-1338
 

BIOASSAY OF
 

IH-BENZOTRIAZOLE
 

FOR POSSIBLE CARCINOGENICITY
 

Carcinogenesis Testing Program
 

Division of Cancer Cause and Prevention
 

National Cancer Institute
 

National Institutes of Health
 

FOREWORD; This report presents the results of the bioassay of
 

IH-benzotriazole conducted for the Carcinogenesis Testing
 

Program, Division of Cancer Cause and Prevention, National Cancer
 

Institute (NCI), National Institutes of Health, Bethesda,
 

Maryland. This is one of a series of experiments designed to
 

determine whether selected chemicals have the capacity to produce
 

cancer in animals. Negative results, in which the test animals
 

do not have a greater incidence of cancer than control animals,
 

do not necessarily mean that the test chemical is not a
 

carcinogen, inasmuch as the experiments are conducted under a
 

limited set of circumstances. Positive results demonstrate that
 

the test chemical is carcinogenic for animals under the
 

conditions of the test and indicate that exposure to the chemical
 

is a potential risk to man. The actual determination of the risk
 

to man from animal carcinogens requires a wider analysis.
 

CONTRIBUTORS: This bioassay of IH-benzotriazole was conducted by
 

EG&G Mason Research Institute, Worcester, Massachusetts,
 

initially under direct contract to NCI and currently under a
 

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

NCI Carcinogenesis Testing Program.
 

The bioassay was conducted under the supervision of Drs. A.
 

Handler1 and E. Smith2, and Mr. G. Wade3. NCI project officers
 

were Drs. E. Weisburger^, T. Cameron^, and N. P. Page^>^. The
 

program manager was Mr. J. Baker3. Ms. A. Good3 supervised the
 

technicians in charge of animal care, and Ms. E. Zepp3 supervised
 

the preparation of the feed mixtures and collected samples of the
 

diets for analysis. Ms. D. Bouthot3 kept all daily records of
 

the test, and Ms. R. Monson3 prepared a report based on these
 

records. Dr. A. Russfield , pathologist, supervised the
 

performance of the necropsies. Histopathologic evaluations were
 

iii
 
performed by Drs. R. Fleischman3

 and A. S. K. Murthy3, and the
 

diagnoses included in this report represent their interpretation.
 

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

Mason Research Institute, Rockville, Maryland^. The statistical
 

analyses were performed by Dr. J. R. Joiner^, using methods
 

selected for the bioassay program by Dr. J. J. Gart^.
 

Chemicals used in this bioassay were analyzed under the direction
 

of Dr. E. Murrill^, and dosed feed mixtures were analyzed by Dr.
 

M. Hagopian-^. The analytical results were reviewed by Dr. S. S.
 

Olin?. The chemical structure was supplied by NCI.
 

This report was prepared at Tracer Jitco? under the direction of
 

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

L. A. Campbell, Director of the Bioassay Program; Dr. S. S. Olin,
 

Deputy Director for Science; Dr. J. F. Robens, toxicologist; Dr.
 

R. L. Schueler, pathologist; Dr. G. L. Miller, Ms. L. A. Waitz,
 

and Mr. W. D. Reichardt, bioscience writers; and Dr. E. W.
 

Gunberg, technical editor, assisted by Ms. Y. E. Presley and Ms.
 

P. J. Graboske.
 

The following other scientists at NCI were responsible for
 

evaluating the bioassay, interpreting the results, and reporting
 

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

J. Fielding Douglas, Dr. Dawn G. Goodman, Dr. Richard A.
 

Griesemer, Dr. Harry A. Milman, Dr. Thomas W. Orme, Dr. Robert A.
 

Squire^, Dr. Sherman Stinson, Dr. Jerrold M. Ward, and Dr.
 

Carrie E. Whitmire.
 

^•Now with Abcor, Inc., 850 Main Street, Wilmington,
 

Massachusetts.
 

^Now with the University of Massachusetts Medical Center, 55 Lake
 

Avenue, Worcester, Massachusetts.
 

iv
 
^EG&G Mason Research Institute, 57 Union Street, Worcester,
 

Massachuset ts.
 

^Carcinogenesis Testing Program, Division of Cancer Cause and
 

Prevention, National Cancer Institute, National Institutes of
 

Health, Bethesda, Maryland.
 

^Now with the Environmental Protection Agency, 401 M Street,
 

S.W., Washington, D.C.
 

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

Rockville, Maryland.
 

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

Maryland.
 

"Mathematical Statistics and Applied Mathematics Section,
 

Biometry Branch, Field Studies and Statistics, Division of
 

Cancer Cause and Prevention, National Cancer Institute,
 

National Instiutes of Health, Bethesda, Maryland.
 

^Midwest Research Institute, 425 Volker Boulevard, Kansas City,
 

Missouri.
 

*ONOW with the Division of Comparative Medicine, Johns Hopkins
 

University, School of Medicine, Traylor Building, Baltimore,
 

Maryland.
 

v
 

SUMMARY
 

A bioassay of IH-benzotriazole for possible carcinogenicity was
 

conducted by administering the test chemical in feed to Fischer
 

344 rats and B6C3F1 mice.
 

Groups of 50 rats of each sex were administered IH-benzotriazole
 

at one of two time-weighted average doses, either 6,700 or 12,100
 

ppm, for 78 weeks. Except for five control and five high-dose
 

rats of each sex, which were killed at week 78, all animals
 

surviving at that time were observed for 26-27 additional weeks.
 

Controls consisted of groups of 50 untreated rats of each sex and
 

were observed for 105-106 weeks. All rats surviving to weeks
 

104-106 were then killed.
 

Groups of 50 mice of each sex were administered IH-benzotriazole
 

at one of two time-weighted average doses, either 11,700 or
 

23,500 ppm, for 104 weeks, then observed for 2 additional weeks.
 

Controls consisted of groups of 50 untreated mice of each sex and
 

were observed for 109 weeks. All mice surviving to weeks 106-109
 

were then killed.
 

Mean body weights of the dosed male and female rats and mice were
 

lower than those of the corresponding controls throughout most of
 

the bioassay. Survival of animals in dosed and control groups of
 

both rats and mice was at least 60%, and sufficient numbers of
 

animals were at risk for development of late-appearing tumors.
 

In male rats, neoplastic nodules of the liver occurred at a
 

statistically significant incidence (P = 0.024) in the high-dose
 

group when compared with the control group (controls 0/48,
 

low-dose 0/46, high-dose 5/45 [11%]). The incidence of this
 

tumor in control Fischer 344 rats used in similar bioassays of
 

other test chemicals at the same laboratory has varied from 0 to
 

11%, with 2/13 historical-control groups having incidences of
 

10-11%. Since the incidence in the high-dose group is no higher
 

than has been observed in some control groups, these tumors 

cannot be clearly associated with administration of the test 

chemical. 

vii
 
Brain tumors occurred in three dosed male rats, in one dosed
 

female rat, and in none of the controls. The occurrence of this
 

rare tumor in dosed animals of each sex is suggestive of, but not
 

considered as sufficient evidence of, carcinogenicity.
 

In female rats, the incidence of endometrial stromal polyps in
 

the low-dose group was significantly higher (P = 0.010) than that
 

in the corresponding controls (controls 2/48, low-dose 10/45,
 

high-dose 8/50). However, the incidence in the high-dose group
 

was not significant, and when the incidences of endometrial
 

stromal polyps and endometrial stromal sarcomas were combined,
 

they were not significant in either the low- or high-dose groups.
 

Thus, these tumors cannot be associated with administration of
 

the chemical.
 

In male mice, no tumors occurred in dosed groups at incidences
 

that were significantly higher than those in controls.
 

In female mice, alveolar/bronchiolar carcinomas occurred at a
 

statistically significant incidence (P = 0.001) only in the
 

low-dose group when compared with the control group (controls
 

0/49, low-dose 9/49 [18%], high-dose 3/49 [6%]). The incidence
 

in the high-dose group was not significant, and the data did not
 

show a dose-related trend. It should be noted that the incidence
 

of these tumors in control B6C3F1 female mice from other
 

bioassays at this laboratory has varied from 0 to 7%, with a mean
 

of 4%. Therefore, the occurrence of this tumor in the female
 

mice cannot be clearly related to the administration of the test
 

chemical.
 

In female B6C3F1 mice there was an increased incidence of
 

alveolar/bronchiolar carcinomas, suggesting a possible
 

carcinogenic effect of iH-benzotriazole. In Fischer 344 rats
 

there was an increased incidence of brain tumors, suggesting a
 

possible carcinogenic effect. However, there was no convincing
 

evidence that under the conditions of this bioassay
 

IH-benzotriazole was carcinogenic in B6C3F1 mice or Fischer 344
 

rats of either sex.
 

viii
 
TABLE OF CONTENTS
 

Page
 

I. Introduction 1
 

II. Materials and Methods 3
 

A. Chemical 3
 

B. Dietary Preparation. 4
 

C. Animals 4
 

D. Animal Maintenance 5
 

£. Subchronic Studies 8
 

F. Chronic Studies 9
 

G. Clinical and Pathologic Examinations 9
 

H. Data Recording and Statistical Analyses 13
 

III. Results - Rats 19
 

A. Body Weights and Clinical Signs (Rats) 19
 

B. Survival (Rats) 19
 

C. Pathology (Rats) 22
 

D. Satistical Analyses of Results (Rats) 24
 

IV. Results - Mice 27
 

A. Body Weights and Clinical Signs (Mice) 27
 

B. Survival (Mice) 27
 

C. Pathology (Mice) 30
 

D. Statistical Analyses of Results (Mice) 31
 

V. Discussion 35
 

VI. Bibliography 39
 

APPENDIXES
 

Appendix A Summary of the Incidence of Neoplasms in
 

Rats Fed IH-Benzotriazole in the Diet 43
 

Table Al Summary of the Incidence of Neoplasms in
 

Male Rats Fed IH-Benzotriazole in the Diet......... 45
 

Table A2 Summary of the Incidence of Neoplasms in
 

Female Rats Fed IH-Benzotriazole in the Diet 49
 

Ix
 
Appendix B
 

Table Bl
 

Table B2
 

Appendix C
 

Table Cl
 

Table C2
 

Appendix D
 

Table Dl
 

Table D2
 

Appendix E
 

Table El
 

Table E2
 

Appendix F
 

Table Fl
 

Page
 

Summary of the Incidence of Neoplasms in
 

Mice Fed IH-Benzotriazole in the Diet 53
 

Summary of the Incidence of Neoplasms in
 

Male Mice Fed IH-Benzotriazole in the Diet 55
 

Summary of the Incidence of Neoplasms in
 

Female Mice Fed IH-Benzotriazole in the Diet 58
 

Summary of the Incidence of Nonneoplastic
 

Lesions in Rats Fed IH-Benzotriazole in the Diet... 63
 

Summary of the Incidence of Nonneoplastic
 

Lesions in Male Rats Fed IH-Benzotriazole in
 

the Diet 65
 

Summary of the Incidence of Nonneoplastic
 

Lesions in Female Rats Fed IH-Benzotriazole
 

in the Diet , 70
 

f
 

Summary of the Incidence of Nonneoplastic
 

Lesions in Mice Fed IH-Benzotriazole in the Diet... 75
 

Summary of the Incidence of Nonneoplastic
 

Lesions in Male Mice Fed IH-Benzotriazole
 

in the Diet 77
 

Summary of the Incidence of Nonneoplastic
 

Lesions in Female Mice Fed IH-Benzotriazole
 

in the Diet 82
 

Analyses of the Incidence of Primary Tumors
 

in Rats Fed IH-Benzotriazole in the Diet 87
 

Analyses of the Incidence of Primary Tumors
 

in Male Rats Fed IH-Benzotriazole in the Diet 89
 

Analyses of the Incidence of Primary Tumors
 

in Female Rats Fed IH-Benzotriazole in the Diet.... 95
 

Analyses of the Incidence of Primary Tumors
 

in Mice Fed IH-Benzotriazole in the Diet 101
 

Analyses of the Incidence of Primary Tumors
 

in Male Mice Fed IH-Benzotriazole in the Diet 103
 

x
 
Table F2
 

Table 1
 

Table 2
 

Figure 1
 

Figure 2
 

Figure 3
 

Figure 4
 

Page 

Analyses of the Incidence of Primary Tumors 

in Female Mice Fed IH-Benzotriazole in the Diet.... 108 

TABLES
 

Chronic Feeding Studies of


in Rats
 

Chronic Feeding Studies of


in Mice
 

FIGURES
 

Growth Curves for Rats Fed


in the Diet
 

 IH-Benzotriazole 

10 

 IH-Benzotriazole 

11 

 IH-Benzotriazole 

20 

Survival Curves for Rats Fed IH-Benzotriazole 

in the Diet 21 

Growth Curves for Mice Fed IH-Benzotriazole 

in the Diet 28 

Survival Curves for Mice Fed

in the Diet 

 IH-Benzotriazole 

29 

xi
 

I. INTRODUCTION
 

IH-Benzotriazole (CAS 95-14-7; NCI
 

C03521), also commonly called
 

1,2,3-benzotriazole, is an anticorrosive chemical used primarily
 

on copper, but also on iron, 

1H-BENZOTRIAZOLE 

steel, cadmium, chromium, zinc, 

and silver-nickel alloys (Sherwin-


Williams Chemicals, 1976).
 

Like other amine anticorrosives, IH-benzotriazole can form
 

covalent and coordinate-covalent linkages with metals, which
 

prevent attack by corrosive agents (Bregman, 1963;
 

Sherwin-Williams Chemicals, 1976).
 

As an anticorrosive, IH-benzotriazole is used in metal working,
 

in art restoration, and in the construction industry as a tarnish
 

remover and a protective coating (Sherwin-Williams Chemicals,
 

1976). It functions as a corrosion inhibitor in water cooling
 

systems such as automobile radiators and boilers (Union Carbide,
 

1966), and in dry cleaning equipment (Levy et al., 1967). It is
 

included in some formulations of automatic dishwasher detergents
 

to prevent tarnishing of metal pots and silverware, and to
 

inhibit the corrosion of metal machine parts (Donaldson, 1971).
 

IH-Benzotriazole is used in synthetic greases, lubricants, and
 

1
 
hydraulic fluids to prevent the oxidation of these materials
 

which is catalyzed by metal ions. In the electronics industry,
 

it is used to treat packing materials for copper electronic parts
 

(Green, 1969), and to extend the life of polymers that are used
 

as insulators for copper wire (Hansen, 1968).
 

Other than as an anticorrosive, IH-benzotriazole is used in
 

electrolytic processing, where the stripping of metals from
 

copper cathodes is eased by pretreatment of the cathode with
 

IH-benzotriazole (Anaconda American Brass, 1965). In photographic processing, IH-benzotriazole acts as an antifogging agent
 

in silver-halide emulsions (Sahyun, 1971), restraining the
 

developer and preventing the blackening or fogging of the image
 

due to overdevelopment (West, 1973).
 

IH-Benzotriazole was selected for study in the Carcinogenesis
 

Testing Program in part because of its use in dishwashing
 

detergents and the possibility that such use could lead to
 

contamination of water supplies.
 

2 
II. MATERIALS AND METHODS
 

A. Chemical
 

Two lots of IH-benzotriazole were obtained from Aldrich Chemical
 

Company, Milwaukee, Wisconsin, and were stored at 4°C. Lot No.
 

122917 was used during the subchronic studies, and Lot No. 030737
 

during the chronic studies. The identity and purity of both lots
 

were determined by analytical procedures. Both lots gave a
 

single homogeneous peak with high-pressure liquid chromatography,
 

and a trace impurity by thin-layer chromatography. Karl Fischer
 

analysis indicated a water content of less than 0.1%. Nonaqueous
 

titration of the amine function with perchloric acid gave a
 

purity of 100.5 + 1.0% for Lot No. 030737. Elemental analyses of
 

both lots (C, H, N) were consistent with Cg^^, the molecular
 

formula of the chemical. The melting point of Lot No. 030737 was
 

97.5-98.5°C, and that of Lot No. 122917 was 96.8-98.9°C; these
 

values were consistent with that of 98-99°C reported previously
 

(Fagel and Ewing, 1951). Infrared, ultraviolet, and nuclear
 

magnetic resonance spectra for both lots were consistent with the
 

structure of IH-benzotriazole and identical to the spectra in the
 

literature (Fagel and Ewing, 1951; Sadtler Standard Spectra,
 

1966). Aldrich Chemical Company specifies this material to have
 

a purity of greater than 99%.
 

3 
B. Dietary Preparation
 

Diets were prepared once per week by first mixing a weighed
 

amount of chemical with an aliquot of ground Wayne® Lab Blox
 

animal feed (Allied Mills, Inc., Chicago, 111.) in a mortar.
 

