NATIONAL TOXICOLOGY PROGRAM
J^chnical Report Series
No. 341

TOXICOLOGY AND CARCINOGENESIS

STUDIES OF
NITROFURANTOIN

(CAS NO. 67-20-9)

IN F344/N RATS AND B6C3Fi MICE

(FEED STUDIES)

U.S. DEPARTMENT OP^ HEALTH AND HUMAN SERVICES

Public Health Service

National Institutes of Health

QP

801

•N55

N67

FOREWORD

The National Toxicology Program (NTP) is made up of four charter agencies of the U.S. Department
of Health and Human Services (DHHS): the National Cancer Institute (NCI), National Institutes of
Health; the National Institute of Environmental Health Sciences (NIEHS), National Institutes of
Health; the National Center for Toxicological Research (NCTR), Food and Drug Administration; and
the National Institute for Occupational Safety and Health (NIOSH), Centers for Disease Control. In
July 1981, the Carcinogenesis Bioassay Testing Program, NCI, was transferred to the NIEHS. The
NTP coordinates the relevant programs, staff, and resources from these Public Health Service agen-
cies relating to basic and applied research and to biological assay development and validation.

The NTP develops, evaluates, and disseminates scientific information about potentially toxic and
hazardous chemicals. This knowledge is used for protecting the health of the American people and for
the primary prevention of disease.

The studies described in this Technical Report were performed under the direction of the NIEHS and
were conducted in compliance with NTP chemical health and safety requirements and must meet or
exceed all applicable Federal, state, and local health and safety regulations. Animal care and use
were in accordance with the Public Health Service Policy on Humane Care and Use of Animals. All
NTP toxicology and carcinogenesis studies are subjected to a comprehensive audit before being pre-
sented for public review.

These studies are designed and conducted to characterize and evaluate the toxicologic potential, in-
cluding carcinogenic activity, of selected chemicals in laboratory animals (usually two species, rats
and mice). Chemicals selected for NTP toxicology and carcinogenesis studies are chosen primarily on
the bases of human exposure, level of production, and chemical structure. Selection per se is not an
indicator of a chemical's carcinogenic potential.

These NTP Technical Reports are available for sale from the National Technical Information Service,
U.S. Department of Commerce, 5285 Port Royal Road, Springfield, VA 22161 (703-487-4650). A list-
ing of printed NTP Technical Reports appears on the inside back cover. Single copies of this Tech-
nical Report are available without charge while supplies last from the NTP Public Information Office,
NIEHS, P.O. Box 12233, Research Triangle Park, NC 27709 (919-541-3991).

Nitrofurantoin, NTP TR 341

NTP TECHNICAL REPORT
ON THE

TOXICOLOGY AND CARCINOGENESIS
STUDIES OF NITROFURANTOIN

(CAS NO. 67-20-9)

IN F344/N RATS AND B6C3Fi MICE

(FEED STUDIES)
^5 John Edgar French, Ph.D., Study Scientist

I^BS NATIONAL TOXICOLOGY PROGRAM

P.O. Box 12233

or

2^^o Research Triangle Park, NC 27709

irn

September 1989

NTP TR 341

NIH Publication No. 89-2597

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U.S. DEPARTMENT OF HEALTH AND HUMAN SERVICES

Public Health Service
National Institutes of Health

CONTENTS

PAGE

ABSTRACT 3

EXPLANATION OF LEVELS OF EVIDENCE OF CARCINOGENIC ACTIVITY 7

CONTRIBUTORS 8

PEER REVIEW PANEL 10

SUMMARY OF PEER REVIEW COMMENTS 11

I. INTRODUCTION 15

II. MATERIALS AND METHODS 23

III. RESULTS 35

RATS 36

MICE 49

IV. DISCUSSION AND CONCLUSIONS 61

V. REFERENCES 67

APPENDIXES

APPENDIX A SUMMARY OF LESIONS IN MALE RATS IN THE TWO-YEAR FEED STUDY

OF NITROFURANTOIN 77

APPENDIX B SUMMARY OF LESIONS IN FEMALE RATS IN THE TWO-YEAR FEED STUDY

OF NITROFURANTOIN 109

APPENDIX C SUMMARY OF LESIONS IN MALE MICE IN THE TWO-YEAR FEED STUDY

OF NITROFURANTOIN 137

APPENDIX D SUMMARY OF LESIONS IN FEMALE MICE IN THE TWO YEAR FEED STUDY

OF NITROFURANTOIN 163

APPENDIX E GENETIC TOXICOLOGY OF NITROFURANTOIN 193

APPENDIX F SENTINEL ANIMAL PROGRAM 201

APPENDIX G FEED AND COMPOUND CONSUMPTION BY RATS AND MICE IN THE TWO-YEAR

FEED STUDIES OF NITROFURANTOIN 205

APPENDIX H INGREDIENTS, NUTRIENT COMPOSITION, AND CONTAMINANT LEVELS IN

NIH 07 RAT AND MOUSE RATION 211

APPENDIX I AUDIT SUMMARY 217

Nitrofurantoin, NTP TR 341

07N^

O

,CH = N>

NH

O

NITROFURANTOIN

CAS No. 67-20-9

C8H6N4O5

Molecular weight 238.2

Synonyms: l-(((5-nitro-2-ruranyl)methylene)amino-2,4-imidazolidinedione);
l-(5-nitro-2-furfurylideneamino)-hydantoin; A'^-(5-nitro-2-furfurylidene)-l-aminohydantoin;
l-((5-nitrofurfurylidene)amino)hydantoin

Trade names: Benkfuran; Berkfurin; Chemiofuran; Cyantin; Dantafur; Furadantin, Furadantine;
Furadantoin; F^uradonin; Furadonine, Furantoin; Furatoin; Furobactina; Ituran; Macrodantin,
Nifurantin; NSC 2107; N-Toin; Orafuran; Parafuran; Urizept; USAF EA-2; Welfurin; Zoofurin

ABSTRACT

Nitrofurantoin was studied and evaluated because of its widespread use as a drug for treating urinary
tract infections in humans, its structural relationship to known carcinogenic 5-nitrofuran com-
pounds, and the lack of adequate studies to assess its carcinogenicity Toxicology and carcinogenesis
studies of nitrofurantoin were conducted by administering nitrofurantoin (greater than 99% pure) in
feed to groups of F344/N rats and B6C3Fi mice of each sex for 14 days, 13 weeks, or 2 years.

Fourteen-Day and Thirteen-Week Studies: None of the rats (at dietary concentrations up to 20,000
ppm) died before the end of the 14-day studies. Rats that received 5,000, 10,000, or 20,000 ppm lost
weight. Four of five male and 4/5 female mice that received 10,000 ppm and 1/5 females that received
5,000 ppm nitrofurantoin died before the end of the studies. Mice that received 5,000 ppm and male
mice that received 10,000 ppm lost weight.

In the 13-week studies, final mean body weights of rats that received 2,500, 5,000, or 10,000 ppm were
10%, 34%, or 47% lower than that of the controls for males and 15%, 31%, or 41% lower for females.
Feed consumption by dosed and control rats was generally similar. Degeneration of the germinal epi-
thelium of the seminiferous tubules of the testis was observed in male rats that received 2,500 to
10,000 ppm nitrofurantoin. Necrosis of the ovarian follicles was observed in 8/10 female rats that re-
ceived 10,000 ppm, in 3/10 females that received 5,000 ppm, and in 1/10 that received 2,500 ppm.

For mice, final mean body weights of the 5,000-ppm groups were 13% lower than that of the controls
for males and 15% lower for females. Two of 10 male mice that received 5,000 ppm and 1/10 males
that received 300 ppm died before the end of the 13- week studies. Estimated feed consumption was
similar for dosed and control groups. Degeneration of the germinal epithelium of the testis was ob-
served in males that received 1,300 to 5,000 ppm; necrosis of the ovarian follicles was observed in fe-
males that received 5,000 ppm but not in the lower dose groups. Necrosis of the renal tubular epithe-
lium was observed in 2/9 males that received 5,000 ppm.

Based on these results, 2-year studies of nitrofurantoin were conducted by feeding diets containing 0,
1,300, or 2,500 ppm nitrofurantoin to groups of 50 male F344/N rats and to groups of 50 male and

Nitrofurantoin, NTP TR 341

female B6C3Fi mice for 103 weeks. Groups of 50 female F344/N rats were fed diets containing 0, 600,
or 1 ,300 ppm nitrofurantoin on the same schedule.

Body Weight and Survival in the Two-Year Studies: Mean body weights and average daily feed con-
sumption of dosed male and female rats were similar to those of the controls throughout the studies.
The average amount of nitrofurantoin consumed per day was estimated to be 60 and 110 mg/kg for
low and high dose male rats and 30 and 60 mg/kg for low and high dose female rats. No significant
differences in the number of rats surviving to the end of the studies were observed between any
groups of rats of either sex (male: control, 24/50; low dose, 27/50; high dose, 26/50; female: 25/50;
26/50; 31/50).

Mean body weights of high dose male and female mice were up to 12% lower than those of the controls
throughout most of the studies. The average daily feed consumption by dosed mice ranged from 93%
to 100% that by controls. The average amount of nitrofurantoin consumed per day was estimated to
be 280-300 mg/kg and 570-580 mg/kg for low and high dose mice. The survival of the control group of
female mice was lower than that of the dosed groups (control, 19/50; low dose, 37/50, high dose, 37/50).
The decrease in survival was most likely related to the increase in microbial infection in the repro-
ductive tract observed in the controls. Groups of male mice had similar survival (28/50; 29/50; 34/50).

t^onneoplastic and Neoplastic Effects in the Two-Year Studies: Organs showing toxicity from nitro-
furantoin exposure identified in the short term studies were the testis in male rats and mice, the
ovary in female rats and mice, and the kidney in male mice. Lesions observed in the 2-year studies
were in the testis in male rats and mice, ovary in female mice, and kidney in male rats.

Chronic nephropathy was observed in nearly all rats, but the severity of the lesions was judged to be
greater in dosed male rats Hyperplasia of the transitional cell epithelium (control, 0/50; low dose,
5/50; high dose, 2/50) and hydronephrosis of the renal pelvis (0/50; 5/50; 2/50) were also observed in
dosed male rats. In the standard single sections of the left and right kidney from each rat, tubular cell
adenomas were observed in one low dose and two high dose males; a tubular cell carcinoma was ob-
served in another high dose male. Because the number of renal tubular cell neoplasms identified by
standard procedures in the dosed male rats was low, additional step-sections of the kidney were eval-
uated. The incidences of tubular cell adenomas derived from the step-sections and original sections
(combined) were significantly increased in dosed male rats (adenomas: 3/50; 11/50; 19/50); tubular
cell carcinomas occurred in two high dose males only.

Lesions considered to be associated with the nephropathy and nitrofurantoin exposure were observed
in male rats and included hyperplasia of the parathyroid glands (3/49; 18/47; 23/49), fibrous osteodys-
trophy of the bone (0/50; 5/50; 5/50), and mineralization of the glandular stomach ( 1/49; 8/50; 14/50).

Atypical cells of the epididymis (0/50; 0/50; 12/50) and degeneration of the testis (0/50; 0/50; 36/50)
were observed in high dose male rats. Fibrinoid necrosis of arterioles (1/50; 8/50; 15/50) and perivas-
cular infiltration of mononuclear cells (3/50; 9/50, 19/50) were also observed in the testis of male rats.
Interstitial cell adenomas of the testis occurred with a negative trend (47/50; 45/50; 21/50), and no
adenomas or carcinomas of the preputial gland were seen in high dose male rats (12/48; 11/50; 0/47).
The incidence of clitoral gland neoplasms was increased in low dose female rats (5/44; 10/38; 4/42).

Osteosarcomas were observed in the bone of one low dose and two high dose male rats. The historical
incidence of osteosarcomas in untreated male F344/N rats is 8/1,937 (0.4%). The incidences of subcu-
taneous tissue neoplasms in dosed male rats were greater than that in the controls (1/50; 7/50; 5/50).

No neoplastic lesions in dosed female rats or male mice were considered to be compound related at the
doses of nitrofurantoin administered.

Nitrofurantoin, NTP TR 341

For female mice, ovarian atrophy was observed in 48/50 low dose and 49/50 high dose mice but not in
controls. Tubular cell adenomas of the ovary (0/50; 0/50; 5/50), benign mixed tumors (tubular and
stromal) (0/50; 0/50; 4/50), and granulosa cell tumors (0/50; 3/50; 2/50) were observed in dosed female
mice. One granulosa cell tumor in the high dose group was malignant. Ovarian abscesses (18/50) and
suppurative inflammation of the uterus ( 1 1/50) were observed in control female mice but not in dosed
female mice and are believed to be related to indigenous microbial infections and most likely were the
cause of early deaths in this group Adenocarcinomas of the uterus were seen in one low dose and in
one high dose mouse.

Testicular aspermatogenesis ( 1/49; 1/49; 16/50), degeneration of the germinal epithelium (0/49; 3/49;
23/50), and atypical cells (0/50; 0/49; 26/50) and depletion ( 1/50; 1/49, 15/50) of the epididymis were ob-
served at increased incidences in high dose male mice.

Spindle cell hyperplasia of the adrenal cortex was observed in dosed female mice (3/50, 41/50, 45/50).
A spindle cell adenoma (adrenal capsule adenoma) was seen in one low dose female mouse, and a
spindle cell carcinoma (adrenal capsule carcinoma) was seen in one low dose male mouse.

Mineralization of the renal medulla (male: 0/50; 0/50; 17/50; female: 0/50; 0/50; 7/50) and dilatation
of the renal tubules (male: 0/50; 0/50; 14/50) were observed in high dose mice.

Hepatocellular neoplasms (adenomas or carcinomas, combined) were observed at an increased inci-
dence in high dose female mice (2/50; 2/50; 8/50) An Ito cell tumor of the liver was observed in one
low dose and one high dose female mouse Malignant lymphomas occurred in female mice (12/50;
19/50; 24/50).

Genetic Toxicology: Nitrofurantoin was mutagenic in Salmonella typhimurium strains TA98 and
TAIOO, with and without metabolic activation, but was not mutagenic for strains TA1535 or TA1537.
Nitrofurantoin induced forward mutations at the TK"^'" locus of L5178Y mouse lymphoma cells in
the absence of metabolic activation (it was not tested with activation). Nitrofurantoin induced in-
creased numbers of sister chromatid exchanges and chromosomal aberrations in cultured Chinese
hamster ovary cells with and without metabolic activation. Results of the sex-linked recessive lethal
assay in Drosophila were negative after administration of nitrofurantoin by feeding or by injection.

Conclusions: Under the conditions of these 2-year feed studies, there was some evidence of carcino-
genic activity* of nitrofurantoin for male [''344/N rats as shown by increased incidences of uncommon
kidney tubular cell neoplasms. Uncommon osteosarcomas of the bone and neoplasms of the subcuta-
neous tissue were observed in dosed male rats. Incidences of interstitial cell adenomas of the testis
and neoplasms of the preputial gland were decreased in the 2,500 ppm group of male rats. There was
no evidence of carcinogenic activity of nitrofurantoin for female F344/N rats fed diets containing 600
ppm or 1 ,300 ppm for 2 years. Female rats may have been able to tolerate higher doses. There was no
evidence of carcinogenic activity of nitrofurantoin for male B6C3Fi mice fed diets containing 1,300
ppm or 2,500 ppm for 2 years. There was clear evidence of carcinogenic activity of nitrofurantoin for
female B6C3Fi mice as shown by increased incidences of tubular adenomas, benign mixed tumors,
and granulosa cell tumors of the ovary.

Nonneoplastic lesions considered related to nitrofurantoin exposure were chronic nephropathy and
associated lesions (hyperplasia of the parathyroid gland, fibrous osteodystrophy of the bone, and
mineralization of the glandular stomach) in male rats and testicular degeneration in male rats and
mice. Ovarian atrophy and hyperplasia of the adrenal cortex spindle cells were observed in dosed fe-
male mice.

*Explanationof Levels of Evidence of Carcinogenic Activity is on page 7.

A summary of the Peer Review comments and the pub he discussion on this Technical Report appears on pages 11-13.

Nitrofurantoin, NTPTK341

SUMMARY OF THE TWO-YEAR FEED AND GENETIC TOXICOLOGY STUDIES OF NITROFURANTOIN

Male F344/N Rats

Female F344/N Rats

Male B6C3F, Mice

Female B6C3F, Mice

Dietary concentrations

1.300 or 2.500 ppm
nitrolurantoin

600 or 1.300 ppm
nitrofurantom

1.300 or 2,500 ppm
nitrofurantoin

1.300 or 2,500 ppm
nitrofurantoin

Survival rates in the 2-year study

24/50; 27/50; 26/50 25/50; 26/50; 3 1/50

Nonneoplastic effects

Chronic nephropathy;
testicular degeneration

None

Neoplastic effects

Renal tubular cell neoplasms None

28/50; 29/50; 34/50

Testicular degeneration

None

19/50; 37/.50; 37/50

Ovarian atrophy; hyperplasia
of adrenal cortex spindle cells

Tubular adenomas, benign
mixed tumors, and granulosa
cell tumors of the ovary

Level of evidence of qiircinogenic activity
Some evidence No evidence

No evidence

Clear evidence

Genetic toxicology

Mutagenic in S. typhimunum strains TA98 and 'I'AIOO with and without metabolic activation; induced forward mutations in
mouse L5178Y lymphoma cells without activation; induced increased numbers of sister chromatid exchanges and chromosomal
aberrations in cultured Chinese hamster ovary cells with and without 89; did not induce sex-linlced recessive lethal mutations
in Drosophila

Nitrofurantoin, NTPTH341

EXPLANATION OF LEVELS OF EVIDENCE OF CARCINOGENIC ACTIVITY

The National Toxicology Program describes the results of individual experiments on a chemical agent and notes the strength of
the evidence for conclusions regarding each study. Negative results, in which the study animals do not have a greater incidence
of neoplasia than control animals, do not necessarily mean that a chemical is not a carcinogen, inasmuch as the experiments are
conducted under a limited set of conditions. Positive results demonstrate that a chemical is carcinogenic for laboratory animals
under the conditions of the study and indicate that exposure to the chemical has the potential for hazard to humans. Other
organizations, such as the International Agency for Research on Cancer, assign a strength of evidence for conclusions based on
an examination of all available evidence including: animal studies such as those conducted by the NTP, epidemiologic studies,
and estimates of exposure. Thus, the actual determination of risk to humans from chemicals found to be carcinogenic in labora-
tory animals requires a wider analysis that extends beyond the purview of these studies.

Five categories of evidence of carcinogenic activity are used in the Technical Report series to summarize the strength of the evi-
dence observed in each experiment: two categories for positive results ("Clear Evidence" and "Some Evidence"); one category
for uncertain findings ("Equivocal Evidence"); one category for no observable elTects ("No Evidence"); and one category for ex-
periments thatbecauseofmajor flaws cannot be evaluated ("Inadequate Study"). These categories of interpretative conclusions
were first adopted in June 1983 and then revised in March 1986 for use m the Technical Reports series to incorporate more
specifically the concept of actual weight of evidence of carcinogenic activity. For each separate experiment (male rats, female
rats, male mice, female mice), one of the following quintet is selected to describe the findings. These categories refer to the
strength of the experimental evidence and not to either potency or mechanism.

• Clear Evidence of Carcinogenic Activity is demonstrated by studies that are interpreted as showing a dose-related
(i) increase of malignant neoplasms, (ii) increase of a combination of malignant and benign neoplasms, or (lii) marked
increase of benign neoplasms if there is an indication from this or other studies of the ability of .-lUch tumors to progress
to malignancy.

• Some Evidence of Carcinogenic Activity is demonstrated by studies that are interpreted as showing a chemically
related increased incidence of neoplasms (malignant, benign, or combined) m w. hich the strength of the response is le.ss
than that required for clear evidence.

• Equivocal Evidence of Carcinogenic Activity is demonstrated by studies that are interpreted as showing a mar-
ginal increase of neoplasms that may be chemically related.

• No Evidence of Carcinogenic Activity is demonstrated by studies that are interpreted as showing no chemically re-
lated increases in malignant or benign neoplasms.

• Inadequate Study of Carcinogenic Activity is demonstrated by studies that because of major qualitative or quanti-
tative limitations cannot be interpreted as valid for showing either the presence or absence of carcinogenic activity.

When a conclusion statement for a particular experiment is selected, consideration must be given to key factors that would ex-
tend the actual boundary of an individual category of evidence. This should allow for incorporation of scientific experience and
current understanding of long-term carcinogenesis studies in laboratory animals, especially for those evaluations that may be
on the borderline between two adjacent levels. These considerations should include:

• The adequacy of the experimental design and conduct;

• Occurrence of common versus uncommon neoplasia;

• Progression (or lack thereof) from benign to malignant neoplasia as well as from preneoplastic to neoplastic lesions;

• Some benign neoplasms have the capacity to regress but others (of the same morphologic type) progress. At present, it
is impossible to identify the difference. Therefore, where progression is known to be a possibility, the most prudent
course is to assume that benign neoplasms of those types have the potential to become malignant;

• Combining benign and malignant tumor incidences known or thought to represent stages of progression in the same or-
gan or tissue;

• Latency in tumor induction;

• Multiplicity in site-specific neoplasia;

• Metastases;

• Supporting information from proliferative lesions (hyperplasia) in the same site of neoplasia or in other experiments
(same lesion in another sex or species);

• The presence or absence of dose relationships;

• The statistical significance of the observed tumor increase;

• The concurrent control tumor incidence as well as the historical control rate and variability for a specific neoplasm;

• Survival-adjusted analyses and false positive or false negative concerns;

• Structure-activity correlations; and

• In some cases, genetic toxicology.

Nitrofurantoin, NTP TR 341

CONTRIBUTORS

The NTP Technical Report on the Toxicology and Carcinogenesis Studies of Nitrofurantoin is based
on the 13-week studies that began in May 1980 and ended in August 1980 and on the 2-year studies
that began in February 1981 and ended in March 1983 at Southern Research Institute (Birmingham,
AL).

National Toxicology Program
(Evaluated Experiment, Interpreted Results, and Reported Findings)

John P^dgar PVench, Ph.D., Study Scientist

John Bucher, Ph.D. Joseph K. Haseman, PhD

Scot L. Eustis, D.V.M., Ph.D. James Huff, Ph.D.

(Discipline Leaders and Principal Contributors)

Jack Bishop, PhD G.N Rao, D.V.M., Ph.D.

Douglas W Bristol, Ph.D. B.A. Schwetz, D. V.M., Ph.D.

R. Chhabra, PhD M. Vernon, PhD

C.W. Jameson, PhD Douglas Walters, Ph.D.
E.P:. McConnell,D.VM.

NTP Pathology Working Group
(Evaluated Slides and Prepared Pathology Report for Rats on 3/13/86)

John Seely, D.V M. (Chair) (PATHCO, Inc.) Peter Millar, M.V.M., MR CVS. (Experimental
Michael p:iwell. D.V.M . Ph D (NTP) Pathology Laboratories, Inc )

Scot L. Eustis, D. V.M., Ph.D. (NTP) Steven Stefanski, D V.M. (NTP)

A.W Macklin, D.V.M., Ph.D. (Burroughs Roger Thompson, D V.M., Ph.D. (Southern
Wellcome Laboratories) Research Institute)

(Flvaluated Slides and Prepared Pathology Report for Mice on 8/28/85)

Roger Alison, M.R.C.V.S. (Chair) (NTP) Kunitoshi Mitsumori, D.V M., Ph. D (NTP)

Gary Boorman, D.V.M., Ph.D. (NTP) Charles Montgomery, D V M. (NTP)

Hershell Giles, D.V.M. (Southern Kenneth Pierce, D.V M., Ph.D. (NTP

Research Institute) (Observer) Guestworker)

Melvin Hamlin, D V.M (Experimental Bernard Sass, V.M.D. (National Cancer Institute)

Pathology Laboratories, Inc.) (Observer) Robert Scully, M D. (Massachusetts General
Robert Maronpot, D.V.M. (NTP) Hospital)

(Performed Supplemental Review of Kidney Lesions on 3/23/88)

Micheal Jokinen, D V M (Chair) (NTP) C.P Peter, D.V.M, Ph.D. (Merck Sharpe &

Gary Boorman, D V M , Ph 1) (NTP) Dohme Research Laboratories)

Michael Elwell, D.V.M., PhD (NTP) Brian Short, D.V.M (Chemical Industry Institute

Scot L. Eustis, D.V.M., Ph.D. (NTP) ofToxicology)

Michael Lipsky, M.D. (University of Maryland)

Nitrofurantoin, NTP TR 341

CONTRIBUTORS (Continued)

Principal Contributors at Southern Research Institute
(Conducted Studies and Evaluated Tissues)

David Prejean, Ph.D. Roger B. Thompson, D. V.M.

Ruby 11. James, B S. H. Giles, D.V M.

Principal Contributors at Experimental Pathology Laboratories, Inc.
(Provided Pathology Quality Assurance)

J Gauchat Melvin Hamlin, D.V. M.

Peter Millar, M.V.M., MR. CVS.

Principal Contributors at Carltech Associates, Inc.
(Contractor for Technical Report Preparation)

William D. Theriault, Ph D. John Warner, M.S.

Abigail C. Jacobs, Ph.D. Naomi Levy, B.A.

Nitrofurantoin, NTP TR 341

PEER REVIEW PANEL

The members of the Peer Review Panel who evaluated the draft Technical Report on nitrofurantoin
on July 14, 1987, and on April 18, 1988, are listed below. Panel members serve as independent scien-
tists, not as representatives of any institution, company, or governmental agency. In this capacity,
Panel members have five major responsibilities: (a) to ascertain that all relevant literature data have
been adequately cited and interpreted, (b) to determine if the design and conditions of the NTP stud-
ies were appropriate, (c) to ensure that the Technical Report presents the experimental results and
conclusions fully and clearly, (d) to judge the significance of the experimental results by scientific cri-
teria, and (e) to assess the evaluation of the evidence of carcinogenicity and other observed toxic
responses.

National Toxicology Program Board of Scientific Counselors
Technical Reports Review Subcommittee

Robert A. Scala, Ph.D. (Chair)

Senior Scientific Advisor, Medicine and P^nvironmental Health Department

Research and Environmental Health Division, Exxon Corporation

East Millstone, N.J

Michael A Gallo, PhD

Associate Professor, Director of Toxicology
Department of Environmental and Community

Medicine, UMDNJ - Rutgers Medical School
Piscataway, NJ

Frederica Perera, Dr. PH.*

Division of P^nvironmental Sciences
School of Public Health
Columbia University
New York, NY

Ad Hoc Subcommittee Panel of Experts

John Ashby, Ph D (Principal Reviewer) William Lijinsky, PhD *

Director, Chemical Carcinogenesis

Imperial Chemical Industries, PLC
Central Toxicology Laboratory
Alderley Park, England

Charles C Capen, D V M , Ph.D.

Department of Veterinary Pathobiology
Ohio State University
Columbus, OH

Vernon M. Chinchilli, Ph.D. (Principal Reviewer)
Department of Biostatistics
Medical College of Virginia
Virginia Commonwealth University
Richmond, VA

Kim Hooper, Ph D.