When this premix appeared homogeneous, it was placed in a
 

Patterson-Kelly twin-shell blender with the remaining feed and
 

mixed for 20 minutes. Diets containing IH-benzotriazole were
 

stored in double plastic bags at 4°C and used within 1 week of
 

preparation.
 

Selected samples from diets containing 10,000 or 20,000 ppm
 

IH-benzotriazole were extracted and analyzed by ultraviolet
 

spectrophotometry at intervals of 2-6 weeks after mixing.
 

Concentrations were all within 25% of the theoretical values, but
 

were consistently low. On the basis of temperature-dependent
 

stability analyses performed at Midwest Research Institute, it
 

was concluded that IH-benzotriazole was stable in feed for 2
 

weeks at 25°C.
 

C. Animals
 

For the subchronic and the chronic studies, .Bale and female
 

Fischer 344 rats and male and female B6C3F1 mice were obtained
 

from Charles River Breeding Laboratories, Inc., Wilmington,
 

Massachusetts. The control rats and mice were received earlier
 

4 
than, and were placed on test prior to, animals in the dosed
 

groups. On arrival at the laboratory, the mice were examined and
 

showed evidence of intestinal parasites. The animals were
 

administered piperazine adipate in the drinking water at 3.0 g/1
 

for two 3-day periods, with an interval of 3 days between the
 

periods. The rats appeared to be normal and were not
 

administered the piperazine adipate. All animals were
 

quarantined for 2 weeks and then were assigned to control or
 

dosed groups so that the means of the weights of the animals in
 

each cage within a particular group were approximately equal.
 

Dosed rats were 42 days of age, control rats were 43 days of age,
 

and both dosed and control mice were 44 days of age when placed
 

on study.
 

D. Animal Maintenance
 

Animal rooms were maintained at temperatures ranging from
 

23-34°C. Air was filtered through Tri-Dek® 15/40 denier dacron
 

filters and air flow was maintained at a velocity permitting six
 

changes of room air per hour. Rooms were illuminated by
 

fluorescent light for 12 hours per day.
 

Rats were housed five per cage in galvanized steel wire-mesh
 

cages (Fenco Cage Products, Boston, Mass.). Cages were suspended
 

over drop trays lined with newspaper; no filters were used.
 

5 
Cages and racks were sanitized every week and paper in the drop
 

trays was replaced daily. After the first 52 weeks on study,
 

rats were transferred to suspended solid polycarbonate cages (Lab
 

Products, Inc., Garfield, N. J.), in which they were also housed
 

five per cage. These cages were equipped with disposable
 

nonwoven fiber filter sheets (Webrex®). The polycarbonate cages
 

were sanitized and supplied with fresh bedding twice per week. A
 

/g\
 

corn cob bedding (Sanicel , Paxton Processing Co., S. Lancaster,
 

®
 

Mass.) or a hardwood chip bedding (Aspen-bed , American
 

Excelsior, Sommerville, Mass.) was used in the rat cages.
 

Mice were housed 10 per cage in polycarbonate cages equipped with
 

nonwoven fiber filter bonnets (Filtek, Lab Products, Inc.,
 

Garfield, N. J.). The number of animals per cage was reduced to
 

five during the last 8 weeks of the chronic studies. Cages and
 

stainless steel perforated lids were sanitized three times per
 

week during the time the mice were housed 10 per cage, and twice
 

per week during the time the mice were housed 5 per cage. Fresh
 

bedding composed of either corn cobs (Sanicel ; Bed-o-Cobs ,
 

Anderson Cob Mills, Inc., Maumee, Ohio) or hardwood chips
 

(Aspen-bed ) was furnished at these times. The filter bonnets
 

were sanitized every 2 weeks. Cage racks for the mice were
 

sanitized every 2 weeks.
 

There was no rotation of the cages within the racks or of the
 

6 
racks within the rooms. All equipment that was sanitized was
 

washed with Dubois Serve Detergent and rinsed at 82°C.
 

Tap water (0.75-1.0 ppm chlorine) was provided in 250-ml glass
 

bottles equipped with rubber stoppers and stainless steel sipper
 

tubes. Bottles were replaced twice per week and were refilled as
 

needed. Sipper tubes and stoppers were cleaned once per week by
 

being soaked in a disinfectant (Environ, Vestal Laboratories, St.
 

Louis, Mo.), and rinsed before use. Controls were fed Wayne® Lab
 

Blox animal feed, and dosed animals were given the same product,
 

which had been mixed with the test chemical. All diets were
 

available ad libitum 7 days per week in Alpine** aluminum feed
 

cups (Curtin-Matheson Scientific, Inc., Woburn, Mass.) or in
 

stainless steel gang hoppers (Scientific Cages, Inc., Bryan,
 

Texas).
 

All control and dosed rats were housed in the same room as rats
 

being administered one of the following compounds:
 

Feed Studies
 

(CAS 53-96-3) fluorenylacetamide
 

(CAS 120-71-8) 5-methyl-o-anisidine
 

(CAS 2438-88-2) 2,3,5,6-tetrachloro-4-nitroanisole
 

(CAS 95-83-0) 4-chloro-o-phenylenediamine
 

(CAS 5131-60-2) 4-chloro-m-phenylenediamine
 

Dosed mice were housed in the same room as rats and mice
 

administered one of the following compounds:
 

7 
Feed Studies
 

(CAS 2243-62-1) 1,5-naphthalenediamine
 

(CAS 1465-25-4) N-1-naphthylethylenediamine dihydrochloride
 

Control mice were housed in the same room as mice administered
 

one of the following compounds:
 

Feed Studies
 

(CAS 122-66-77) hydrazobenzene
 

(CAS 2438-88-2) 2,3,5,6-tetrachloro-4-nitroanisole
 

(CAS 126-72-7) tris(2,3-dibromopropyl)phosphate
 

(CAS 1465-25-4) N-1-naphthylethylenediamine dihydrochloride
 

(CAS 615-66-7) 2-chloro-p-phenylenediamine sulfate
 

(CAS 142-04-1) aniline hydrochloride
 

E. Subchronic Studies
 

Subchronic feeding studies were conducted with Fischer 344 rats
 

and B6C3F1 mice to estimate the maximum tolerated doses of
 

IH-benzotriazole, on the basis of which two concentrations
 

(hereinafter referred to as "low doses" and "high doses") were
 

determined for the chronic studies. IH-Benzotriazole was
 

administered in the diet 7 days per week for 8 weeks at doses of
 

300, 1,000, 3,000, 10,000, or 30,000 ppm. Five males and five
 

females of each species were tested at each dose, and five males
 

and five females of each species served as untreated controls.
 

At the end of the studies, all animals were killed by C02
 

inhalation and necropsied.
 

In comparison with the controls, mean weight depressions in male
 

8 
and female rats receiving the chemical were no greater than 12%
 

at each dose ranging from 300 to 10,000 ppm; mean weight depressions increased sharply to 40 and 34% for males and females,
 

respectively, at 30,000 ppm. No deaths occurred before
 

termination of the study. The low and high doses for the chronic
 

studies using rats were set at 10,000 and 20,000 ppm.
 

In the mice, no effects on mean body weights were seen at doses
 

up to 10,000 ppm, but a slight weight depression of approximately
 

5% was seen in both sexes at 30,000 ppm. No deaths occurred in
 

the mice before termination of the study. The low and high doses
 

for the chronic studies using mice were set at 20,000 and 40,000
 

ppm.
 

F. Chronic Studies
 

The test groups, doses administered, and times on study of the
 

chronic feeding studies are shown in tables 1 and 2. For both
 

rats and mice, dosage changes were based on weight differences
 

between the dosed and control groups and not on clinical signs or
 

mortality.
 

G. Clinical and Pathologic Examinations
 

All animals were observed twice daily for signs of toxicity.
 

Body weights were recorded every 2 weeks for the first 12 weeks
 

9 
Table 1. Chronic Feeding Studies of IH-Benzotriazole in Rats
 

IH-Benzo-


Sex and Initial triazole
 

Test No. of in Dietb
 

Group Animals3

 (ppm)
 

Male
 

Controld 50 0
 

Low -Dose 50 10,000
 

5,000
 

High-Dose 50 20,000
 

5,000
 

20,000
 

10,000
 

Female
 

Controld 50 0
 

Low-Dose 50 10,000
 

5,000
 

High-Dose 50 20,000
 

5,000
 

20,000
 

10,000
 

Time on Study Time -Weigh ted
 

Dosed Observed Average Dosec
 

(weeks) (weeks) Cppm)
 

105-106
 

27
 

51 26-27 6,700
 

1
 

7
 

19
 

51 27 12,100
 

106
 

27
 

51 27 6,700
 

1
 

7
 

19
 

51 27 12,100
 

a

Rats were 42 or 43 days of age when placed on study.
 

"Diets were available ad libitum 7 days per week.
 

c

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

Z(no. of weeks receiving each dose)
 

"Controls were placed on study 4 weeks earlier than dosed animals.
 

10
 
Table 2. Chronic Feeding Studies of iH-Benzotriazole in Mice
 

m— Rpm^ JJ\ZlL£i\J
 n—
 

Sex and Initial triazole Time on Study Time-Weighted 

Test No. of in Dietb Dosed Observed Average Dosec 

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

Male 

Controld 50 0 109 

Low-Dose 50 20,000 18 

10,000 86 2 11,700 

High-Dose 50 40,000 18 

20,000 86 2 23,500 

Female 

Controld 50 0 109 

Low-Dose 50 20,000 18 

10,000 86 2 11,700 

High-Dose 50 40,000 18 

20,000 86 2 23,500 

a

Mice were 44 days of age when placed on study.
 

"Diets were available ad libitum 7 days per week.
 

c

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

£(no. of weeks receiving each dose)
 

"Controls were placed on study 14 weeks earlier than dosed animals.
 

11
 
and every month thereafter. Clinical observations were recorded
 

every month. Animals that were moribund and those that survived
 

to the termination of the bioassay were killed using C02
 

anesthesia and necropsied.
 

The pathologic evaluation consisted of gross and microscopic
 

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

from killed animals and from animals found dead. The following
 

tissues were examined microscopically whenever possible: tissue
 

masses, abnormal regional lymph nodes, skin, maridibular lymph
 

nodes, mammary gland, salivary gland, thigh muscle, sciatic
 

nerve, bone marrow, costochondral junction (rib), ttiymus, larynx,
 

trachea, lungs and bronchi, heart, thyroid, parathyroid,
 

esophagus, stomach, duodenum, jejunum, ileum, colon, mesenteric
 

lymph nodes, liver, gall bladder (mice), pancreas, spleen,
 

kidney, adrenal, bladder, seminal vesicles/prostate/testis
 

(males), ovary/uterus (females), nasal cavity, brain, pituitary,
 

eyes, external and middle ear, and spinal cord. The different
 

tissues were preserved in 10% buffered formalin, embedded in
 

paraffin, sectioned, stained with hematoxylin and eosin. Special
 

staining techniques were utilized when indicated for more
 

definitive diagnosis.
 

A few tissues from some animals were not examined, particularly
 

from those animals that died early. Also, some animals may have
 

12
 
been missing, cannibalized, or judged to be in such an advanced
 

state of autolysis as to preclude histopathologic evaluation.
 

Thus, the number of animals from which particular organs or
 

tissues were examined microscopically varies, and does not
 

necessarily represent the number of animals that were placed on
 

study in each group.
 

H. Data Recording and Statistical Analyses
 

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

System (Linhart et al., 1974). The data elements include
 

descriptive information on the chemicals, animals, experimental
 

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

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

verification of data transcription and for statistical review.
 

These data were analyzed using the statistical techniques
 

described in this section. Those analyses of the experimental
 

results that bear oh 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
 

13
 
censored as of the time that they died of other than natural
 

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

causes were not statistically censored. Statistical analyses for
 

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

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

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

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

departure from linearity test, which is only reported when its
 

two-tailed P value is less than 0.05.
 

The incidence of neoplastic or nonneoplastic lesions has been
 

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

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

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

the denominators included only those animals for which that site
 

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

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

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

consist of the numbers of animals necropsied.
 

The purpose of the statistical analyses of tumor incidence is to
 

determine whether animals receiving the test chemical developed a
 

significantly higher proportion of tumors than did the control
 

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

14
 
exact test (Cox, 1970) was used to compare the tumor incidence of
 

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

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

compared simultaneously with those for a control group, a
 

correction to ensure an overall significance level of 0.05 may be
 

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

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

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

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

where the Fisher exact P values are shown.
 

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

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

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

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


tailed 0.05 level of significance. Unless otherwise noted, the
 

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

from linear trend.
 

A time-adjusted analysis was applied when numerous early deaths
 

resulted from causes that were not associated with the formation
 

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

first tumor was observed were excluded by basing the statistical
 

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

15
 
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 an animal died naturally or was sacrificed
 

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

methods of comparing these curves were used for two groups;
 

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

groups. The statistical tests for the incidence of tumors which
 

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

noted, in the direction of a positive dose relationship. Significant departures from linearity (P < 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 the exact interval on the odds ratio (Gart, 1971). The
 

relative risk is defined as p^/Pc where pt is the true binomial
 

16
 
probability of the incidence of a specific type of tumor in a
 

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

spontaneous incidence of the same type of tumor in a control
 

group. The hypothesis of equality between the true proportion of
 

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

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

of unity represent the condition of a larger proportion in the
 

dosed group than in the control.
 

The lower and upper limits of the confidence interval of the
 

relative risk have been included in the tables of statistical
 

analyses. The interpretation of the limits is that in
 

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

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

control group would be within the interval calculated from the
 

experiment. When the lower limit of the confidence interval is
 

greater than one, it can be inferred that a statistically significant result (P < 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. RESULTS - RATS
 

A. Body Weights and Clinical Signs (Rats)
 

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

than those of the corresponding controls throughout most of the
 

bioassay. A dose-related effect was generally indicated by the
 

data for each sex, except for the last 24 weeks in the females
 

when the mean body weights of both the low- and high-dose groups
 

gradually rose toward those of the controls (figure 1).
 

Fluctuation in the growth curve may be due to dose changes.
 

Eye discoloration or inflammation or both were found in one low-


dose male, one high-dose male, two high-dose females, and two
 

control males. One high-dose male had inflammation of the
 

posterior ventral surface. Alopecia was noted in one control
 

male. None of these clinical signs could be clearly related to
 

administration of the test chemical.
 

B. Survival (Rats)
 

The Kaplan and Meier curves estimating the probabilities of
 

survival for male and females rats fed IH-benzotriazole in the
 

diet at the doses of this bioassay, together with those of the
 

controls, are shown in figure 2.
 

In male rats, the result of the Tarone test for dose-related
 

19
 
TIME ON STUDY (WEEKS) 

Figure 1. Growth Curves For Rats Fed 1H-Benzotriazole In The Diet 

20 
TIME ON STUDY (WEEKS) 

Figure 2. Survival Curves For Rats Fed IH-Benzotriazole In The Diet 

21 
trend in mortality is not significant. In females, the result of
 

the Tarone test is significant (P = 0.049), but in a negative
 

direction.
 

In males, 36/50 (72%) of the high-dose group, 34/50 (68%) of the
 

low-dose group, and 32/50 (64%) of the controls were alive at the
 

end of the bioassay. In females, 43/50 (86%) of the high-dose
 

group, 40/50 (80%) of the low-dose group, and 36/50 (72%) of the
 

controls were alive at the end of the bioassay. In each sex,
 

five animals in the high-dose and control groups were killed at
 

week 78. Sufficient numbers of rats of each sex were at risk for
 

the development of late-appearing tumors.
 

C. Pathology (Rats)
 

Histopathologic findings on neoplasms in rats are summarized in
 

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

are summarized in Appendix C, tables Cl and C2.
 

Hepatic lesions observed in male and female rats consisted of
 

neoplastic nodules of the liver in five males and two females
 

from the high-dose groups. No animals in the control or low-dose
 

groups had neoplastic nodules. Morphologic features of the
 

neoplastic nodules were similar to those described in the
 

literature (Squire and Levitt, 1975). Neoplastic nodules were
 

spherical lesions up to several liver lobules in size composed of
 

22
 
plates of hepatocytes which were sharply demarcated and compressed the adjacent normal parenchyma. Hepatocytes within these
 

nodules showed varying degrees of cytoplasmic change, including
 

clear cell, eosinophilic, and basophilic alterations. Mitotic
 

figures and varying degrees of nuclear atypia including multinucleation and hyperchromasia were noted.
 

Brain lesions observed in rats as early as week 21 consisted of
 

an oligodendroglioma in one low-dose male and gliomas in two
 

low-dose males and one high-dose female. These tumors were not
 

present in the control animals.
 

In female rats a non-dose-related increase in the incidence of
 

C-cell adenomas and carcinomas of the thyroid was noted. The
 

incidence in the controls was 0/43, the low-dose group 5/43, and
 

the high-dose group 3/50.
 

Most of the remaining neoplasms occurred randomly in control and
 

dosed groups, and their incidences and types, with few
 

exceptions, were similar to those observed historically in
 

Fischer 344 rats. With the possible exception of the liver in
 

male rats and the brain in male and female rats, no particular
 

organ or system seemed to be the target of this chemical. One
 

high-dose female had a rare and unusual neoplasm which appeared
 

to be a primary nonchromaffin paraganglioma of the heart (Scotti,
 

23
 
1971). The mass was visible microscopically in the wall of the
 

left ventricle and was composed of sheets of cells arranged in
 

small organoid-like packets. The cells had large oval and round
 

vesicular nuclei, high nuclear cytoplasmic ratios, and a light
 

gray-blue cytoplasm. The endocrine-like appearance of the mass
 

suggested it might be a chemodectoma, since all efforts to
 

demonstrate chromaffin granules were negative.
 

A variety of degenerative, proliferative, and inflammatory
 

lesions were observed in the control and dosed rats, but none
 

were believed to be related to administration of IH-benzotriazole.
 

Although administration of this chemical may be associated with
 

neoplastic nodules of the liver in male Fischer 344 rats, there
 

was no convincing evidence, based on the histopathologic
 

examination, that administration of IH-benzotriazole was
 

carcinogenic under the conditions of this bioassay.
 

D. Statistical Analyses of Results (Rats)
 

Tables El and E2 in Appendix E contain the statistical analyses
 

of the incidences of those primary tumors that occurred in at
 

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

in one or more than one group.
 

24
 
In male rats, the result of the Cochran-Armitage test for
 

positive dose-related trend in the incidence of neoplastic
 

nodules of the liver is significant (P = 0.008), and the results
 

of the Fisher exact test show that the incidence in the high-dose
 

group (5/45, 11%) is significantly higher (P = 0.024) than that
 

in the controls (0/48). At this laboratory, 2/13 historical-


control groups have shown incidences of 10-11% of this neoplasm,
 

although the average incidence in the controls is 2%. No other
 

type of liver tumor is observed.
 

In females, the results of the Cochran-Armitage test for positive
 

dose-associated trend are not significant for any of the incidences of tumors listed. The results of the Fisher exact test
 

show that the incidence of endometrial stromal polyps in the
 

low-dose group is significantly higher (P = 0.010) than that in
 

the corresponding controls. The incidence in the high-dose group
 

is not significant. When the incidences of endometrial stromal
 

polyps and of endometrial stromal sarcomas are combined for
 

analyses, the results of the Fisher exact test are no longer
 

significant in either the low- or high-dose groups. The
 

incidence of C-cell adenomas or carcinomas of the thyroid is
 

higher in the low-dose group than in the control group, but the
 

probability level of 0.028 resulting from the Fisher exact test
 

is above the 0.025 level required for significance when multiple
 

25
 
comparison is considered. The incidence in the high-dose group
 

is not significant.
 

Significant results in the negative direction are observed in the
 

incidences of pituitary and adrenal tumors in female rats, due to
 

higher incidences in the controls than in the dosed groups.
 

26
 
IV. RESULTS - MICE
 

A. Body Weights and Clinical Signs (Mice)
 

Mean body weights of the dosed male and female mice were lower
 

than those of the corresponding controls throughout most of the
 

bioassay, and a dose-related effect was indicated by the data for
 

each sex (figure 3). Fluctuation in the growth curve may be due
 

to mortality; as the size of a group diminishes, the mean body
 

weight may be subject to variation.
 

Alopecia was noted in 1 low-dose female, 2 high-dose females, 27
 

control males, and 30 control females. One control male and one
 

control female showed distention of the stomach. A low-dose male
 

had swelling and inflammation in the anal region, and one high-


dose male had an abscess in the same area. One high-dose female
 

exhibited poor balance in the month before terminat Lon of the
 

study. None of these clinical signs could be clearly related to
 

administration of the test chemical.
 

B. Survival (Mice)
 

The Kaplan and Meier curves estimating the probabilities of
 

survival for male and female mice fed IH-benzotriazole in the
 

diet at the doses of this bioassay, together with those of the
 

controls, are shown in figure 4.
 

27
 
TIME ON STUDY (WEEKS) 

Figure 3. Growth Curves For Mice Fed 1H-Benzotriazole In The Diet 

28 
TIME ON STUDY (WEEKS) 

Figure 4. Survival Curves For Mice Fed 1H-Benzotriazole In The Diet 

29 
In male mice, the result of the Tarone test for dose-related
 

trend in mortality is not significant. In females, the result of
 

the Tarone test is significant (P = 0.002), but in a negative
 

direction.
 

In males, 42/50 (84%) of the high-dose group, 36/50 (72%) of the
 

low-dose group, and 33/50 (66%) of the controls were alive at the
 

end of the bioassay. In females, 47/50 (94%) of the high-dose
 

group, 39/50 (78%) of the low-dose group, and 30/50 (60%) of the
 

controls were alive at the end of the bioassay. Sufficient
 

numbers of mice of each sex were at risk for development of late-


appearing tumors.
 

C. Pathology (Mice)
 

Histopathologic findings on neoplasms in mice are summarized in
 

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

are summarized in Appendix D, tables Dl and D2.
 

In female mice, a non-dose-related increase in the incidence of
 

alveolar/bronchiolar adenomas and carcinomas was noted. The
 

incidence in the controls was 0/49, the low-dose group 10/49, and
 

the high-dose group 4/49.
 

The remaining neoplasms occurred randomly in control and dosed
 

groups, and their incidence and type, with few exceptions, were
 

30
 
similar to those observed historically in B6C3F1 mice. No
 

particular organ or system seemed to be the target of this
 

chemical.
 

A variety of degenerative, proliferative, and inflammatory
 

lesions were seen in the control and dosed mice.
 

Based on the histopathologic examination, there was no evidence
 

for the carcinogenicity of IH-benzotriazole in the B6C3F1 mouse
 

under the conditions of this bioassay.
 

D. Statistical Analyses of Results (Mice)
 

Tables Fl and F2 in Appendix F contain the statistical analyses
 

of the incidences of those primary tumors that occurred in at
 

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

in one or more than one group.
 

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

positive dose-related trend and those of the Fisher exact test
 

for higher incidence in the dosed groups than in the control
 

group are not significant.
 

In female mice, the results of the Cochran-Armitage test for
 

positive dose-associated trend are not significant for any of the
 

incidences of tumors listed. The results of the Fisher exact
 

test show that the incidences of alveolar/bronchiolar carcinomas
 

31 
and of the combination of alveolar/bronchiolar adenomas or
 

carcinomas in the low-dose group are higher than those in the
 

corresponding controls (P = 0.001 in each case); however, the
 

incidences in the high-dose group are not significant. The
 

incidence of alveolar/bronchiolar carcinomas in historical-


control female mice in this laboratory is 10/275 (3.6%). The
 

incidence of this tumor in historical-control groups has varied
 

from 0-7%. The association of these lung tumors with
 

administration of the chemical is equivocal, in the absence of a
 

significant incidence in the high-dose female group and in both
 

of the dosed groups of male mice.
 

Significant results in the negative direction are observed in the
 

combined incidence of lymphomas in male mice, due to the higher
 

incidence of malignant lymphomas in the controls than in either
 

dosed group. A negative result is also observed in the incidence
 

of hepatocellular carcinomas in male mice, but when the
 

incidences of hepatocellular adenomas or carcinomas in male mice
 

are combined, statistical results are not significant. In female
 

mice, a negative result is observed in the incidences of
 

malignant lymphomas, but when all lymphomas are combined, results
 

are no longer significant.
 

In some of the 95% confidence intervals of relative risk, shown
 

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

32
 
significant results. It should also be noted that some of the
 

intervals have upper limits greater than one, indicating the 

theoretical possibility of the induction of tumors by 

IH-benzotriazole, which could not be detected under the 

conditions of this test. 

33
 

V. DISCUSSION
 

The toxicity of IH-benzotriazole for Fischer 344 rats and B6C3F1
 

mice was shown by consistently lowered mean body weights of all
 

dosed groups when compared with corresponding control groups. No
 

other clinical signs were observed that were related to administration of the test chemical. Survival was not decreased in any
 

of the dosed groups compared with respective control groups, and
 

in the female rats or mice, survival in the dosed groups was
 

slightly higher than in the control groups. The survival in the
 

dosed and control groups of both rats and mice was at least 60%,
 

and sufficient numbers of animals were at risk for development of
 

late-appearing tumors.
 

In male rats, neoplastic nodules of the liver occurred at a
 

statistically significant incidence (P = 0.024) in the high-dose
 

group when compared with the control group (controls 0/48, low-


dose 0/46, high—dose 5/45 [11%]), and the data, showed a dose-


related trend (P = 0.008). The incidence of this tumor in
 

control Fischer 344 rats used in similar bioassays of other test
 

chemicals at the same laboratory has varied from 0 to 11%, with
 

2/13 historical-control groups having incidences of 10-11%.
 

Since the incidence in the high-dose group is no higher than has
 

been observed in some control groups, these tumors cannot be
 

clearly associated with administration of the test chemical.
 

35
 
Brain tumors occurred in 3/44 (7%) low-dose males, as compared
 

with 0/46 controls. This tumor was also present in 1/50 (2%)
 

high-dose females, as compared with 0/50 controls. The
 

occurrence of brain tumors in dosed males and females is
 

suggestive of, but not considered as sufficient evidence of,
 

carcinogenicity. The suggestion of carcinogenicity is supported
 

by the absence of these lesions in the concurrent controls and by
 

the historical record of the low incidence of brain tumors (0/250
 

males and 1/249 females) in control Fischer 344 rats at this
 

laboratory.
 

In female rats, the incidence of endometrial stromal polyps in
 

the low-dose group was significantly higher (P = 0.010) than that
 

in the corresponding controls (controls 2/48, low-dose 10/45,
 

high-dose 8/50). However, the incidence in the high-dose group
 

was not significant, and when the incidences of endometrial
 

stromal polyps and endometrial stromal sarcomas were combined,
 

they were not significant in either the low- or high-dose groups.
 

Thus, these tumors cannot be associated with administration of
 

the chemical.
 

In male mice, no tumors occurred in dosed groups at incidences
 

that were significantly higher than those in controls.
 

In female mice, alveolar/bronchiolar carcinomas occurred at a
 

36
 
statistically significant incidence (P = 0.001) only in the
 

low-dose group when compared with the control group (controls
 

0/49, low-dose 9/49 [18%], high-dose 3/49 [6%]). The incidence
 

in the high-dose groups was not significant and the data did not
 

show a dose-related trend. It should be noted that the
 

incidence of these tumors in control B6C3F1 female mice from
 

other bioassays at this laboratory has varied from 0 to 7%, with
 

a mean of 4%. Therefore, the occurrence of this tumor in the
 

female mice cannot be clearly related to the administration of
 

the test chemical.
 

IH-Benzotriazole has been reported not to be carcinogenic when
 

total doses of 900-1,000 mg were administered to Wistar rats by
 

weekly subcutaneous injection at a level of 0.1 g/kg body weight
 

in a vehicle consisting of colloidal Infusin or species-


nonspecific serum (Vasil'eva, 1970); when a total dose of 92 mg
 

was similarly administered to hybrid (C57BL x CBA)F1 mice,
 

leukemia was induced at an incidence (13.7%) that was significantly higher than that (3.4%) in untreated controls, but not higher
 

than that (11.1%) in vehicle controls.
 

In female B6C3F1 mice there was an increased incidence of
 

alveolar/bronchiolar carcinomas, suggesting a possible
 

carcinogenic effect of IH-benzotriazole. In Fischer 344 rats
 

there was an increased incidence of brain tumors, suggesting a
 

37
 
possible carcinogenic effect. However, there was no convincing
 

evidence that under the conditions of this bioassay
 

IH-benzotriazole was carcinogenic in B6C3F1 mice or Fischer 344
 

rats of either sex.
 

38
 
VI. BIBLIOGRAPHY
 

Anaconda American Brass, Stripping electrodeposited materials
 

from the cathode. British Patent 993,392, May 26, 1965;
 

U. S. Application, May 10, 1963. Chem. Abstract 63, 7909,
 

1965.
 

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

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

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

Panel on Carcinogenicity of the Cancer Research Commission
 

of UICC, Vol. _2, International Union Against Cancer, Geneva,
 

1969.
 

Bregman, J. I., Cooling water - Other systems. Corrosion
 

Inhibitors, The Macmillan Co., New York, 1963, p. 151.
 

Cox, D. R., Regression models and life tables. _J._ R_^ Statist.
 

Soc. B 34(2):187-220. 1972.
 

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

1970, pp. 48-52.
 

Donaldson, R., Composition for inhibiting the corrosion of
 

articles of copper-based alloys. British Patent 1,235,468,
 

June 16, 1971. Chem. Abstract 75. 79774k, 1971.
 

Fagel, J. and Ewing, G. J., The ultraviolet absorption of
 

benzotriazole. J. Am. Chem. Soc. 73;4360-4362, 1951.
 

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

significance tests, confidence limits and adjustments for
 

stratification. Rev. Int. Stat. Inst. 39^(2): 148-169, 1971.
 

Green, F. 0. , Vapor-phase inhibitor impregnated packaging 

material. German. Patent 1,298,385, June 26, 1969. U.S. 

Application February 20, 1961. Chem. Abstract 7_5_, 72174y, 

1969. 

Hansen, R. H. , Benzotriazole and its derivatives and related
 

compounds as inhibitors for oxidation of polypropylene
 

electrical insulation in the presence of copper. U.S. Patent
 

3,367,907 February 6, 1968. Chem. Abstract 68. 69766a, 1968.
 

39
 
Kaplan, E. L. and Meier, P., Nonparametric estimation from
 

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

1958.
 

Levy, S. G., Baker, D. A., and Monroe, R. F., Noncorrosive
 

dry-cleaning compositions. U.S. Patent 3,337,471 August 22,
 1967. Chem. Abstract 67. 101324x, 1967.
 

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

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

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

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

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

Sadtler Research Laboratories, Sadtler Standard Spectra, IR No.
 

1631, NMR No. 6410, Sadtler Research Laboratories,
 Philadelphia, 1966.
 

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

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

induced by different numbers of administrations of benzo(a)
 

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

Sahyun, M. R. V. , Interaction of benzotriazole with development
 

and fog centers. Photogh. Sci. Eng. 15(l);48-53, 1971.
 

Scotti, T., Heart. In: Pathology, Anderson, W. A. D., ed., C. V.
 

Mosby, St. Louis, 1971, pp. 687-690.
 

Sherwin-Williams Chemicals. Technical Bulletin 531. CobratecR.
 

Corrosion inhibitors for copper, brass, bronze, and
 

multi-metal systems. Chicago, Sherwin-Williams Chemicals,
 1976.
 

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

classification of specific hepatocellular lesions in rats.
 

Cancer Res. 35:3214-3223. 1975.
 

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

j52:679-682, 1975.
 

Union Carbide Corporation, Free-flowing collapsible capsule
 

corrosion and leakage inhibitors. Netherlands Application
 

6,605,917, October 31, 1966; U.S. Application, April 30,
 1965. Chem. Abstract 66, 67671a, 1967.
 

40
 
Vasil'eva, N. N., Comparative evaluation of the toxic and
 

possible carcinogenic action of benzatriazole and phenidone.
 

Gig. Tr. Prof. Zabol _14 (3): 55-56, 1970.
 

West, W., Photography. In: The Encyclopedia of Chemistry, 3rd
 

edition, Hampel, C. A. and Hawley, G. G., eds, Van Nostrand
 

Reinhold, New York, 1973, pp. 840-842.
 

41
 

APPENDIX A
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

RATS FED IH-BENZOTRIAZOLE IN THE DIET
 

43
 

TABLE A1. 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS 

FED 1H-BENZOTRIAZOLE IN THE DIET 

CONTROL LOW DOSE HIGH DOSE 

ANIMAL S INITIALLY IN STUDY

ANIMAL S NECROPSIED

ANIMAL S EXAHINE D HISTOP AT HOLOGICALLY

 50

 48

 48

 50

 50

 46

 50 

 50 

 46 

INTEGUMENTAR Y SYSTE M 

*SKIN

PAPILLOMA ,

FIBROM A

LIPOM A

 N3S

 (48) (50)

 2 (4X )

 2 (4«) 

 (50) 

1

 1

 (2%) 

(2%) 

*SUBCUT TISSOF.