Hazard Evaluation System and

Information Services
Department of Health Services
State of California
Berkeley, CA

Donald H.Hughes, Ph D.

Scientific Coordinator, Regulatory Services

Division, The Procter and Gamble Company
Cincinnati, OH

P'rederick Cancer Research Facility
Frederick, MD

Franklin E Mirer, Ph D **

Director, Health and Safety Department
International Union, United Auto
Workers, Detroit, MI

James A. Popp, D V.M., Ph D (Principal
Reviewer) Head, Department of
Experimental Pathology and Toxicology

Chemical Industry Institute of Toxicology

Research Triangle Park, NC

Andrew Sivak, Ph D.

Vice President, Biomedical Science
Arthur D. Little, Inc.
Cambridge, MA

•Unable to attend July 14. 1987.mppling
'♦Unable to attend April 18, 1988, meeting

Nitrofurantoin, NTP TR 341

10

SUMMARY OF PEER REVIEW COMMENTS
ON THE TOXICOLOGY AND CARCINOGENESIS STUDIES OF

NITROFURANTOIN

On July 14, 1987, the draft Technical Report on the toxicology and carcinogenesis studies of nitrofu-
rantoin received peer review by the National Toxicology Program Board of Scientific Counselors'
Technical Reports Review Subcommittee and associated Panel of Flxperts. The review meeting was
held at the National Institute of Environmental Health Sciences (NIEHS), Research Triangle Park,
NC.

Dr. J.E. French, NIEHS, introduced the studies by reviewing the experimental design, results, and
proposed conclusions (some evidence of carcinogenic activity for male rats, no evidence of carcino-
genic activity for female rats, no evidence of carcinogenic activity for male mice, clear evidence of car-
cinogenic activity for female mice).

Dr. Popp, a principal reviewer, agreed with the conclusions for female rats and male and female mice;
he said that the Panel should discuss the concomitant ovarian toxicity in female mice Dr. Popp
opined that the results for male rats, a slight increase in renal tubular neoplasms (control, 0/50; low
dose, 1/50; high dose, 3/50) coupled with a corresponding lack of an increase in renal tubular hyper-
plasia, more closely supported equivocal evidence of carcinogenic activity. Dr. French acknowledged
that the lack of hyperplasia must be considered but felt that the presence of a carcinoma was evidence
supporting progression and, although the numbers of renal tubular cell tumors were relatively low,
there was a twentyfold difference between the high dose and mean historical control incidences. Dr.
Hughes pointed out that there was one study in the historical control data base with two tubular neo-
plasms. Dr. J. Haseman, NIEHS, reported that for the most recent 73 corn oil gavage and feed stud-
ies, 57 had a zero incidence of tubular neoplasms in controls, 15 had an incidence of one, and 1 had an
incidence of two.

As a second principal reviewer, Dr. Ashby agreed with the conclusions for male and female rats and
male mice, while suggesting that the conclusion for female mice be changed to some evidence of carci-
nogenic activity. He stated that two of the three types of ovarian tumors were observed only in the
high dose groups. He questioned whether those tumors could be combined for assessment. Also con-
founding the interpretation was the presence of ovarian atrophy in almost all of the exposed animals.
Dr. French remarked that both ovarian tubular adenomas and benign mixed tumors were uncommon
and histogenetically it was considered appropriate to combine them.

As a third principal reviewer. Dr. Chinchilli agreed with the conclusions for male and female rats and
male mice, noting that osteosarcomas of the bone and subcutaneous tumors observed in male rats are
uncommon. For female mice, he questioned why statistical analyses for the ovary were based on a
sample of 50 when tissues from only 39 control mice were available for microscopic evaluation Dr. S.
Eustis, NIEHS, explained that ovaries from all 50 control female mice were examined; however, ovar-
ian abcesses had destroyed much of the tissues from 1 1 animals. In his opinion, the examination was
sufficient to determine whether a tumor was present. Dr. Haseman commented that whether the de-
nominator was 39 or 50, the differences in tumor incidences were highly significant and quite
striking. Dr. Chinchilli inquired if a statistical comparison test using historical control data could be
used in analysis of uncommon tumors. Dr Haseman agreed that rare or uncommon tumors might be
the one instance in which a formal analysis incorporating historical data should be considered, al-
though lack of agreement as to which test is most appropriate was still a problem

Dr. William H. Butler, of the British Industrial Biological Research Association and representing
Norwich Eaton Pharmaceuticals, Inc , presented a review of his observations from an examination of
the slides containing ovary sections from the female mice. He contended that the occurrence of ovari-
an abcesses in a number of controls obviated a proper analysis. He also suggested that the tubular

11 Nitrofurantoin, NTPTR 341

SUMMARY OF PEKR REVIEW COMMENTS (Continued)

cell adenomas might have resulted from hormonal stimulation due to ovarian atrophy and that the
existence of other negative studies supported equivocal evidence of carcinogenic activity. Dr. Butler
opined further that there was no evidence of carcinogenic activity in male rats because the incidence
of renal tubular cell neoplasms was low and within the expected historical range, because there was
no evidence of similar lesions in female rats, because there was no increase in hyperplasia, and be-
cause there was a high incidence of chronic nephropathy. Dr. E. McConnell, NIEHS, emphasized that
both increases and decreases in hyperplasia are considered in the evaluations In the case of the renal
tumors in male rats, the lack of hyperplasia was noteworthy but did not necessarily offset the in-
crease in an uncommon tumor.

Dr. Popp moved that the conclusion for male rats be changed to equivocal evidence of carcinogenic ac-
tivity and that the conclusion for female rats, no evidence of carcinogenic activity, be accepted as
written. Dr. Ashby seconded the motion, which was defeated by four votes (Dr. Ashby, l)r Chinchilli,
Dr. Hooper, and Dr. Mirer) to three (Dr. Gallo, Dr Popp, and Dr. Sivak), with two abstentions (Dr.
Capen and Dr. Hughes). Dr. Hooper moved that the conclusions be accepted as written for male rats,
some evidence of carcinogenic activity, and for female rats, no evidence of carcinogenic activity Dr
Ashby seconded the motion, which was approved by four votes (Dr. Ashby, Dr. Chinchilli, Dr. Hooper,
and Dr. Mirer) to three (Dr. Gallo, Dr Popp, and Dr. Sivak), with two abstentions (Dr. Capen and Dr.
Hughes). Dr. Popp moved that the conclusions for male mice, no evidence of carcinogenic activity,
and for female mice, clear evidence of carcinogenic activity, be accepted as written. Dr. Chinchilli
seconded the motion, which was approved by five votes to two (Dr. Ashby and Dr. Gallo), with two ab-
stentions (Dr. Capen and Dr. Hughes)

Update and Reevaluation of Further I'athology on Kidneys from Male Rats
(April 18, 1988)

At the Peer Review meeting on April 18, 1988, Dr French summarized the discussion from the Peer
Review meeting on July 14, 1987, when an important portion of the discussion focused on the tubular
cell neoplasms in the kidney of dosed male rats and the level of evidence for carcinogenic activity rec-
ommended by the staff. The level of evidence selected for male rats (some evidence of carcinogenic ac-
tivity) was based on: dose-related, albeit marginally increased, incidences of uncommon neoplasms of
the tubular cells in the kidney (0/50, 1/50; 3/50) (see Table 1 1 ), the possibility of progression to malig-
nancy as evidenced by a tubular cell carcinoma in the kidney in a high dose male rat, and comparison
with historical controls.

Dr. French went on to explain that because of the microscopic size of the majority of these tumors and
questions concerning the dose-response relationship, additional histologic sections of the kidneys
were prepared for evaluation. The purpose was to obviate the possibility of bias due to chance and to
determine if the number of tumors found in each group would increase proportionally in relation to
dose. The data derived only from the additional step-sections are shown in Table 12. All of the addi-
tional kidney tumors were observed microscopically, and there was an increase in the number of mul-
tiple adenomas observed in the high dose male rats The composite results of both data sets are shown
in Table 13 The incidences of tubular cell neoplasms in the kidney of male rats were as follows: con-
trol, 3/50; low dose, 11/50; high dose, 20/50. The low dose incidence was statistically different from
that in the controls at the 0.05 level, and the high dose incidence was statistically different at the
0.001 level.

Dr. French said that the data indicate that male rats receiving 0, 1,300, or 2,500 ppm nitrofurantoin
in feed at for 2 years developed compound related tubular cell neoplasms and that these data support
the conclusion previously written in the Technical Report and approved by the Panel. Summary

Nitrofurantoin, NTPTR 341 12

SUMMARY OF PKER REVIKW COMMENTS (Continued)

tables and representative photomicrographs of selected tumors were included in the Results section of
the Report, and the Discussion section was modified to reflect the additional findings

Discussion among the Panel members and the staff centered around several issues: the size and rela-
tive numbers of lesions in the recut sections vs. the original histologic sections (it was noted that all of
the adenomas were quite small and that there were no concomitant increases in hyperplasia and car-
cinomas in the recut sections); which sets of numbers could be compared with, or added to, the his-
torical control data base (it was agreed that only the original incidences could be compared or added);
whether the Panel should move to affirm or change the level of evidence in male rats; and the generic
issue of when and why the NTP should return to a study and do additional sections (there was agree-
ment that this would not be done routinely).

Dr. Hooper moved that the conclusion for male rats be changed to clear evidence of carcinogenic activ-
ity. Dr. Perera seconded the motion, which was defeated by five votes (Dr. Ashby, Dr. Capen, Dr.
Gallo, Dr. Popp, and Dr Sivak) to four ( Dr Chinchilli, Dr. Hooper, Dr. Lijinsky, and Dr. Perera), with
one abstention (Dr Hughes). Dr. Popp moved that the Panel concur with the staffs original recom-
mendation, some evidence of carcinogenic activity The motion was seconded and approved by five
votes (Dr. Ashby, Dr. Capen, Dr Gallo, Dr. Popp, and Dr Sivak) to four (Dr. Chinchilli, Dr. Hooper,
Dr. Lijinsky, and Dr Perera), with one abstention (Dr. Hughes).

13 Nitrofurantoin, NTP TR 341

Nitrofurantoin, NTP TR 341 14

I. INTRODUCTION

Use, Production, and Exposure
Absorption, Metabolism, and Excretion
Acute Toxicity

Cellular and Subcellular Toxicity
Epidemiology and Systemic Toxicity
Reproductive and Developmental Toxicity
Long-Term Toxicity and Carcinogenicity
Genetic Toxicology
Study Rationale

15 Nitrofurantoin, NTP TR 341

I. INTRODUCTION

o

0?N~

,CH^N-

NITKOFURANTOIN

CASNo. 67 20-9

NH

\

O

C8H6N4O5

Molecular weight 238.2

Synonyms: l-(((5-nitro-2-furanyl) methylene )amino-2, 4- imidazolidinedione);
l-(5-nitro-2-furfurylideneamino)-hydantoin; jV-(5-nitro-2-furfurylidene)-l-aminohydantoin;
l-((5-nitrofurfurylidene)amino)hydantoin

Trade names: fienkfuran; Bcrkfurin; Cheiiiiofiiran; Cyantin; Dantafur; Furadanlin; Furadant ine,
F^uradantoin; Furadonin, Furadonine, Furantoin; Furatoin; Furobactina; Itiiran; Macrodantin;
Nifurantin, NSC 21U7; N Toin, Orafuran, Parafuran, Urizept; USAF RA-2, Welfurin; ZooCurin

Use, Production, and Exposure

Nitrofurantoin is used extensively in the treat-
ment of urinary tract infections in humans
(D'Arcy, 1985) A derivative of 5-nitrofuran, ni-
trofurantoin is structurally related to furan and
to nitrofurazone, the first 5-nitrofuran described
by Dodd and Slillman (1944) to be an effective
broad-spectrum antibiotic, as well as to many
other nitrofurans (Bryan, 1978) The 5-nitro-
furan derivatives have been used extensively in
topically and parenterally administered antisep-
tics in humans and animals and as antitumor
agents, food preservatives, and feed additives for
food production animals.

The starting material for synthesis of 5-nitrofu
rans with antimicrobial properties is 2-furalde-
hyde (Ichikawa, 1978). 2-Furaldehyde is con-
verted by air oxidation and metal catalysts to
furoic acid and is thermally decarboxyiated to
furan. Preferential eiectrophilic substitution of
the furan ring occurs at the 2-position How
ever, nitration of the furan nucleus in the 2 and
5 positions is favored under conditions of fuming
nitric acid and acetic anhydride (containing the
active species CH3C02"N02"^). The 2,5-dini-
trofuran is converted to 5-nitrofuran by weak
bases, which eliminate acetic acid.

Clinical use of nitrofurantoin began after World
War II, between 1953 and 1984, an estimated
121,430,000 courses of therapy were given, ac-
cording to data from one manufacturer (D'Arcy,
1985). Nitrofurantoin treatment for infections
may occur over periods of up to 30 months
(Simonian et al., 1977) Recent production fig-
ures are not available, but in 1974, commercial
production was reported to the International
Trade Commission (USITC, 1976) (implying
that production was greater than 1,000 lb/year)
and was listed with the U.S. Environmental
Protection Agency TSCA in 1980 (NIOSH,
1983). In 1986, 9,300 kg of nitrofurantoin in
various preparations was purchased by drug-
stores and hospitals (U.S. Pharmaceutical Mar-
ket Data Base, 1986). Exposure to nitrofuran-
toin in the United States has been estimated at
8,900 kg/year (NCI/SRI, 1978).

Absorption, Metabolism, and Excretion

After oral or parenteral administration, nitrofu-
rantoin is rapidly absorbed and is excreted pri-
marily unchanged in the urine and bile of hu-
mans (Conklin and Ilailey, 1969; Conklin,
1972a,b), rats (Paul, HE., et al., 1960; Buzard et
al., 1961; Veronese et al., 1974; Wierzba et al.,
1982), mice (Maiti and Banerjee, 1978), and dogs

Nitrofurantoin, NTP TR 341

16

I. INTRODUCTION

(Conklin and Wagner, 1971). Sites of optimal
absorption of nitrofurantoin in the gastrointes-
tinal tract may vary between humans (duo-
denum) (Conklin and Hailey, 1969) and rodents
(mice, ileum) (Maiti and Banerjee, 1978).

After a single oral administration of 50 or 150
mg nitrofurantoin to healthy male volunteers
(19-43 years old, 63-96 kg), the terminal elimi-
nation half-life was 1 .2 or 1.7 hours, respectively
(Liedtke et al., 1980). Intravenous administra-
tion of 50 mg nitrofurantoin (male, 25-35 years
old, 62-80 kg) resulted in a half-life value of 58
± 15 minutes, and 47% ± 13% and 12% ± 0 3%
were excreted unchanged as parent compound
and aminofurantoin, respectively, in the urine
(Floener and Patterson, 1981). These half-life
values are significantly longer than those in
earlier reports that suggested approximately
50% of the administered dose was excreted in
humans within 20-30 minutes (Reckendorf et
al., 1962; Paul and Paul, 1964; Schirmeister et
al, 1965; Sachs et al., 1968; Conklin, 1972a;
Bron et al , 1979). Liedtke et al. (1980) sug-
gested that this discrepancy in half-life is due to
improved analytical methods (high-performance
liquid chromatography vs. spectrophotometry).
A decrease in urine pH increased the half-life,
which suggests that changes in pH may influ-
ence dissolution, bioavailability, and/or the rate
of excretion in humans (Bron et al., 1979). Ab-
sorption of nitrofurantoin is increased with the
presence of food in the gastrointestinal tract
(Bates et a!., 1974; Rosenberg and Bates, 1976;
Hoener and Patterson, 1981). Absorption of ni-
trofurantoin in humans (male, 21-32 years old)
was also influenced by the presence and size of
the macrocrystals in the formulation (Bates et
al., 1974; Meyer et al., 1974). During conditions
of renal impairment, nitrofurantoin excretion is
greatly diminished (Kunin, 1972).

Under aerobic conditions, the reduction of nitro-
furantoin by the addition of an electron to the 5-
nitrofuran ring via a nitroreductase, NADPH,
and a flavoprotein has been reported to occur in
vitro in hepatic and/or lung microsomes from
rats (male, CD, 160-180 g. Mason and Holtzman,
1975a; male, HLA-SD, 150 g, Boyd et al., 1979a;
male, Sprague Dawley, 135-140 g, Sasame and
Boyd, 1979), chickens (Leghorn, 8 days old.

Peterson et al., 1982a), guinea pigs (age and sex
not specified, 400-600 g, Leskovac and Popovic,
1980), or Erhlich ascites tumor cells (Biaglow et
al., 1977). This results in a transient nitroaro-
matic anion radical that may react with molecu-
lar oxygen, producing superoxide anion free
radical, and possibly hydrogen peroxide and the
regeneration of nitrofurantoin. Oxidative me-
tabolism of the nitrofurantoin side chain has
also been reported to occur (Pugh et al., 1972).
Jonen and Kaufman (1980) reported that in rats
(male, Sprague Dawley, 250-300 g, age not spe-
cified), 3-methylcholanthrene and P-nitrofia-
vone, but not phenobarbital, pretreatment in
creased the clearance of napthofurantoin from
the isolated perfused liver and increased the
formation of a polar metabolite, similar to a hy-
droxylated furan derivative ( l-[[(5-ac(-nitro-4,5-
dihydro-4-oxo-2-furanyl)-methylene]amino|-2,4-
imidazolidinedione) (Olivard et al., 1976)

Reductive metabolism of nitrofurantoin under
anaerobic conditions has been described in both
rodents and bacteria. Without oxygen, nitrofu-
rantoin is believed to be permanently reduced to
nitroso and/or hydroxy lamine forms (Mason and
Holtzman, 1975b, Biaglow et al , 1977; Leskovac
and Popovic, 1980). Aufrere et al. ( 1978) studied
the reductive metabolism of nitrofurantoin un-
der anaerobic conditions with young male
Sprague Dawley rats (60 g) and reported that
the metabolism of nitrofurantoin was greatest in
homogenates of cecum and colon contents of
germ-free acclimatized and control rats but not
germ-free rats and in liver, small intestine
walls, and kidney (in decreasing order of activi-
ty) in all groups. Nitrofurantoin was reduced
under these conditions to two metabolites, l-[((3-
cyano-l-oxopropyD-methylenel-amino 1-2,4- imi-
dazolidinedione (major) and l-[[(5-amino-2-fu-
ranyl)methylene|-amino|-2,4-imidazolidinedi-
one (aminofurantoin) (minor). Two other path-
ways were reported by Olivard et al. (1962) to oc-
cur in the gastrointestinal tract: reduction of ni-
trofurantoin to the 5-aminofuran and acetyla-
tion to form the 5-acetamidofuran or 5-diacetyl-
aminofuran, and acid hydrolysis of the azo-
methine bridge to produce 5-nitro-2-furanocar-
boxaldehyde, which may be excreted as 5-nitro-
2-furoic acid or as a hydrazine derivative, which
may be acetylated and excreted.

17

Nitrofurantoin, NTP TR 34 1

I. INTRODUCTION

Nitrofurantoin is excreted rapidly in adult rats
(male and female, Wistar, at least 33 days old),
but not in young rats (male and female, Wistar,
5-15 days old) due to greater renal tubular reab-
sorption rates in young rats (Braunlich et al.,
1978). Wierzba et al. (1982) reported that nitro-
furantoin excretion is age dependent in both hu-
mans and rats. Patients under 2 years of age
(sex unspecified) with a urinary tract infection
and normal renal function excreted 25% ± 5.7%
of their first dose (oral) after 12 hours at an ini-
tial excretion rate of 0.68 ± 0.23 mg/hour Pa-
tients older than 2 years of age (sex and age
range not given, same clinical conditions) ex-
creted 44% ± 16% at an initial rate of 4.55 ±
2.64 mg/hour. In comparison, Wierzba et al. re-
ported that intravenous administration of nitro-
furantoin (20 mg/kg) to 2-week-old or 2- to
3-month old rats (Wistar, sex unspecified) re
suited in half-life values of 0.95 and 0.41 hours,
respectively.

After a single oral dose of nitrofurantoin (ga-
vage, 25 mg/kg) to female albino rats (strain,
age, and sex unspecified), 52% and 2.6% nitrofu-
rantoin (percentage total dose) were recovered in
the urine and feces, respectively (Paul, M.F , et
al., 1960). When administered intravenously to
rats (strain, age and sex unspecified) to specific
organ sites of the digestive tract (25 or 100
mg/kg), nitrofurantoin was absorbed rapidly via
the small intestine, metabolized by liver, intes-
tine, and kidney, and excreted (half-life of 25
minutes) in the urine (50% recovered as nitrofu-
rantoin) (Buzard et al., 1961). Veronese et al.
(1974) reported that after intravenous adminis-
tration of nitrofurantoin to rats (male, Sprague
Dawley, 150-200 g), 16%-30% of the total dose
was recovered in the urine The proportion of ni-
trofurantoin or metabolite recovered was in-
versely related to dose; relative urinary excre-
tion of nitrofurantoin decreased with increasing
dose. Statham et al. (1985) compared the phar
macokinetics between control and vitamin
R-deficient male Sprague Dawley rats (age un-
specified, 200 g) administered nitrofurantoin
subcutaneously (15 mg/kg) and found that ni-
trofurantoin was rapidly absorbed and cleared
from blood, lung, liver, and kidney in a biphasic
manner. Metabolism of nitrofurantoin occurred
in control animals, but there were increased
levels of unchanged nitrofurantoin in vitamin

E-deficient rats. Urinary excretion was 68% of
the total dose administered in control rats and
35% in vitamin R-deficient rats.

Intravenous administration of nitrofurantoin
(1.5-24 mg/kg) to adult male beagles (10-16 kg)
stimulated bile secretion, and nitrofurantoin
was excreted in bile (at 6 mg/kg, 22.6% ± 4.7%
total dose) and urine (24.1% ± 4.7%) (Conklin
and Wagner, 1971). In these studies, carbon
tetrachloride administration was found to im
pair bile How and nitrofurantoin excretion Ni-
trofurantoin (after intravenous administration)
is excreted in bile, reabsorbed, and enterohepat-
ically recirculated (Conklin et al , 1973).

Oral administration of nitrofurantoin also may
result in the excretion of nitrofurantoin in the
milk of lactating humans ( Varsano et al., 1973),
rats, and dogs (Paul, M.F , et al , 1960). Admin-
istration of nitrofurantoin (oral, 100 or 200 mg)
to lactating women with normal glucose-6-phos-
phate dehydrogenase levels who had stopped
nursing resulted in excretion of nitrofurantoin
in their milk. The milk to serum ratio was ap-
proximately 0 29 in those with detectable levels.
Sixteen hours after being dosed, rats (age, sex,
and strain unspecified, 100 mg/kg) excreted 5
mg/liter, and dogs (age, sex, and strain unspeci-
fied, 20 mg/kg) excreted 2.33 mg/liter.

Acute Toxicity

Acute toxicity varies somewhat between rats
(LD5o = 112 mg/kg by intraperitoneal injection;
604 mg/kg by gavage; vehicle, 5%-15% acacia in
water; male, Sprague Dawley, 180 g; Preti,
1970) and mice (LD5o=150 mg/kg by intraperi-
toneal injection; 360 mg/kg by gavage; vehicle,
age, and sex unspecified; NIOSH, 1983). The
oral TD[^„ for humans is 80 mg/kg. Dietary de-
ficiencies in both selenium (Burk and Lane,
1983) and vitamin E (Boyd et al , 1979b) in-
creased the acute toxicity of nitrofurantoin to
rats (male, Holtzman and Sprague Dawley,
respectively).

Cellular and Subcellular Toxicity

In vitro studies indicate that under aerobic con-
ditions, reduction of nitrofurantoin stimulates
consumption of oxygen and production of super-
oxide anion free radical and hydrogen peroxide

Nitrofurantoin, NTP TR 341

18

I. INTRODUCTION

in avian liver and mammalian liver, lung, small
intestine, kidney, and gastrointestinal contents,
which may result in cytotoxicity and localized
injury in vivo to cellular membranes (Mason and
Holtzman, 1975a; Biaglow et al., 1977; Aufrere
et al., 1978; Boyd et al., 1979a; Sasame and
Boyd, 1979; Leskovac and Popovic, 1980; Peter-
son et al., 1982a) and to microbial organisms
(Hassan and Fridovich, 1979). Using rat (age,
sex, and strain unspecified) lung explants treat-
ed in culture with 10 3 M nitrofurantoin, Mar-
tin (1983) found significant lung cell injury,
which was increased with increased oxygen ten-
sion and decreased in the presence of superoxide
dismutase, catalase, a-tocopherol, and other an-
tioxidants. Rose et al (1982) reported that 100
mg/kg nitrofurantoin administered intraperito-
neally to rats (Wistar, age and sex not reported)
for 7 consecutive days caused changes in (5-glu-
curonidase and P-galactosidase activity in nerve
homogcnates, which indicated significantly in
creased enzyme activity and nerve degeneration.

Under anaerobic conditions, lung and liver mi-
crosomal and soluble fractions (male, HLA-SD,
150 g) mediated the covalent binding of ["C|ni-
trofurantoin-derived radioactivity to acid-pre-
cipitated macromolecules (Boyd et al., 1979a).
Covalent binding of ["Cjnitrofurantoin activity
was greatest in the kidney, liver, ileum, lung,
and heart of rats. Reduced glutathione was re-
ported to decrease covalent binding of ['^Cjnitro-
furantoin-derived radioactivity. Olive and
McCalla (1977) reported that nitrofurantoin, ni-
trofurazone, and other 5-nitrofurans are toxic to
L cells in culture under aerobic conditions but
that toxicity and DNA damage increase as oxy-
gen content in the culture decreases, Russo et
al. (1982) reported liver DNA damage in rats
(male, Sprague Dawley, 100-200 g) 72-96 hours
after administration of a single oral dose of 56 or
1 12 mg/kg of nitrofurantoin <gavage, 0.9% saline
with 1% carboxymethylcellulose). Nitrofura-
zone, a related 5 nitrofuran, has been shown to
bind to nucleic acids and proteins in vivo and in
vitro (Tatsumietal., 1977).

Under aerobic and anaerobic conditions in vitro,
nitrofurantoin depletes human erythrocyte glu-
tathione, according to Dershwitz and Novak
(1982). Minimal binding of nitrofurantoin or
metabolites occurs to erythrocyte macromole-
cules. Nitrofurantoin was reported to increase

the rate of superoxide anion radical formation
under aerobic conditions from oxyhemoglobin.
Under reduced oxygen tension, nitrofurantoin is
reduced and requires both an NADPH-depend-
cnt fiavoprotein and hemoglobin for superoxide
anion radical formation to induce erythrocyte
toxicity.

Peterson et al. (1982b) reported that a potential
mechanism of detoxication of nitrofurantoin (or
its reaction products) is by a selenium-depen-
dent glutathione peroxidase In selenium-defi-
cient chicks, the LD50 value is 53 mg/kg, where-
as in normal chicks, the LD50 value is 148
mg/kg; toxicity can be counteracted by adding
selenium, but not vitamin E, back into the diet.
Nitrofurantoin in the diet initially decreased
glutathione peroxidase activity but not liver glu-
tathione, catalase, or superoxide dismutase con-
tent, except at highly toxic doses over time. In
selenium deficient rats (male, Holtzman) but
not in controls, nitrofurantoin (100 mg/kg in
feed, 2-5 months after they were weaned) caused
renal tubular necrosis and an associated in-
crease in serum glutamic-pyruvic transferase
activity (Burk and Lane, 1983).

Epidemiology and Systemic Toxicity

Adverse reactions to the administration of nitro-
furantoin in the treatment of infections in hu-
mans have been reported (Delaney et al., 1977;
Penn and Griffin, 1982; D'Arcy, 1985). Adverse
effects reported included allergic (dermatologic),
gastrointestinal, hematologic, hepatic, pul-
monary, and neurologic reactions with varying
degrees of incidence, time of treatment to onset,
and severity of symptoms. The overall incidence
of adverse reactions (all types) to nitrofurantoin
administration reported worldwide between
1953 and 1984 was 0.0028% (D'Arcy, 1985). Al-
lergic reactions, acute lung reactions, peripheral
neuropathologic effects, and gastrointestinal (in-
cluding liver) reactions (in descending order of
occurrence) were the most frequently reported
(Penn and Ctriffin, 1982; D'Arcy, 1985).

Nitrofurantoin-induced acute pulmonary reac-
tions are characterized by development of fever,
cough, and shortness of breath within hours to
days after initiation of therapy in humans, ac-
cording to Whitcomb and Domby (1978). Eosino-
philia and diffuse alveolar or interstitial

19

Nitrofurantoin, NTP TR 341

I. INTRODUCTION

infiltrates may be present. Chronic interstitial
lung disease may also result after nitrofurantoin
therapy of 6 months or longer, according to these
authors. Development of progressive dyspnea
and diffuse interstitial infiltration was stated to
be characteristic. Both acute and chronic forms
are usually reversible after discontinuation of
nitrofurantoin therapy or with discontinuation
of nitrofurantoin and corticosteroid treatment.
Rats (male, Sprague Dawley, 350 g) adminis-
tered 300-500 mg/kg nitrofurantoin by subcuta-
neous injection developed lung injury (severe
respiratory distress, pulmonary edema, and
hemorrhage) (Boyd et al., 1979b). Rats raised on
a vitamin E-deficient diet or fed enriched vita-
min E diets and exposed in an oxygen-rich envi-
ronment were stated to have increased suscep-
tibility to nitrofurantoin toxicity Selenium-
deficient or control rats (male, Holtzman, 2 5
months old) administered 100 mg nitrofuran-
toin/kgdid not have lung injury (Burk and Lane,
1983).

Anttinen et al. (1982) reported a focal nodular
hyperplasia of the liver which developed in a
young girl after 7 months of nitrofurantoin
treatment for infection. No epidemiologic stud-
ies were found on the use of nitrofurantoin, other
than for adverse reactions.

Behar et al. (1965) reported that rats (male and
female, Sabra, age unspecified) administered ni-
trofurantoin orally (0, 20, 50, or 100 mg/kg, two
times per day) developed structural and func-
tional changes in the sciatic nerve. Nitrofuran-
toin plasma levels were dependent on the dose
and the duration of administration. The degree
of axonal degeneration was time related but not
dose related. Decreases in the conduction ve-
locity of sciatic nerve transmission were time
and dose related, but clinical neurologic changes
were not observed. Toole et al. (1968) prospec-
tively examined volunteers (male and female,
age 22-58 with normal renal and hematologic
function) who received 100 mg nitrofurantoin,
four times per day for 14 consecutive days. This
treatment was associated with impaired nerve
conduction velocity (normal renal function); 8/14
subjects had no side effects. Five subjects com-
plained of epigastric discomfort, nausea, or ano-
rexia, and two had clinical vasomotor changes
(ataxia) that were reversed after treatment was

stopped. In an evaluation of four case reports,
Yiannikas et al. (1981) stated that neuropatho-
logic effects in all the patients were directly
associated with nitrofurantoin treatment and
were characterized by acute, severe axonal de-
generation: they concluded that a direct neuro-
toxic effect was responsible. Hepatic injury
(hepatocellular and cholestatic) (Goldstein et al.,
1974), granulomatous formation (Strohscheer
and Wegener, 1977), and chronic active hepatitis
and necrosis (Sharp et al., 1980) have been asso-
ciated with nitrofurantoin therapy.

Glucose-6-phosphate dehydrogenase deficiency
and nitrofurantoin administration have been as-
sociated with the development of megaloblastic
anemia (Pritchard et al., 1965; Toole et al.,
1968). Glucose-6-phosphate dehydrogenase is
required for the NADPH-dependent reduction of
glutathione, which is required for erythrocyte
protection from peroxides. Initiation of nitrofu-
rantoin administration may result immediately
in dizziness, weakness, headache, anorexia, and
intermittent vomiting. Loss of erythrocytes,
oxygen deficiency, and possibly renal problems
are associated with lysis of erythrocytes and
methemoglobinemia.

Reproductive and Developmental Toxicity

Paul and Harrington (1967) found that nitro-
furantoin did not inhibit maternal or neonatal
liver or kidney glucuronyl transferase in rabbits
(male and female, strain unspecified; neonatal,
1-2 days old; adult, age unspecified) or rats (male
and female, strain unspecified; neonatal, 2 hours
to 3 days old; adult, 11 months old) There were
no differences in nitrofurantoin metabolism be-
tween adult or neonatal rabbit liver or kidney
and rat liver, but neonatal rat kidney metabo-
lized nitrofurantoin significantly more slowly
than did adult rat kidney.

Oral administration of nitrofurazone, a 5-nitro-
furan analog of nitrofurantoin, has been re-
ported to inhibit spermatogenesis at the sperma-
tocyte or spermatid stage in rats (sex, strain, and
age unspecified), which under long-term admin-
istration results in testicular atrophy. Gener-
ally, this effect was reversible upon discontinua-
tion of drug administration. A similar effect was
also reported in mice, along with interstitial cell
hyperplasia and seminal vesicle hypertrophy

Nitrofurantoin, NTPTR341

20

I. INTRODUCTION

(Prior and Ferguson, 1950; Nissim, 1957;
Montemurro, 1969).

In a continuous breeding study conducted by the
National Toxicology Program (NTP) on nitro-
furazone, timed-pregnant CD®-1 mice were fed
dietary concentrations of nitrofurazone ranging
from 38 to 500 ppm. Exposure was from gesta-
tional age of 6-15 days, and observations were
continued through day 17. No teratogenic ef-
fects were seen in fetuses evaluated on day 17.
Selective embryotoxicity, expressed as an in-
creased incidence of late fetal death and intra-
uterine growth retardation, was observed at ex-
posure concentrations that were only marginally
toxic to the exposed dams (Price et al., 1985).

Long-Term Toxicity and Carcinogenicity

Female Holtzman rats (age not reported) given
3,000 ppm nitrofurantoin in feed for either 36 or
44.5 weeks (survival and body weights were not
reported) did not develop any compound-related
tumors (Morris et al., 1969). Similarly, tumors
were not induced in young female Sprague
Dawley rats (40-72 g) fed nitrofurantoin at 1,870
ppm from week 0 to 16, at 1,000 ppm from week
16 to 75, and control diets from week 75 to 80
(Cohen et al., 1973). Body weights of dosed ani-
mals were stated to be reduced, but not signifi-
cantly, relative to those of the controls The data
indicate that 19/36 nitrofurantoin-dosed rats de-
veloped mammary gland tumors (fibroadenomas
or adenocarcinomas, combined), whereas 12/30
control animals developed these tumors. Six of
36 dosed and 6/30 control rats were diagnosed
with mammary gland adenocarcinomas. No de-
tails were given on survival of the dosed
animals.

Ito et al. (1983) reported that nitrofurantoin ad-
ministered in feed to BDFi mice (C57BL/6N X
DBA/2N)Fi (50 males and females per dose
group, from 9 weeks to 2 years of age) at 0, 750,
or 3,000 ppm caused no differences in survival
but that the high dose group body weight was
significantly lower than that of controls
(P<0 05) in each sex. They concluded that there
were no differences in tumor incidence between
dosed and control animals. However, they
stated that negative differences did occur in
male mice for liver adenomas (control. 6; low

dose, 1; high dose, 0); the number of animals ex-
amined in each group was not given. In female
mice, uterine tumors (reticulum cell sarcoma
type A) occurred at incidences of 7, 5, and 12.
Metastases from the uterine tumors were re-
ported to have been found in the peritoneal cavi-
ty, lymph nodes, liver, and lungs at incidences of
2, 3, and 5. The earliest uterine tumor was
found in the high dose group at week 44.

Sutton et al. ( 1987a, b) reported in abstracts on
studies in which nitrofurantoin was fed to
Sprague Dawley rats (50 males and 50 females
per dose; 0, 15, 27, or 49 mg/kg per day) and
Swiss mice (Crl:CD®-l[ICR]BR; 50 males and 50
females per dose; 0, 50, 100, or 200 mg/kg per
day). No evidence of compound-related tumor
incidences was reported for either rats or mice.
The increased number of deaths in high dose
male mice indicates that the maximum tolerated
dose may have been exceeded in their studies.

Genetic Toxicology

Results from a variety of bacterial and fungal as-
says have shown that nitrofurantoin is a muta-
gen in vitro. Growth inhibition due to DNA
damage was noted in Bacillus subtilis
(McCarroll et al., 1981; Suter and Jaeger, 1982)
and in Escherichia coli (Yahagi et al., 1974;
McCalla and Voutsinos, 1974) following expo-
sure to nitrofurantoin The mutagenic activity
of nitrofurantoin has been demonstrated in nu-
merous bacterial gene reversion tests with E.
coli (Yahagi et al., 1974; McCalla and Voutsinos,
1974; Simmon and Eckford, 1978, Lu et al.,
1979; Olive, 1979a,b) and Salmonella typhi-
murium. particularly strains TA98 and TAIOO,
which initiate error-prone DNA repair processes
(Wang and Lee, 1976. Goodman et al., 1977;
Simmon and Eckford, 1978; De Flora, 1979;
Ebringer and Bencova, 1980; Zeiger et al., 1981 ).
By comparing the results of nitrofurantoin expo-
sure of Salmonella strain TAIOO with those of
the nitro reductase-deficient strain TAIOO-FRI,
Rosenkranz and Speck (1976) demonstrated that
reduction of the nitro group is required to pro-
duce a mutagenic response in the absence of
metabolic activation. However, they observed
significant gene reversion in both strains of
Salmonella treated with nitrofurantoin in the

21

Nitrofurantoin, NTP TR 341

I. INTRODUCTION

presence of rat liver S9, indicating that mamma-
lian liver enzymes are also capable of metabo-
lizing nitrofurantoin to a mutagenic interme-
diate. In numerous independent NTP-sponsored
tests on chemical mutagenicity in Salmonella
with a preincubation protocol, nitrofurantoin
was consistently mutagenic to strains TA98 and
TAIOO both with and without metabolic ac-
tivation by Aroclor 1254-induced male Sprague
Dawley rat or Syrian hamster liver 89. No sig-
nificant increase in mutant colonies was ob-
served in strains TA1535 or TA1537, which, un-
like TAIOO and TA98, do not exhibit enhanced
error prone repair of damaged DNA (Haworth et
al., 1983). A representative sample of these tests
as conducted by one laboratory is presented in
Table P:i

Exposure to nitrofurantoin induced formation of
white (lacking chlorophyll) colonies of Kuglenu
gracilis strain z (McCalla, 1962, 1965, p:bringer
et a!., 1978); this bleached condition is heritable,
but whether it is the result of direct interaction
of the nitrofurantoin with chloroplast DNA is
unknown. Mitotic recombination in Saccharo-
myces cerevtsiae (Siebert et al., 1979; Callen,
1981) and mitotic nondisjunction in Aspergillus
nidulans (Bignami et al., 1974) were observed
after treatment with nitrofurantoin.

Mutagenicity results from assay systems that
use cultured mammalian cells are variable.
Without S9, single-strand breaks in the DNA of
cultured mouse L cells were detected after incu-
bation with 430 pM nitrofurantoin for 1 hour
(Olive and McCalla, 1977), and resistance to 6-
thioguanine was induced in Chinese hamster
V79 spheroids after treatment with 100-300
pg/ml nitrofurantoin (Olive, 1981). Induction of
forward mutations at the TK locus of mouse
L5178Y lymphoma cells was observed after
treatment with 5-500 pg/ml nitrofurantoin in
the absence of S9; it was not tested with S9 (Ta-
ble E2). Incubation of Chinese hamster ovary
cells with 40 pM nitrofurantoin for 1 hour pro-
duced a 74% increase in sister chromatid ex-
changes over the baseline frequency (Shirai and
Wang, 1980). Chromosomal aberrations were
observed in 19% of cultured Chinese hamster fi-
broblast cells incubated with 0 062 mg/ml nitro-
furantoin (Ishidate et al., 1978) In NTF studies.

nitrofurantoin induced increased numbers of sis-
ter chromatid exchanges and chromosomal aber-
rations in cultured Chinese hamster ovary cells
with and without metabolic activation (Ta-
bles E3 and P>4). Results of other investigations
with in vitro assays that use mammalian sys-
tems showed no effect of nitrofurantoin treat-
ment on sister chromatid exchange frequencies
(Sasaki et al., 1980), chromosomal aberrations
(Tonomura and Sasaki, 1973), and unscheduled
DNA synthesis (Tonomura and Sasaki, 1973).
No meiotic chromosomal abnormalities were ob-
served in mice (Fonatsch, 1977), induction of mi-
cronuclei was not observed in rats (Setnikar et
al , 1976, Goodman et al., 1977), sperm mor-
phologic abnormalities were absent in dosed
male mice (Topham, 1980), and dominant lethal
mutations were not detected in two strains of ex-
posed male mice (Epstein et al , 1972; Setnikar
etal., 1976).

In tests by Kramers (1982), nitrofurantoin fed to
adult male Drosophila, strain Oregon R, at a
concentration of 5 mM produced a marginally
significant increase in the frequency of sex-
linked recessive lethal mutations, but considera-
ble inconsistency occurred between tests, leav-
ing the question of nitrofurantoin's mutagenic
activity in this assay unresolved In an NTI^
Drosophila sex-linked recessive lethal assay, no
increase in mutations was observed in adult
male Canton-S flies after administration of ni-
trofurantoin orally (2,000 ppm in sucrose for 3
days) or by injection (10,000 ppm in saline) (Zim-
mering et al., 1985; Table E5).

Study Rationale

Nitrofurantoin was nominated and selected for
study by the National Cancer Institute as a re-
sult of a review of International Agency for Re-
search on Cancer chemicals, because it had the
largest production volume and was the most
widely used 5-nitrofuran drug, and because it is
structurally similar to other 5-nitrofuran deriv-
atives reported to be carcinogenic in rodent
studies. Administration of nitrofurantoin in
feed was chosen to obtain exposure by the oral
route, which is the primary route for administra-
tion of the drug in humans.

Nitrofurantoin, NTP TR 341

22

II. MATERIALS AND METHODS

PROCUREMENT AND CHARACTERIZATION OF

NITROFURANTOIN
PREPARATION AND CHARACTERIZATION OF

FORMULATED DIETS
FOURTEEN-DAY STUDIES
THIRTEEN-WEEK STUDIES

TWO-YEAR STUDIES
Study Design

Source and Specifications of Animals
Animal Maintenance
Clinical Examinations and Pathology
Statistical Methods

23 Nitrofurantoin, NTPTR 341

II. MATERIALS AND METHODS

PROCURKMKNT AND
CHARACTKRIZATION OF
NITROFURANTOIN

Nitrofurantoin was obtained in one lot (lot no.
03540) IVoin Norwich Katon Pharmaceuticals
(Norwich, NY). Purity and identity determina-
tions were conducted by Midwest Research Insti-
tute (MRU (Kansas City, MO). MRI reports on
analyses performed in support of the nitrofuran-
toin studies are on file at the National Institute
of Environmental Health Sciences.

Lot no. 03540 was obtained as a yellow, micro-
crystalline powder with a melting point of 251°-
255° C. The identity of nitrofurantoin was con-
firmed by infrared (P'igure 1), ultraviolet/visible,
and nuclear magnetic resonance (F'igure 2) spec-
troscopy. The infrared and nuclear magnetic
resonance spectra were consistent with litera
ture spectra (Analytical Profiles of Drug Sub
stances, 1976).

The purity of nitrofurantoin was determined by
elemental analysis, water analysis, titration of
the imide group, thin-layer chromatography,
and high-performance liquid chromatography.
Cumulative data indicated that lot no. 03540
was greater than 99% pure Results of elemen-
tal analyses for carbon, hydrogen, and nitrogen
agreed with the theoretical values. Water con-
tent by Karl Fischer titration was less than
0.02%. Titration of the imide group with tetra-
butylammonium hydroxide indicated a purity of
99.6%. Thin layer chromatography on silica gel
plates with either a cyclohexane acetone:
methanol:acetic acid (45:45:5:5) or a toluene:
2-butanone:acetic acid (40:60:1) solvent system
detected a single spot with visualization by

ultraviolet and visible light and a sodium hy-
droxide-saturated methanol spray. No impuri-
ties with a peak area greater than 0.1% of the
major peak area were detected by high-perform-
ance liquid chromatography on a pBondapak
Ci8 column with a water:acetonitrile (88:12) mo-
bile phase at a fiow rate of 1 ml/minute and
ultraviolet detection at 365 nm Analysis of lot
no 03540 by the same high-performance liquid
chromatographic system (with a slightly dif
ferent solvent ratio) did not detect nitrofurazone,
5-nitro-2-furaldehyde, or 3-(f(5-nitro-2-furanyl)-
methylene|amino|-2,4-imidizolidinedione at
concentrations equal to or greater than the
minimal detectable concentrations (0.03%, 2.4%,
andO.2% w/v).

Stability studies performed by high-performance
liquid chromatography with a pBondapak Cih
column, a wateriacetonitrile (70:30) mobile
phase at a flow rate of 1.5 ml/minute, and ultra-
violet detection at 254 nm indicated that nitro-
furantoin was stable for 2 weeks at tempera-
tures up to 60° C. Further confirmation of the
stability of the bulk chemical (stored at 5° C)
during the toxicity studies was obtained by titra-
tion with tetrabutylammonium hydroxide and
high-performance liquid chromatography with a
Hewlett-Packard RP-8 column or a Perkin-
Elmer ODS Sil-X column, ultraviolet detection
at 365 nm, and a wateracelonitrile mobile phase
at a flow rate of 1 ml/minute. The acetonitrile
concentration was increased from 30% to 50%
over 20 minutes or from 5% to 45% over 15 min-
utes. No degradation of the bulk chemical was
seen over the course of the studies. Identity of
the chemical at the study laboratory was con-
firmed by infrared spectroscopy.

Nitrofurantoin, NTPTR341

24

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Nitrofurantoin, NTP TR 341

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Nitrofurantoin, NTP TR 341

26

II. MATERIALS AND METHODS

PREPARATION AND
CHARACTERIZATION OF
FORMULATED DIETS

Formulated diets were prepared by adding a dry
premix (approximately equal amounts of feed
and nitrofurantoin) to the appropriate amount of
feed (Table 1). The mixture was blended for 15
minutes. The homogeneity of diet mixtures for-
mulated at the analytical chemistry and study
laboratories was evaluated by extracting feed
samples (taken from three positions in the blen-
der) with acetonitrile and determining the ab-
sorption at 365 nm (analytical chemistry labora-
tory) or from 360 to 380 nm (study laboratory).
Further studies indicated that nitrofurantoin
was stable in feed at 2,000 ppm when stored for 2
weeks in the dark at temperatures up to 45" C.
In these studies, samples were extracted with
acetonitrile:acetic acid (99:1) and analyzed by
high-performance liquid chromatography with a

TABLE 1

pBondapak Cig column, a water/acetonitrile
(80:20) mobile phase, and ultraviolet detection
at 254 nm. In the 13- week and 2-year studies,
formulated diets were stored at 5° C for no longer
than 14 days.

Periodic analyses of feed mixtures by the same
analytical methods used for the homogeneity
studies were conducted by the study and analyti-
cal chemistry laboratories to determine if the
formulated diets contained the correct concen-
trations of nitrofurantoin. Formulated diets
were analyzed once during the 13-week studies
The results ranged from 93% to 103% of the tar-
get concentrations (Table 2) During the 2-year
studies, feed mixtures were analyzed every 1-2
months, and concentrations varied from 80% to
11 3% of the target concentrations (Table 3). Be-
cause 78/83 formulated diets analyzed were
within 10% of the target concentrations, it is es-
timated that the formulated diets were prepared

PREPARATIO.N AND STORAGE OF FORMULATED DIETS IN THE FEED STUDIES OF

NITROFURANTOIN

Fourteen-Day Studies

Thirteen-Week Studies

TwoYear Studies

Preparation

Premix prepared by mixing weighed amounts
of nitrofurantoin and feed in a specimen cup
for 1 min; remaining feed mixed with premix
in a 16-qt blender for 15 min

Maximum Storage Time

14d

Storage Conditions

Room temperature in the dark

Same as 14-d studies

14d

5° C in the darlc

Same as 14-d studies

14d

5° C in the dark before use and
then at room temperature

TABLE 2. RESULTS OF ANALYSIS OF FORMULATED DIETS IN THE THIRTEEN WEEK FEED

STUDIES OF NITROFURANTOIN (a)

Target Concen

tration

Dctermi

ined Concentration (b)

<ppni)

(ppm)

300

(c)287

600

560

1,300

1,340

2,500

2,520

5,000

4,990

10,000

(c 19,723

Determined as a
Percent of Target

95.7
93.3
103.1
100.8
99.8
97.2

(a) Mix dale; 4/29/80

(bl Results of duplicate analysis

(c) Average of values obtained from three locations in the blender

27

Nitrofurantoin, NTPTR341

TABLE 3.

RESULTS OF ANALYSIS OF FORMULATED DIETS IN THE TWO-YEAR FEED STUDIES OF

NITROFURANTOIN

Date Mixed

600

Concenlratlon of Nitrofurantoin in Feed
for Target Concentration (ppm) (a)

1,300

2,500

02/11/81
02/18/81

03/18/81
04/15/81

05/13/81
05/18/81
05/19/81
06/10/81

07/08/81
08/05/81

09/02/8 1
09/30/81

1 0/28/K 1
10/;iO/Hl
ll/lH/81

12/16/81
02/10/82

02/12/82
04/07/82

06/02/82
07/28/82
09/22/82
11/17/82
01/12/83
01/18/83

602
600

(b)680
(c)490
(dl596

608
630
593

605
541

578
570
602
609
587
618

1,300

2,480

1,310

2,450

1,316

2,500

1,320

1 ,320

2,580

1,300

2,620

1.410

1.430

2,350

1,340

2,540

1,350

1,340

2,480

1,370

2,450

1,230

1 ,280

2,320

(e) 1,190

2,560

1,330

1 .200

(b) 2,000

Ul)2,310

1 ,230

2,300

1 .240

1 .250

2,510

1.180

2,450

(b) 1.140

2,370

1 .220

(dl 1.290

1 .240

2,380

1.210

2,350

1.240

1 ,220

2,460

1 ,280

2,440

1,310

1,300

2.410

1 ,300

2,410

1.310

1.290

2,490

1.260

2.350

1.250

1.240

2,490

1,280

2,510

1,300

1.410

2,740

1.330

(b) 2,810

(bl 1.470

(d) 1.380

(d) 2,650

1,289

2,457

69.6

146.5

5.4

6.0

1,140-1,470

2,000-2,810

41

28

Mean (ppm)

Standard deviation

CoetTicient of variation (percent)

Range (ppm)

Number of samples

602
31.4
5.2
541-680
14

(a) Results of duplicate analysis

(b)Out of specifications; not used in study.

(c) Remix out of specifications; not u.sed in study or included in the mean.

(dl Remix; not included in the mean,

(e) Result of triplicate analysis

within specifications 94*^ of the time Referee
analyses were performed periodically by the
analytical chemistry laboratory. Good agree-

ment was generally found between the labora-
tories (Table 4).

Nitrofurantoin, NTP TR 341

28

TABLE i. RESULTS OF REFEREE ANALYSIS OF FORMULATED DIETS IN THE TWO YEAR FEED

STUDIES OF NITROFURANTOIN

Date IVIixed

Target Concentration
(ppm)

Determined Concentration (ppm)

Study Laboratory (a)

Referee Laboratory (b)

02/11/81
10/28/81
06/02/82
11/17/82

1,300

«00

2,500

1 .300

1,300

593

2,460

1 ,240

1 ,270

640

2,650

1 ,290

(a) Results of duplicate analysis

(b) Results of triplicate analysis

FOURTEEN-DAY STUDIES

Pour- to five-week-old F344/N rats and 4- to 6-
week-old B6C3Fi mice were obtained from
Charles River Breeding Laboratories. Rats were
observed for 21 days and mice for 14 days before
being placed on study. Groups of five rats of
each sex were fed diets containing 0, 1,300,
2,500, 5,000, 10,000, or 20,000 ppm nitrofuran-
toin for 14 consecutive days Groups of five mice
of each sex were fed diets containing 0, 600,
1,300, 2,500, 5,000, or 10,000 ppm. Animals
were housed five per cage Feed and water were
available ad libitum. Further details of animal
maintenance are given in Table 5.

THIRTEEN-WEEK STUDIES

Thirteen-week studies were conducted to evalu-
ate the cumulative toxic effects of repeated expo-
sure to nitrofurantoin and to determine the con-
centrations to be used in the 2-year studies.

Five- to six-week-old F344/N rats and B6C3Fi
mice of each sex were obtained from Charles
River Breeding Laboratories 18 days before be-
ing placed on study. Groups of 10 rats of each
sex were fed diets containing 0, 600, 1,300,
2,500, 5,000, or 10,000 ppm nitrofurantoin for 13
weeks. Groups of 10 mice of each sex were fed
diets containing 0, 300, 600, 1,300, 2,500, or
5,000 ppm. Animals were housed five per cage
Feed and water were available ad libitum.

Animals were observed two times per day; mori-
bund animals were killed. Feed consumption
was measured once per week by cage Individual
animal weights were recorded once per week.

At the end of the 13-week studies, survivors
were killed. A necropsy was performed on all
animals except those excessively autolyzed or
cannibalized. Liver weights were determined at
necropsy. Groups and tissues examined are
listed in Table 5

TWO-YEAR STUDIES

Study Design

Groups of 50 female rats were fed diets contain-
ing 0, 600, or 1,300 ppm nitrofurantoin for 103
weeks Groups of 50 male rats and 50 mice of
each sex were fed diets containing 0, 1,300, or
2,500 ppm for 103 weeks. On January 8, 1982,
eight cages of control male rats were in-
advertently fed diets containing 600 ppm
nitrofurantoin.

Source and Specifications of Animals

The male and female F344/N rats and B6C3Fi
(C57BL/6N, female X C3H/HeN MTV", male)
mice used in these studies were produced under
strict barrier conditions at Harlan Industries.
Breeding stock for the foundation colonies at the
production facility originated at the National In-
stitutes of Health Repository. Animals shipped
for study were progeny of defined microflora-
associated parents that were transferred from
isolators to barrier-maintained rooms. Rats
were shipped to the study laboratory at 4-5
weeks of age and mice at 6-7 weeks of age. The
animals were quarantined at the study labora-
tory for 16 days. Thereafter, a complete necrop-
sy was performed on five animals of each sex and

29

Nitrofurantoin, NTP TR 341

TABLE 5.