"IBROMA

"TBROSARCOM A

 (48 )

 3 (6^) 

2 ( H%) 

 (5D ) (50) 

PESPIRATOR Y SYSTE M 

IL'JNG

ALV^OHR/BPONCHIO L AR ADENOM A

ALVEOLAR/BRONCHIOLA R CARCINOM A

"IBROSARCOMA , METASTATIC

 (48 )

 3 (6%) 

1 ( 2%) 

 (46)

 1 (2%)

 (46) 

2 (455) 

HEMATOPOIETIC SYSTEM 

*MULT1PL':' ORGAN S

MALIGNAN T LYMPHOMA , NOS

UNDIFPSRFNTIATK D LEUKEMI A

M YELOM3N3CYTIC LEUKEMI A

 (48)

 1

 2

 (2%) 

( 4!5) 

 (53)

 5 (1 OX)

 (50) 

3 (6%) 

#SPL?EN

MY'-ILOMONOCYTIC LEUKEMI A

 (!»8)

 4 (8%) 

 (44) (46) 

CIRCULATOR Y SYSTEM 

NON S 

# NUMBE R

* NUMBE R

 0° ANIMAL S

 OF ANIMAL S

 WIT H TISSUE

 NECROPSIED 

 EXAHINE D MICR3 SCDPIC ALLY 

45 
TABLE A1. MALE RATS: NEOPLASMS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

DIGESTIVE SYSTEM 

ILIVFR (48) (45) (45) 

N'OPLASTIC NODULE 5 (11*) 

URINAR Y SYSTE M 

#KIDNEY (<*8) (U5) (46)

 

LIPOMA 1 (2«)

 

ENDOCRIN F SYSTEM 

SPITUITAR Y (45) (40) (45) 

ADENOMA , NO S 10 (22%) 5 (13X) 4 (9X) 

«D?ENAL (U6 ) (44) (46) 

PHEOCHROMOCYTOMA 4 (9%) 3 (7«) 3 (7X) 

#THYPOIC (43) (40) (tt) 

FOLIIC'J LAB-CELL CARCINOM A 1 (2%) 

C-CELL ADENOH A 3 (7%) 1 (255) 

C-CELL CARCINOM A 2 (5S) 1 (3X) 1 (2X) 

f PARATHYROI D (25) (21) (8)

 

ADENOMA , NO S 1 (4%)

 

*PANCREATIC ISLETS (44) (44) (46) 

TSLET-~ FLI ADENOM A 3 (7*)

 

REPRODUCTIV E SYSTEM 

*PP'PUTIAL GLAN D (48) (50) (50) 

CARCINOMA , NOS 1 (2«) 

ITESTIS (48) (43) (46) 

INTERSTITIAL-CFL L TUMO R 37 (77%) 38 (88S) 38 (83%) 

NERVOUS SYSTEM 

#RRAI N (46 ) (44) (46)

 

OSTEOSARCOMA, . METASTATIC 1 i2%l 

# NUMBER 0" ANIMAL S WITH TISSUE EXA1INED MICROSCOPICALLY 

* NUMBE R OF ANIMAL S NSCROPSIED 

46 
CONTROL LOW DOSE HIGH DOSE 

GLIOMA, NOS

OLIGODKNDROGLIOM A

 2

 1

 (5«) 

 (2X) 

SPECIAL SENSE ORGAN S 

NON E 

MUSCULOSKELETA L SYSTEM 

*SKULL

OSTEOSARCOM A

 (48)

 1 (2*) 

 (50) (50) 

BODY CAVITIES 

*BODY CAVITIES

HESOTHELIOHA, NOS

 (48)

 1 (2%) 

 (50) (50) 

&LL OTHER SYSTEMS 

S3NF 

ANIMA L DISPOSITION SUMMAR Y 

ANIMAL S INITIALLY IN STUD Y

NATURA L DEATH 9

MORIBUND SACRIFICE

SCHEtULE D SACBIFICE

ACCIDENTALLY KILLED 

TERMINA L SACRIFICE

ANIHA L MISSING 

 50

 8

 5 

5

 32

 50

 11

5 

 34

 50 

5 

4 

5 

 36 

3 INCLUDES AUTOLYZE D 4NIMALS __ _ _ _._ _ 

# NUMBER OF ANIMALS

* NUMBER OF ANIMAL S

 HITH TISSUE

 NECHOPSIED 

 EXAMINED MICROSCOPICALLY 

47 
TABLE A1.MALE RATS: NEOPLASMS (CONTINUED) 

Ct ONTROL LOW DOSE HIGH DOSE 

TUMO R SUMMAR Y 

TOTAL ANIMALS WITH PRIMAR Y

TOTAL PRIMAR Y TUMORS 

 TUMORS* 42 

74 

43 

64 

40 

61 

TOTAL ANIMAL S WITH BENIGN

TOTA L BENIG N TUMORS 

 TUMORS 40 

61 

42 

54 

40 

48 

T3TAL ANIMAL S WIT H MALIGNAN T

TOTAL MALIGNANT TUMORS 

 TUMORS 11 

12 

10 

10 

8 

8 

TOTAL ANIMALS WITH SECONDARY

TOTA L SECONDAR Y TUMORS 

 TUMORS* 2 

2 

TOTAL ANIMAL S WITH TUMORS

BENIG N O R MALIGNAN T 

TOTAL UNCERTAI N TUMORS 

 UNCERTAIN1 

1 

5 

5 

TOTAL ANIMALS 9 ITH TUMORS

PRIMAR Y OR METASTATTC 

TOTAL UNCBRTAIN TUMORS 

 UNCERTAIN­

* PRIMAR Y TUMORS : ALL TUMORS EXCEPT SBC3NDA8Y TUMORS 

* SECONDAR Y TUMORS: BETASTATIC TUS3RS 38 TUSOJS INVASIVE INTO AN ADJACENT ORGAN 

48 
TABLE A2.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE RATS 

FED 1H-BENZOTRIAZOLE IN THE DIET 

CONTROL LOW DOSE 

ANIMALS INITIALLY IN STUDY 50 50 

ANIMAL S NECROPSIED 50 48 

ANIMAL S EXAMINE D HISTOP AT HOLOGICALLY 50 48 

INTEGUMENTAR Y SYSTEM 

*SKIN (50)- (48) 

SQUAMOU S CELL CARCINOMA 

*SUBC0T TISSUE (50) (48) 

SARCOMA , NOS 1 (2%) 

RESPIRATOR Y SYSTE M 

#LUN G (50) (48) 

ADENOCAHCINOMA , NOS, METASTATIC 

HKMATOPOITIC SYSTFM 

*MULTIPLE ORGAN S (50) (48) 

MALIGNAN T LYMPHOMA , NOS 2 (4% ) 

MYELOHOTTOCYTIC LEUKEMI A 2 (4%) 

tSPLEE N (50) (48) 

MYELOMONOCYTIC LEUK5MI A 2 (4S) 

SLIVER (50) (48) 

LEUKEMIA , NOS 1 (2%) 

CIRCULATOR Y SYSTEM 

• HEAR T (50) (47) 

PARAGANGLIOMA , NO S 

DIGESTIV E SYSTEM 

#LIVER (50) (48) 

A DENOCARCINOMA , NOS,_ METASTATIC 

# NUMBE R OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALL Y 

* NUMBE R OF UNIMAL S NECBOPSIED 

HIGH DOSE 

50 

50 

50 

(50) 

1 (2%) 

(50) 

(50) 

1 (2X) 

(50) 

1 (2%) 

(50) 

(50) 

(50) 

1 (2%) 

(50) 

1 12*L 

49 
TABLE A2. FEMALE RATS: NEOPLASMS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

NEOPLASTIC NODDLE 2 (4*) 

#STOMAC H (U9 ) (48) (50) 

PAPILLOBA. NOS 1 (2«) 

ADENOCARCINOMA, NOS, BETASTATIC 1 <2X) 

ICOLON (HO) («6) (50) 

ADENOCARCINOHA, NOS, HETASTATIC 1 (2%) 

URINAR Y SYSTEM 

#OPINARY BLADDER (1*7) (47) (49) 

TRANSITIONAL-CEL L PAPILLOM A 1 (2X) 

ENDOCRINE SYSTEM 

#PITflITAPY (40) (46) (47) 

ADENOMA , NOS 16 (U0%) 9 (20*) 8 (17%) 

CHROMOPHOBE ADENOM A 1 (3%) 

tADBINA L C»8) (48) (50) 

PHEOCHROBOCYTOMA 6 (13%) 2 (4%) 1 (2X) 

#THYROIt C*3) (1*3) (50) 

FOLLICd LAB-CELL CAPCINOM A 1 (2X) 

C-CELL ADENOM A i* (9%) 

C-CELL CARCINOM A 1 (2%) 3 (6%) 

• PANCREATIC ISLETS (1*6) (48) (49) 

ISLfT-CELL CARCINOM A 1 (2X) 

REPRODUCTIVE SYSTEM 

*MAMMAR Y GLAN D (50) (48) (50) 

ACINCMA , NOS 2 (4X) 

ADENOCARCINOMA , NOS 1 (2%) 1 (2*) 

"IBRD ADENOM A 6 (12%) 2 (4«) 2 (4X) 

*CLITORAL GLAN D (50) (48) (50) 

ADENOMA , NOS 1 (2%) 

tUTERU S (1*8) (45) (50)

 

A DENOCARCINOMA. NOS 1 12«L _ _.

 

# NUMBER OF ANIMAL S WITH TISSUE EXAHINE1 ) MICROSCOPICALL Y 

* NUMBER OF ANIMAL S NECBOPSIED 

50 
TABLE A2. FEMALE RATS: NEOPLASMS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

FIBROMA 2 (ttX) 

ENDOMETRIA L STROMA L POLYP 2 ( «*) 10 (22%) 8 (16X) 

ENDOMETRIA L STROHAL SARCOMA 2 ( «•*) 2 {<•«) 

tUT^RUS/ENDOMETRIUM (»8) («5) (50) 

A OENOCARCINOMA , HOS 1 (2%) 

tOVARY/PAROVARIA N (48) (l»5) (50) 

ADENOCARCINOHA , HOS, BETASTATIC 1 <2X) 

NRRVOOS SYSTEM 

tBHAI N (50) (l»7) (50) 

GLIOBA, NOS 1 <2X) 

SPECIAL SENSE OR3ANS 

NONE 

BUSCUIOSKELETAL SYSTEM 

NONE 

BODY CAVITIES 

*PERITONEOM (50) (48) (50) 

ADENOCARCINOMA , NOS, HETASTATIC 1 (2X) 

ALL OTHER SYSTEMS 

THORAX 

SARCOMA. NOS 1

 

t NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* BOMBER OF ANIMAL S NECBOPSIED 

51 
TABLE A2. FEMALE RATS: NEOPLASMS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

ANIMA L DISPOSITION SUMMARY 

ANIMAL S INITIALLY IN STUDY 50 50 50

 

NATURA L DEATHS 7 5

 

MORIBUN D SACRIFICE 2 5 2 

SCHECriLE D SACBIFICE 5 5

 

ACCIDENTALLY KILLED 

TERMINA L SACRIFICE 36 U O U3

 

ANIMA L MISSING 

3 INCLUDES AOTOLYZE D ANIMAL S 

TUMOR SUMMAR Y 

TOTA L ANIMAL S WIT H PRIMARY TUMORS* 27 27 30 

TOTAL PRIMAR Y TUMORS 42 36 35 

TDTAL ANIMAL S WITH BENIGN TUMORS 2 3 23 20 

TOTAL BENIG N TUMORS 33 30 21 

TOTAL ANIMALS HITH MALIGNAN T TUMORS 9 6 10 

TOTAL MALIGNAN T TUMORS 9 6 11 

TOTAL A N I M A LA N I M A LS W I T H SECONDAR SECONDAR Y T U M O R ST U M O R S* 1

TOTAL SECONDAR SECONDARY TUMORS 66 

TOTAL A N I M A LA N I M A LS »ITIT H TtJBORS UNCERTAIN UNCERTAIN-­

BENIGN ORR MALIGNAN MALIGNAN TT 33 

TOTAL UNCERTAI UNCERTAI N TUMOR TUMORSS 33 

TOTAL A N I M A LA N I M A L S WITH TUMORS UNCERTAIH UNCERTAIH-­

P H I M A RP H I M A RY OR METASTATI METASTATIC 

TOTAL UNCERTAI UNCERTAIN T U M O RT U M O RS 

* P K I M A RP K I M A RY TTJHOHS: ALL TUMORS EXCEPT SECON DABY TUMOBS 

t SECONDAR SECONDARY TUMORS: METASTATI METASTATIC TUHDBS 38 TUMOJS INVASIVE INTO AN ADJACEN ADJACENT ORGAN 

52 
APPENDIX B
 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN
 

MICE FED IH-BENZOTRIAZOLE IN THE DIET
 

53
 

TABLE B1.

 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE 

FED 1H-BENZOTRIAZOLE IN THE DIET 

CONTROL LOW DOSE 

ANIMAL S INITIALLY IN STUDY 50 50 

ANIMALS HISSING 2 2 

ANIMAL S NEC80PSIED 39 45 

ANIMALS fXAMINED HISTOPATHOLOGICALL Y 39 44 

INTEGUMENTAR Y SYSTEM 

NONE 

PRSPIBATOR Y SYSTEM 

fLON G (39) (43) 

HEPATOCFLLtJLAP CARCINOMA , METAST 2 ( 5X) 3 (7X) 

ALVEOLAR/BRONCHI OLA R ADENOM A 2 (5%) 2 (5X) 

ALVEOLAR/BRONCHIOLA R CARCINOM A 2 ( 5X) 5 (12X) 

HEMATOPOIETIC SYSTEM 

* MULTIPLE ORGANS (39) (45) 

MALIGNAN T LYMPHOMA , NOS 11 (28%) 1 (2X) 

MALIG.LYMPHOHA , LYMPHOCYTIC TYPE 1 (2*) 

MALIG.LYMPHOBA , HISTIOCYTIC TYPE 

#SPLEEN (38) (43) 

ANGIOSARCOM A 1 (2X) 

MALIGNAN T LYMPHOMA , NOS 1 (3%) 

MALIGNAN T LYMPHOMA , MIXED TYPE 

IMESENTEPIC L. NODE (36) (35) 

ANGIOSARCOM A 

MALIG . LYMPHOMA , HISTIOCYTIC TYPE 1 (3X) 1 (3«) 

MALIGNAN T LYMPHOMA, MIXED TYPE 1 (3X) 

#LIVEP (39) (43) 

MALIG . LYMPHOMA , HISTIOCYTIC TYPE 

tSMALL INTESTINE (37) (42) 

MALIG. LYMPHOMA , HISTIOCYTIC TYPE 

CIRCULATORY SYSTEM 

N3N E 

* NUMBE R 3F ANIMALS WIT H TISSUE EXAMINE D HICROSC3PICALLY 

* NUMBER OF ANIMAL S NECROPSIED 

55 

HIGH DOSE 

50 

43 

48 

(46) 

5 (11«) 

(48) 

1 (2%) 

(46) 

2 C»X) 

1 (2X) 

(43) 

1 (2X) 

1 (2X) 

(47) 

2 (4X) 

(45) 

1 (2X) 
TABLE B1. MALE MICE: NEOPLASMS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

DIGESTIVE SYSTEM 

#LIVER (39) (43) (*»7) 

HEPATOCELLULA R ADENOH A 1 (2%) 2 (4%) 

HEPATOCELLOLAR CARCINOMA 12 (31%) 11 (26X) 5 (1 1 X) 

ANGIOSARCOH A 1 (2X) 

tsrOHACH (37) (42) (45) 

S QUA MOOS CELL PAPILLOH A 1 (2X) 

URINAR Y SYSTEM 

NONF 

'ND3CRIN E SYSTE M 

#PITUITAR Y (27) (39) (30) 

ADENOHA , NOS 1 ( 3S) 

PEPRODUCTIVF SYSTEM 

#T"STIS (38) (42) (47) 

INTEBSTITIAL-CELL TUMOS 1 (2%) 

HEMANGIOMA 1 (2%) 

NERVOU S SYSTE M 

NON E 

SPECIAL SENSE ORGAN S 

NON E 

MUSCHLOSKEL^TAL SYSTEM 

*SKEtETAL MUSCLE (39) (45) (48) 

PHABCOHYOSAHCOM A 1 (2%) 

BODY CAVITIES 

NON E 

* NUMBE R 3* ANIMALS WIT H TISSUE EXAMINE D MICROSC3PICALLY 

* NUMBEB OF ANIMALS NECROPSIF.B 

56 
TABLE B1.MALE MICE; NEOPLASMS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

ALL OTHER SYSTEMS 

NON1 

ANIHAL DISPOSITION SUMMAR Y 

ANIMALS INITIALLY IN STUDY 50 50 50 

NATURAL DEATH9 13 8 8

 

MORIBUND SACRIFICE 2 » 

SCHECULE D SACRIFICE 

ACCIDENTALLY KILLED 

TERMINA L SACRIFICE 33 36 42

 