EXPERIMENTAL DESIGN AND MATERIALS AND METHODS IN THE FEED STUDIES OF

NITROFURANTOIN

Fourteen-Day Studies

Thirteen-Week Studies

Two-Year Studies

EXPERIMENTAL DESIGN

Size of Study Groups

5 males and 5 females of each species 10 males and 10 females of each species 50 males and 50 females uf each species

Doses

Rals--0. 1,300,2,500,5.000. 10.000, or Rats--O,fi00. I,:i00, 2,500, 5,000, or

20,000 ppm nitrofurantoin in feed; 10,000 ppm nitrofurantoin in feed;

mice 0,600, 1,300, 2,500, 5,000, or mice-O. 300, HOO, 1 ,300, 2,500, or

10,000 ppm 5,000 ppm

Date of First Dose

Rats--2/19/80; mice-3/r2/80

5/5/80

Rat.s-male: 0, 1 ,300. or 2„500 ppm nitro-
furantoin in feed; female: 0, 600, or
1 ,300 ppm; mice -0, 1 ,300, or 2,,5()0 ppm

Rats-2/26/81; mice -2/19/81

Date of Last Dose

RaLs--3/3/80; mice -3/27/80

8/3/80

Rats-2/16/83; mice -2/9/83

Duration of Dosing

14 consecutive d

13 wk

103 wk

Type and Frequency of Oliservation

Observed 2 x d; weighed initially Observed 2 X d; weighed 1 X wk

and 1 X wk thereafter

Observed 2 X d; weighed initially, 1 x wk
for 12 wk (rats) or 13 wk (micel, and then
1 X mo; palpated at weighing starting at
w k 44 or earlier if masses were noted

Necropsy and Histologic E.xaniinations

Necropsy performed on all animals;
histologic exams not performed

Necropsy performed on all animals;
the following tissues examined histo-
logically for control and high dose
groups: adrenal glands, brain, cecum,
colon, esophagus, heart, kidneys,
liver, lungs and bronchi, mammary
gland, mandibular and mesenteric
lymph nodes, pancreas, parathyroid
glands, pituitary gland, prostate/tesles/
seminal vesicles or ovaries/uterus,
regional lymph nodes, salivary glands,
skin, small intestine, spleen, sternum
including marrow, stomach, thigh
muscle, thymus, thyroid gland,
trachea, and urinary bladder. Liver
weights recorded at necropsy

Necropsy and histologic exams performed
on all animals; the following tissues
examined histologically: adrenal glands,
brain, cecum, colon, duodenum, esoph-
agus, gallbladder (mice), gross lesions,
heart, ileum, jejunum, kidneys, larynx,
liver, lungs and mainstem bronchi, mam-
mary gland, mandibular and mesenteric
lymph nodes, nasal cavity and turbinates,
pancreas, parathyroid glands, pituitary
gland, preputial or clitoral gland, pros-
tate/testes/seminal vesicles/epididymis
or ovaries/uterus, rectum, regional lymph
nodes, salivary glands, skin, spleen, ster-
nebrae or femur or vertebrae including
marrow, stomach, thymus, thyroid gland,
ti.ssue masses, trachea, and urinary
bladder

ANIMALS AND ANIMAL MAINTENANCE

Strain and Species

F344/N rats; B6C3Fi mice

Animal Source

Rats-Charles River Breeding
Laboratories (Portage, MI);
mice-Charles River Breeding
Laboratories ( Kingston, NY )

Study Laboratory

Southern Research Institute

F344/N rats; B6C3F, mice

Charles River Breeding Laboratories
(Kingston, NY)

Southern Research Institute

F344/N rats; B6C3F, mice

Harlan Industries (Indianapolis, IN)

Southern Research Institute

Method of Animal Identification

Ear marked with poultry punch

Ear marked with poultry punch

Ear marked with poultry punch

Nitrofurantoin, NTP TR 341

30

TABLE 5. EXPERIMENTAL DESIGN AND MATERIALS AND METHODS IN THE FEED STUDIES OF

NITROFURANTOIN (Continued)

Fourteen-Day Studies

Thirteen-Week Studies

Two-Year Studies

ANIMALS AND ANIMAL MAINTENANCE (Continued)

Time Held Before Study

Rats- 21 d;mice -14d

18d

16d

Age Wlien Placed on Study
Rals--7-8 wlc; mice-6-8 wlc

Rats-8 wk; mice-8-9 wk

Rats 6-7 wk; mice -8-9 wk

Age When Killed

Rats--9- 10 wk; mice-8-10 wk

Rats-22-23 wk; mice-22-24 wk

RaU -110- 1 12 wk; mice -112-114 wk

Necropsy Dates

Rals--3/5/80-3/7/80;
mice--3/27/80-3/28/80

Rats-8/6/80-8/15/80;
mice-8/4/80-8/14/80

Rats -2/25/83-3/2/83;
mice-2/18/83-2/23/83

Method of Animal Distribution

Animals distributed to weight
classes and assigned to cages accord-
ing to one table of random numbers;
cages assigned to groups according to
another table of random numbers

Same as 14-d studies

Same as 14-d studies

Feed

NIH 07 Rat and Mouse Ration
(Zeigler Bros.. Inc., Gardners, PA);
available ad libitum

Same as 14-d studies

Same as 14-d studies

Bedding

Beta Chip® hardwood chips
(Northeastern Products Corp.,
Warrensburg, NY)

Same as 14-d studies

Same as 14-d studies

Water

Automatic watering system
(Edstrom Industries, Waterford,
Wl ); available ad libitum

Same as 14d studies

Same as 14-d studies

Cages

Polycarbonate (Lab Products,

Inc., Garfield, NJ)

Same as 1 4-d studies

Same as 14-d studies

Cage Filters

Reemay spun-bonded polyester
fillers (Snow Filtration,
Cincinnati, OH)

Same as 1 4-d studies

Same as 14-d studies

Animals per Cage

5

Other Chemicals on Study in the Same Room

None None

Animal Room Environment

Temp -22°-23°C;hum-41%-52%
(rats), 41%-49% (mice); fluorescent
light 12 h/d; 15 room air changes/h

Temp-22°-24° C; hum--44%-70%;
fluorescent light 12 h/d; 15 room
air changes/h

None

Temp-17.2°-31.1°C;hum-50% ± 10%;
fluorescent light 12 h/d; 15 room air
changes/h

31

Nitrofurantoin, NTP TR 341

II. MATERIALS AND METHODS

species to assess their health status. Rats were
placed on study at 6-7 weeks of age and mice at
8-9 weeks of age. The health of the animals was
monitored during the course of the studies ac-
cording to the protocols of the NTP Sentinel Ani-
mal Program (Appendix F).

Animal Maintenance

All animals were housed five per cage. F'eed and
water were available ad libitum. Further de-
tails of animal maintenance are given in Ta-
ble 5.

Clinical Examinations and J'athology

All animals were observed two times per day.
Body weights were recorded once per week for
the first 12 (rats) or 13 (mice) weeks of the stud
ies and once per month (hereafter Mean body
weights were calculated for each group Ani-
mals found moribund and those surviving to the
end of the studies were humanely killed A nec-
ropsy was performed on all animals, including
those found dead. Some tissues were excessively
autolyzed or cannibalized, and thus, the number
of animals from which particular organs or tis-
sues were examined microscopically varies and
is not necessarily equal to the number of ani-
mals that were placed on study.

During necropsy, all organs and tissues were ex-
amined for grossly visible lesions. Tissues were
preserved in 10% neutral buffered formalin, em
bedded in paraffin, sectioned, and stained with
hematoxylin and eosin. Tissues examined mi-
croscopically are listed in Table 5.

When the pathology evaluation was completed,
the slides, paraffin blocks, and residual wet tis-
sues were sent to the NTP Archives for inven-
tory, slide/block match, and wet tissue audit.
The slides, individual animal data records, and
pathology tables were sent to an independent
quality assessment laboratory. The individual
animal records and tables were compared for ac-
curacy, slides and tissue counts were verified,
and histotechnique was evaluated. All tumor di-
agnoses, all target tissues, and all tissues from a
randomly selected 10% of the animals were eval-
uated by a quality assessment pathologist. The
quality assessment report and slides were sub-
mitted to the Pathology Working Group (PWG)

Chairperson, who reviewed all target tissues
and those about which there was a disagreement
between the laboratory and quality assessment
pathologists.

Representative slides selected by the Chairper-
son were reviewed by the PWG, which included
the laboratory pathologist, without knowledge of
previously rendered diagnoses. When the con-
sensus diagnosis of the PWG differed from that
of the laboratory pathologist, the laboratory pa-
thologist was asked to reconsider the original di-
agnosis. This procedure has been described, in
part, by Maronpot and Boorman (1982) and
Boorman et al. (1985). The final diagnoses rep-
resent a consensus of contractor pathologists and
the NTP Pathology Working Group. For subse-
quent analysis of pathology data, the diagnosed
lesions for each tissue type are combined accord-
ing to the guidelines of McConnell et al ( 1986)

Slides/tissues are generally not evaluated in a
blind fashion (i.e., without knowledge of dose
group) unless the lesions in question are subtle
or unless there is an inconsistent diagnosis of le-
sions by the laboratory pathologist. Nonneo-
plastic lesions are not examined routinely by the
quality assessment pathologist or PWG unless
they are considered part of the toxic effect of the
chemical

Additional histologic sections of the right and
left kidney of male rats in each dose group and
the controls were prepared and reviewed by a
special PWG. Three or four longitudinal sec-
tions were prepared from the remaining half of
each of the right and left kidney at approxi-
mately 1-mm intervals by standard procedures.
All lesions observed during this special PWG re-
view were evaluated in a "blind" fashion.

Statistical Methods

Data Recording: Data on body weight and feed
consumption for this experiment were recorded
in the Carcinogenesis Bioassay Data System
(Linhart et al., 1974). Other data elements were
recorded in the Toxicology Data Management
System. The data elements include animals, ex-
perimental design, survival, and individual
pathologic results, as recommended by the Inter-
national Union Against Cancer (Berenblum,
1969).

Nitrofurantoin, NTP TR 341

32

II. MATERIALS AND METHODS

Suruiual Analyses: The probability of survival
was estimated by the product-limit procedure of
Kaplan and Meier (1958) and is presented in the
form of graphs. Animals were censored from the
survival analyses at the time they were found to
be missing or dead from other than natural
causes; animals dying from natural causes were
not 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) life table test for a dose-
related trend. When significant survival differ-
ences were detected, additional analyses using
these procedures were carried out to determine
the time point at which significant differences in
the survival curves were first detected. All re-
ported P values for the survival analysis are
two-sided.

Calculation of Incidence: The incidence of neo-
plastic or nonneoplastic lesions is given as the
ratio of the number of animals bearing such le-
sions at a specific anatomic site to the number of
animals in which that site was examined In
most instances, the denominators include only
those animals for which the site was examined
histologically. However, when macroscopic ex-
amination was required to detect lesions (e.g ,
skin or mammary tumors) prior to histologic
sampling, or when lesions could have appeared
at multiple sites (e.g., lymphomas), the denomi-
nators consist of the number of animals on which
a necropsy was performed.

Analysis of Tumor Incidence: Three statistical
methods are used to analyze tumor incidence
data: life table tests, logistic regression, and
Fisher exact/Cochran-Armitage trend analyses.
Tests of significance include pairwise compari-
sons of high dose and low dose groups with con-
trols and tests for overall dose-response trends.
For studies in which administration of the study
compound has little effect on survival, the re-
sults of the three alternative analyses will
generally be similar. When differing results are
obtained by the three methods, the final inter-
pretation of the data will depend on the extent to
which the tumor under consideration is regarded
as being the cause of death Continuity-

corrected tests are used in the analysis of tumor
incidence, and reported P values are one-sided.
The procedures described below also were used
to evaluate selected nonneoplastic lesions.

Life Table Analyses-This method of analysis as-
sumes that all tumors of a given type observed in
animals dying before the end of the study were
"fatal"; i.e , they either directly or indirectly
caused the death of the animal. According to
this approach, the proportions of tumor-bearing
animals in the dosed and control groups were
compared at each point in time at which an ani-
mal died with a tumor of interest. The denomi-
nators of these proportions were the total num-
ber of animals at risk in each group. These re-
sults, including the data from animals killed at
the end of the study, were then combined by the
Mantel-Haenszel method (1959) to obtain an
overall P value This method of adjusting for in-
tercurrent mortality is the life table method of
Cox (1972) and of Tarone (1975). The underly-
ing variable considered by this analysis is time
to death due to tumor If the tumor is rapidly
lethal, then time to death due to tumor closely
approximates time to tumor onset. In this case,
the life table test also provides a comparison of
the time-specific tumor incidences.

Logistic Regression Analyses-This method of
analysis assumes that all tumors of a given type
observed in animals that died before the end of
the study were "incidental"; i e., they did not al-
ter the risk of death and were discovered merely
as the result of death from an unrelated cause.
According to this approach, tumor prevalence
was modeled as a logistic function of dose and
time. Both linear and quadratic terms in time
were incorporated initially, and the quadratic
term was eliminated if it did not significantly
enhance the fit of the model. The dosed and con-
trol groups were compared on the basis of the
likelihood score test for the regression coefficient
of dose. This method of adjusting for intercur-
rent mortality is the prevalence analysis of
Dinse and Lagakos (1983), further described and
illustrated by Dinse and Haseman (1986). If the
tumor type is nonlethal, prevalence analyses
and incidence analyses are equivalent.

33

Nitrofurantoin, NTP TR 341

II. MATERIALS AND METHODS

Fisher Exact/Cochran-Armitage Trend Analy-
ses-In addition to survival-adjusted methods,
the results of the Fisher exact test for pairwise
comparisons and the Cochran-Armitage linear
trend test (Armitage, 1971; Gart et al., 1979) are
given in the appendixes containing the analyses
of tumor incidence. These two tests are based on
the overall proportion of tumor-bearing animals
and do not adjust for survival differences.

Historical Control Data: Although the concur-
rent control group is always the first and most
appropriate control group used for evaluation,
there are certain instances in which historical
control data can be helpful in the overall assess-
ment of tumor incidence. Consequently, control
tumor incidences from the NTP historical con-
trol data base (Haseman et al., 1984, 1985) are
included for those tumors appearing to show
compound-related effects.

Nitrofurantoin, NTP TR 341

34

III. RESULTS

RATS

FOURTKEN-DAY STUDIES

THIRTEEN-WEEK STUDIES

TWO-YEAR STUDIES

Body Weights, Feed Consumption, and Clinical Signs

Survival

Pathology and Statistical Analyses of Results

MICE

FOURTEEN-DAY STUDIES

THIRTEEN-WEEK STUDIES

TWO-YEAR STUDIES

Body Weights, Feed Consumption, and Clinical Signs

Survival

Pathology and Statistical Analyses of Results

35 Nitrofurantoin, NTP TR 341

III. RESULTS: RATS

FOURTEEN-DAY STUDIES

None of the rats died before the end of the stud-
ies (Table 6). Rats that received 5,000. 10,000.
or 20,000 ppm lost weight, and those that re-
ceived 2,500 ppm gained notably less than did
the controls. Feed consumption by rats that

received 5,000, 10,000, or 20,000 ppm was
notably less than that by controls during week 1.
Compound-related clinical signs included inac-
tivity, rough hair coats, sunken eyes, bright yel-
low urine, and/or yellow fur. Blue discoloration
of the joints was observed in rats that received
20,000 ppm.

TABLE 6. SURVIVAL, MEAN BODY WEIGHTS, AND FEED CONSUMPTION OF RATS
FOURTEEN DAY FEED STUDIES OF NITROFURANTOIN

N THE

Survival

•Mean

Body We

ij-hts

iKrams)

Final Weight Relative
to Controls

Feed
sumpt

C<
ion

)n-

Concentration

Initial (b)

Final

Change (ct

, (d)

(ppm)

(a)

(percent)

Week 1

Week 2

MALE

0

5/5

190 ± 8

249 ±

5

-1-59 ± 5

18

17

1,300

5/5

208 + 7

258 ±

8

-1-50 ± 4

104

19

20

2.500

5/5

187 ± 8

225 ±

7

■(-38 ±4

90

15

16

,5.0()()

5/5

182 ± 3

176 ±

4

-6 + 2

71

8

9

10,000

5/5

187 ± H

147 ±

4

-40 ± 2

59

13

17

20,000

5/5

18fi ± fi

117 ±

5

-69 ± 3

47

11

14

FEMALE

0

5/5

127 ± 5

1.54 ±

4

-1-27 ± 1

11

12

1.300

5/5

135 ± 4

1.57 ±

4

-(-22 ± 1

102

11

11

2.500

5/5

127 ± 4

143 ±

3

-(•16 ± 1

93

9

11

5,000

5/5

124+1

122 ±

H

- 2 ± 5

79

5

11

10.000

5/5

137 ± 3

103 ±

2

-34 ± 3

67

8

15

20,000

5/5

130 ± 4

84 ±

3

-46 ± 3

55

5

12

(a) Number surviving/number initially in the group

(bl Initial group mean body weight ± standard error of the mean

(c) Mean body weight change of the group ± standard error of the mean

(d) Grams of feed consumed per animal per day; average of daily determinations; not corrected for scatter.

Nitrofurantoin, NTPTR341

36

III. RESULTS: RATS

THIRTEEN-WEEK STUDIES

One of 10 female rats that received 10,000 ppm
died before the end of the studies (Table 7). The
final mean body weights of rats that received
2,500, 5,000, or 10,000 ppm were 10%, 34%, or
47% lower than that of the controls for males
and 15%, 31%, or 41% lower for females. Feed
consumption by dosed and control groups was
generally similar. The urine of dosed rats was
bright yellow. Minimal-to-mild degeneration of
the germinal epithelium of the seminiferous tu-
bules of the testis with aspermatogenesis was
observed in 29/30 males that received 2,500 ppm

or more. Minimal-to-mild necrosis of the ovari-
an follicles was observed in 8/10 females that
received 10,000 ppm, and minimal necrosis was
observed in 3/10 females that received 5,000
ppm and 1/10 females that received 2,500 ppm
The liver weight to body weight ratios for rats
that received 5,000 or 10,000 ppm were signif-
icantly greater than those of controls (Table 8)

Dose Selection Rationale: Because of lower body
weight gain at higher concentrations, dietary
concentrations selected for rats for the 2-year
studies were 1,300 and 2,500 ppm nitrofurantoin
for males and 600 and 1 ,300 ppm for females

TABLE 7. SURVIVAL, MEAN BODY WEIGHTS, AND FEED CONSl'MPTION OF RATS IN THE
THIRTEEN-WEEK FEED STUDIES OF NITROFURANTOIN

Survival

(a)

Mean B(

ody We

iyhts

((jranisl

Final Weight Relative
to Controls
(percent)

Feed C<
sumption

)n-

Concentration

Initial (l>)

Final

Change

(c)

(d)

(ppm)

Week 4

Week 12

MALE

0

10/10

130 ± 2

344 ±

8

+ 214 ±

8

17

16

600

10/10

129 ± 2

366 ±

6

-1-237 ±

6

106

17

16

1,300

10/10

127 ±2

334 ±

7

-1-207 ±

8

97

17

16

2,500

10/10

129 ± 1

311 ±

6

-1-182 ±

6

90

15

16

5.000

10/10

130 ± 2

228 ±

6

-1-98 ±

5

66

15

13

10.000

10/10

128 ± 1

182 ±

3

-1-54 ±

3

53

18

20

FEMALE

0

10/10

109 ± 2

198 ±

3

-t-89 +

2

U

12

600

10/10

109 ± 2

189 ±

2

-^80 ±

2

95

13

11

1,300

10/10

111 ± 1

191 ±

1

-(-80 ±

1

96

12

12

2,500

10/10

107 ± 1

168 ±

3

+ e,\ ±

3

85

10

9

5,000

10/10

110 ± 1

136 ±

2

+ 26 ±

•>

69

12

11

10,000

ie)9/10

110 ± 1

116 ±

4

+ 6 ±

4

59

16

22

(a) Number surviving/number initially in the group

(b) Initial group mean body weight ± standard error ofthe mean. Subsequent calculations are based on those animals
surviving to the end of the study.

(cl Mean body weight change of the survivors ± standard error of the mean

(d) Grams of feed consumed per animal per day; not corrected for scatter.

(e) Week of death: 12

37

Nitrofurantoin, NTP TR 341

TABLE 8.

ABSOLUTE AND RELATIVE LIVER WEIGHTS OF RATS IN THE THIRTEEN-WEEK FEED
STUDIES OF NITROFURANTOIN (a)

Concentration
(ppm)

Number of

Animals

Necropsy
Body Weight

(grams)

Liver Weight
(mg)

Liver Weight/Necropsy
Body Weight

(mg/g)

MALE

0
600

1,300

2,500

5,000

10,000

FEMALE

0

600

1.300

2,500

5,000

10,000

10

348

+

7.8

13,191

+

447

38.0

± 1.26

10

367

+

6.5

12.893

+

660

35.1

± 1.64

10

337

±

7.3

12,261

+

295

36.4

± 0.80

10

ib>3l3

±

6.1

12,3.50

+

337

39.4

± 0.98

10

ibi246

±

5.6

(b) 10,772

+

345

(C143.8

± 0.59

10

(bil82

±

3.6

(b) 8,972

±

250

(b)49.4

± 1.91

10

198

±

3.5

6,698

+

229

33.8

± 0.90

10

192

±

2.2

6,945

+

131

36.1

±0.53

10

197

±

1.7

ibi 7,950

+

162

40.4

±0.71

10

lb) 172

±

3.0

6,227

+

177

36.3

± 1.13

10

( b ) 1 46

±

16

6.837

+

226

(b>46.8

± 1,53

9

(b)llH

±

44

11)15.270

+

549

(b)44.7

± 4.35

(a) Mean ± standard error; P values vs. the controls by Dunnett's test (Dunnett. 1955)

(b)P<0.01

(c)P<0.05

TWO YEAR STUDIES

Body Weights, Feed Consumption, and
Clinical Signs

Mean body weights of high dose male rats were
6%-9% lower than those of the controls from
week 1 to week 11 and thereafter were within
5% of the control weights (Table 9 and Figure 3).
Mean body weights of dosed female rats were

within 6% of those of the controls throughout the
studies The average daily feed consumption per
rat by low dose and high dose rats was 99% and
95% that by controls for males (Table Gl) and
101% and 98% for females (Table G2) The
average amount of nitrofurantoin consumed per
day was estimated to be 60 and 110 mg/kg for
low and high dose male rats and 30 and 60
mg/kg for low and high dose female rats. Dosed
rats had bright yellow urine.

Nitrofurantoin, NTP TR 341

38

TABLE 9.

MEAN BODY WEIGHTS AND SURVIVAL OF RATS IN THE TWO-YEAR FEED STUDIES OF

NITROFURANTOIN

Weeks

Con

itrol

Low Dose

High Dose

on

Av. Wt.

No. of

Av. Wt.

Wt. (percent of

No. of

Av. Wt.

Wt. (percent of

No. of

Study

(grams)

Survivors

(grams)

controls)

Survivors

(grams)

controls)

Survivors

MALE

1,300 ppm

2,500 ppm

0

117

SO

119

102

50

116

99

50

1

159

50

159

100

.50

149

94

50

2

190

50

192

101

.50

177

:w

SO

3

218

50

217

100

.50

198

91

50

4

236

50

238

101

,50

214

91

50

5

254

50

256

101

50

231

91

SO

6

264

50

268

102

50

240

91

SO

7

280

SO

286

102

SO

257

92

50

8

293

50

296

101

50

268

91

SO

»

306

50

311

102

50

283

92

SO

10

312

50

316

101

50

291

93

50

11

321

50

327

102

.50

300

93

50

Vi

329

50

334

102

SO

312

95

50

17

363

50

365

101

50

140

94

50

■21

381

.50

.381

100

.50

161

95

50

•iS

.193

50

391

99

50

174

:i5

50

T.t

404

50

406

100

50

185

95

SO

.14

416

50

416

100

50

402

97

50

.■w

422

50

426

101

50

414

98

.50

44

430

50

427

99

SO

420

98

50

48

443

50

448

101

49

4.18

99

SO

Vi

449

50

452

101

49

446

99

50

r>5

445

50

450

101

49

439

99

49

ryj

452

49

452

100

49

448

99

47

ti4

455

49

451

99

49

456

100

47

fi9

4.54

48

445

98

49

453

100

47

74

442

47

439

99

47

444

100

46

7S

446

46

440

99

47

442

99

45

82

441

44

434

98

45

437

99

44

86

440

41

428

97

45

435

99

44

'.111

440

37

432

98

42

430

98

43

1)4

436

33

426

98

41

425

97

37

■J8

435

30

422

97

38

413

95

35

1(14

412

27

401

97

■J9

191

95

28

FEMALE

600 ppm

1,300 ppm

0

98

50

98

100

50

96

98

50

I

121

50

121

100

50

117

97

SO

2

135

SO

138

102

.50

136

101

SO

3

14«

50

151

102

SO

148

100

SO

4

157

50

160

102

SO

156

99

50

S

166

50

170

102

50

163

98

50

6

170

50

174

102

SO

167

98

50

7

177

50

181

102

.50

174

98

SO

8

180

50

184

102

SO

176

98

50

9

1^5'

SO

191

103

SO

184

99

50

10

189

50

194

103

SO

187

99

SO

11

193

50

198

103

50

191

99

SO

12

195

50

201

103

50

194

99

50

17

210

.50

216

103

50

206

93

SO

21

214

50

223

104

50

212

99

SO

25

225

50

231

103

50

217

96

SO

29

228

50

239

105

SO

223

98

50

34

237

50

248

105

50

233

98

50

38

245

50

256

104

50

237

97

SO

44

256

'.0

271

106

50

246

96

SO

48

267

50

279

104

50

256

96

50

52

283

50

294

104

50

270

95

SO

55

288

50

300

104

50

277

96

SO

59

30.1

50

310

102

49

291

96

48

64

316

49

325

103

47

302

96

48

69

328

48

339

103

4fi

314

96

46

74

332

47

343

103

44

318

96

44

78

340

47

347

102

44

324

95

44

82

343

46

355

103

41

.131

97

43

86

345

46

359

104

41

339

98

42

90

352

44

367

104

39

344

98

41

94

353

42

368

104

37

346

98

40

98

361

37

375

104

34

352

98

36

104

352

28

359

102

28

351

100

31

39

Nitrofurantoin, NTP TR 341

500 0

450.0-

<

ct:

O 350.0

X JOO.O

o

>-
Q
O

m

z
<

250.0

200 0-

150 0

100.0-

50.0

. ^ s « ® '^ g

4^

^ i.

» A
A

r 500 0

450 0

C- 400.0

- 350.0

300.0

A

fl

500 0

<
a:
o

X

o

>-

Q
O

CD

Z
<

UJ

3

450 0-

400.0

350 0

300 0-

250.0

200 0-

150 0

100 0

50.0

MALE RATS

■

= UNTREATEO

o

= 1.300 PPM

A

= 2.500 PPM

—I —
IS

— I —
30

— I —
45

— I —
60

— I —
75

— 1 —
90

250 0

200 0

150 0

100 0

50.0

105

WEEKS ON STUDY

O

■

A

n ^ ° -
:-" 1

8-

8

\ o 2

8 X ^

Q ■ A

■ :

a;

— I —

JO

— I —
45

— I —
60

FEMALE RATS
■ = UNTREATED
0=600 PPM
A= 1,300 PPM

— I —
75

— I —
90

500 0

450 0

400 0

350 0

300 0

250.0

200 0

150.0

100 0

50 0

lOS

WEEKS ON STUDY

FIGURE 3. GROWTH CURVES FOR RATS FED DIETS CONTAINING
NITROFURANTOIN FOR TWO YEARS

Nitrofurantoin, XTP TR 341

40

III. RESULTS: RATS

Survival

Estimates of the probabilities of survival for
male and female rats fed diets containing nitro-
furantoin at the concentrations used in these
studies and for controls are shown in Table 10
and in the Kaplan and Meier curves in Figure 4.
No significant differences in survival were ob-
served between any groups of rats of either sex.

Pathology and Statistical Analyses of
Results

This section describes the statistically signifi-
cant or biologically noteworthy changes in the

incidences of rats with neoplastic or nonneo-
plastic lesions of the kidney, parathyroid glands,
glandular stomach, bone, subcutaneous tissue,
testis, epididymis, preputial gland, clitoral
gland, mammary gland, and eye.

Summaries of the incidences of neoplasms and
nonneoplastic lesions, individual animal tumor
diagnoses, statistical analyses of primary tu-
mors that occurred with an incidence of at least
5% in at least one animal group, and historical
control incidences for the neoplasms mentioned
in this section are presented in Appendixes A
and B for male and female rats, respectively.

TABLE 10. SURVIVAL OF RATS IN THE TWO-YEAR FEED STUDIES OF NITROFURANTOIN

Control

fiOO pptii

1,300 ppm 2,500 ppm

MALE (a)

Animals initially in study

Nonaccidental deaths before termination (bl

Killed at termination

Died during termination period

Survival P values (c)

FEMALE (a)

Animals initially in study

Nonaccidental deaths before termination (bl

Killed at termination

Survival P values (c)

50

50

50

26

23

24

23

27

26

1

0

0

0.668

0.512

0.732

50

50

50

25

24

19

25

26

31

0.446

0.951

0.489

(a) First day of termination period: 730

(b) Includes animals killed in a moribund condition

(c)The result of the life table trend test is in the control column, and the results of the life table pairwise comparisons with the
controls are in the dosed columns.

41

Nitrofurantoin, NTP TR 341

1.0

0 9

<
>

o

>■ 0.7

00

<

00

O 0.6

cc

CL

0.5

0.4

wlALE RATS

■

Control

o

1500 ppm

A

2500 ppm

—\ —
15

X

30 *5 60 75

WEEKS ON STUDY

— I —

90

105

120

1.0

0 9-

<
>

^ 0.8
(/I

0.7-

0.4

m
<

O 0 6-

ct

Q.

0.5-

FEMALE RATS

■ Control

O 600 ppm
A 1300 ppm

15 30 45 60 75

WEEKS ON STUDY

90

i

105

120

FIGURE 4. KAPLAN-MEIER SURVIVAL CURVES FOR RATS FED DIETS
CONTAINING NITROFURANTOIN FOR TWO YEARS

Nitrofurantoin, NTP TR 341

42

III. RESULTS: RATS

Kidney: Chronic nephropathy occurred in
nearly all rats. This spontaneous disease was
characterized by varying degrees of tubular de-
generation with atrophy of the epithelium and
dilatation of the tubules, regeneration of tubular
epithelium, thickening of the tubular basement
membranes, interstitial fibrosis, chronic inflam-
mation, and glomerulosclerosis. The severity of
this disease in each rat was judged on a scale of
1 = minimal, 2 = mild, 3 = moderate, 4 =
marked. The mean severity of the nephropathy
was somewhat increased in dosed male rats
relative to controls but was decreased in dosed
female rats (male: control, 50/50 [severity, 3.1];
low dose, 48/50 [3.31; high dose, 48/50 [3.5); fe-
male: 44/50 [2.41; 40/50 [2.2|; 48/50 [2.0|). Hy-
perplasia of the transitional epithelium lining
the renal pelvis and hydronephrosis were also
observed in some dosed male rats (transitional
epithelium hyperplasia: 0/50; 5/50; 2/50; hydro-
nephrosis: 0/50; 5/50; 2/50).

A single section of the left and right kidney of
each rat was examined microscopically as a
standard procedure during the histopathologic
evaluation. With this procedure, renal tubular
cell adenomas were seen in 1/50 low dose and

2/50 high dose male rats, and a tubular cell car-
cinoma was observed in 1/50 high dose male
rats; none was seen in controls (Table 11). Tu-
bular cell hyperplasia was seen in all groups of
males, including controls.

Because the number of renal tubular cell neo-
plasms identified by standard procedures in the
dosed male rats was low, the marginally in-
creased incidence was not statistically signifi-
cant relative to concurrent controls. Since tubu-
lar cell neoplasms are often late appearing and
are seen only during microscopic examination in
2-year-old rats (i.e., they are often not seen
grossly at necropsy), step-sections of kidney
were made to provide additional data. The re-
maining half of the right and left kidney from
each male rat was embedded, and three or four
additional step-sections from each half kidney
were made at approximately 1-mm intervals.
These were examined microscopically, and addi-
tional tubular cell neoplasms were observed in
all groups (Tables 12 and 13). The tubular cell
adenomas occurred with a significant positive
trend, and the incidences in the low and high
dose groups were significantly greater than that
in the controls.

TABLE 11. RENAL TUBULAR CELL LESIONS IN MALE RATS IN THE TWO-YEAR FEED STUDY OF

NITROFURANTOIN: ORIGINAL EVALUATION (a)

Control

1,300 ppm (b)

2,500 ppm (b)

Hyperplasia

Overall Rates

Adenoma

Overall Rates

Carcinoma

Overall Rates

Adenoma or Carcinoma (c)

Overall Rates
Adjusted Rates
Terminal Rates
Day of First Observation
Life Table Tests
Logistic Regression Tests

2/50(4%)

2/50 (4%)

1/50(2%)

0/50(0%)

1/50(2%)

2/50(4%)

0/50(0%)

0/50(0%)

1/50(2%)

0/50(0%)

1/50(2%)

3/50(6%)

0.0%

3.7%

10.9%

0/24(0%)

1/27(4%)

2/26(8%)

730

719

P = 0.068

P = 0.523

P = 0.134

P = 0.066

P = 0.523

P = 0.134

(a) The statistical analyses used are discussed in Section II (Statistical Methods) and Table A3 (footnotes).

(b)The estimated dose in milligrams per kilograms per day is given in Section III (Body Weights, Feed Consumption, and

Clinical Signs) and in Appendix G.

(c) Historical incidence at study laboratory (mean ± SD): 2/439(0.5% ± 0.9%); historical incidence in NTP studies: 8/1,929

(0.4% ± 0.9%)

43

Nitrofurantoin, NTP TR 341

TABLE 12. RENAL TUBULAR CELL LESIONS IN MALE RATS IN THE TWO-YEAR FEED STUDY
OF NITROFURANTOIN: ADDITIONAL STEP SECTIONS (a)

Lesion

Control

1,300 ppm

2.500 ppm

Hyperplasia
Hyperplasia, cystic
Adenoma
Carcinoma

9/50(18%)
1/50(2%!
3/50(6%)
0/50(0%)

9/50(18%)
5/50(10%)
10/50(20%)
0/50(0%)

7/50(14%)
3/50(6%)
17/50(34%)
1/50(2%)

(a) Hyperplasia and adenomas observed in one control and three low dose males; hyperplasia, adenoma, and carcinoma
observed in one high dose male.

TABLE 13. RENAL TUBULAR CELL LESIONS IN MALE RATS IN THE TWO-YEAR FEED STUDY

OF NITROFURANTOIN: COMPOSITE RESULTS

Lesion

Control

1,300 ppm

2.500 ppm

Hyperplasia

Hyperplasia, cystic

Adenoma

Carcinoma

Adenoma and carcinoma (combined)

Overall Rates

Terminal Rates

Day of First Observation

Lo(;istic Regression Tests

10/50(20%)

11/50(22%)

8/50(16%)

1/50(2%)

.5/50(10%)

3/50(6%)

3/50(6%)

11/50(22%)

19/50(38%)

0/50(0%)

0/50(0%)

2/50(4%)

3/50(6%)

11/50(22%)

20/50(40%)

2/21(10%)

5/23(22%)

1/1(100%)

723

489

619

P< 0.001

P = 0.026

P<0.001

Tubular cell hyperplasia, adenoma, and carci-
noma occurred in the cortex of the kidney and
appeared to encompass a morphologic continu-
um. Tubular cell hyperplasia generally was
characterized by one or two cross-sections of a
normal-to-slightly enlarged tubule with strati-
fied epithelium that partially or completely oc
eluded the tubular lumen. The cells were often
enlarged and contained nuclei with prominent
nucleoli. Adenomas were circumscribed masses
of epithelial cells usually larger than the cross-
sectional diameter of three tubules. The epithe-
lium formed a solid sheet of cells within the
mass or was arranged in packets separated by
basement membrane. The cells were generally
uniform in appearance and similar to those in
hyperplasia. The tubular cell carcinomas were
larger than the adenomas and exhibited stjme
cellular atypia or pleomorphism. The carcino-
mas did not metastasize to other organs.

Characteristic differences between tubular cell
hyperplasia and adenomas in control and dosed
male rats are shown in f^igures 5 through 8 The
malignant character of the tubular cell carci-

noma in the high dose male rat is shown in
Figures 9 and 10.

Parathyroid Glands, Glandular Stomach, and
Bone: Hyperplasia of the parathyroid glands oc-
curred at increased incidences in dosed male rats
(male: control, 3/49; low dose, 18/47; high dose,
23/49; female: 0/49; 0/50; 1/47) This lesion fre-
quently accompanies severe renal disease, and
the increased incidence reflects the increased se-
verity of the nephropathy in dosed male rats.
.Mineralization of the glandular stomach (male:
1/49; 8/50; 14/50; female: 0/50; 2/48; 3/50) and fi-
brous osteodystrophy of the bone also occurred
with increased incidences in dosed male rats
(male: 0/50; .5/50; 5/50; female: none observed)
and were probably the result of the calcium-
phosphate imbalance that accompanies severe
renal disease.

Osteosarcomas were observed in 1/50 low dose
and 2/50 high dose male rats. The overall his-
torical incidence of osteosarcomas in untreated
control male F344/N rats is 8/1,937 (0.4%). The
highest incidence observed in an untreated
control group is 2/50

Nitrofurantoin, NTPTR341

44

>i; "'41 -^^^^i^C

31.3»J

- ^'"^^

V i-T» V

Af Si""

FigTjre 9 Tubular cell carcinoma (arrows) in kidney of a high dose male rat. Note the necrosis in the renter
ot'the large mass.

r/«^ /-.r

^1: 7*>

Figure 10. Higher magnification of the tubular cell carcinoma shown m Figure 9 Note the pleomorphic
cells with large vesicular nuclei and prominent nucleoli and cell in mitosis (arrow).

III. RESULTS: RATS

Subcutaneous Tissue: The incidences of fibro-
mas and fibromas or fibrosarcomas (combined)
in low dose male rats were significantly greater
than those in controls (Table 14).

Testis and Epididymis: Aspermatogenesis and
atrophy are frequently observed in the testis of
2-year-old F344/N rats and are usually associ-
ated with the occurrence of interstitial cell tu-
mors. Although these lesions were present in
control and dosed male rats, the incidence and
severity were higher in dosed rats. Degenera-
tion of the spermatogenic (germinal) epithelium,
fibrinoid necrosis of arterioles, and perivascular
infiltration of mononuclear infiammatory cells
also occurred at increased incidences in dosed
rats (Table 15). The degeneration of the

spermatogenic epithelium was characterized by
a decrease in the number of cells, nuclear pykno-
sis (necrosis), cytoplasmic vacuolization, forma-
tion of spermatid giant cells, and accumulation
of cellular debris in the tubular lumens. The
fibrinoid necrosis involved small arteries and ar-
terioles and consisted of the deposition of hya-
line material within the intima and media. This
was usually accompanied by perivascular ac-
cumulations of lymphocytes, plasma cells, and
macrophages. Atypical cells occurred in the epi-
didymis of dosed rats.

Interstitial cell adenomas of the testis occurred
with a significant negative trend, and the inci-
dence in the high dose group was significantly
lower than that in the controls (Table 16).

TABLE 14. SUBCUTANEOUS TISSUE TUMORS IN MALE RATS [N THE TWO YEAR FEED STUDY OF

NITROFURANTOIN

Control

1,300 ppni

2,500 ppm

Fibroma

Overall Rates
Adjusted Rates
Terminal Rates
Day of First Observation
Life Table Tests
Logistic Regression Tests

Fibrosarcoma

Overall Rates

Fibroma or Fibrosarcoma (a)

Overall Rates
Adjusted Rates
Terminal Rates
Day of First Observation
Life Table Tests
Logistic Regression Tests

0/50 10% 1

5/50(10%)

4/50(8%)

0.0%

16.2%

15.4%

0/24(0%)

3/27(11%)

4/26(15%)

644

730

P = 0.077

P = 0.047

P = 0.071

P = 0.076

P = 0.042

P = 0.071

1/50(2%)

2/50(4%)

1/50(2%)

1/50(2%)

7/50(14%)

5/50(10%)

2.7%

21.9%

17.5%

0/24(0%)

4/27(15%)

4/26 (15%>)

633

644

636

P = 0.127

P = 0.054

P = 0.128

P = 0.109

P = 0.039

P = 0.109

(a) Historical incidence of fibromas, fibrosarcomas, sarcomas, neurofibromas, or neurofibrosarcomas (combined) at study
laboratory (mean ± SD): 28/439(6% ± 4%); historical incidence in NTP studies: 144/1,937(7% ± 4%)

45

Nitrofurantoin, NTP TR 341

TABLE 15. NUMBER OF RATS WITH SELECTED LESIONS OF THE EPIDIDYMIS, PREPUTIAL GLAND,
TESTIS, OR CLITORAL GLAND IN THE TWO-YEAR FEED STUDIES OF NITROFURANTOIN

Site/Lesion Control 600 ppm 1,300 ppm 2,500 ppm

MALE

Number examined microscopically (a) 50 50 50

0 12

Epididymis

Atypical cells

0

Preputial gland

Adenoma

(b)6

Carcinoma

(b)6

Testis

Degeneration

0

Fibrinoid necrosis of arterioles

1

Perivascular infiltration of mononuclear cells

3

Interstitial cell adenoma

47

5

(ciO

6

(clO

0

36

8

15

9

19

45

21

FEMALE

Number examined microscopically 44 38 42

Clitiiral Kland

Adenoma 17 4

Carcinoma 4 3 0

(a) Unless otherwise specified

(bl Forty-eight animals were examined.

(cl Forty-seven animals were examined.

TABLE 16. TESTICULAR INTERSTITIAL CELL ADENOMAS IN MALE RATS IN THE TWO-YEAR FEED

STUDY OF NITROFURANTOIN (a)

Control 1,300 ppm 2,500 ppm

Overall Rates 47/50(94%) 45/50(90%! 21/50(42%)

Adjusted Rates 100.0% 100.0% 61.8%

Terminal Rates 24/24(100%) 27/27(100%) 14/26(54%)

Day of First Observation 455 548 559

Life Table TesU P<0.001N P = 0.162N P<0.001N

Logistic Regression Tests P<0.001N P = 0.221N P<0.001N

(a) Historical incidence of interstitial cell tumors at study laboratory (mean ± SO): 384/439(87% ± 8%); historical incidence
in NTP studies: 1,681/1,909(88% ± 7%)

Nitrofurantoin, NTP TR 341 46

III. RESULTS: RATS

Preputial and Clitoral Glands: Adenomas, carci-
nomas, and adenomas or carcinomas (combined)
of the preputial gland in male rats occurred with
significant negative trends; none was seen in the
high dose group (Table 17). The incidence of
adenomas or carcinomas (combined) of the

clitoral gland in low dose female rats, although
not significantly greater than that in the con-
trols (control, 5/44; low dose, 10/38, high dose,
4/42) (see Table 15), was greater than the high-
est incidence in untreated historical control fe-
male F344/N rats (6/49).

TABLE 17.

PREPUTIAL GLAND TUMORS IN MALE RATS IN THE TWO-YEAR FEED STUDY OF

NITROFURANTOIN

Control

1.300 ppm

2,500 ppm

Adenoma

Overall Rates
Adjusted Rates
Terminal Rates
Day ul'Firsl Observation
Lite Table Tests
[.DKistic Regression Tests

Carcinoma

Overall Rates
Adjusted Rates
Terminal Rates
Day of First Observation
Life Table Tests
Logistic Regression Tests

Adenoma or Carcinoma (a)

Overall Rates
Adjusted Rates
Terminal Rates
Day ol'First Observation
Life Table Tests
Logistic Regression Tests

6/48(13%)

5/50(10%)

0/47(0%)

21.9%

15.2%

0.0%

4/22(18%)

3/27(11%)

0/26(0%)

533

630

P = (I0IIN

P = 0.388N

P = 0.013N

P = 0.018N

P = 0.461N

P = 0.018N

6/48(13%)

6/50(12%)

0/47(0%)

18.3%

17.3%

0.0%

2/22(9%)

2/27(7%)

0/26(0%)

455

320

P = 0.019N

P = 0.522N

P = 0.015N

P = 0.038N

P = 0.603

P = 0.028N

12/48(25%)

11/50(22%)

0/47(0%)

37.6%

30.6%

0.0%

6/22(27%)

5/27(19%)

0/26(0%)

455

320

P<0.001N

P = 0.352N

P<0.001N

P = 0.001N

P = 0.494N

P<0.001N

3); 23/439(5% ± 4%);

historical incidence in NTP studies: 1 23/1 ,937

47

Nitrofurantoin, NTP TR 341

III. RESULTS: RATS

Mammary Gland: Adenocarcinomas in female
rats occurred with a significant negative trend;
none occurred in the high dose group (Table 18).

Eye: Cataracts and retinal degeneration were

observed at increased incidences in high dose
male and low dose female rats (cataracts--male:
control, 1/4; low dose, 0/6; high dose, 14/22; fe-
male: 0/1; lfi/19; 2/6; retinal degeneration-
male: 2/4; 0/6, 17/22; female: 0/1; 17/19; 3/6).

TABLE 18. MAMMARY GI.ANU LESIONS IN FEMALE RATS IN THE TWO YEAR FEED STUDY OF

NITROFURANTOIN

Control

600 ppm

1,300 ppm

Hyperplasia, Cystic or Lobular

Overall Rale.s

Adenocarcinoma (a)
Overall Rates
Adjusted Rates
Terminal Rates
Day Dt'Kirst Ohservation
Life Tal)le Tests
Logistic Regre.ssion Tests

10/49(20%)

24/50(48%)

14/50(28%)

(5/50(12%)

5/50(10%)

0/50(0%)

19.4%

17.2%

0.0%

3/25(12%)

3/26(12%)

0/31 (0%)

(i61

687

P = 0.013N

P = 0.497N

P = ().015N

P = 0.01HN

P = 0.5,S3N

P = 0.019N

(a) Historical incidence of adeiKjinasor adenocarcinomas(combined)at study laboratory (mean ± SDi: 17/439(4% ± 3%);
historical incidence in NTP studies: 64/1,984(3% ± 3%)

Nitrofurantoin, NTP TR 341

48

III. RESULTS: MICE

FOURTEEN-DAY STUDIES

Four of five male and 4/5 female mice that re-
ceived 10,000 ppm and 1/5 females that received
5,000 ppm nitrofurantoin died before the end of
the studies (Table 19). Mice that received 5,000
and male mice that received 10,000 ppm lost

weight. Final mean body weights of other dosed
groups were similar to those of the controls. Es-
timated feed consumption by the groups that re-
ceived 10,000 ppm was notably higher than that
by the controls during the second week of the
studies. Mice that received 10,000 ppm were in-
active, had sunken eyes, and walked on tiptoe.

TABLE 19. SURVIVAL. MEAN BODY WEIGHTS. AND FEED CONSUMPTION OF MICE IN THE
FOURTEEN-DAY FEED STUDIES OF NITROFURANTOIN

Survival

Mean

Body Weights

(grams)

Final Weight Relative
to Controls

Feed
sumpti

Con-

Concentration

Initial (b)

Final

Change (c)

on (d)

(ppm)

(a)

(percent)

Week

1

Week 2

MALE

0

5/5

24.8 ± 0.6

27.2 ± 1.2

-1-2.4 ±0.6

6

6

600

5/5

24.2 ± 0.4

28.6 ± 0.4

^-4.4 ± 0.2

105.1

6

6

1,300

5/5

24.0 ± 0.8

27.0 ± 0.8

-^3.0 ±0.0

99.3

7

6

2,500

5/5

25.0 ± 0.0

27.4 ± 0.2

-1-2.4 ± 0.2

100.7

7

6

5,000

5/5

24.6 ± 0.5

20.2 ± 0.7

-4.4 ± 1.2

74.3

6

6

10,000

(e)l/5

24.6 ± 0.2

19.0

-6.0

69.9

7

13

FEMALE

0

5/5

18.6 ±0.4

21.0 ±0.5

-t-2.4±0.2

6

7

600

5/5

19.0 ± 0.5

22.0 ± 0.3

-1-3.0 ± 0.3

104.8

7

7

1.300

5/5

18.8 ± 0.4

21.4 ± 0.4

-1-2.6 ±0.2

101.9

8

6

2,500

5/5

18.6 ± 0.5

21.0 ± 0.5

+ 2A ±0.4

100.0

7

6

5,000

If) 4/5

19.0 ± 0.3

17.5 ± 1.0

-1.5 ± 1.2

83.3

7

6

10,000

(gll/5

18.8 ± 0.5

20.0

-t-2.0

95.2

6

12

(a) Number surviving/number initially in the group

(b) Initial group mean body weight ± standard error of the mean. Subsequent calculations are based on those animals
surviving to the end of the study.

(c) Mean body weight change of the survivors ± standard error of the mean

(d) Grams of feed consumed per animal per day; average of daily determinations; not corrected for scatter,
(el Day of death: 6,6,6,15

(f) Day of death: 12

(g) Day of death: 6,7,8,9

49

Nitrofurantoin, NTP TR 341

III. RESULTS: MICE

THIRTEEN-WEEK STUDIES

Two of 10 male mice that received 5,000 ppm
and l/IO males that received 300 ppm died be-
fore the end of the studies (Table 20). The final
mean body weight of mice that received 5,000
ppm was 13% lower than that of the controls for
males and 15% lower for females. Estimated
feed consumption by dosed groups was similar
to that by controls. The urine of mice that re-
ceived 5,000 ppm was bright yellow. Inactivity,
hypothermia, and sunken eyes were observed
in mice that received 5,000 ppm but not in
those that received 2,500 ppm. Minimal-to
mild degeneration of the germinal epithelium
of the testis (accompanied by aspermatogene-

sis) was observed in all males that received
1,300, 2,500, or 5,000 ppm; necrosis of the ovar-
ian follicle was observed in 8/10 females that
received 5,000 ppm but not in those that re-
ceived lower doses. Minimal-to-mild necrosis of
the kidney epithelium was observed in 2/9
males that received 5,000 ppm. The liver
weight to body weight ratios were not affected
by administration of nitrofurantoin (Table 21).

Dose Selection Rationale: Because of lower
mean body weight gain in males and females at
higher concentrations and kidney necrosis and
deaths in males at 5,000 ppm, dietary concen-
trations selected for mice in the 2-year studies
were 1,300 and 2,500 ppm nitrofurantoin.

TABLE 20.

SURVIVAL

, MEAN BODY

WEIGHTS, AND FEED CONSUMPTION OF MICE

IN THE

THIRTEENWEEK FEED STUDIES OF MTROFURANTOLV

ition

Surviv,

(a)

Mean

Body Weights (

grams)

Final

1 Weight Relative
to Controls
(percent)

Feed Co
sumption

n-

Concentra

al Initial (b)

Final

Change (c)

(d)

(ppm)

Week

4

Week 12

MALE

0

10/10

25.2

± 0.4

35.0 ± 0.5

-1-9.8 ± 0.6

7

8

300

(el9/10

25.4

±0.5

35.3 ± 0.8

-t-lO.l ±0.8

100.9

7

9

600

10/10

25.5

± 0.3

36.2 ± 0.8

-1-10.7 ± 1.0

103.4

8

7

1.300

10/10

25.1

± 0.4

35.3 ± 0.7

-1-10.2 ± 0.5

100.9

6

8

2,500

10/10

25.6

± 0.3

34.3 ± 0.7

-1-8.7 ± 0.6

98.0

7

8

5,000

if) 8/10

24.8

±0.6

30.3 ± 0.9

+ 5.B ±0.8

86.6

9

10

FEMALE

0

10/10

19.3

±0.3

27.4 ± 0.5

-^8.1 ±0.5

5

6

300

10/10

19.1

± 0.3

27.9 ± 0.9

-t-8.8 ± 0.8

101.8

7

8

600

10/10

19.1

± 0.2

28.2 ± 0.3

-t-9.1 ± 0.4

102.9

7

9

1,300

10/10

19.3

± 0.2

28.2 ± 0.5

-1-8.9 ± 0.5

102.9

7

8

2,500

10/10

18.7

± 0.2

26.6 ± 0.6

-1-7.9 ± 0.6

97.1

7

8

5,000

10/10

19.0

± 0.4

23.3 ± 0.3

-(-4.3 ± 0.4

85.0

7

6

(a) Number surviving/number initially in the group

(b) Initial group mean body weight ± standard error of the mean. Subsequent calculations are based on those animals
surviving to the end of the study.

(c) Mean body weightchange ofthe survivors ± standard error of the mean

(d) Grams of feed consumed per animal per day; not corrected for scatter.
(e)Weel( of death: 11

(fiWeelt of death: 2,2

Nitrofurantoin, NTP TR 341

50

TABLE 21.

ABSOLUTE AND RELATIVE LIVER WEIGHTS OF MICE IN THE THIRTEEN-WEEK FEED
STUDIES OF NITROFURANTOIN (a)

Necro

psy

Liver Weight/Necropsy

Concentration

Number of

Body

Weight

Liver

Weight

Body Weight

(ppm)

Animals

(grams)

(mg)

(mg/g)

MALE

0

10

35.8

+

0.22

1,801

± 48

50.3 ±

1.06

300

9

35.7

+

0.91

(b

1,599

± 70

{b)44.9 ±

1.82

600

10

36.6

+

0.93

Ic

1,529

± 26

(c)42.0 ±

1.20

1,300

10

35.4

+

0.79

1,668

± 67

47.2 ±

1.69

2,500

10

35.4

+

0.82

1,635

± 51

46.3 ±

1.33

5,000

8

(C130.5

+

0.93

(c

1,490

± 56

48.8 ±

0.92

FEMALE

0

10

27.8

+

0.71

1 ,263

± 32

45.5 ±

0.83

300

10

30.3

+

1.17

1,220

±50

ic)40.3 ±

0.81

600

10

29.0

+

0.88

1,253

± 50

43.3 ±

1.32

1 ,300

10

27.7

+

0.58

1 ,244

± 26

45.0 ±

0.94

2,500

10

26.8

+

0.59

1,266

± 49

47.2 ±

1.24

5,000

10

(c)23.5

+

0.22

(b

1,101

± 19

46.9 ±

0.90

(a) Mean ± standard error; P values vs. the i

(b)P<0.05

(c)P<0.01

iintrolsby Dunnett's test i Dunnett, 1955).