ANIHR L MISSING 2 2 

9 INCLUDES AUTOLYZED ANIMALS 

TUMOF SUMMAR Y 

TOTAL ANIMALS HITH PRIMAR Y TUMORS* 24 23 18

 

TOTAL PRIMAR Y TUMORS 29 27 24

 

TOTAL ANIMALS WITH BENIGN TUMORS 2 5 4

 

TOTAL BENIGN TUMORS 2 5 4 

TOTAL ANIMALS WITH MALIGNANT TUMORS 22 20 15 

TOTAL MALIGNAN T TUMORS 27 22 20 

T O P A L A N I M A LA N I M A LS il ITH SECONDAR SECONDARY TUMORSf 2 3 

TOTAL SECONDAR SECONDARY TUMORS 22 33 

TOTAL ANIMALS WITH TUMORS UNCERTAIN UNCERTAIN-­

BENIGN OR MALIGNAN MALIGNANT 

TOTAL UNCERTAI UNCERTAIN TUMORS 

TOTAL A N I M A LA N I M A L S »ITI H TUHOBS UNCERTAIN UNCERTAIN-­

P R I M A RP R I M A R Y OR METASTATI METASTATIC 

TOTAL UNCERTAI UNCERTAIN TUMORS 

* PRIMARY TUMORS: ALL TUMORS EXCEPT SECONDAS SECONDASY T U M O RT U M O RS 

* SECONDAR SECONDARY TUMORS TUMORS:: flETASTATI flETASTATICC TUH3B TUH3BS DDB TOH01 TOH01S IHViSIV IHViSIVE IMTO AN ADJACEN ADJACENT OBGA OBGAN N

57 
TABLE B2. 

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICE 

FED 1H-BENZOTRIAZOLE IN THE DIET 

CONTROL LOW DOSE HIGH DOSE 

ANIMALS INITIALLY IN STUD Y

ANIMALS NECROPSIED

ANIMALS EXAMINE D HISTOPATHOLOGIC ALLY

 50 

 49 

 49 

50 

49 

49 

50 

50 

50 

INTEGUMENTAR Y SYSTEM 

*SKIN

"IBROSARCOMA 

 (49)­ (49) 

2 (4X) 

(50) 

RESPIRATOR Y SYSTEM 

#LUNG

ALVEOLAR/BRONCHIOLA R ADENOM A 

ALVEOLAR/BRONCHIOLAR CARCINOMA 

LIPOSARCOM A 

 (49) (49) 

1 (2X) 

9 (18X) 

(49) 

1 (2%) 

3 (6X) 

1 (2X) 

HEHATOPOIETIC SYSTEM 

"MULTIPLE ORGAN S

MALIGNAN T LYMPHOMA , NOS

H ALIG.LYMPHOSA, LYMPHOCYTIC TYPE 

MALIG.LYHPHOMA , HISTIOCYTIC TYPE

MALIGNAN T LYBPHOMA, MIXE D TYPE 

 (49) 

 10

 1

 (20X) 

 (2t) 

(49) 

4 (8X) 

2 (4X) 

(50) 

1 (2X) 

fSPLEE N

HEMANGIOSARCOM A

ANGIOSARCOMA 

MALIG.LYHPHOMA ,

MALIG.LYHPHOMA ,

 LYMPHOCYTIC

 HISTIOCYTIC

 TYPE 

 TYPE 

 (45) 

1 (2X) 

(47) 

2 (4X) 

(50) 

1

2

 (2X) 

 (4X) 

f PANCREATIC L.NODE

MALIGNANT LYHPHOHA, NOS

 (44) 

1 (2X) 

(42) (44) 

tMESENTEHIC L. NODE

MALIG. LYHPHOHA, HISTIOCYTIC TYPE 

 (44) (42) (44) 

1 <2X) 

tLIVER

MALIGNANT LYHPHOHA. NOS

 (46) 

1 (2X) 

(48) (49) 

1 (2%) ., 

t NUMBER

* NUHBER

 OF ANIMALS

 OF ANIHALS

 WITH TISSUE EXAHINED

 NECROPSIED 

 MICROSCOPICALLY 

58 
TABLE B2. FEMALE MICE: NEOPLASMS (CONTINUED)

 

CONTROL LOW DOSE

MALIG. LYHPHOMA. HISTI OCYTIC TYPE 1 (2X) 

f JEJUNUM (»2) (47)

MALIG-LYMPHOMA, HI STIOCI TIC TYPB 

MALIGNANT LYBPHOHA, MIXED TYPE 1 (2K) 

CIRCULATORY SYSTEM 

NON E 

DIGESTIVE SYSTEM 

tLIVER (U6) (48)

HEPATOCELLULA R ADENOM A 

HBPATOCELLULAR CARCINOMA 1 (2*) 2 (4!t) 

URINAR Y SYSTEM 

tKIDNE Y (U6) (48)

TUBULAR-CEL L ADENOM A 1 (2*) 

ENDOCRINE SYSTEM 

fPITUITARY (34) (36)

ADENOMA . NO S 3 (9°5) 2 (6X) 

tADRENA L (46) (45)

PHEOCHROMOCYTOMA 3 (7%) 

PHEOCHROMOCYTOMA, MALIGNAN T 1 (235) 

tTHYROIC C"») (42)

FOLLICULAR-CELL ADENOM A 2 (5X) 1 (2X)

FOLLICULAR-CELL CARCINOMA 2 (5X) 

REPPODOCTIVE SYSTEM 

*MAMMAR Y 3LAN D (49) (49)

PAPILLARY ADENOCARCINOMA 1 (2JC) 

#UTEROS (U4 ) (46)

ENDOMFTRIAL STROMAL POLYP 1 ±2%}_ 1 .(2X)

# NUMBER OF ANIMAL S WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBE R OF ANIMAL S NECROPSIED 

 HIGH DOSE 

 (49) 

1 (2X) 

 (49) 

1 (2X) 

 (50) 

 (38) 

 (46) 

 (38) 

1 (3«) 

1 (3X) 

 (50) 

 (46) 

 ... _ , 

59 
TABLE B2. FEMALE MICE: NEOPLASMS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

ENDOMETRIA L STROMAL SARCOM A

ANGIOSAHCOM A

 1 (2X) 

1 (2X) 

#OVAR Y

OAPILLARY CYSTADENOCARCINOMA,NOS

TOBOLA R ADENOM A

 (44 )

 1 (2*) 

 (44)

 1 (2X) 

 (45) 

NERVOU S SYSTEM 

NON F 

SPFCIAL SENSE ORGAN S 

NONE 

MUSrrrTLOSKELETA L SYSTEM 

NONE 

BODY CAVITIE S 

N3N ' 

ALL OTHER SYSTEMS 

NON T 

ANIMAL DISPOSITION SUMMAR Y 

ANIMAL S INITIALL Y IN STUDY

NATURA L DEATHS

MORIBUN D SACRIFICE

SCHECULE D SACRIFICE 

ACCIDENTALL Y KILLE D

TERMINA L SACRIFICE

ANIMA L MISSIN G 

 50

 17

 3

 30

 50

 6

 5 

 39

 50 

2 

1 

 47 

5 INCLUDE S AUTOLYZE D ANIMAL S 

f NUHBE R

* NUMBE R

 OP ANIMAL S WIT H TISSUE

 OF ANIMAL S NECROPSIED 

 FXAMINE D MICROSCDPICftLL Y 

60 
TABLE B2. FEMALE MICE: NEOPLASMS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

TUMO P SUMMAR Y 

T3TAL ANIMAL S WIT H PRIMAR Y TUMORS *

TOTAL PRIMAR Y TUMOR S

 2 1 

 28 

25 14 

33 15 

TOTA L ANIMAL S WIT H BENIG N

TOTAL BENIGN TUMOR S

 TUMORS 9 

 10 

6 3 

6 3 

TOTAL ANIMAL S WITH MALIGNAN T

TOTA L MALIGNAN T TUMORS

 TUMORS 1 6 

 18 

23 1 1 

27 12 

T3TAL ANIMAL S WITH SECONDAR Y TUMORSt 

TOTAL SECONDAR Y TUMORS 

TOTAL ANIMALS WITH TUMORS

BENIG N O R MALIGNAN T 

TOTAL UNCERTAI N TUMORS 

 UNCERTAINTOTA L ANIMAL S fflT H TUHOBS

PRIMAR Y O R HFTASTATIC 

TOTAL UNCERTAI N TUMORS 

 UNCERTAIN­

* PRIMAR Y TUMORS : ALL TUMOBS EXCEPT

# SECONDAR Y TUMORS : MSTASTATIC TUM3RS 38

 SECO TU

 TUM03S

NDAR Y MORS 

 INVASIV E INTO AN 1DJACENT OFGAN 

61 

APPENDIX C
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS
 

IN RATS FED IH-BENZOTRIAZOLE IN THE DIET
 

63
 

TABLE C1.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS 

FED 1H-BENZOTRIAZOLE IN THE DIET 

CONTROL LOW DOSE HIGH D lOSE 

ANIMAL S INITIALLY I N STUD Y 5 0 50 50 

ANIMAL S NECBOPSIED 48 50 50 

AtJIIAL S EXAMINF D HISTOP AT HOLOGICALLY US 46 46 

INT^GUM^NTAP Y SYSTE M 

*SKT N (48) (50) (50) 

EPIDERMA L INCLUSION CYST 1 (2X ) 

HEMORRHAGI C CYST 1 (2X) 

INFLAMMATION , FOCAL 1 (2X) 

ABSCESS, NOS 1 (2«) 

HYPERKE R ATOSIS 1 (2X) 

ACANTHOSI S 1 (2X) 

*SUBCUT TISSOE (48) (50) (50)

 

ABSCESS, NOS 3 (6«) 

RESPIRATOR Y SYSTEM 

*NASA L TtlRBINATE (48) (50) (50)

 

INFLAMMATION , SOPPU HA TI VE 1 ( 2») 

#TRACHE A (29) (43) (45)

 

INFLAMMATION , CHRONIC 2 (5X)

 

f LUNG/BRONCHUS (48) (45) (46) 

BRONCHI ECTASIS 3 (7X) 1 (2%) 

INFLAMMATION , CHRONIC 1 (2X) 

INFLAMMATION , CHRONIC FQCAL 2 (4X) 

tLUN G (48) (46) (46) 

HEMORRHAG E 2 (4X) 

BRONCHOPNEDMONIA , FOCAL 1 (2X) 

INFLAMMATION , FOCAL 1 (2X) 1 (2X) 

BRONCHOPNEOM3NIA , ACOTE 1 (2X) 

PNBOMONIA , CHRONIC MURIN E 1 (2«) 

INFLAMMATION , CHRONIC 1 (2X) 

PERIVASCOLITIS 1 12 XL 

t NOHBER OF ANIMAL S WITH TISSOE EXAHINE D HICROSCOPICALLY 

* NTJMBEH OF ANIMAL S NECROPSIED 

65 
TABLE C1.MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

HYPERPLASIA , EPITHELIAL 

HBHATOPOIETIC SYSTEM 

#BON E MARRO W 

HYPOPLASIA , HEMATOPOIETIC 

fSPLEEN 

INFABCT , NOS 

ATROPHY. NOS 

#MANDIBOLA R L. NODE 

HYPERPLASIA , LYMPHOI D 

#PANCREATI C L.NOD E 

H^MORPHA G E 

ATROPHY , NOS 

tTHYMU S 

HEMORRHAS V 

ATROPHY, NOS 

HYPERPLASIA , NOS 

CIRCnLATOR Y SYSTEM 

KBYOCARCIO M 

INFLAMMATION , CHRONIC 

"IBROSIS 

FIBROSIS, FOCAL 

FIBROSIS, DISUSE 

DEGENERATION , NOS 

#ENDOCARDIO M 

INFLAMMATION , FOCAL 

* AOPTA 

INFLAMMATION , ACUTE/CHRONIC 

DIGESTIV E SYSTE M 

#LIV*R 

FIBROSIS. FOCAL 

* NUMBER OF ANIMAL S WITH TISSUE

* N17MBF.R OF ANIMAL S NECHOPSIED 

CONTROL LOW C>OSE 

C»6) C»5)

1 (2*) 

(48) (1»4) 

1 (2%) 1 (2*) 

1 (2%) 

C»3) (<*3) 

(1*3) (t*3) 

1 (2«) 

1 (2X) 

(32) (1*1) 

1 (2X) 

4 (10*) 

1 (3X) 

(48) (46) 

1 (2X) 

1 (2X) 

1 12%) 

(1*8) (46) 

1 (2%) 

(48)

 (50) 

1 (2%) 

(48) (46) 

1 J2«l 

 EXAMINED MICROSCOPICALLY 

HIGH 

1 

(45) 

(46) 

(43) 

1 

(43) 

(40) 

(45) 

1 

1 

(45) 

(50) 

2 

(45) 

DOSE 

(2«) 

(2%) 

(2%) 

(2%) 

(4-5) 

66 
TABLE C1.MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

CIRRHOSIS, PORTAL 1 (2%)

 

NECROSIS, FOCAL 2 (4X) 1 (2X)

 

METAMORPHOSIS TATTY 2 (H%) 1 (2X)

 

NUCLEAR ALTERATION 1 (2X)

 

BASOPHILIC CYTO CHANG E 13 (28X) 11 (24%)

 

EOSIN3PHILIC CYTO CHANG E 5 (11X) 11 (24 X)

 

CLEAR- CELL CHANGE U (9*) 6 (13X)

 

HYPERPLASIA, FOCAL 3 (6«) 

ANGIECTASIS 1 (2X) 

#LIVER/CENTRILOBULA R (48) («6) (45)

 

METAMORPHOSIS FATTY 1 (2%)

 

*BILE EUCT («8) (50) (50)

 

HYPERPLASIA , NOS 1 (2X) 

#PANCREA S (HH) (»i») (46) 

INFLAMMATION , NOS 2 (5«) 

INFLAMMATION , FOCAL 2 (5X) 

INFLAMMATION , CHRONIC 2 (4X) 

INFLAMMATION, CHRONIC FOCAL 1 (2X) 3 (7%) 

#PERIF.SOPHAGEAL TISSU C»2) (39) (35) 

CYST, NOS 1 (3X) 

tSTOMACH (48) (it 5) (44) 

INFLAMMATION , NOS 1 (2X) 

PERIARTERITIS 1 (2«) 

NECROSIS, FOCAL 1 (2«) 

HYPERPLASIA , BASAL CELL 1 (2X) 

#GASTRIC SUBMUCOS A C»8) (U5) (44 )

 

FDEMA , NO S 1 (2X)

 

iCOLON («2) (44 ) (42) 

PARASITIS M 2 (5%) 6 (14S) 

URIYAR Y SYSTEM 

#KIDN*Y (48) (45) (46) 

CYST, NOS 1 (2X) 

HEMORRHAG F 1 (2%) 

FIBROSIS, FOCAL 1 (2X) 

NEPHROPATH Y 35 173 %L _ 

# NUMBE R OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMB58 OF ANIMAL S NECROPSIED 

67 
TABLE C1.MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

CONTROL LOW DOSE 

NEPHROSIS, NOS i»0 (89JS) 

#"ENA L FAPILL A 

MINERALIZATIO N 

HEMORRHAGIC CYST 

(U8 ) (45) 

1 (2%) 

1 (2X) 

ENDOCRIN E SYSTEM 

#PITUITARY (45) (40) 

CYST, NOS 2 (5X) 

HEMORRHAG E 2 (5X) 

HYPERPLASIA, FOCAL 1 (2X) 2 (5X) 

tADRENA L MEDULL A C»6) (44) 

HYPERPLASIA , NOS 1 <2X) 

HYPERPLASIA , FOCAL 3 (7X) 

f THYROI D (i»3) (40) 

HYPFRPLASIA , C-CELL 3 (8X) 

#PANCREATIC ISLETS (44) (4U) 

HYPF.RPLASIA, HOS 1 (2%) 

REPRODUCTIVE SYSTEM 

•PROSTATE (45) (43) 

INFLAMMATION, ACUTE 3 (7X) 

IN PL AMB AT ION, ACOTE FOCAL 21 (49S) 

INFLAMMATION , ACUTE/CHRONIC 

*TESTIS (48) (43) 

MINERALIZATIO N 1 (2%) 

GRANULOMA , SPERMATIC 1 (2X) 

DEGENERATION , NOS 

ATROPHY, NOS i* (8«) 1 (2X) 

HYPERPLASIA , INTERSTITIAL CELL 3 (6S) 1 (2X) 

*FJPIDIDYMIS C*8) (50) 

ABSCESS, NOS 1 (2«) 

NERVOU S SYSTEM 

#BRAI N (46) (44) 