TWO-YEAR STUDIES

Body Weights, Feed Consumption, and
Clinical Signs

Mean body weights of high dose male mice
were 5%-17% lower than those of the controls
after week 1 (Table 22 and Figure 11). Mean
body weights of high dose female mice were 4%-
14% lower than those of the controls from week
11 to the end of the study. The average daily

feed consumption by low dose and high dose
male mice was 100% and 95% that by controls
(Table G3) and by low dose and high dose
female mice, 93% and 96% that by controls
(Table G4). The average amount of nitrofuran-
toin consumed per day was estimated to be 300
mg/kg and 570 mg/kg for low and high dose
male mice and 280 mg/kg and 580 mg/kg for
low and high dose female mice. Dosed mice had
bright yellow urine.

51

Nitrofurantoin, NTP TR 341

TABLE 22.

MEAN BODY WEIGHTS AND SURVIVAL OF MICE IN THE TWO-YEAR FEED STUDIES OF

NITROFURANTOIN

Weeks

on
Study

Control

Av. Wt.

(grams)

No. of
Survivors

1,300 ppm

2,500 ppm

Av. Wl. Wt. (percent of No. of Av. Wt. Wt. (percent of No. of
(grams) controls) Survivors (grams) controls) Survivors

MALE

0

M.2

50

24.5

101

.50

235

97

I

2S.8

50

25.9

100

.50

22,0

85

2

26.0

50

26.1

100

.50

23 7

91

3

26.7

50

24.9

93

50

22.2

83

4

27.6

50

27.6

100

.50

25.3

92

5

29.2

SO

28.8

99

50

26.9

92

6

29.9

50

29.1

97

50

27 4

92

7

30.0

50

29.7

99

.50

27.8

93

8

30.9

49

29.7

96

49

28,5

92

»

31.4

47

30.8

98

49

29.3

93

10

31.6

47

30.8

97

49

29.6

94

11

31.9

47

31.3

98

49

29 7

93

12

31.8

46

31.5

99

49

30 I

95

13

33.4

46

31.7

95

49

31.0

93

18

.34.9

45

13.9

97

49

32.0

92

22

36.0

45

35.11

97

49

.33 1

92

•-'6

37.2

45

353

95

48

34 1

92

.10

37.4

44

36.4

97

47

34 1)

91

a.'i

38.0

44

36.8

97

46

34 9

92

40

38.1

44

36.2

95

46

354

93

4!>

39.1

44

37.9

97

46

.160

92

49

40.0

44

37.B

94

45

36.6

92

M

41.4

44

.39.0

94

45

37 6

91

S7

41.5

44

39.6

95

44

.•17 8

91

61

41.3

43

39 1

95

44

37 7

91

65

41.4

42

402

97

44

.•17 9

92

69

41.1

39

396

96

44

.•17 5

91

74

40.7

39

.38.0

93

43

3r, 4

89

78

40.4

38

38.2

95

42

.16 3

90

82

41.0

38

.38.6

94

41

370

90

86

39.6

38

37.2

94

39

36 3

92

90

40.0

37

38.fi

97

36

363

91

94

40.1

35

.38.5

96

35

36 0

90

98

.399

34

38.9

97

33

36 3

91

104

39 B

■ZH

39.1

99

29

,16 5

92

FEMALE

0

18 I

50

18.1

100

50

17 3

96

1

18.4

50

18.0

98

SO

17 7

9B

2

18.7

50

19.4

104

SO

18 9

101

3

20.1

50

19.9

99

50

196

98

4

21.0

50

21.4

102

SO

21 1

100

5

22.1

50

22.8

103

50

21 9

99

6

22.7

50

22.7

100

50

22 1

97

7

23.1

50

23.1

100

SO

22 6

98

8

23.5

50

23.9

102

SO

23 2

99

9

23.8

50

23.9

100

SO

23 2

97

10

24.1

50

24.0

100

50

•J36

98

11

24.7

50

24.1

98

50

■23.6

96

12

25.1

50

24.4

97

50

236

94

13

25.4

50

25.0

98

50

243

96

18

26.6

50

26.2

98

50

25 3

95

22

28.4

49

27.3

96

50

26 1

92

26

29.2

49

28.2

97

50

27 0

92

30

29.6

49

28.8

97

50

26 8

91

35

30.8

49

29.6

96

.50

21 7

90

40

32.1

49

29.8

93

50

•28.4

88

45

32.7

49

31 0

95

50

28 B

87

49

.343

49

32.4

94

50

•-'9 6

86

53

354

49

33 a

115

50

;i 1 1)

K8

57

35.2

49

34.6

9B

50

.1 1 3

89

fil

36.1

49

35.1

97

50

32 1

89

65

37.0

47

365

99

50

34 2

92

69

37.1

45

37.1

100

49

:14 7

94

74

36.4

44

36.7

101

49

.14 4

95

78

36.8

41

36.9

100

49

:14 9

95

82

38.1

40

38.7

102

48

.36 6

96

86

37.7

36

387

103

47

35.7

95

90

390

34

40.2

103

45

36 4

93

94

398

30

40.5

102

45

36 6

92

98

405

29

41.0

101

44

37 0

91

104

41 7

■-'1

4ns

'.*7

17

37 0

89

50
50
50
50
50
50
50
50
50
50
49
49
49
49
49
49
49
49
49
49
49
49
48
48
48
48
48
48
47
46
44
4:i
42
,18

50
50
50
50
50
50
50
50
50
50
50
50
50
50
50
50
50
50
50
50
50
50
50
50
50
.50
50
.50
50
49
49
47
44
42
40

Nitrofurantoin, NTP TR 341

52

50 0

45.0

2

< 40.0

o

>- 35.0
O

UJ

>_ 30.0

Q

O

m

z

< 25.0

20 0-

15.0

A

IaA

■ • n
0 o

. O Q ^
O A 4i

A
A

9 A

!• O'^O ■■ .♦■■ -J

OoaAaAa °.o

A A A 4i A A A

— 1 —

MALE MICE

■

= UNTREATEO

o

= 1.300 PPM

A

= 2.500 PPM

30 45 60

WEEKS ON STUDY

— r —
75

— I —
90

50 0

45.0

40 0

35.0

30 0

25.0

- 20.0

15 0

105

<

o

z

I
o

>-
Q

o
m

z
<

50 0

45 0

40.0

35.0

30 0

25 0-

20.0

15.0

- 45 0

9 S

got

■

A A A

c

A A

: A

FEMALE MICE

■ =

UNTREATED

o=

1.300 PPM

A =

2.500 PPM

— I —
90

50.0

40 0

35 0

30 0

- 25.0

20.0

15.0

105

WEEKS ON STUDY

FIGURE 11. GROWTH CURVES FOR MICE FED DIETS CONTAINING
NITROFURANTOIN FOR TWO YEARS

53

Nitrofurantoin, NTP TR 341

III. RESULTS: MICE

Survival

Estimates of the probabilities of survival for
male and female mice fed diets containing nitro-
furantoin at the concentrations used in these
studies and for controls are shown in Table 23
and in the Kaplan and Meier curves in Fig-
ure 12. The survival of the control group of fe-
male mice was significantly lower than that of
both the low and high dose groups. No sig-
nificant differences in survival were observed
between any groups of male mice.

Pathology and Statistical Analyses of
Results

This section describes the statistically signifi-
cant or biologically noteworthy changes in the
incidences of mice with neoplastic or nonneo-
plastic lesions of the ovary, hematopoietic
system, uterus, liver, testis, epididymis, kidney,
and adrenal glands.

Summaries of the incidences of neoplasms and
nonneoplastic lesions, individual animal tumor
diagnoses, statistical analyses of primary tu-
mors that occurred with an incidence of at least
5% in at least one animal group, and historical
control incidences for the neoplasms mentioned
in this section are presented in Appendixes C
and D for male and female mice, respectively.

TABLE 23. SURVIVAL OF MICE IN THE TWO-YEAR FEED STUDIES OF NITROFURANTOIN

Control

1,300 ppm

2,500 ppm

MALE (a)

Animals initially in study

Nonaccidental deaths before termination (b)

Killed at termination

Died during termination period

Survival P values (c)

FEMALE (a)

Animals initially in study

Nonaccidental deaths before termination (bl

Killed at termination

Died during termination period

Survival P values (c)

50

50

22

21

28

29

0

0

0.173

0.897

50

50

31

13

19

36

0

1

<0.00I

< 0.001

50
16
33

1

0.197

50

13

37

0

<0.001

(a) First day of termination period: 730

(b) Includes animals killed in a moribund condition

(c)The result of the life table trend test is in the control column, and the results of the life table pairwise comparisons with
.the controls are in the dosed columns.

Nitrofurantoin, NTPTR 341

54

1.0

0.9-

> 0.8
>

z>

0.7

o

^ 0 6
CD

<

CD

o
a:
a. 0.5

0.4

O.J

MALE MICE

■ Control
O 1300 ppm
A 2500 ppm

— r-
15

30

— I —
45

60

WEEKS ON STUDY

— I —

75

— I —
90

105

120

45 60 75

WEEKS ON STUDY

120

FIGURE 12. KAPLAN-MEIER SURVIVAL CURVES FOR MICE FED DIETS
CONTAINING NITROFURANTOIN FOR TWO YEARS

55

Nitrofurantoin, NTP TR 341

III. RESULTS: MICE

Ovary: Ovarian abscesses were observed in
18/50 control female mice but in none of the
dosed mice (Table 24). Atrophy, characterized
by the absence of graafian follicles and corpora
lutea, occurred in 48/50 low dose and 49/50 high
dose female mice but not in controls. Uncommon
ovarian tumors including tubular adenomas, be-
nign mixed tumors, and granulosa cell tumors
occurred only in dosed female mice (Table 25).
Tubulostromal tumors form a continuous mor-
phologic spectrum and typically consist of com-
plex branching tubules originating from the
surface mesothelium and varying numbers of in-
tertubular cells derived from the ovarian stro-
ma. Those tumors with a minimum of stromal

cells were classified as tubular adenomas,
whereas those with a prominent stromal cell
component were classified as benign mixed tu-
mors (Figures 13 and 14). The granulosa cell tu-
mors were characterized by the predominant
component of typical granulosa cells arranged in
a variety of patterns. The tumor diagnosed as a
neoplasm, NOS, is an extremely uncommon tu-
mor in B6C3Fi mice. The pattern of growth and
cellular morphology were characteristic of the
Sertoli cell tumor that occurs in the testis. Al-
though of unusual morphologic pattern, this tu-
mor is of sex cord origin similar to that of the
granulosa cell tumors.

TABLE 24.

NUMBER OF FEMALE MICE WITH OVARIAN LESIONS IN THE TWO-YEAR FEED STUDY

OF NITROFURANTOIN

Lesion

Control

1.300 ppm

2,500 ppm

Number examined microscopically

Abscess
Atrophy

Tubular adenoma
Cystadenoma
Papillary cystadenoma
Granulosa cell tumor, benign
Granulosa cell tumor, malignant
Mixed tumor, benign
Neoplasm, NOS

50

50

0
48
0
0
1
3
0
0
0

50

0
49
5
1
0
1
1
4
1

Nitrofurantoin, NTP TR 341

56

Figure 13. Benign mixed tumor obliterating ail normal tissue in ovary of a high dose female mouse

Figure 14 Higher magnification of ovanan mixed tumor shown in Figure 13. The tumor consists of gonadal
stromal relis and tubules lined by cuboidal epithelium. The tubules appear to be adowngrowthofthe modified
mesotheiium covenng the surface of the ovary

TABLE 25. OVARIAN TUMORS IN FEMALE MICE IN THE TWO-YEAR FEED STUDY OF

NITROFURANTOIN (a)

Control

1,300 ppm (b)

2,500 ppm (b)

Tubular Adenoma (c)

Overall Elates

0/50(0%)

0/50(0%)

5/50(10%)

Adjusted Elates

0.0%

0.0%

13.0%

Terminal Elates

0/19(0%)

0/37(0%)

4/37(11%)

Day of First Observation

729

Life Table Tests

P = 0.019

(d)

P = 0.127

Logistic Regression Tests

P = 0.018

(d)

P = 0.ir2

Mixed Tumor, Benign (e)

Overall Rates

0/50(0%)

0/50(0%)

4/50(8%)

Adjusted Elates

0.0%

0.0%

10.0%

Terminal Rates

0/19(0%)

0/37 (0%>)

3/37(8%)

Day of First Observation

630

Life Table TesU

P = 0.035

(d)

P = 0.162

Logistic Regression Tests

P = 0.018

(d)

P = 0.084

Tubular Adenoma or Mixed Tumor, Benign

Overall Rates

0/50(0%)

0/50(0%)

9/50(18%)

Adjusted Rates

0.0%

0.0%

22.6%

Terminal Elates

0/19(0%)

0/37 (0%)

7/37(19%)

Day of First Observation

630

Life Table Tests

P = 0.001

(d)

P = 0.028

Logistic Regression Tests

P<0.001

(d)

P = 0.010

Granulosa Cell Tumor, Benign

Overall Rates

0/50(0%)

3/50(6%)

1/50(2%)

Adjusted Elates

0.0%

8.1%

2.7%

Terminal Rates

0/19(0%)

3/37(8%)

1/37(3%)

Day of First Observation

730

730

Life Table Tests

P = 0.564

P = 0.260

P = 0.633

Logistic Regression Tests

P = 0.564

P = 0.260

P = 0.633

Granulosa Cell Tumor, Benign or Malignant (0

Overall Rates

0/50(0%)

3/50(6%)

2/50(4%)

Adjusted Elates

0.0%

8.1%

5.1%

Terminal Elates

0/19(0%)

3/37(8%)

1/37(3%)

Day of First Observation

730

729

Life Table Tests

P = 0.375

P = 0.260

P = 0.404

Logistic Regression Tests

P = 0.362

P = 0.260

P = 0.374

(a) The statistical analyses used are discussed in Section II (Statistical Methods) and Table D3 (footnotes).

(b)The equivalent dose in milligrams per kilograms per day is given in Section III (Body Weights, Feed Consumption, and

Clinical Signs) and in Appendix G.

(c) Historical incidence in NTP studies (mean): 4/1,858(0.2%)

(d) No P value is reported because no tumors were observed in the 1 ,300-ppm and control groups.

(e) Historical incidence in NTP studies (mean): 1/1,858 ( <0.1%)

(f) Historical incidence of granulosa cell tumors or luteomas (combined) in NTP studies (mean): 8/1 ,858 (0.4%)

57

Nitrofurantoin, NTP TR 341

III. RESULTS: MICE

Hematopoietic System: Lymphomas in female
mice occurred with a significant positive trend
by the logistic regression test; the incidence in
the high dose group was not significantly
greater than that in the controls (Table 26).

Uterus: Suppurative inflammation was ob-
served in 11/50 control mice but in none of the
dosed animals. Adenocarcinomas were seen in
1/50 low dose and in 1/50 high dose mice. The
highest observed incidence of uterine adenomas
or adenocarcinomas (combined) in untreated his-
torical control female B6C3Fi mice is 1/47; the
overall historical incidence is 6/2,010 (0.3%).

Liver: An Ito cell tumor was observed in 1/50
low dose and 1/50 high dose female mice. These
tumors consisted of well-differentiated fat cells
and varying amounts of collagen-containing
mesenchymal tissue. Previously, these tumors
have been classified as lipomas or liposarcomas
because of their component of fat cells. A lipoma
has been diagnosed in 1/2,033 (<0.1%) un-
treated historical control female B6C3Fi mice.
Hepatocellular adenomas and hepatocellular
adenomas or carcinomas (combined) in female
mice occurred with significant positive trends by
the logistic regression test; the incidences in the
dosed groups were not significantly different
from those in the controls (Table 27).

TABLE 26.

MAUGNANT LYMPHOMAS LN FE.MALE MICE I.\ THE TWO-YEAR FEED STUDY OF

NITROFURANTOIN (a)

Control

1,300 ppm

2,500 ppm

Overall Rates
Adjusted Rates
Terminal Rates
Day of First Observation
Life Table Tests
Logistic Regression Tests

12/50(24%)

19/50(38%)

24/50(48%)

50.2%

43.4%

52.7%

8/19(42%)

13/37(35%)

16/37(43%)

631

596

568

P = 0.352

P = 0.447N

P = 0.449

P = 0.038

P = 0.295

P = 0.076

(a) Historical incidence of lymphomasor leukemia (combined)atstudy laboratory (mean ± SD): 104/448(23% ± 7%);
historical incidence in NTP studies: 616/2,041(30% ± 12%)

TABLE 27. HEPATOCELLULAR TUMORS IN FEMALE MICE IN THE TWO-YEAR FEED STUDY OF

NITROFURANTOIN

Control

1,300 ppm

2,.500 ppm

Adenoma

Overall Rates
Adjusted Rates
Terminal Rates
Day of First Observation
Life Table Tests
Logistic Regression Tests

Carcinoma

Overall Rates

Adenoma or Carcinoma (a)
Overall Rates
Adjusted Rates
Terminal Rates
Day of First Observation
Life Table Tests
Logistic Regression Tests

1/50(2%)

1/50(2%)

7/50(14%)

2.7%

2.7%

18.1%

0/19(0%)

1/37(3%)

6/37(16%)

603

730

660

P = 0.042

P = 0.658N

P = 0.147

P = 0.016

P = 0.758

P = 0.054

1/50(2%)

1/50(2%)

2/50(4%)

2/50(4%)

2/50(4%)

8/50(16%)

6.6%

4.9%

20.0%

0/19(0%)

1/37(3%)

6/37(16%)

603

670

660

P = 0.093

P = 0.531N

P = 0.217

P = 0.029

P = 0.704N

P = 0.079

(a) Historical incidence at study laboratory (mean ± SD): 36/447(8% ± 5%); historical incidence in NTP studies: 177/2,033
(9% ± 5%)

Nitrofurantoin, NTP TR 341

58

III. RESULTS: MICE

Testis: Aspermatogenesis and degeneration of
the germinal epithelium were observed at in-
creased incidences in high dose male mice
(aspermatogenesis: control, 1/49; low dose, 1/49;
high dose, 16/50; degeneration of the germinal
epithelium. 0/49; 3/49; 23/50).

Epididymis: Atypical cells and depletion were
observed in high dose male mice (atypical cells:
control, 0/50; low dose, 0/49; high dose, 26/50;
depletion: 1/50; 1/49; 15/50).

Kidney: Mineralization of the medulla was ob-
served in high dose mice (male: control, 0/50;
low dose, 0/50; high dose, 17/50; female: 0/50;
0/50; 7/50). Dilatation of the tubules was
observed in high dose male mice (male: 0/50;
0/50; 14/50; female: 0/50; 1/50; 1/50).

Adrenal Glands: Cortical spindle cell hyper-
plasia was observed at increased incidences in
dosed female mice (male: control, 5/50; low dose,
3/49; high dose, 4/50; female: 3/50; 41/50; 45/50).
A spindle cell (adrenal capsule) adenoma was
seen in 1/50 low dose female mice, and a spindle
cell (adrenal capsule) carcinoma was seen in
1/49 low dose male mice. The historical inci-
dences of these tumors are unknown because
neither has been entered into the data base as a
category separate from adenoma, NOS. Pheo-
chromocytomas or malignant pheochromo-
cytomas (combined) in male mice occurred with
a significant negative trend; the incidences in
the dosed groups were significantly lower than
that in the controls (Table 28).

TABLE 28.

ADRENAL MEDULLARY LESIONS IN MALE MICE IN THE TWO-YEAR FEED STUDY OF

NITROFURANTOIN

Control

1.300 ppm

2,500 ppm

Hyperplasia

Overall Rates

Pheochromocytoma

Overall Rates
Adjusted Rates
Terminal Rates
Day of First Observation
Life Table Tests
Logistic Regression Tests

Malignant Pheochromocytoma

Overall Rates

Pheochromocytoma or Malignant Pheochromocytoma (a)

Overall Rates
Adjusted Rates
Terminal Rates
Day of First Observation
Life Table Tests
Logistic Regression Tests

4/49(8%)

6/48(13%)

5/49(10%)

4/49(8%)

0/48(0%)

1/49(2%)

13.3%

0.0%

3.0%

3/28(11%)

0/28(0%)

1/33(3%)

688

730

P = 0.062N

P = 0.065N

P = 0.138N

P = 0.059N

P = 0.061N

P = 0.132N

2/49(4%)

0/48(0%)

0/49(0%)

•toma (a)

6/49(12%)

0/48(0%)

1/49(2%)

19.7%

0.0%

3.0%

4/28(14%)

0/28(0%)

1/33(3%)

688

730

P = 0.010N

P = O.OiaN

P = 0.038N

P = 0.009N

P = 0.016N

P = 0.034N

(a)Historical incidence atstudy laboratory (mean ± SD): 4/437(1% ± 1%); historical incidence in NTP studies: 25/1,962
(1% ± 2%)

59

Nitrofurantoin, NTP TR 341

Nitrofurantoin, NTP TR 341 60

IV. DISCUSSION AND CONCLUSIONS

61 Nitrofurantoin, NTPTR 341

IV. DISCUSSION AND CONCLUSIONS

Nitrofurantoin was studied and evaluated be-
cause of its widespread use as a drug for treating
urinary tract infections in humans, its struc-
tural relationship to known carcinogenic 5-ni-
trofuran compounds (lARC, 1974; Cohen, 1978),
and the lack of adequate studies to assess its car-
cinogenicity. Toxicology and carcinogenesis
studies of nitrofurantoin were conducted by ad-
ministering USP-grade nitrofurantoin (greater
than 99% pure) in feed to groups of F344/N rats
and B6C3Fi mice of each sex for 14 days, 13
weeks, or 2 years. In the 2-year studies, nitrofu-
rantoin was administered in feed at 0, 1,300, or
2,500 ppm to male rats and male and female
mice and at 0, 600, or 1 ,300 ppm to female rats.

In the 13-week studies, only one rat (a high dose
female) died. Mean body weights relative to
those of controls were similar for dosed male and
dosed female rats, but because of the lower rela
tive mean body weight of females in the 2,500-
ppm group, the dietary concentrations selected
for the 2-year studies were lower for females
than for other groups. P'or mice, two deaths oc-
curred in the high dose male group, but final
mean body weights relative to those of controls
were similar for males and females. Organs af-
fected in the 13- week studies were the testis or
ovary in rats and mice and the kidney in male
mice.

In the 2-year studies, there were no significant
differences in survival between dosed and con-
trol groups of rats of either sex or male mice (see
Tables 10 and 23). Regarding female mice, the
survival of the control group was lower than that
of both the low and high dose groups. Ovarian
abscesses and suppurative infiammation of the
uterus were observed only in control female
mice. These infections are believed to be indige-
nous and were absent in dosed mice, most likely
due to the therapeutic activity of nitrofurantoin.
Treatment at the doses used in these studies
would be expected to achieve the minimally ef-
fective dose level (-30 pg/ml urine) against a
broad spectrum of bacteria (Paul, M.F., et al.,
1960; Buzard et al., 1961; Veronese et al., 1974;
Liedtke et al., 1980; Hoener and Patterson,
1981).

Generally, body weights and estimated feed con-
sumption values indicate that no or minimal
overt toxicity or feed palatability problems were

encountered in these studies except for female
mice. Absorption, metabolism, and excretion of
nitrofurantoin are rapid (Paul, M.F , et al., 1960;
Buzard et al., 1961; Conklin and Hailey, 1969;
Conklin, 1972a,b; Veronese et al., 1974; Maiti
and Banerjee, 1978; Wierzba et al., 1982) and
change with age in both humans and rats
(Braunlich et al., 1978; Wierzba et al., 1982). Af-
ter an initial decrease relative to controls, body
weight differences between dosed and control
rats leveled off. This adaptive response may be
due to an age dilTerence in susceptibility to ni-
trofurantoin, toxicity, or possible metabolic
adaptation (enzyme induction) to chemical expo-
sure. High dose male mice demonstrated similar
differences in body weight relative to those of
controls throughout the study. The exception
may be the high dose female mice, whose body
weights were decreased relative to those of the
controls. Interpretation is made more difficult
because of the decreased survival of control fe-
male mice. Survival of dosed female mice was
not affected, and body weight differences may
have been due to palatability of nitrofurantoin
or to induced neoplasia.

The oral (gavage) toxicity of nitrofurantoin var-
ies somewhat between rats (LD5() = 604 mg/kg)
and mice (LD5o = 360 mg/kg) (Preti, 1970;
NIOSH, 1983). The toxicity of nitrofurantoin
administered in feed over longer daily time
periods is different. The absorption and urinary
excretion of macrocrystalline nitrofurantoin (ad-
ministered in a capsule) were greater in non-
fasting than in fasting volunteers (Bates et al.,
1974); a much smaller difference was observed
for the microcrystalline form (given as a tablet).
In the current NTP studies, the doses of nitro-
furantoin consumed by rats (up to 110 mg/kg per
day) would not be expected to be overtly toxic.
In mice, the doses (300 or 570 mg/kg, males; 280
or 580 mg/ kg, females) might have been lethal if
given as a single bolus but apparently were well
tolerated given over a period of 24 hours. The
recommended dosage of nitrofurantoin for hu-
mans is 50-100 mg, four times a day for 2 weeks
(Penn and Griffin, 1982). However, treatment
may continue for longer periods (6-30 months)
and at higher doses (Simonian et al., 1977; Penn
and Griffin, 1982); for an individual weighing 70
kg, this is equivalent to approximately 3-6 mg/kg
per day up to 10 mg/kg per day.

Nitrofurantoin, NTP TR 341

62

IV. DISCUSSION AND CONCLUSIONS

Metabolically, under aerobic conditions, the re-
duction of nitrofurantoin stimulates tlie con-
sumption of oxygen and tlie production of super-
oxide anion free radical and hydrogen peroxide
in avian liver and in mammalian liver, lung,
small intestine, kidney, and gastrointestinal
contents which may result in toxicity and lo-
calized injury (Mason and Holtzman, 1975a;
Biaglow et al., 1977; Aufrere et al., 1978; Boyd et
al., 1979a; Sasame and Boyd, 1979; Leskovac
and Popovic, 1980; Peterson et al., 1982a). Un-
der anaerobic conditions, nitrofurantoin is per-
manently reduced to nitroso and/or hydroxyl-
amine forms (Mason and Holtzman, 1975a;
Biaglow et al., 1977; Leskovac and Popovic,
1980), which may result in binding to cellular
macromolecules (DNA and protein) (Boyd et al.,
1979b). The covalent binding to macromolecules
is apparently greatest in the kidney, liver, ile-
um, lung, and heart of rats (Aufrere et al., 1978).
Toxicity and DNA damage may increase as
oxygen tension decreases (Russo et al., 1982).

The kidney is a primary organ of metabolism
and excretion and is the site of chemical-related
toxicity. The severity of chronic nephropathy
was greatest in high dose male rats. This spon-
taneous disease occurs in nearly all laboratory
rats, and the onset is generally earlier and the
effects are more severe in males than in females
(Chennekatu et al., 1986). Proteinuria begins
when the male rat is several months old and in-
creases progressively as the animal ages, which
indicates progressive impairment of some renal
functions. The reason for the apparent selective
toxicity of nitrofurantoin to the kidney of male
rats may be related to the fact that the kidneys
receive up to 20% of the cardiac output of blood,
have a large endothelial and epithelial surface
area that is exposed to the blood or glomerular
filtrate containing the chemical, perform diverse
metabolic functions, have a high concentrating
function for excreted and absorbed metabolites,
and have an age-related deterioration in kidney
function. The nonneoplastic lesions observed in
this study (parathyroid gland hyperplasia, fi-
brous osteodystrophy of the bone, and minerali-
zation of the glandular stomach) are character-
istic of renal secondary parathyroidism and are
believed to be secondary to the chronic nephrop-
athy (Burns, 1979).

The kidney was identified as a target organ in
mice in the 13-week studies in which mineral-
ization of the renal medulla in dosed male and
female mice and dilatation of the renal tubules
in dosed male mice were observed. The original
evaluation of the kidney by standard procedures
(i.e., microscopic examination of single longitu-
dinal sections of the left and right kidney) iden-
tified small numbers of tubular cell neoplasms
in dosed male rats but not in controls (control,
0/50; low dose, 1/50; high dose, 3/50). The inci-
dences in dosed rats were not statistically
greater than that in concurrent controls, but tu-
bular cell neoplasms occur rarely in untreated
historical controls (8/1,929, 0.4%) (Table A4a).
Thus, an informal comparison of the incidences
in dosed male rats with historical controls sug-
gested that the neoplasms may be chemical
related.

Kurokawa et al. (1983) compared results of
examination of single vs. multiple sections of
kidney and found that incidences were greater
with multiple sections. Therefore, the NTP pre-
pared step-sections of the remaining right and
left halves of the kidney to provide additional
data and to clarify the potential relationship of
the tubular cell neoplasms to the administration
of nitrofurantoin. The results of this subsequent
evaluation unequivocally demonstrated a dose-
related and significantly increased incidence of
renal tubular cell adenomas in male rats given
nitrofurantoin (low dose, P<0.05; high dose,
P< 0.001). The data are considered to represent
some, rather than clear, evidence of carcinogenic
activity for the following reasons: standard
histologic procedures (single sections of kidney)
showed only small numbers of tubular cell neo-
plasms in dosed male rats; the tubular cell neo-
plasms in dosed rats were predominantly ade-
nomas; the adenomas were small, microscopic
tumors; some were difficult to distinguish from
hyperplasia; and the biologic potential of many
of the small adenomas is uncertain.

The liver has been identified as a major site of
metabolism, a minor site of excretion, and a
potential target organ Hepatocellular neo-
plasms (adenomas or carcinomas, combined) in
female mice occurred with a positive trend (see
Table 27). An Ito cell tumor of the liver was

63

Nitrofurantoin, NTPTR 341

IV. DISCUSSION AND CONCLUSIONS

observed in one low dose and in one high dose fe-
male mouse (Table Dl). Although these Ito cell
neoplasms are relatively uncommon, they were
considered to be unrelated to nitrofurantoin
administration.

Osteosarcomas observed in the bone (Table Al)
of dosed male rats are also of marginal incidence
but are rare in control animals (8/1,937, 0.4%).
The incidences of subcutaneous fibromas or fi-
brosarcomas (combined) were greater in dosed
male rats than in controls (see Table 14).

Effects on the testis in male rats and mice in the
13-week studies included aspermatogenesis and
degeneration. Nitrofurazone, an analog of nitro-
furantoin, inhibits spermatogenesis in rats,
which results in testicular atrophy after long-
term administration (Prior and P'erguson, 1950;
Nissim, 1957; Montemurro, 1969). A similar
effect was reported in mice, together with inter-
stitial cell hyperplasia and seminal vesicle hy-
pertrophy. In the current 2-year studies, admin-
istration of nitrofurantoin was associated with
the induction of atypical cells of the epididymis,
testicular degeneration, and a decrease in the in-
cidence of interstitial cell adenomas of the testis
in rats (see Table 15) and an increase in the in-
cidence of atypical cells and depletion of the epi-
didymis in high dose male mice (Table C5). In
high dose male mice, testicular aspermato-
genesis and degeneration of the germinal epithe-
lium were observed. No reports have been pub-
lished on whether this effect has been observed
or studied in humans. Other compound-related
changes in the reproductive tissues of dosed
male animals relative to those of controls were
decreases in adenomas or carcinomas (combined)
of the preputial gland in high dose male rats (see
Table 17).

No neoplastic lesions in dosed female rats or
dosed male mice were considered to be compound
related at the doses of nitrofurantoin admin-
istered in these 2-year studies. The absence of
any observed toxicity-related effects suggests
that female rats might have been able to toler-
ate higher doses. Only the incidence of clitoral
gland neoplasms in low dose female rats (Tables
Bl and B4a) gave any indication of a potential
compound-related effect; this effect was not sup-
ported by a similar observation in the higher
dose group.

Ovarian atrophy was associated with increased
incidences of tubular adenomas of the ovary, be-
nign mixed tumors, and granulosa cell tumors in
dosed mice (see Table 25). Ovarian follicular ne-
crosis was associated with nitrofurantoin admin-
istration in the 13- week studies. Biskind and
Biskind (1944) reported the influence of gonado-
tropic hormones on the biologic behavior of
ovarian tumors. Ovarian atrophy is recognized
as an event that is common to and associated
with the development of ovarian tumors. In a
model developed for studying ovarian tumori-
genesis. Murphy (1972) reported that B6C3Fi-
W'^IW" mice, (C57BL/6J X C3H/HeJ)Fi-WVWy
hybrids, develop spontaneous complex tubular
adenomas (mesothelial adenomas) (95%-100%
incidence). Homozygous recessive W allele mu-
tants are sterile, develop macrocytic anemia,
and lack hair pigmentation. Ovaries of these
hybrid mice contain less than 1% of the normal
complement of oocytes. Tumor development is
associated with loss of oocytes and follicular cells
and increased levels of pituitary gonadotropic
hormones (luteinizing hormone and follicle stim-
ulating hormone, two to four times normal
values) (Murphy, 1972; Murphy and Beamer,
1973). Prolonged stimulation of the ovary by go-
nadotropins apparently induces tubular adeno-
mas of the ovary. More recently, Tennent and
Beamer (1986) and Beamer and Tennent (1986)
reported that gonadotropins are necessary for
normal follicular atresia and stromal leuteiniza-
tion following oocyte death (x-irradiation) but
were not sufficient to induce adenomas in hypo-
gonal ihpglhpg) mice, which retain follicular
structure in the absence of oocytes and are de-
ficient in gonadotropin-releasing hormone.

Ovarian atrophy and loss of follicular cells result
in increased gonadotropin stimulation. Pro-
longed stimulation may promote hyperplasia of
ovarian cells, resulting in benign tumors; how-
ever, under certain conditions (e.g., oocyte death
due to irradiation or possibly to chemical toxic-
ity), prolonged stimulation alone is insufficient
to induce complex tubular adenomas. Increased
gonadotropin stimulation may promote tumor
mass by hypertrophy and/or hyperplasia. The
initiating events are not clear; however, genetics
and age may influence the progression of events.
Although the majority of ovarian tumors
observed in the current studies were not

Nitrofurantoin, NTP TR 341

64

IV. DISCUSSION AND CONCLUSIONS

considered to be malignant (tubular cell adeno-
mas or mixed tumors), granulosa cell tumors
may progress to malignancy in mice (Beamer et
al., 1985). Progression from complex tubular
adenomas or granulosa cell tumors to malignant
neoplasms may occur in B6C3Fi mice as in other
strains (Murphy and Russell, 1963; Alison et al.,
1987). Nitrofurantoin toxicity observed in the
gonads of female mice in these studies is difficult
to interpret; it may initiate genetic events lead-
ing to tumor development, or it may upset hor-
monal balance between the pituitary gland and
the gonads and indirectly cause or enhance tu-
mor development as described in studies with
7,12-dimethylbenz[alanthracene (DMBA) (Ta-
guchi et al., 1988). In another study of a 5-nitro-
furan (nitrofurazone), there were dose-related
increased incidences of ovarian tumors of these
same types (NTP, 1988).

Ovarian abscesses and suppurative inflamma-
tion of the uterus were observed in control but
not in dosed female mice (Table D5) and are be-
lieved to be related to indigenous microbial in-
fections. Adenocarcinomas of the uterus were
seen in two dosed mice (Table Dl); uterine ade-
nomas and adenocarcinomas are observed rarely
in historical controls (6/2,010, 0.3%).

Spindle cell ("Type A" cell) hyperplasia of the
adrenal gland was observed in dosed female
mice. A spindle cell adenoma was seen in one
low dose female mouse, and a spindle cell carci-
noma was seen in one low dose male mouse.
Since gonadectomy of mice is known to result in
hyperplasia and neoplasia of the "Type A" cells
in the adrenal cortex (Turusov, 1979), the
spindle cell hyperplasia in female mice given ni-
trofurantoin is likely related to the ovarian atro-
phy and disruption of normal hormone levels.

Nitrofurantoin was mutagenic for Salmonella
strains TA98 and TAIOO, with and without
metabolic activation, but not for strains TA1535
or TA1537 (Table El). Nitrofurantoin induced
forward mutations at the TK locus of mouse
L5178Y lymphoma cells in the absence of met-
abolic activation (Table E2), induced increased
numbers of sister chromatid exchanges (SCEs)
(Table E3) and chromosomal aberrations in cul-
tured Chinese hamster ovary (CHO) cells with
and without metabolic activation (Table E4),
and was negative for sex-linked recessive lethal

mutations in Drosophila (Table E5). The mu-
tagenicity of nitrofurantoin has been attributed
to a reduced nitre group on the furan ring as a
result of the metabolic action of either a bac-
terial nitroreductase or a eukaryotic cell enzyme
system. In bacteria, anaerobic conditions favor-
ing rapid action of the nitroreductase enzyme
system have been shown to enhance the muta-
genicity of nitrofurantoin (Rosenkranz and
Speck, 1976). Availability of at least one alter-
nate nitroreductive pathway has been demon-
strated in Salmonella, in that the nitroreductase
deficient strain TAIOO-FRI is mutated by nitro-
furantoin in the absence of oxygen but not in its
presence. Kramers (1982) has also shown that
Drosophila are able to carry out a presumably
similar metabolism. Several compounds with
nitro groups which are known to be Salmonella
mutagens have also been shown to induce muta-
tions in germ-free (lacking gut fiora) Drosophila.
Hence, the files were able to appropriately me-
tabolize nitro compounds to active intermedi-
ates. Further, extracts from germ-free files en-
abled a nitroreductase deficient Escherichia coli
strain to reduce the related 5-nitrofuran, nitro-
furazone, to a mutagenic form.

Olive (1980) demonstrated that the reduction po-
tential (electron affinity) of nitroheterocyclic
compounds including nitrofurantoin is directly
correlated with the mutagenic activity of these
chemicals in hamster V79 spheroids. Further,
Shirai and Wang (1980) investigated the rela-
tionship between the strength of the mutagenic
response of eight different nitrofurans, including
nitrofurantoin, in Salmonella and their ability
to induce SCEs in cultured CHO cells. The mag-
nitude of the responses in both test systems cor-
related well; i.e., weak Salmonella mutagens
were weak inducers of SCEs, and vice versa.
Thus, nitrofurantoin is mutagenic in cultured
mammalian cells as well as in bacteria, both of
which apparently have the capacity to transform
these chemicals into mutagenic intermediates.

The reductive metabolism of nitrofurantoin in
mammalian cell systems is mediated by
NADPH-cytochrome c reductase and xanthine
oxidase. The presumed active intermediate is
the hydroxylamino compound that has been iso-
lated by some investigators but is extremely
oxygen-labile and difficult to detect. Another

65

Nitrofurantoin, NTP TR 341

IV. DISCUSSION AND CONCLUSIONS

potential metabolic reaction could involve hy
drolysis of the carbon-nitrogen bond, resulting
in production of a furfuraldehyde and an imida-
zole. Aldehydes, hydrazines, semicarbazides,
imidazoles, and related compounds have clasto-
genic potential. However, nitrofurantoin has
not demonstrated mutagenic activity in any of
the in vivo somatic and germ cell test systems in
which it has been studied.

The experimental and tabulated data for the
NTP Technical Report on nitrofurantoin were
examined for accuracy, consistency, complete-
ness, and compliance with Good Laboratory
Practice regulations. As summarized in Appen-
dix I, the audit revealed no major problems with
the conduct of the studies or with collection and
documentation of the experimental data. No dis-
crepancies were found that influenced the final
interpretation of the results of these studies.

Under the conditions of these 2-year feed stud-
ies, there was some evidence of carcinogenic
activity* of nitrofurantoin for male F344/N rats
as shown by increased incidences of uncommon
kidney tubular cell neoplasms. Uncommon

osteosarcomas of the bone and neoplasms of the
subcutaneous tissue were observed in dosed
male rats. Incidences of interstitial cell adeno-
mas of the testis and neoplasms of the preputial
gland were decreased in the 2,500-ppm group of
male rats. There was no evidence of carcinogenic
activity of nitrofurantoin for female F344/N rats
fed diets containing 600 ppm or 1,300 ppm for 2
years. Female rats may have been able to tol-
erate higher doses. There was no evidence of car-
cinogenic activity of nitrofurantoin for male
B6C3Fi mice fed diets containing 1,300 ppm or
2,500 ppm for 2 years. There was clear evidence
of carcinogenic activity of nitrofurantoin for fe-
male B6C3P'i mice as shown by increased inci-
dences of tubular adenomas, benign mixed tu-
mors, and granulosa cell tumors of the ovary.

Nonneoplastic lesions considered related to ni-
trofurantoin exposure were chronic nephropathy
and associated lesions (hyperplasia of the para-
thyroid gland, fibrous osteodystrophy of the
bone, and mineralization of the glandular stom-
ach) in male rats and testicular degeneration in
male rats and mice. Ovarian atrophy and hy-
perplasia of the adrenal cortex spindle cells were
observed in dosed female mice.

•Explanation of Levels of Evidence of Carcinogenic Activity is on page 7.

A summary of the Peer Review comments and the public discussion on this Technical Report appears on pages 1113.

Nitrofurantoin, NTP TR 34 1

66

V. REFERENCES

67 Nitrofurantoin, NTPTR 341

V. REFERENCES

1. Alison, RU . Morgan, K T , Haseman, J.K ;
Boorman, G.A. (1987) Morphology and classifi
cation of ovarian neoplasms in F344 rats and
(C57BIV6 X C3H)Fi mice. J. Natl. Cancer Inst.
78:1229-1243.

11 Biaglow, J.E ; Jacobson, B.E.; Nygaard, O.F
(1977) Metabolic reduction of 4-nitroquinoline-
/V-oxide and other radical-producing drugs to
oxygen-reactive intermediates. Cancer Res.
37:3306-3313.

2. Analytical Profiles of Drug Substances (1976)
Vol 5. Florey, K., Ed. New York: Academic
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evaluation of different pairs of DNA repair-defi-
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strains for rapid detection of chemical mutagens
and carcinogens. Mutat Res. 97:1-18.

130. Sutton, ML; Graham, T.C.; Levin, R.A.;
Denine, E.P. (1987a) Oncogenicity study of
Macrodantin^ in mice The Toxicologist 7:102.

132. Taguchi, O.; Michael, S.D., Nishizuka, Y.
(1988) Rapid induction of ovarian granulosa cell
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neonatally estrogenized mice. Cancer Res.
48:425-429.

133 Tarone. RE. (1975) Tests for trend in life
tabic analysis. Biometrika 62:679-682.

134. Tatsumi, K . Kitamura, S., Yoshimura, II.
(1977) Binding of nitrofuran derivatives to nu-
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2948-2952.

135. Tennent, B J ; Bcamer, W G. (1986) Ovari-
an tumors not induced by irradiation and gonad-
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prod. 34:751-760.

136. Tonomura, A.; Sasaki, M. (1973) Chromo-
some aberrations and DNA repair synthesis in
cultured human cells exposed to nitrofurans.
Jpn. J. Genet. 48:291 294

137. Toole, J.F.; Gergen, J. A.; Hayes, D M ,
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138. Topham, J C. (1980) The detection of car-
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139 Turusov, V.S., Ed. (1979) Pathology of Tu-
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140. U.S. International Trade Commission
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141. U.S. Pharmaceutical Market Data Base
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Nitrofurantoin, NTP TR 341

74

V. REFERENCES

142. Varsano, I.; Fischl, J.; Shochet, SB (1973)
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143. Veronese, M.; Salvaterra, M.; Barzaghi, D.;
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75

Nitrofurantoin, NTP TR 341

Nitrofurantoin, NTF TR 341 76

APPENDIX A

SUMMARY OF LESIONS IN MALE RATS IN

THE TWO-YEAR FEED STUDY OF NITROFURANTOIN

PAGE

TABLE Al SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS IN THE

TWO-YEAR FEED STUDY OF NITROFURANTOIN

TABLE A2 INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF MALE RATS IN THE TWO-YEAR

FEED STUDY OF NITROFURANTOIN

TABLE A3 ANALYSIS OF PRIMARY TUMORS IN MALE RATS IN THE TWO-YEAR FEED

STUDY OF NITROFURANTOIN

78

82

94

TABLE A4a HISTORICAL INCIDENCE OF KIDNEY TUBULAR CELL TUMORS IN MALE F344/N

RATS RECEIVING NO TREATMENT 99

TABLE A4b HISTORICAL INCIDENCE OF SKELETAL SYSTEM TUMORS IN MALE F344/N

RATS RECEIVING NO TREATMENT 100

TABLE A4c HISTORICAL INCIDENCE OF SUBCUTANEOUS TISSUE TUMORS IN MALE

F344/N RATS RECEIVING NO TREATMENT 101

TABLE A4d HISTORICAL INCIDENCE OF TESTICULAR TUMORS IN MALE F344/N RATS

RECEIVING NO TREATMENT 102

TABLE A4e HISTORICAL INCIDENCE OF PREPUTIAL GLAND TUMORS IN MALE F344/N

RATS RECEIVING NO TREATMENT 103

TABLE A5 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS

IN THE TWO-YEAR FEED STUDY OF NITROFURANTOIN 104

77

Nitrofurantoin, NTP TR 341

TABLE Al. SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS IN THE TWO-YEAR FEED

STUDY OF NITROFURANTOIN

Untreated Control

Low Dose

High Dose

Animals initially in study

50

50

50

Animals removed

50

50

50

Animals examined histopathologically

50

50

50

ALIMENTARY SYSTEM

Intestine large, colon

(50)

(50)

(48)

Polyp adenomatous

1

(2%)

Muscularis, leiomyosarcoma

1

(2%)

Intestine small, ileum

(50)

(49)

(47)

Leukemia mononuclear

1

(2%)

Intestine small, jejunum

(50)

(50)

(48)

Cystadenocarcinoma

1

(2%)

Leukemia mononuclear

I

(2%)

Liver

(50)

(50)

(50)

Fibrosarcoma, metastatic, skin

1

(2%)

Hepatocellular carcinoma

1

(2%)

Leukemia mononuclear

23

(46%)

14

(28%)

15

(30%)

Mesothelioma malignant

1

(2%)

Neoplastic nodule

1

(2%)

2

(4%)

Neoplastic nodule, multiple

1

(2%)

Mesentery

♦(50)

*(50)

♦(50)

Mesothelioma malignant

3

(6%)

1

(2%)

Mesothelioma malignant, multiple

1

(2%)

Pancreas

(50)

(50)

(49)

Leukemia mononuclear

3

(6%)

1

(2%)

1

(2%)

Mesothelioma malignant

1

(2%)

2

(4%)

Acinus, adenoma

2

(4%)

Stomach

(49)

(50)

(50)

Serosa, mesothelioma malignant

1

(2%)

2

(4%)

Stomach, forestomach

(49)

(50)

(49)

Papilloma squamous

1

(2%)

CARDIOVASCULAR SYSTEM

Heart

(50)

(50)

(50)

Leukemia mononuclear

1

(2%)

2

(4%)

1

(2%)

ENDOCRINE SYSTEM

Adrenal gland, cortex

(50)

(50)

(50)

Adenoma

1

(2%)

Leukemia mononuclear

9

(18%)

4

(8%)

Adrenal gland, medulla

(50)

(50)

(50)

Leukemia mononuclear

5

(10%)

5

(10%)

1

(2%)

Pheochromocytoma malignant

3

(6%)

1

(2%)

2

(4%)

Pheochromocytoma benign

18

(36%)

16

(32%)

21

(42%)

Pheochromocytoma benign, multiple

5

(10%)

4

(8%)

2

(4%)

Islets, pancreatic

(50)

(50)

(49)

Adenoma

2

(4%)

2

(4%)

1

(2%)

Carcmoma

2

(4%)

Pituitary gland

(50)

(50)

(50)

Leukemia mononuclear

4

(8%)

2

(4%)

Pars distalis, adenoma

10

(20%)

12

(24%)

13

(26%)

Pars distalis, carcinoma

1

(2%)

2

(4%)

3

(6%)

Thyroid gland

(50)

(50)

(50)

Leukemia mononuclear

1

(2%)

C-cell, adenoma

2

(4%)

7

(14%)

6

(12%)

CcelL adenoma, multiple

1

(2%)

C-cell, carcinoma

3

(6%)

1

(2%)

Follicular cell, adenoma

2

(4%)

Follicular cell, carcinoma

1

(2%)

Nitrofurantoin, NTP TR 341

78

TABLE Al. SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS IN THE TWO-YEAR FEED

STUDY OF NITROFURANTOIN (Continued)

Untreated Control

Low Dose

High Dose

GENERAL BODY SYSTEM
None

GENITAL SYSTEM

Epididymis

150)

(50)

(50)

Mesothelioma malignant

2

(4%l

5

(10%)

Preputial gland

(48)

(50)

(47)

Adenoma

6

(13%)

4

(8%)

Adenoma, multiple

1

(2%)

Carcinoma

6

(13%)

6

(12%)

Leukemia mononuclear

1

(2%)

Prostate

(50)

(50)

(49)

Leukemia mononuclear

3

(6%)

Seminal vesicle

(50)

(50)

(50)

Leukemia mononuclear

1

(2%)

Mesothelioma malignant

2

(4%)

Testes

(50)

(50)

(50)

Mesothelioma malignant

I

(2%)

Interstitial cell, adenoma

13

(26%)

4

(8%)

15

(30%)

Interstitial cell, adenoma, multiple

34

(68%)

41

(82%)

6

(12%)

Tunic, mesothelioma benign

1

(2%)

2

(4%)

Tunic, mesothelioma malignant

2

(4%)

5

(10%)

1

(2%)

Tunic, sarcoma

1

(2%)

HEMATOPOIETIC SYSTEM

Blood

*(50)

•(50)

*(50)

Leukemia mononuclear

16

(32%)

11

(22%)

10

(20%)

Bone marrow

(50)

(50)

(50)

Leukemia mononuclear

16

(32%)

7

(14%)

4

(8%)

Lymph node

(50)

(50)

(50)

Fibrosarcoma, metastatic, skin

1

(2%)

Bronchial, carcinoma, metastatic, Zymbal

1 gland

1

(2%)

Bronchial, leukemia mononuclear

1

(2%)

Deep cervical, leukemia mononuclear

1

(2%)

Iliac, leukemia mononuclear

1

(2%)

1

(2%)

Inguinal, leukemia mononuclear

4

(8%)

2

(4%)

1

(2%)

Mediastinal, leukemia mononuclear

9

(18%)

4

(8%)

1

(2%)

Pancreatic, leukemia mononuclear

3

(6%)

2

(4%)

2

(4%)

Renal, leukemia mononuclear

1

(2%)

1

(2%)

1

(2%)

Lymph node, mandibular

(46)

(50)

(47)

Leukemia mononuclear

8

(17%)

7

(14%)

3

(6%)

Lymph node, mesenteric

(49)

(48)

(49)

Leukemia mononuclear

8

(16%)

4

(8%)

3

(6%)

Spleen

(50)

(50)

(50)

Hemangiosarcoma

1

(2%)

Leukemia mononuclear

23

(46%)

13

(26%)

13

(26%)

Mesothelioma malignant

1

(2%)

2

(4%)

Sarcoma

1

(2%)

Thymus

(40)

(36)

(34)

Leukemia mononuclear

3

(8%)

INTEGUMENTARY SYSTEM
Mammary gland
Adenoma
Fibroadenoma

(46)

1 (2%)

2 (4%)

(49)
4 (8%)

(46)
2 (4%)

79

Nitrofurantoin, NTP TR 341

TABLE Al. SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS IN THE TWO-YEAR FEED

STUDY OF NITROFURANTOIN (Continued)

Untreated Control

Low Dose

High Dose

INTEGUMENTARY SYSTEM (Continued)
Skin

Basal cell carcinoma

Carcinoma

Keratoacanthoma

Keratoacanthoma, multiple

Papilloma squamous

Sebaceous gland, carcinoma

Subcutaneous tissue, basosquamous tumor benign

Subcutaneous tissue, fibroma

Subcutaneous tissue, fibrosarcoma

Subcutaneous tissue, fibrosarcoma, multiple

Subcutaneous tissue, hemangioma

Subcutaneous tissue, hemangiosarcoma

Subcutaneous tissue, leukemia mononuclear

Subcutaneous tissue, neoplasm, NOS

Subcutaneous tissue, schwannoma, NOS

Subcutaneous tissue, sebaceous gland, adenoma

(50)
3

(6%)

4 (8%)

(2%)
(2%)

1 (2%)

(4%)
(2%)

3)

(50)

3 (6%)

1 (2%)

5 (10%)

4 (8%)

1 (2%)

1 (2%)

2 (4%)

1 (2%)

5 (10%)

4 (8%)

1 (2%)

1 (2%)

1 (2%)

1 (2%)
1 (2%)

1 (2%)

1 (2%)

MUSCULOSKELETAL SYSTEM
Bone

Fibrosarcoma, metastatic, skin
Femur, osteosarcoma
Vertebra, osteosarcoma
Skeletal muscle

Abdominal, mesothelioma malignant
Diaphragm, mesothelioma malignant

(50)

1 (2%)

*(50)

1 (2%)
1 (2%)

(50)

1 (2%)

•(50)

1 (2%)

(50)

1 (2%)
1 (2%)
•(50)

NERVOUS SYSTEM
Brain

Astrocytoma malignant
Leukemia mononuclear

(50)

1 (2%)

(50)

1 (2%)

(50)

1 (2%)

RESPIRATORY SYSTEM
Lung

Carcinoma, metastatic, skin
Carcinoma, metastatic, Zymbal gland
Fibrosarcoma, metastatic, skin
Leukemia mononuclear
Osteosarcoma, metastatic, bone
Bronchus, squamous cell carcinoma
Pleura, mediastinum, alveolar/bronchiolar
carcinoma, metastatic, uncertain primary
site
Nose

Submucosa, leukemia mononuclear

(50)

21 (42%)

1 (2%)
(50)

1 (2%)

)0)

(50)

1 (2%)

1

(2%)

1 (2%)

10 (20%)

10

(20%)

1

(2%)

1 (2%)

(49)

(48)

SPECIAL SENSES SYSTEM
Ear

Pinna, schwannoma malignant
Harderian gland

Leukemia mononuclear
Zymbal gland

Carcinoma

•(50)

I (2%)
♦(50)

1 (2%)
•(50)

2 (4%)

♦(50)
•(50)
•(50)

•(50)

•(50)

•(50)

1 (2%)

Nitrofurantoin, NTP TR 341

80

TABLE Al.