INFLAMMATION f NOS 1 (2XJ 

* BOMBER OF ANIMAL S WITH TISSUF. EXAMINED HICROSCOPICAL1Y 

* NUMBE R OF ANIMAL S NECBOPSIED 

HIGH DOSE 

36 (78X) 

(46) 

(45) 

5 (11%) 

(46) 

(44) 

2 (5X) 

(46) 

(45) 

9 (20%) 

12 (27 X) 

1 (2X) 

(46) 

2 (4X) 

4 (9X) 

(50) 

C»6) 

68 
TABLE C1.MALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

CONTROL LOW DOSE 

INFAPCT, NOS 

CORPOR A AMYLACE A 

SPECIA L SENSE ORGAN S 

*FYE (48) (50) 

SYNECHIA , ANTERIO R 1 (2X) 

CATARACT 1 ( 2*) 1 (2X) 

*FY "/RETINA (1(8) (50) 

ATROPHY, NOS 2 (4%) 

MUSCOLOSKELETA L SYSTEM 

HON E 

BODY CAVITIES 

*NEDIASTINUN (48) (50) 

INFLAMMATION . ACUTE/CHRONIC 

PERIARTERITI S 

*MESENTERY (48) (50) 

PERIARTERITIS 1 (2«) 

A LL OTHER SYSTEMS 

ADIPOSE TISSUE 

INFLAMMATION, ACOTE/CHHONIC 

INFLAMMATION , CHRONIC 1 

SPECIAL MORPHOLOG Y SUMMAR Y 

NO LESION REPOBTED 

ADTO/NECROPSY/HISTO PER? 1 1 

AUTO/NECROPSY/NO HISTO 1 

AUTOLYSIS/NO NECROPSY 2 

# NUHBBB OF ANIMALS WIT H TISSUE EXAMINED MICBOSCOPICALLY 

* NUMBER OP ANIMAL S NECROPSIED 

HIGH DOSE 

1 (2%) 

1 (2%) 

(50) 

(50) 

(50) 

1 (2%) 

1 (2X) 

(50) 

1 

1 

4 

69 
_ _ 

TABLE C2. 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS 

FED 1H-BENZOTRIAZOLE IN THE DIET 

CONTROL LOW DOSE HIGH DOSE 

ANIMAL S INITIALLY IN STUD Y 5 0 50 50 

ANIMALS NECHOPSIED 50 1*8 50 

ANIMALS EXAMINE D HISTOP AT HOLOGICALLY 50 48 50 

INTEGUMFNTAR Y SYSTEM 

NONF 

PESPIFHTCP Y SYSTEM 

*NASA L TURBINAT E (50) (48) (50) 

INFLAMMATION , NO S 1 (255) 

#T,'JNG /BRONCHU S (50) (UB) (50) 

BRONCH I ECTASI S 5 (10X) 

INFLAMMATION , ACUTE FOCAL 2 (<*«) 

«LUNG/BRONCHIOL F (50) (48) (50) 

INFLAMMATION , ACUTE FOCAL 1 (2X) 

• LUN G (50) (1*8) (50) 

HEMORRHAG E 2 (4X) 

INFLAMMATION , NOS 1 (2?) 

BRONCHOPNEOM3NIA , FOCAL 1 (2X) 

INFLAMMATION , FOCAL 1 (2%) 

INFLAMMATION , INTERSTIPIAL 1 (2%) 

BRONCHOPNEUN3NIA , ACUTE 1 (2«) 

INFLAMMATION , ACUTE FOCAL 2 (4X) 

ABSCESS, NOS 2 (U%) 1 (2X) 

PNEUMONIA , CHRONIC MURIN E 1 ( 2X) 

GRANULOMA , NOS 1 (2%) 3 (6X) 

GRANULOMA , FOREIG N EODY 1 (2H) 

INFLAMMATION , NECRO GRA N 1 (2X) 

HYPERPLASIA , EPITHELIA L 1 (2X) 2 (4X) 

fTJJN G /ALVEOLI (50) (48) (50) 

INFLAMMATION , ACUTE 1 (2%) 

* NUMBE R OF ANIMAL S WITH TISSUE EXAMIN3 D MICROSCOPICALL Y 

* NUMBE R OF ANIMAL S NECEOPSIED 

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

CONTROL LOW DOSE HIGH DOSE 

H5HATOPOIETIC SYSTEM 

#SPLEEN 

ATROPHY , NO S 

HEMftTOPOIESIS 

fMESENTFBIC L. NODE 

DILATATION, NOS 

ATROPHY , NOS 

#THYMU S 

ATROPHY , NO S 

CIPC0LATOR Y SYSTEM 

f ENDOCARDIU M 

INFLAMMATION , CHRONI C FOCAL 

tCARDIA C VALV F 

INFLAMMATION , CHRONIC 

*AORTA 

PFPIARTERITI S 

*PULMONAR Y ARTER Y 

MINERALIZATIO N 

DIGESTIVE SYSTF M 

fLIVE P 

ECTOPIA 

NECROSIS, PO^A L 

METAMORPHOSI S FATT Y 

NUCLEA R ALTERATIO N 

BASOPHILIC CYTO CHANG E 

ROSIN3PHILIC CYTO CHANG E 

CLEAP-CSLL CHANG E 

HVPEPPLASIA , FOCAL 

ANGIECTASI S 

*BILE DUCT 

INFLAMMATION^ . VOCAL 

# NUMBER OF ANIMAL S WITH TISSUE

* NUMBE R OF ANIMAL S NEC80PSIED 

(50) (48) 

7 (14%) 

(43) (47) 

2 (4X) 

(31) (42) 

(50) (47) 

1 <2X) 

(50) (47 ) 

(50) (48) 

(50) (48) 

(50) (48) 

1 (2X) 

4 (8*) 1 (2%) 

28 (58X) 

1 (2X) 

3 (6X) 

9 (18%) 

(50) (48) 

2 (4X1 

 EXAMINE D MICROSCOPICALLY 

(50) 

1 (2X> 

(48) 

1 (2X) 

1 (2X) 

(38) 

1 (3X) 

(50) 

(50) 

2 (4X) 

(50) 

1 (2X) 

(50) 

1 (255) 

(50) 

1 (2*) 

1 (2«) 

37 (74%) 

4 (8X) 

1 (2%) 

(50) 

71 
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

# PANCREA S (46 ) (48) (49)

 

INFLAMMATION , CHRONIC FOCAL 4 (8%)

 

PI8ROSIS, FOCAL 1 (2X)

 

tSTOMACH C»9) (48) (50)

 

HYPERPLASIA , PAPILLAR Y 1 (255)

 

tCOLON (40) (45) (50) 

PARASITIS M 2 (4%) 

URINAR Y SYSTEM 

• KIDNE Y (49) (48) (50) 

HYCRONEPHROSI S 1 (2X) 

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

GLOMERULONEPHRITI S , NOS 1 (2%) 

NFPHROPATH Y 18 (37*) 

NEPHBOSIS, NOS 16 (33*) 17 (34X) 

IRENAL PAPILLA (49) (48) (50) 

MINERALIZATIO N 1 (2%) 

HYPERPLASIA, EPITHELIAL 1 (2S) 

#KIPN£Y/P1*LVIS (49) (48) (50) 

MINERALIZATIO N 7 (15*) 3 (6%) 

HYPERPLASIA, EPITHELIAL 4 (S%) 13 (26%) 

HYPERPLASIA , CYSTIC 1 (2%) 

ENDOCRIN E SYSTEM 

tPirniTARY (40) (46) (47) 

MINERALIZATIO N 1 (3%) 

HEMORRHAGE 1 (2%) 

HYPERPLASIA , FOCAL 1 (2%) 1 (2%) 

#ADR?NA L (48) (48) (50)

 

n^AMORPHOSIS FATTY 1 (2X)

 

#ADRENA L CORTEX (48) (48) (50) 

HYPEBPLASIA, FOCAL 1 (2%) 

#THYROIE (43) (43) (50) 

HYPERPLASIA. C-C5LL 1 ilJl. 1_L2*L 1 12%) 

# NUMBER OF ANIMAL S WITH TISSUE EXABINED MICROSCOPICiLir 

* NUMBE R OF ANIMAL S NECROPSIED 

72 
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

#PAR A THYROI D 

HYPEPPLASIA . NODULA R 

REPRODUCTIV E SYSTEM 

*MAMNAR Y GLAN D 

SALACTOCELE 

HYPERPLASIA, NOS 

*CLITORAL GLAN D 

ABSCESS, NOS 

#UTEPUS 

HYCROHETR A 

HEMORRHAG E 

ABSCESS, NOS 

NECROSIS, NOS 

AMYLOIDOSIS 

fUr ERUS/ENDOMETRIUM 

INFLAMHATION , NOS 

INFLAMMATION , ACUTE 

HYPERPLASIA , CYSTIC 

f OVARY/OVIDUCT 

INFLAMMATION , ACUTE 

#OVARY/PftROVARIA N 

CYST, NOS 

tOVARY 

CYST, NOS 

INFLAMMATION , NOS 

DEGENERATION , CYSTIC 

NERVOUS SYSTEM 

tBRAI N 

INFARCT, FOCAL 

SPECIA1 SENSE OR3ANS 

*FYE 

SYNECHIA, POSTERI08 

# NUMBER OF ANIMAL S WITH TISSUE

* NO MB EH OF ANIMALS NECBOPSIED 

CONTROL LOW DOSE 

(31) (15) 

1 (3%) 

(50) (48) 

2 (<IX ) 1 (2S) 

1 (251) 4 (8%) 

(50) (48) 

(48) (45) 

4 (9«) 

1 (2K ) 

1 (2*) 

(48) (45) 

1 (2%) 

11 (24* ) 

<* (9X) 

(148) (45) 

4 (9X) 

(48) (U5 ) 

2 («) 

(U9) (45) 

1 (2X) 

1 (2X) 

2 (H%) 

(50) (47) 

1 (2X) 

(50) (48) 

EXAMINED MICROSCOPICALLY 

HIGH DOSE 

(14 ) 

(50 ) 

3 (6X) 

1 (25E) 

(50) 

1 <2X) 

(50) 

4 (8X) 

1 (2%) 

1 (2%) 

(50) 

12 (24%) 

4 (8X) 

(50) 

2 (4X) 

(50) 

1 (2JS) 

(50) 

(50) 

(50) 

1 (( 2X2X) )

73 
TABLE C2. FEMALE RATS: NONNEOPLASTIC LESIONS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

* EY^ /CORNEA (50) (48) (50) 

INFLAMMATION, CHRONIC 1 (2X) 

*EY13/IRIS (50) (U8 ) (50) 

INFLAMMATION , ACUTE 1 (2%) 

*FYE/CRYSTALLINE LENS (50) (1*8) (50) 

CATARAC T 1 (2%) 

Mt!SCOLOSKELF.TAL SYSTEM 

NONE 

BODY CAVITIES 

NON E 

A LL OTHER SYSTEMS 

ADIPOSE TISSUE 

INFLAMMATION , CHRONIC 2 

SPECIAL KORPHOLOGY SUMMAR Y 

NO LESION REPORTE D 11 2 2 

AUTO/NECPOPSY/HISTO PERF 1 

AUTOLYSIS/NO NECROPS Y 2 

# NUMBE R OF ANIMAL S WITH TISSUE EXASINE D MICROSCOPICALLY 

* NUMBE R OF ANIMAL S N3CROPSISD 

74 
APPENDIX D
 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS
 

IN MICE FED IH-BENZOTRIAZOLE IN THE DIET
 

75
 

TABLE 01.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE MICE 

FED 1H-BENZOTRIA20LE IN THE DIET 

CONTROL LOW DOSE HIGH DOSE 

ANIMALS INITIALLY IN STUDY 50 50 50 

ANIMAL S MISSING 2 2 

ANIMALS NECROPSIED 39 45 48 

ANIMAL S EXAMINE D HI STOP ATHOLOGICALL Y 39 44 48 

INTEGUMENTAR Y SYSTEM 

*S!CIN (39) (U5) (48) 

EPIDERMA L INCLUSION CYST 1 (3«) 

INFLAMMATION , DIFFUSE 1 (2*) 

INFLABMATION , ACUTE/CHR ONIC 1 (2X) 

INFLAMMATION , CHRONIC 1 (355) 

"IBROSIS 1 (3%) 

FIBROSIS, FOCAL 1 (2X) 1 (2%) 

RESPIRATORY SYSTEM 

•TPACHEA (38) (41) (38)

 

HYPERPLASIA , EPITHELIAL 1 (2X)

 

#LUNG/ERONCHU S (39) 0*3) (46) 

INFLAMMATION , FOCAL 1 (2%) 

INFLAMMATION , ACUTE FOCA L 1 (2«) 

INFLAMMATION , ACUTE/CHRONIC 1 (2K) 

#L(TNT (39) (43) (46) 

HEMORRHAG " 1 (2%) 5 (11%) 

HYPERPLASIA , EPITHELIA L 1 (2X) 

HYPEPPLASIA , ALVEOLA R EPITHELIU M U (9X) 

fLUNG/ALVSOLI (39) (43) (46)

 

INFLAMMATION , FOCAL 1 (2X)

 

H?,MA70POIFTI" SYSTFM 

*BONE MARROW (36) (43) (46)

 

HYPERPLASIA , H EM ATOPOIETIC 1 (2X)

 

fSPLEE N (38) (43) (46)

 

ATROPHY , NOS J-I2SL _ 

* NUMBE R OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBE R OF ANIMAL S NECROPSIBD 

77 
TABLE 01. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

CONTROL LOW DOSE 

HYPBRPLASIA, HEHATOPOIETIC 

HYPE5PLASIA, LYMPHOID 1 (3*) 3 (7X) 

HE H ATOP 0 IE SIS 1 (3X)

 

BHYTHRDPOIF.SIS 3 (8X) 

tHANDIBULA R L. NODE (36) (35) 

HYPERPLASIA , PLASHA CEIL 1 OX)

 

#PANCHEATIC L.NODE (36) (35) 

INFLAHHATION, ACUTE 1 (3X) 

NECROSIS. TCCAL 1 (3X) 

HYPEPPLASIA , LYHPHOID 1 (3X) 

*MESENT5RIC L. NODE (36) (35) 

THROMBOSIS. NOS 1 (3X) 

HEMORRHAG E 13 (37X) 

NECROSIS, NOS 

HYPERPLASIA, NOS t (11«> 

HYPERPLASIA, LYHPHOID 4 (11X) 

HE M ATOP 0 IE SIS 

CIRCULATORY SYSTEH 

tHEART (39) (43) 

PEHIARTERITIS 

*CORONARY ARTERY (39) (US)

 

HYPERTROPHY, NOS 

*HESFNTF.RIC ARTERY (39) (US)

 

HYPERTROPHY, NOS 

DIGESTIVE SYSTEM 

tLIVER (39) (43) 

HEMORRHAG E 

HEHORRHAGIC CYST 

INFLABHATION, ACUTE/CHRONIC 2 (5X» 

ABSCESS, CHRONIC 1 (2X) 

NECROSIS, NOS 1 12%) 

NECROSIS, FOCAL 

HETAHORPHOSIS FATTY 

BASOPHILIC CYTO CHANGE, _ 

t NUHBER OF ANIMALS WITH TISSUE EXAMINED HICROSCOPICALLY 

* NUMBER OF ANIMALS NECBOPSIED 

HIGH DOSE 

1

1

(U3) 

(U3) 

(<*3) 

13

1

1

(«»)

2

(48) 

1

(*8) 

1

(47) 

1

1

2

1

1

 (2%) 

 (2X) 

 (3 OX) 

 <2t) 

 (2X) 

 (5X) 

 (2X) 

 (2X) 

 (2X) 

 (2X) 

 <«X) 

 (2X) 

 C2X) : 

78 
TABLE 01. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

HYPERPLASIA, FOCAL 

HEM ATOP DIES IS 

#STOHACH 

GRAW7LOMA , FOREIGN BODY 

HYPERPLASIA , EPITHELIAL 

#J5JUNU« 

AMYLOIDOSI S 

HYPERPLASIA , LYMPHOID 

#ILEU M 

A MYLOIDOSIS 

# COT, ON 

PARASITISM 

URINAR Y SYSTEM 

#KIDNEY 

GLOMFROLOHEPHRITIS, CHRONIC 

NEPHROSIS, NOS 

GLOMEROLOSCLEROSIS , NOS 

#KIDNEY/CORTE!t 

SCA R 

#KIDNI?Y/GLONEBUL us 

AMYLOIDOSIS 

ENDOCRINE SYSTEM 

fPITUITARY 

HYPEHPLASIA, FOCAL 

#ADRENA L 

AMYLOIDOSIS 

tADRENA L COBTEX 

HYPERPLASIA, FOCAL 

fTHYROI D 

CYSTIC FOLLICLES 

# NUMBER OF ANIMAL S WITH TISSUE

* NUMBER OF ANIMAL S NBC80PSIBD 

79 

CONTROL LOW DOSE 

3 (8X) 1 (2X) 

(37) (12) 

1 (2X) 

(37) (U2) 

1 (3X) 

1 (2X) 

(37) (t2) 

2 (5*) 

(37) (i»1) 

« (10X) 

(39) (43) 

21 (49X) 

3 (8%) 

(39) (43) 

1 (3%) 

(39) C»3) 

2 (5X) 

(27) (39) 

(36) (39) 

2 (6X) 

(36) (39) 

1 (3X) 

(38) (39) 

1 (3X) 

 EXAHINED MICROSCOPICALLY 

HIGH DOSE 

1 (2X) 

(45) 

1 (2X) 

(45) 

(45) 

(44) 

2 (5X) 

(47) 

1 (2X) 

2 (4X) 

(47) 

(47) 

(30) 

1 (3X) 

(44) 

(44) 

(36) 
TABLE D1. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

ABYLOIDOSIS 1 (3X) 

HYPERPLASIA , FOLLICUL AR-CELL 2 ( 5X) 1 (3*) 

tPARATHYROI D (28) (15) (18)

 

HYP3RPLASIA, NOS 1 (4X) 

#PANCREATIC ISLETS (36) CM) (I'M

 

HYPERPLASIA , NOS 1 (3X) 

REPRODUCTIVE SYSTEM 

ITBSTIS (38) (t2) (47) 

HYPSRPLASIA, INTERSTITIAL CELL 1 (2X) 

NERVOU S SYSTEM 

NDN E 

SPECIAL SENSE ORGANS 

NONE 

MUSCULOSKELETAL SYSTEM 

NONE 

B3DY CAVITIES 

*MEDIASTINUM (39) (45) (48) 

INFLAMMATION , FOCAL GPANULOMATO U 1 (2X)

 

*PBRITONF_!JM (39) (45) (48) 

HYPERPLASIA , MESOTHELIAL 1 (2X) 

ALL OTHEF SYSTEMS 

*MfILTIPLE OR3AN S (39) (45) (48) 

PERIARTERITIS 2 (4X)

 

ABYLOIDOSIS 1 13%L 

# NUMBE R OF ANIMAL S WIT H TISSUE EXAMINE D MICROSCOPICALLY 

* NUMBE R OF ANIMAL S NECROPSIED 

80 
TABLE 01. MALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

SPECIAL MORPHOLOGY SUMMAR Y 

NO LESION REPORTED 5 1 10 

ANIMA L MISSING/N O NECBOPSY 2 2 

AUTO/NECROPSY/HISTO PERF 1 1 

AUTO/NECROPSY /NO HI STO 1 

AUTOLYSIS/NO NECROPSY 9 3 2 

t NUMB?R OF ANIMAL S WITH TISSUE EXAMINE D MICROSCOPICALL Y 

* NUMBE R OF ANIMAL S NECROPSIED 

81 
TABLE D2.

 

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE 

FED 1H-BENZOTRIAZOLE IN THE DIET 

CONTROL LOW DOSE 

ANIMAL S INITIALLY IN STUDY 5 0 50 

ANIMALS NECEOPSIED 49 49 

ANIMAL S EXAMINE D HISTOP AT HOLOGICALLY 49 49 

INTEGUMENTAR Y SYSTEM 

*SKIN (49) (49)

 

INFLAMMATION , ACUTE 1 (2%) 

*SUBCUT TISSUE (49) (49) 

ABSCESS, CHRONIC 1 (2%) 

RESPIRATOR Y SYSTEM 

#LUNG /BRONCHUS (49) (49)

 

PFRIVASCOLITIS 

tLUNG (49) (49) 

EDEMA, INTERSTITIAL 1 (2%) 

HEMORRHAG E 

INFLAMMATION, NOS 1 (2%) 

INFLAMMATION , FOCAL 3 (6X) 

"IBROSIS, FOCAL 1 (2*) 

HYPERTROPHY , FOCAL 

HYPERPLASIA, ALVEOLAR EPITHELIUM 1 (2X) 

HEMATOPOIFTIC SYSTEM 

tBON E MARRO H (40) (47) 

MYELOFIBROSIS 21 (45X) 

tSPLEEN (45) (47) 

NECRCSIS, NOS 1 (2%) 

HYPERPLASIA, LYMPHOID 1 (2«) 5 (11X) 

HEM ATOP DIE SIS 1 (2X) 

ERYTHRDPOIESIS 4 (9%) 

#LYMPH NODE OF THORAX (44) (42)

 

HYPF.BPLASIA. NOS _ 1 (2£t 

# NUMBER OF ANIMALS WITH TISSUE EXAMINED HICBOSCOPICALLY 

* NUHBEB OF ANIMAL S NECROPSIED 

HIGH DOSE 

50 

50 

50 

(50) 

(50) 

(49) 

1 (2*) 

(49) 

2 (4%) 

3 (6X) 

1 (2X) 

1 (2X) 

(48) 

13 (27X) 

(50) 

(44) 

82 
TABLE 02. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

CONTROL LOW DOSE 

# PA NCR FA TIC L.NOD E (44 ) (42)

 

H'MATOPOIESIS 1 (2%) 

#LUMBA R LYMP H NODE (44) (42) 

HYPEKPLASIA, NOS 1 (2*) 

fMESENTERIC L. MODE (44) (42) 

HEMORRHAG E 9 (21X) 

HYPERPLASIA , NOS 1 12%) 

HYPBRPLASIA, LYMPHOID 1 (2X ) 

HEH ATOPOIESIS 1 (2X) 

#RBNA L LYMPH NODE (44) (42) 

HYPERPLASIA, NOS 1 (2X) 

HEMATOPOIESIS 1 (255) 

tTHYMU S (19) (37)

 

CYST, NOS 

CIRCULATORY SYSTEM 

fMYOCARDIUM (49) (49) 

DEGENERATION, NOS 1 (2%) 

* AORTA (49) (49) 

MINERALIZATIO N 1 (2X) 

*PniMONA8Y ARTEHY (49) (49) 

HYPERTROPHY, FOCAL 

HYPERPLASIA, NOS 1 (2«) 

*PANCREATIC ARTERY, (49) (49)

 

HYPERPLASIA, NOS 

*MESENTEBIC ARTER Y (49) (49) 

HYPERPLASIA, NOS 

DIGESTIVE SYSTEM 

#LIVKR (46) (48)

 

METAMORPHOSIS FATTY 

HYPERPLASTIC NODDLE 1 1221

 

* NUMBER OF ANIMALS WITH TISSUE EXAMINED MICROSCOPICALLY 

* NUMBER OF ANIMALS NECROPSIED 

HIGH DOSE 

(44) 

(44) 

(44) 

4 (9X) 

(44) 

(40) 

1 (3%) 

(50) 

(50) 

(50) 

1 (2%) 

(50) 

1 (2X) 

(50) 

1 (2%) 

(49) 

1 (2X) 

83 
TABLE D2. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED)

 

HYPERPLASIA. RETICOLUM CELL 

HEBATOPOIESIS 

f PANCREA S 

CYSTIC DUCTS 

#PANCBEATIC DUCT 

INFLAMMATION , CHRONIC 

#PANCREATIC ACINUS 

ATROPHY, NOS 

#STOMACH 

HYPFRPLASIA , FOCAL 

•JEJUNU M 

HYPERPLASIA, LYMPHOID 

#ILEOM 

HYPERPLASIA , LYMPHOI D 

tCOLON 

PARASITISM 

URINAR Y SYSTEM 

fKIDNS Y 

MULTIPLE CYSTS 

N5PHROSIS, NOS 

GLOMERULOSCLEROSIS, NOS 

#nBINARY BLADDER 

PERIARTERITI S 

ENDOCRINE SYSTEM 

• ADRENA L CORTEX 

METAMORPHOSI S FATTY 

*THYROIC 

CYSTIC FOLLICLES 

INFLAMMATION , ACUTE FOCAL 

HYPERPLASIA ^ FOLLICULftR-C ELL 

# NUMBE R OF ANIMAL S WITH TISSUE

* NUMBE R OF ANIMAL S NECROPSIED 

CONTROL LOW DOSE 

1 (2%) 

1

1

 (2*) 

 (2*) 

(38) (U7 ) 

(38) (1*7) 

(38) (K7 ) 

(«H)' (46) 

1 (2X) 

(U2 ) (1*7) 

1 (2%) 

(U2) (47) 

(«0) (47) 

1 (2%) 

(U6 ) (48) 

1 (2«) 

25 (52X) 

2 (4X) 

(U3) (44) 

(it 6) (45) 

2 (IX) 

(4H) (1*2) 

1 (2%) 

1 (2%) 

2 15*L . _ 

 EXAMINE D MICROSCOPICALLY 

84 

HIGH DOSE 

1 (2%) 

(46) 

2 (4X) 

(46) 

1 (2%) 

(46) 

1 (2«) 

(49) 

(49) 

1 (2%) 

(49) 

1 (2X) 

(47) 

2 (4%) 

(50) 

3 (6*) 

(48) 

2 (4%) 

(46) 

(38) 
TABLE D2. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

REPRODUCTIVE SYSTEM 

IUTERUS (44) (46) (46) 

HY CRONSTRA 12 (26%) 9 (20%) 

PYOHETR A 1 (2*) 

*OTERUS/ENDOMETRIUM (44) (46) (46) 

INFLAMMATION , ACUTE/CHRONIC 1 (2«) 

HYPERPLASIA, CYSTIC 30 (63%) 17 (3755) 7 (15%) 

#OVAP Y (44) (44) (45) 

MINERALIZATIO N 1 (2«) 1 (2%) 

CYST, NOS 7 (16%) 2 (5«) 4 (9X) 

HEMORBHAG E 1 (2%) 

HFMORRHAGI C CYST 4 (9*, 

ABSCESS, NOS 1 (2%) 

INFLAMMATION , CHRONIC 2 { 5%) 

NE"VO'JS SYSTEM 

*BRHI N (45 ) (46) (49) 

EPIDERMA L IN-LUStON CYST 1 (2«) 

SP^riAL SENSE OR3AN S 

NON E 

IflSCULDSKELETA L SYSTE M 

*SKELSTAL MUS:LE (49) (49) (50) 

ABSCESS, NOS 1 ( 2%) 

BODY CAVITIES 

NOS E 

ALL OTHER SYSTEMS 

MULTIPLE ORGAN S (49) (49 ) (50) 

PERIARTERITIS. . _ _1 12*1 

* NfJMEE R 0" ANIMAL S »ITH TISSUE EXAMINE D MICR3 SCDPICALLY 

* NUMBER OF ANIMAL S NECROPSIED 

85 
TABLE D2. FEMALE MICE: NONNEOPLASTIC LESIONS (CONTINUED) 

CONTROL LOW DOSE HIGH DOSE 

AMYLOIDOSIS 1 (2X) 

impose TISSCJE 

STEATITIS 1 

NECROSIS, FAT 1 

OMENTU M 

INFLAMMATION , CHRONIC "OCAL 

NECROSIS, FO~AL 

1 

1 

SPECIAL MORPH0103Y SUMMAR Y 

NO LESION REPORTED 3 2 9 

AUTO/NECBDPSY/HISTO PEHF

^UTOLYSIS/NO NECROPSY

 3 

1 

2 

1 

* NUMBE R 3P ANIMALS WITH TISSUE EXAMINED MICROSCOPIC ALLY 

* NUMBER 0? ANIMAL S NECHOPSIED 

86 
APPENDIX E
 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS
 

IN RATS FED IH-BENZOTRIAZOLE IN THE DIET
 

87
 

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

Fed IH-Benzotriazole in the Diet3
 

89

 

Topography: Morphology


Integumentary System: Fibroma^


P Values0 »d


Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Lung: Alveolar/Bronchiolar Adenoma
 

or Carcinomab


P Values0 »d


Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


 Control
 

 3/48 (6)
 

 N.S.
 

 105
 

 3/48 (6)
 

 N.S.
 

 105
 

Low
 

Dose
 

2/50 (4)
 

N.S.
 

0.640
 

0.055
 

5.345
 

89
 

1/46 (2)
 

N.S.
 

0.348
 

0.007
 

4.143
 

104
 

High
 

Dose
 

1/50 (2)
 

N.S.
 

0.320
 

0.006
 

3.822
 

83
 

2/46 (4)
 

N.S.
 

0.696
 

0.060
 

5.792
 

105
 
90

 

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

Fed IH-Benzotriazole in the Diet3
 

(continued)
 

Low High
 

Topography: Morphology Control Dose Dose
 

Hematopoietic System: Lymphoma or
 

Leukemiab 7/48 (15) 5/50 (10) 3/50 (6)
 

P Valuesc»d N.S. N.S. N.S.
 

Relative Riskf 0.686 0.411
 

Lower Limit 0.184 0.072
 

Upper Limit 2.334 1.687
 

Weeks to First Observed Tumor 80 92 101
 

Liver: Neoplastic Noduleb 0/48 (0) 0/46 (0) 5/45 (11)
 

P Values0 »d P = 0.008 N.S. P = 0.024
 

Relative Riskf Infinite
 

Lower Limit 1.348
 —
 Upper Limit Infinite
 —
 

Weeks to First Observed Tumor — 101
 —
 
91

 

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

Fed IH-Benzotriazole in the Diet3
 

(continued) 

Low High 

Topography: Morphology Control Dose Dose 

Pituitary: Adenoma, NOSb 10/45 (22) 5/40 (13) 4/45 (9)
 

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

Relative Risk^ 0.563 0.400
 

Lower Limit 0.164 0.098
 

Upper Limit 1.639 1.273
 

Weeks to First Observed Tumor 103 85 105
 

Adrenal: Pheochromocytoma° 4/46 (9) 3/44 (7) 3/46 (7)
 

P Values0 »d N.S. N.S. N.S.
 

Relative Riskf 0.784 0.750
 

Lower Limit 0.121 0.116
 

Upper Limit 4.367 4.186
 

Weeks to First Observed Tumor 78 72 105
 
92

 

Table El.


(continued)
 

Topography: Morphology
 

Thyroid: C-cell Carcinomab
 

P Values0 »d
 

Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Thyroid: C-cell Adenoma or
 

Carcinoma"
 

P Values0 »d
 

Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

 Analyses of the Incidence of Primary Tumors in Male Rats
 

Fed IH-Benzotriazole in the Diet3
 

Low 

Control Dose 

2/43 (5) 1/40 (3) 

N.S. N.S. 

0.538 

0.009 

9.907 

95 104 

5/43 (12) 1/40 (3)
 

N.S. N.S.
 

0.215
 

0.005
 

1.806
 

95 104
 

High
 

Dose
 

1/44 (2)
 

N.S.
 

0.489
 

0.008
 

9.035
 

105
 

2/44 (5)
 

N.S.
 

0.391
 

0.039
 

2.242
 

105
 
Table El. Analyses of the Incidence of Primary Tumors in Male Rats
 

Fed IH-Benzotriazole in the Dieta
 

93

 

(continued) 

Topography: Morphology Control 

Pancreatic Islets:

Adenomab

 Islet-cell 

 0/44 (0) 

P Values0 >d N.S. 

Departure from Linear Trend6 P = 0.013 

Relative Riskf 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor — 

Testis: Interstitial-cell Tumorb 37/48 (77) 

P Values0 >d N.S. 

Relative Riskf 

Lower Limit 

Upper Limit 

Weeks to First Observed Tumor 78 

Low
 

Dose
 

3/44 (7)
 

N.S.
 

Infinite
 

0.604
 

Infinite
 

100
 

38/43 (88)
 

N.S.
 

1.146
 

0.925
 

1.354
 

89
 

High
 

Dose
 

0/46 (0)
 

N.S.
 

—
 

—
 

—
 

—
 

38/46 (83)
 

N.S.
 

1.072
 

0.856
 

1.318
 

78 
__ 

94

 

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

Fed IH-Benzotriazole in the Diet3
 

(continued) 

Low High 

Topography ; Morphology Control Dose Dose 

Brain: Glioma, NOS, or
 

Oligodendrogliomab 0/46 (0) 3/44 (7) 0/46 (0)
 

P Values0 »d N.S. N.S. N.S.
 

Departure from Linear Trend6 P = 0.012
 

Relative Riskf Infinite
 

Lower Limit 0.631
 —
 Upper Limit Infinite
 —
 

Weeks to First Observed Tumor 21
 —
 

a

Dosed groups received time-weighted average doses of 6,700 or 12,100 ppm.
 

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

c

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


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

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

comparison of that dosed group with the control group when P < 0.05; otherwise, not
 

significant (N.S.) is indicated.
 

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

e

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

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

control group.
 
95

 

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

Fed IH-Benzotriazole in the Diet3
 

Low High
 

Topography; Morphology Control Dose Dose
 

Hematopoietic System: Lymphoma
 

or Leukemia*5 4/50 (8) 3/48 (6) 1/50 (2)
 

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

Relative Riskf 0.781 0.250
 

Lower Limit 0.120 0.005
 

Upper Limit 4.374 2.411
 

Weeks to First Observed Tumor 102 101 97
 

Pituitary: Adenoma, NOSb 16/40 (40) 9/46 (20) 8/47 (17)
 

P Values0 >d P = 0.010 (N) P = 0.032 (N) P = 0.016 (N)
 

Relative Riskf 0.489 0.426
 

Lower Limit 0.218 0.179
 

Upper Limit 1.039 0.936
 

Weeks to First Observed Tumor 101 65 105
 
96

 

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

Fed IH-Benzotriazole in the Diet3
 

(continued) 

Low High 

Topography: Morphology Control Dose Dose 

Pituitary: Adenoma, NOS, or
 

Chromophobe Adenoma"3 17/40 (43) 9/46 (20) 8/47 (17)
 

P Values0 »d P = 0.005 (N) P = 0.019 (N) P = 0.008 (N)
 

Relative Risk^ 0.460 0.401
 

Lower Limit 0.208 0.171
 

Upper Limit 0.962 0.867
 

Weeks to First Observed Tumor 101 65 105
 

Adrenal: Pheochromocytoma^ 6/48 (13) 2/48 (4) 1/50 (2)
 

P Values0 >d P = 0.025 (N) N.S. N.S.
 

Relative Riskf 0.333 0.160
 

Lower Limit 0.034 0.004
 

Upper Limit 1.754 1.249
 

Weeks to First Observed Tumor 106 105 78
 
97

 

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

Fed IH-Benzotriazole in the Diet3
 

(continued) 

Low High 

Topography: Morphology Control Dose Dose 

Thyroid: C-cell Carcinomab 0/43 (0) 1/43 (2) 3/50 (6)
 

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

Relative Riskf Infinite Infinite
 

Lower Limit 0.054 0.519
 

Upper Limit Infinite Infinite
 

Weeks to First Observed Tumor — 105 105
 

Thyroid: C-cell
 

Adenoma or Carcinoma^ 0/43 (0) 5/43 (12) 3/50 (6)
 

P Values0 »d N.S. P = 0.028 N.S.
 

Relative Riskf Infinite Infinite
 

Lower Limit 1.268 0.519
 

Upper Limit Infinite Infinite
 

Weeks to First Observed Tumor — 105 105
 
98

 

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

Fed IH-Benzotriazole in the Diet3
 

(continued)
 

Low High
 

Topography: Morphology Control Dose Dose
 

Mammary Gland: Fibroadenomab

 6/50 (12) 2/48 (4) 2/50 (4)
 

P Values0 »d N.S. N.S. N.S.
 

Relative Riskf 0.347 0.333
 

Lower Limit 0.036 0.034
 

Upper Limit 1.829 1.758
 

Weeks to First Observed Tumor 106 89 105
 

Uterus: Endometrial Stromal Polypb 2/48 (4) 10/45 (22) 8/50 (16)
 

P Values0 >d N.S. P = 0.010 N.S.
 

Relative Riskf 5.333 3.840
 

Lower Limit 1.222 0.818
 

Upper Limit 47.758 35.654
 

Weeks to First Observed Tumor 106 91 105
 
99

 

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

Fed IH-Benzotriazole in the Diet3
 

(continued)
 

Low High
 

Topography: Morphology Control Dose Dose
 

Uterus: Endometrial Stromal
 

Polyp or Sarcoma"3 4/48 (8) 10/45 (22) 9/50 (18)
 

P Value sc'd N.S. N.S. N.S.
 

Relative Riskf 2.667 2.160
 

Lower Limit 0.835 0.651
 

Upper Limit 10.849 9.012
 

Weeks to First Observed Tumor 106 91 105
 

a

Dosed groups received time-weighted average doses of 6,700 or 12,100 ppm.
 

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

c

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


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

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

comparison of that dosed group with the control group when P < 0.05; otherwise, not
 

significant (N.S.) is indicated.
 
Table E2. Analyses of the Incidence of Primary Tumors in Female Rats
 

Fed IH-Benzotriazole in the Dieca
 

(continued)
 

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

e

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

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

control group.
 

100

 
APPENDIX F 

ANALYSES OF THE INCIDENCE OF PRIMARY TUMORS 

IN MICE FED 1H-BENZOTRIAZOLE IN THE DIET 

101 

103

 

Table Fl.


Topography: Morphology
 

Lung: Alveolar/Bronchiolar
 

Carcinoma*3
 

P Values0>d
 

Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Lung: Alveolar/Bronchiolar
 

Adenoma or Carcinoma"
 

P Values0 »d
 

Relative Risk^
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

 Analyses of the Incidence of Primary Tumors in Male Mice
 

Fed IH-Benzotriazole in the Dieta
 

Low High
 

Control Dose Dose
 

2/39 (5) 5/43 (12) 5/46 (1 1)
 

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

2.267 2.120
 

0.398 0.371
 

22.762 21.333
 

109 106 95
 

4/39 (10) 7/43 (16) 5/46 (1 1)
 

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

1.587 1.060
 

0.440 0.246
 

6.878 5.005
 

109 106 95
 
__ 

104

 

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

Fed IH-Benzotriazole in the Diet3
 

(continued)
 

Low High
 

Topography; Morphology Control Dose Dose
 

Hematopoietic System: Malignant
 

Lymphoma, NOS° 12/39 (31) 1/45 (2) 0/48 (0)
 

P Values0 »d P < 0.001 (N) P < 0.001 (N) P < 0.001 (N)
 

Departure from Linear Trend6 P = 0.015
 

Relative Riskf 0.072 0.000
 

Lower Limit 0.002 0.000
 

Upper Limit 0.453 0.220
 

Weeks to First Observed Tumor 101 53
 

Hematopoietic System: Malignant
 

Lymphoma, Histiocytic Type'3 1/39 (3) 1/45 (2) 4/48 (8)
 

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

Relative Riskf 0.867 3.250
 

Lower Limit 0.011 0.340
 

Upper Limit 66.545 156.521
 

Weeks to First Observed Tumor 109 106 106
 
Table Fl. Analyses of the Incidence of Primary Tumors in Male mice
 

Fed IH-Benzotriazole in the Diet3
 

(continued)
 

Topography: Morphology


Hematopoietic System: All Lymphomasb


P Valuesc»d


Departure from Linear Trend6


Relative Risk^
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


All Sites: Angiosarcomab


P Values0 »d


Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


 Control
 

 13/39 (33)
 

P = 0.011 (N)
 

P = 0.043
 

 101
 

 0/39 (0)
 

 N.S.
 

—
 

Low
 

Dose
 

4/45 (9)
 

P = 0.006 (N)
 

0.267
 

0.069
 

0.782
 

53
 

2/45 (4)
 

N.S.
 

Infinite
 

0.258
 

Infinite
 

99
 

High
 

Dose
 

6/48 (13)
 

P = 0.019 (N)
 

0.375
 

0.130
 

0.953
 

106
 

3/48 (6)
 

N.S.
 

Infinite
 

0.492
 

Infinite
 

106
 

105

 
106

 

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

Fed IH-Benzotriazole in the Diet3
 

(continued)
 

Low High
 

Topography: Morphology Control Dose Dose
 

Liver: Hepatocellular Carcinoma*5 12/39 (31) 11/43 (26) 5/47 (11)
 

P Values0 »d P = 0.016 (N) N.S. P = 0.019 (N)
 

Relative Riskf 0.831 0.346
 

Lower Limit 0.379 0.105
 

Upper Limit 1.816 0.955
 

Weeks to First Observed Tumor 86 71 106
 

Liver: Hepatocellular Adenoma
 

or Carcinoma"3 12/39 (31) 12/43 (28) 7/47 (15)
 

d
 P Values0 » N.S. N.S. N.S.
 

Relative Riskf 0.907 0.484
 

Lower Limit 0.426 0.180
 

Upper Limit 1.941 1.200
 

Weeks to First Observed Tumor 86 71 106
 

a

Dosed groups received time-weighted average doses of 11,700 or 23,500 ppm.
 

"Number of tumor-bearing animals/number of animals examined at site (percent)
 
Table Fl. Analyses of the Incidence of Primary Tumors in Male Mice
 

Fed IH-Benzotriazole in the Dieta
 

(continued)
 

c

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

comparison of that dosed group with the control group when P < 0.05; otherwise, not
 

significant (N.S.) is indicated.
 

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

e

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

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

control group.
 

107

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

Fed IH-Benzotriazole


Topography: Morphology


Lung: Alveolar/Bronchiolar Carcinoma*5


P Values0 »d


Departure from Linear Trend6


Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Lung: Alveolar/Bronchiolar Adenoma
 

or Carcinomab


P Valuesc'd


Departure from Linear Trend6


Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


 Control
 

 0/49 (0)
 

 N.S.
 

 P = 0.002
 

—
 

 0/49 (0)
 

 N.S.
 

 P = 0.002
 

—
 

 in the Dieta
 

Low
 

Dose
 

9/49 (18)
 

P = 0.001
 

Infinite
 

2.631
 

Infinite
 

20
 

10/49 (20)
 

P = 0.001
 

Infinite
 

2.976
 

Infinite
 

20
 

High
 

Dose
 

3/49 (6)
 

N.S.
 

Infinite
 

0.602
 

Infinite
 

80
 

4/49 (8)
 

N.S.
 

Infinite
 

0.928
 

Infinite
 

80
 

108

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

109

 

(continued)
 

Topography ; Morphology
 

Hematopoietic System: Malignant
 

Lymphoma, NOS°
 

P Values0 »d
 

Departure from Linear Trend6
 

Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Hematopoietic System: Malignant
 

Lymphoma, Histiocytic Type^
 

P Values0 »d
 

Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Fed IH-Benzotriazole in the Diet3
 

Low
 

Control Dose
 

12/49 (24) 0/49 (0)
 

P < 0.001 (N) P < 0.001 (N)
 

P = 0.007
 

0.000
 

0.000
 

0.272
 

58
 

1/49 (2) 5/49 (10)
 

N.S. N.S.
 

5.000
 

0.589
 

231.287
 

109 20
 

High
 

Dose
 

1/50 (2)
 

P = 0.001 (N)
 

0.082
 

0.002
 

0.518
 

89
 

4/50 (8)
 

N.S.
 

3.920
 

0.407
 

188.989
 

106
 
__ 

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

Fed IH-Benzotriazole in the Dieta
 

(continued)
 

Topography : Morphology


Hematopoietic System: Malignant
 

Lymphoma, Mixed Typeb


P Values0 »d


Departure from Linear Trend6


Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Hematopoietic System: All Lymphomasb


P Valuesc»d


Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor


Low 

 Control Dose 

 0/49 (0) 3/49 (6) 

 N.S. N.S. 

P = 0.013 

Infinite 

0.602 

Infinite 

— 106 

 13/49 (27) 8/49 (16) 

 N.S. N.S. 

0.615 

0.243 

1.451 

 58 20 

High
 

Dose
 

0/50 (0)
 

N.S.
 

—
 

—
 

—
 

7/50 (14)
 

N.S.
 

0.528
 

0.195
 

1.295
 

110

 

89 
Table F2. Analyses of the Incidence of Primary Tumors in Female Mice
 

Ill

 

(continued)
 

Topography; Morphology
 

Pituitary: Adenoma, NOSb
 

P Values0 »d
 

Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Adrenal: Pheochromocytoma or
 

Pheochromocytoma Malignant"
 

P Values0 »d
 

Relative Riskf
 

Lower Limit
 

Upper Limit
 

Weeks to First Observed Tumor
 

Fed IH-Benzotriazole in the Diet3
 

Low 

Control Dose 

3/34 (9) 2/36 (6) 

N.S. N.S. 

0.630 

0.055 

5.162 

109 106 

4/46 (9) 0/45 (0)
 

P = 0.015 (N) N.S.
 

0.000
 

0.000
 

1.099
 

68
 

High
 

Dose
 

0/38 (0)
 

N.S.
 

0.000
 

0.000
 

1.472
 

—
 

0/46 (0)
 

N.S.
 

0.000
 

0.000
 

1.076
 

—
 
112

 

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

Fed IH-Benzotriazole in the Dieta
 

(continued) 

Low High 

Topography: Morphology Control Dose Dose 

Thyroid: Follicular-cell Carcinomab 2/44 (5) 0/42 (0) 1/38 (3)
 

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

Relative Riskf 0.000 0.579
 

Lower Limit 0.000 0.010
 

Upper Limit 3.524 10.653
 

Weeks to First Observed Tumor 107 106
 —
 

Thyroid: Follicular-cell Adenoma
 

or Carcinomab 4/44 (9) 1/42 (2) 2/38 (5)
 

P Values0 »d N.S. N.S. N.S.
 

Relative Riskf 0.262 0.579
 

Lower Limit 0.005 0.055
 

Upper Limit 2.505 3.789
 

Weeks to First Observed Tumor 80 106 106
 

a

Dosed groups received time-weighted average doses of 11,700 or 23,500 ppm.
 

"Number of tumor-bearing animals/number of animals examined at site (percent),
 
Table F2. Analyses of the Incidence of Primary Tumors in Female Mice
 

Fed IH-Benzotriazole in the Dieta
 

(continued)
 

c

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

comparison of that dosed group with the control group when P < 0.05; otherwise, not
 

significant (N.S.) is indicated.
 

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

e

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

•"-The	 95% confidence interval of the relative risk between each dosed group and the
 

control group.
 

113

 

Review of the Bioassay of l#-Benzotriazole* for Carcinogenicity
 

by the Data Evaluation/Risk Assessment Subgroup
 

of the Clearinghouse on Environmental Carcinogens
 

April 26, 1978
 

The Clearinghouse on Environmental Carcinogens was
 

established in May, 1976, in compliance with DHEW Committee
 

Regulations and the Provisions of the Federal Advisory
 

Committee Act. The purpose of the Clearinghouse is to
 

advise the Director of the National Cancer Institute (NCI)
 

on its bioassay program to identify and to evaluate chemical
 

carcinogens in the environment to which humans may be
 

exposed. The members of the Clearinghouse have been drawn
 

from academia, industry, organized labor, public interest
 

groups, State health officials, and quasi-public health and
 

research organizations. Members have been selected on the
 

basis of their experience in carcinogenesis or related
 

fields and, collectively, provide expertise in chemistry,
 biochemistry, biostatistics, toxicology, pathology, and
 

epidemiology. Representatives of various Governmental
 

agencies participate as ad hoc members. The Data Evaluation/
 

Risk Assessment Subgroup of the Clearinghouse is charged
 

with the responsibility of providing a peer review of
 

reports prepared on NCI-sponsored bioassays of chemicals
 

studied for carcinogenicity. It is in this context that the
 

below critique is given on the bioassay of 15-Benzotriazole
 

for carcinogenicity.
 

The primary reviewer agreed with the conclusion in
 

the report that, under the conditions of test, there was
 

no clear evidence demonstrating the carcinogenicity of
 

l#-Benzotriazole in rats or mice. After a brief description
 

of the experimental design, he said that the weight gain
 

and survival among the animals were acceptable. A few
 

gliomas were observed among treated rats, a brain tumor
 

rarely found in control animals. He suggested that if a
 

follow-up study is warranted, it should be conducted in
 

new-born animals, since they are particularly susceptible
 

to the induction of CNS tumors. Although an increased
 

incidence of lung tumors were found in one sex of mice at
 

the low dose level, the primary reviewer opined that the
 

results could be discounted since these tumors usually are
 

induced in both sexes and in a dose-related fashion.
 

115
 
The secondary reviewer commented on the lower incidence
 

of pituitary tumors in treated rats as compared to control
 

animals. A Subgroup member added that the number of lymphomas
 

also were lower among treated rats of both sexes.
 

A motion was made that the report on the bioassay of
 

Iff-Benzotriazole be accepted as written. It was further
 

moved that l#-Benzotriazole probably poses no carcinogenic
 

risk to man. The motion was seconded and approved unanimously
 

Members present were:
 

Michael Shimkin (Acting Chairman), University of California
 

at San Diego
 

Joseph Highland, Environmental Defense Fund
 

George Roush, Jr., Monsanto Company
 

Louise Strong, University of Texas Health Sciences Center
 

John Weisburger, American Health Foundation
 

Subsequent to this review, changes may have been made
 

in the bioassay report either as a result of the review
 

or other reasons. Thus, certain comments and criticisms
 

reflected in the review may no longer be appropriate.
 

116
 

4U.S. GOVERNMENT PRINTING OFFICE: 1978 260-899/3143 1-3 

DHEW Publication No. (NIH) 78-1338