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE RATS IN THE TWO-YEAR FEED
STUDY OF NITROFURANTOIN (Continued)

Untreated Control

Low Dose

High Dose

URINARY SYSTEM

Kidney (50)

Carcinoma, metastatic, Zymbal gland
Fibrosarcoma, metastatic, skin

Leukemia mononuclear 4 (8%)

Mesothelioma malignant
Osteosarcoma, metastatic, bone
Pelvis, transitional epithelium, carcinoma
Renal tubule, adenoma
Renal tubule, carcinoma
Urinary bladder (50)

Leukemia mononuclear
Mesothelioma malignant 1 (2%)

(50)

1 (2%)
3 (6%)
1 (2%)

1 (2%)

(50)

1 (2%)
1 (2%)

(50)

1 (2%)

1 (2%)

1 (2%)

1 (2%)

2 (4%)
1 (2%)

(50)

1 (2%)

SYSTEMIC LESIONS
Multiple organs

Leukemia mononuclear
Mesothelioma malignant
Hemangiosarcoma
Mesothelioma benign
Hemangioma

(50)

•(50)

23 (46%)

14 (28%)

3 (6%)

5 (10%)

2 (4%)

1 (2%)

1 (2%)

(50)
15 (30%)

1 (2%)

2 (4%)
1 (2%)

ANIMAL DISPOSITION SUMMARY
Animals initially in study
Moribund
Terminal sacrifice
Dead

50

24

23

3

50

22

27

1

50

21

26

3

TUMOR SUMMARY

Total animals with primary neoplasms** 49

Total primary neoplasms 151

Total animals with benign neoplasms 49

Total benign neoplasms 102

Total animals with malignant neoplasms 37

Total malignant neoplasms 49

Total animals with secondary neoplasms*** 2

Total secondary neoplasms 2

Total animals with malignant neoplasms-
uncertain primary site 1
Total animals with neoplasms-
uncertain benign or malignant
Total uncertain neoplasms

50

156

47

116

31

39

2

5

48

120

40

84

30

35

2

5

• Number of animals receiving complete necropsy examination; all gross lesions including masses examined microscopically.

*• Primary tumors: all tumors except secondary tumors

•** Secondary tumors: metastatic tumors or tumors invasive into an adjacent organ

81

Nitrofurantoin, NTP TR 341

TABLE A2.

INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF MALE RATS IN THE TWO-YEAR FEED
STUDY OF NITROFURANTOIN: UNTREATED CONTROL

WEEKS ON
STUDY

OOOOOOOOOOOOOOOOOOOOIl 1 1 I
5677888888889999999900000
6537003556990123457703455

CARCASS
ID

1000000000000000000000001
05332993568971 13697721820
11 12112321135124241223232

ALIMENTARY SYSTEM
Esophagus
Intestine largo
Intestine large, cecum
Intestine large, colon
Intestine large, rectum
Intestine small
Intestine smail. duodenum
Intestine small, ileum

Leukemia mononuclear
Intestine small, jejunum

Cystadenocarcmoma

Leukemia mononuclear
Liver

Leukemia mononuclear

Mesothelioma malignant

Neoplastic nodule
Mesentery

Mesothelioma malignant
Pancreas

Leukemia mononuclear

Mesothelioma malignant

Acinus, adenoma
Salivary glands
Stomach

Serosa, mesothelioma mahgnant
Stomach, forestomach
Stomach, glandular

M-t-+- + + + + -f-*- + + + + + -t--t--t-+--t--t- + 4 + + -»-
■(- + + + + + + + + + + + + + + + t + -f + + -t- + + +
+ + + + + + + + + + -*- + + + + ■»- + + + + + + ■*- + +
-♦- + + + + + + + + + + + -(-•(- + -♦-+ ■*--*- + -*-■»- + + +
■♦- + -»--t- + -f + + + + + + + + + + + + + + +- + + -(- +
■*- + -♦-•»--♦- + + + + + + + ■t--»- + -f + + -+--t-+- + + 4 +
M+ + + + + + + + + + + + + + + + -t- + -(- + + +-+- +
+ + + + + + + + + + + + + + -t- + + -f-t--i--f-h-t- + -t-

X
+ + + + + + + -*-■♦- + + + + + + -»- + + ■*--*--*- + + + +
X

X
+ + -f-ff4 + + -»- + + + + + 4-t- + + -(- + + + + -(■ +

X XXXX XXXXXXXX X X
X

+ + •(- +
X X
-f + + + + + + + + + + + -f + + + + -»--l- + + + + + +
X XX
X

X

444--t- + + -(--*- + + + + -»- + -*-M+-F-l- + -(- + -t- + +
+ + + + + + + -(-4- + + + + + -t--*- + -(- + + -t- + + + -(-

X

+ + -t- + + + + + + + + + + -t- + -*- + -(- + + + + -t- + +-
+ + -f + f-t- + + + + -(- + + + + + + -t- + + + + 4-t--(-

CARDIOVASCULAR SYSTEM
Heart
Leukemia mononuclear

+ + -*- + + + + -*- + + + + + + + + + + + -(- + + + + +

X

ENDOCRINE SYSTEM
Adrenal gland
Adrenal gland, cortex

Leukemia mononuclear
Adrenal gland, medulla

Leukemia mononuclear

Pheochromocytoma malignant

Pheochromocytoma benign

Pheochromocytoma benign, multiple
Islets, pancreatic

Adenoma
Parathyroid gland
Pituitary gland

Leukemia mononuclear

Pars distalis. adenoma

Pars distalis, carcinoma
Thyroid gland

C-cell. adenoma

C-celt. adenoma, multiple

+ + + + + + + + + -l- + + -t--l- + -t- + + + + + + + -(- +
+ + + + + -t- + + + + + + + + + + + + + + + -t-4 + +

XXX XX X X
+ + + + + + -l- + + + + + + + + + + + + + + -f + + +

XXX XX

X
X X XXXX

X
+ -*- + -t- + -t- + + -f + + + + + -l- + + + + + + + + +- +

-t--t-M + + + + + + + -*- + +- + +--(-+ +- + -»■+■ + +■•(- +
+ + + + + + + + + + + + + + -t--t- + -t- + + -l- + + + +

XX X
X XX XX XX

+ + + + + -(--*- + -t--*- + + -f + + -t- + -t--t--*--(--«- + -t-+-

GENERAL BODY SYSTEM
None

GENITAL SYSTEM
Coagulating gland
Epididymis

Mesothelioma malignant
Preputial gland

Adenoma

Carcinoma
Prostate

Leukemia mononuclear
Seminal vesicle
Testes

Mesothelioma malignant

Interstitial cell, adenoma

Interstitial cell, adenoma, multiple

Tunic, mesothelioma malignant

+ -t-+--|- + + + + + -l- + + + + -f + +--fH- + + + + + +

X X

+ + + + + + + + + + + + + + -l- + +--t--t--f + -f + + +
X X
X XX X

+ f-t--t- + + + -»--(- + + + -*--*- + -»- + -t- + + + -t--t- + +

XX X
^- + -^-f + +-■♦- + + >- + ■^ + ^- + + + -^- + + -^-^- + + +
+ f + -»- + -)--i- + + + + + + -t-t' + + -(- + + -»--t- + + +

XXX XXXX XX X

X XXX XXXXXXXX
X X

-*- : Tissue examined microscopically

Not examined
— : Present but not examined microscopically
I; Insufficient tissue

Missing

Autolysis precludes examination

Incidence of listed morphology

Nitrofurantoin, NTP TR 341

82

TABLE A2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF MALE RATS:

(Continued)

UNTREATED CONTROL

WEEKS ON
STUDY

CARCASS
ID

ALIMENTARY SYSTEM

Esopha^s
Intestine large
Intestine large, cecum
Intestine large, colon
Intestine large, rectum
Intestine small
Intestine small, duodenum
Intestine small, ileum

Leukemia monoDUclear
Intestine small, jejunum

CystadGnocamnoma

Leukemia mononuclear
Liver

Leukemia mononuclear

Mesothelioma malignant

Neoplastic nodule
Mesentery

Mesothelioma malignant
Pancreas

Leukemia mononuclear

Mesothelioma malignant

Acinus, adenoma
Salivary glands
Stomacn

Serosa, mesothelioma malignant
Stomach, forestomach
Stomach, glandular

CARDIOVASCULAR SYSTEM

Heart
Leukemia mononuclear

ENDOCRINE SYSTEM

Adrenal gland
Adrenal giand, cortex

Leukemia mononuclear
Adrenal gland, medulla

Leukemia mononuclear

Pheochromocytoma malignant

Pheochromocytoma benign

Pheochromocytoma benign, multiple
Islets, pancreatic

Adenoma
Parathyroid gland
Pituitary gland

Leukemia mononuclear

Pars distalis, adenoma

Pars distalis. carcinoma
Thyroid gland

C-cell. adenoma

C-cell, adenoma, multiple

GENERAL BODY SYSTEM
None

GENITAL SYSTEM

Coagulating gland
Epididymis

Mesothelioma malignant
Preputial gland

Adenoma

Carcinoma
Prostate

Leukemia mononuclear
Seminal vesicle
Testes

Mesothelioma malignant

Interstitial cell, adenoma

Interstitial cell, adenoma, multiple

Tunic, mesothelioma malignant

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

n

n

n

0

n

0

0

0

n

n

0

0

n

0

0

1)

0

0

0

0

U

0

U

0

U

5

5

5

5

5

5

5

5

5

5

5

5

s

5

5

5

5

5

5

b

5

5

5

b

b

I

n

n

n

n

0

0

0

n

(1

0

0

0

0

0

0

0

0

a

0

«

U

U

1

1

0

4

fi

9

1

1

2

2

3

4

4

4

4

5

5

5

6

6

7

7

8

8

8

0

0

3

1

3

5

4

5

4

5

5

2

3

4

5

3

4

5

4

5

3

4

3

4

5

4

5

+ + + +
X X

+ + +
■(- + ■*-

+ ■*-■!-■(- +

+ ■♦-■»-■(- +

I + + + +■

+ + I + + +
-(- + I + + +

+ -t- +
+ + -(-

+ -(- + +

+

+

+

+

+

+

+

+

4-

+

+

+

+

+

+

+

■1-
X

+

+

+

+

+

+

•1-

+

+

+

■t-

+

+

+
X

+

+

+

4-

+

+

+

+

+

+

+

X

X

X

X

X

X

X

X

X

X

X

X

+

+

+

+

-)-

+

+

+

-+-

+

+

+

+

+

■1- -(-
+ +

X

X X

+ + + + 4- + + 4- + + + 4-4-

+ +

X

X

■<- 4-

+ ■(-
X

TOTAL;
TISSUES
TUMORS

X

+ +

+ + + + +
X

+ + + + + + + +

■!- + + -*- + -(- + ■»-

X

+ +
X

-*- M

+

+

+

+

-t-

+

+■

+

■f

+

^

+

+

+

+

+

+

+

4-

+

+

+

4-

4-

4-

-»-

+

4-

■f

"*"

X

X

+
X

+

+

+

+■

•1-

+

+

■»-

+
X

+

+

4-

4-

t

4-

4-

4-

4-

X

X

X

X

X

X

X

X

X

X

X

X

X

X

X

X

X

X

X

X

X

X

49

50

50

50

50

50

49

50

1

50

1

1

50

23

1

1

6

3

50

3

1

2

4S

49

1

49

49

50

1

50
50

9
50

5

3
18

5
50

2
49
50

4
10

1
50

2

I

1
50

2
48

6

6
50

3
50
50

1
13
34

2

83

Nitrofurantoin, NTP TR 341

TABLE A2.

INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF MALE RATS: UNTREATED CONTROL

(Continued)

WEEKS ON
STUDY

00000000000000000000111 11
5677888888889999999900000
6537003556990123457703455

CARCASS
ID

1000000000000000000000001
0533 2 993568971136977218 2 0
11121123 2 1135124241223232

HEMATOPOIETIC SYSTEM

Blood
Leukemia mononuclear

Bone mam>w
Leukemia mononuclear

Lymph node
Bronchial, leukemia mononuclear
Iliac, leukemia mononuclear
IngTjmal. leukemia mononuclear
Mediastinal, leukemia mononuclear
Pancreatic, leukemia mononuclear
Renal, leukemia mononuclear

Lymph node, mandibular
Leukemia mononuclear

Lymph node, mesentenc
Leukemia mononuclear

Spleen
Leukemia mononuclear
Mesothelioma malignant
Sarcoma

Thymus
Leukemia mononuclear

+ + + M+ + + + -t- + + +
X XX XXXXXX

+ + + +--f4--f + +- + + + + -f + + + + +- + + + + + +

XX XXXXXXX X X

+ +--f + + + + -K + + -t- + + -»- + 4- + + + + + + + + +

X
X

XX X

XX XXXX
X X

M+ + + + +M+ + -»--t- + -t--t-4- + + + -t- + -t- + + + +
X X XXX

+ + -*--(- + + -f-*- + -f + + -t- + + + + + + +- + + + + +
XX XX XX

+ + + + + + + f + + 4- + -t-f + + + -»--t--*--*--(--t-4- +

X XXXX XXXXXXXX X X
X

X

+ + + + +M++M+-t- + + + + M + MM+ + +- + + 4-

X

INTEGUMENTARY SYSTEM
Mammary gland

Adenoma

Fibroadenoma

Skin
Basal cell carcinoma
Keratoacanthoma
Papilloma squamous
Sebaceous gland, carcinoma
Subcutaneous tissue, fibrosarcoma
Subcutaneous tissue, hemangiosarcoma
Subcutaneous tissue, leukemia
mononuclear

+ + -+- + + M+ + +--f-t- + + + + + + -t- + + + + -t-^--*-

+ + + + + + + 4- + + -f-f + + + + + + + + -(- + + + +

X
XX X

X

X

X
X X

X

MUSCULOSKELETAL SYSTEM

Bone
Fibrosarcoma, metastatic, skm

Skeletal muscle
Abdominal, mesothelioma malignant
Diaphragm, mesothelioma malignant

+ -«-4- + + + + + -(--(- + 4--»- + + + + + + + + + + + +

X

-t-
X
X

NERVOUS SYSTEM
Brain
Astrocytoma malignant

■t--(--4- + + + + + + + -t-+- + -t--(--f+- + T + + + + + +-

X

RESPIRATORY SYSTEM
Lung
Leukemia mononuclear
Pleura, mediastinum,
alveolar/'bronchiolar carcinoma.
metastatic, uncertain pnmary site
Nose

Submucosa, leukemia mononuclear
Trachea

+ + + + + +- + +- + + -f + 1--t- + -t- + + -l-+--t-4-4- + -(-

X XXXX XXX XXXX X X

X

+ + + + + -t-+--t- + + + + + +- + + + +-^-t-+- + + -t- +

X

+ + + + 4- + -*- + + + + + + +- + + + + +- + -t- + + -t- +

SPECIAL SENSES SYSTEM
Ear

Pinna, schwannoma malignant
Eye
Hardeman gland

Leukemia mononuclear
Lacnmal gland
Zymbal gland

Carcinoma

+ + + + + +
X

f

+
X

4-

■(- +
X X

URINABY SYSTEM
Kidney

Leukemia mononuclear
Unnary bladder

Mesothelioma malignant

+ + +- + +- + + +- + + + +■ + +■ + + + + + + + + + + ■»-
X X XX

+ -+--t- + -t- + + -t--f + -l-+- + -)-+f + + + + + + + + -t-
X

Nitrofurantoin, NTP TR 341

84

TABLE A2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF MALE RATS: UNTREATED CONTROL

(Continued)

WEEKS ON
STUDY

CARCASS

ID

HEMATOPOIETIC SYSTKM
Blood

Leukemia mononuclear
Bone marrow

Leukemia mononuclear
Lymph node

Bronchial, leukemia mononuclear

lUac. leukemia mononuclear

Inguinal, leukemia mononuclear

Mediastinal, leukemia mononuclear

Pancreatic, leukemia mononuclear

Renal, leukemia mononuclear
Lymph node, mandibular

Leukemia mononuclear
Lymph node, mesenteric

Leukemia mononuclear
Spleen

Leukemia mononuclear

Mesothelioma malignant

Sarcoma
Thymus

Leukemia mononuclear

INTEGUMENT ABY SYSTEM

Mammary gland
Adenoma
Fibroadenoma

Skin
Basal cell carcinoma
Keratoacanthoma
Papilloma squamous
Sebaceous gland, carcinoma
Subcutaneous tissue, fibrosarcoma
Subcutaneous tissue, hemangiosarcoma
Subcutaneous tissue, leukemia
mononuclear

MUSCULOSKELETAL SYSTEM

Bone
Fibrosarcoma, metastatic, skin

Skeletal muscle
Abdominal, mesothelioma malignant
Diaphragm, mesothelioma malignant

NERVOUS SYSTEM
Brain
Astrocytoma malignant

RESPIRATORY SYSTEM
Lung
Leukemia mononuclear
Pleura, mediastinum,
alveolar/bronchiolar carcinoma,
metastatic, uncertain pnmary site
Nose

Submucosa, leukemia mononuclear
Trachea

SPECIAL SENSES SYSTEM
Ear

Pinna, schwannoma malignant
Eye
Hardenan gland

Leukemia mononuclear
Lacnmal gland
Zymbal gland

Carcinoma

URINARY SYSTEM
Kidney

Leukemia mononuclear
Unnary bladder

Mesothelioma malignant

1 1 1 I 1
0 0 0 0 0
5 5 5 5 5

1 1 1
0 0 0
5 5 5

1 1
0 0
5 5

1 1
0 0
5 5

1 1
0 0
5 5

1 1
0 0
5 5

"D — D — 0"
4 6 9
1 3 5

"ij — 0 — D — ir

112 2
4 5 4 5

"D — tr

3 4
5 2

"0 JT

5 5
4 5

13 !r

6 6
4 5

1! — D — 0 I r

8 8 8 0 0
3 4 5 4 5

TOTAL:
TISSUES
TUMORS

X X

+ +
X X

X

X X
X

X

+ + -»- +
X

+ + + +
XX X

M + M M
X

X XX

^ + + + + + +
X X

M M

+ -t-
X

X

■t- +

X

+ ■(-
+ M
+ +

+ + + +

+ + +

+ + +

+ + ■»- +
XX X

4- + +

4- + + -F

+ + + -t-

+ -f -t- +
+ + ■»■ +

+ + + +

18

16

50

16

50

1

1

4

9

3

1

46

50

23

1

1

40

3

46
1
2

50
3
4
1
1
1
2

50
1

1
1
1

50

1

50
21

1
50

1
50

16
1
4
1
1
1
2
2

50

4

50

1

85

Nitrofurantoin, NTP TR 341

TABLE A2. INDIVIDUAL AOTMAL TUMOR PATHOLOGY OF MALE RATS IN THE TWO-YEAR FEED

STUDY OF NITROFURANTOIN: LOW DOSE

WEfeKS ON
STUDY

OOOOOOOOOOOOOlllIl ILlll 11
4677888999999000000000000
6909166024679002234444455

CARCASS
ID

'i 2 2 2 2 '2 'i 2 3 3 3 2 2 2 'i 'J 'i 2 2 'i 2 '2 2 2 2
6759614800068275728861913
11112 11112332122323452 2 31

ALIMENTARY SYSTEM
EsophapiS
Intestine large
Intestine large, cecum
Intestine large, colon

Polyp adeoomatous
Intestine large, rectum
Intestine small
Intestine small, duodenum
Intestine small, ileum
Intestine small, jejunum
Liver

Fibrosarcoma, metastatic, slun

Hepatocellular carcinoma

Leukemia mononuclear

Neoplastic nodule

Neoplastic nodule, multiple
Mesentery

Mesothelioma malignant

Mesothelioma malignant, multiple
Pancreas

Leukemia mononuclear

Mesothelioma malignant
Salivary glands
Stomacn

Serosa, mesothelioma malignant
Stomach, forostomach
Stomach, glandular

+ 44 + + + + + + + + + + + + + + + + + + + + + +
444 + + + + + + + + 444444444 + + + + +
+ 44444 + + + + + + + + + + + + + + + + + + +
+ + + + + 444 + + + + + + + + + + + + + + + + +

X

M+ + +M4 + + 444444 + 4 + 44 + + + 4 + +
444444 + + ^-4 + + + + + + + + + + + + + + +
444 + + 4 + + + + + 4 + + 4444 + + + + + + +
+ M + + + + + + 444444 + 4 + 444 + + + + +
444444 + + + + + + + + + + + + + + + + + + +
+ + + + 44+ + + + + + + + + + + + + + + +. + + +

XXX XX XX
X

4 4 4+ +

X

X

+ + + + 444 + + + + + + + + + + + + + + + + + +

X

X X

44 + + + + + + + + 4444 + + + + + + + + + + +
444444 + + + 44 + + + + + + + + + + + + + +

X X

4 + + + + + + + + + + + + + + + + + + + + + + + +
4 + + + + + + + + 4444 + + + + + + + + + + 44

CARDIOVASCULAR SYSTEM
Blood vessel
Heart
Leukemia mononuclear

+
4 + 4 + + + + + + + + + + + + + + + + + + + + + +

X X

ENDOCRINE SYSTEM
Adrenal gland
Adrenal gland, cortex

Leukemia mononuclear
Adrenal gland, medulla

Leukemia mononuclear

Pheochromocytoma malignant

Pheochromocytoma benign

Pheochromocytoma benign, multiple
Islets, pancreatic

Adenoma
Parathyroid gland
Pituitary gland

Pars distahs. adenoma

Pars distahs, carcinoma
Thyroid gland

Leukemia mononuclear

C-cell. adenoma

C-cell. carcinoma

Follicular cell, adenoma

Follicular cell, carcinoma

444 + + + + + + + + + + + + + + + + + + + + + +
444 + 44 + + + + + + + + + + + + + + + + + + +

X X

+ + + + 44 + + + + + 44 + + 4 + + + + + + + + +

XX X

X
X XX

X XX

+ + + + + 444 + + + -!- + + + + + + + + + + + ^ +

X

444 + 44444M4 + 44 + 4M44 + + + + + +
444 + 44 + + + + + + + + + + + + + + + + + + +

XXX XX XX X
X

444444 + + + + + + + + + + + + + + + + + + +

X
X XX
XX X

X

GENERAL BODY SYSTEM
None

OENITAL SYSTEM
Coagulating gland
Epididymis

Mesothelioma malignant
Preputial gland

Aaenoma

Adenoma, multiple

Carcinoma

Leukemia mononuclear
Prostate
Seminal vesicle

Leukemia mononuclear

Mesothelioma malignant
Testes

Interstitial cell, adenoma

Interstitial cell, adenoma, multiple

Tunic, mesothelioma benign

Tunic, mesothelioma malignant

+

44444444 + + + 44444 + + fr44 + + +

XX XX

44 + + + + + + + + + + + + + + + + + + + + + + +

X X

X XXX

X

4 + + + + + + + + + + ^. + + + + + + + + + f + + +
444 + + + + + + + 4 + 4 + + + + + 4444444

X
X X

+ + + + + + 4 + + + + + 4 + + + + + + + + + + + +

X X
XX XX XXXXXXXXXX XX XX

X

XX XX

Nitrofurantoin, NTP TR 341

86

TABLE A2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF MALE RATS:

(Continued)

LOW DOSE

WEEKS ON
STUDY

CARCASS
ID

ALIMENTARY SYSTEM
Esopha^Tus
Intestine large
Intestine large, cecum
Intestine large, colon

Polyp adenomatous
Intestine large, rectum
Intestine small
Intestine small, duodenum
Intestine small, ileum
Intestine small, jejunum
Liver

Fibrosarcoma, metastatic, skin

Hepatocellular carcinoma

Leukemia mononuclear

Neoplastic nodule

Neoplastic nodule, multiple
Mesentery

Mesothelioma malignant

Mesothelioma malignant, multiple
Pancreas

Leukemia mononuclear

Mesothelioma malignant
Salivary glands
Stomacn

Serosa, mesothelioma malignant
Stomach, forestomach
Stomach, glandular

CARDIOVASCULAR SYSTEM

Blood vessel
Heart

Leukemia mononuclear

ENDOCRINE SYSTEM
Adrenal gland
Adrenal gland, cortex

Leukemia mononuclear
Adrenal gland, medulla

Leukemia mononuclear

Pheochromocytoma malignant

Pheochromocytoma benign

Pheochromocytoma benign, multiple
Islets, pancreatic

Adenoma
Parathyroid gland
Pituitary gland

Pars distahs, adenoma

Pars distahs, carcinoma
Thyroid gland

Leukemia mononuclear

C-cell. adenoma

C-cell, carcinoma

Follicular cell, adenoma

Follicular ceil, carcinoma

GENERAL BODY SYSTEM
None

GENITAL SYSTEM
Coagulating gland
Epididymis

Mesothelioma malignant
Preputial gland

Adenoma

Adenoma, multiple

Carcinoma

Leukemia mononuclear
Prostate
Seminal vesicle

Leukemia mononuclear

Mesothelioma malignant
Testes

Interstitial cell, adenoma

Interstitial cell, adenoma, multiple

Tunic, mesothelioma benign

Tunic, mesothelioma malignant

1 1 I 1 1
0 0 0 0 0
5 5 5 5 5

1111
0 0 0 0
5 5 5 5

1 1
0 0
5 5

"3 — 5 — 2 — T
0 112

4 4 5 3

"2 — T
5 5

4 5

"2 — T
6 7
4 4

~2 — 3"
9 0
5 5

■t- +
+ +
+ +

+ + + + ^-

+ + +- 4- +

X XXX XX

+ + + + + + + -•-

X
XXX

+ + +M+ + + +

+ + + f + + +- +

XXX

+ + + + i-i-*'i~

X X

X

+ ■*- + +• + + + + + + + ■♦■
■t- + + + + f + + + + + +
X
X
-*--»-■*- + + ■)- + -(--(- + + +

X

TOTAL:
TISSUES
TUMORS

+

+

+■

+

+

*■

+

+

+

+

+

+

-t-

+

+

+

-

-t-

+

X

+

+■

+

+

+

+

+
X

+

4-

i-

X

+

+

+

+

+

+

+

+

+

X

-t-

X

+

+

+
X

+

+

+

+■

+

+

+

+

-♦-

-*-

+

+

+

+

+-

+

+

+

+

+

+

+

^■

+-

+

+

+

+

+

+

^•

+

4-

+-

+

+

+

+

+

+

+

+

+

-f

"•"

■•"

+

-)■

+

+

+

+

+

-

■1-

+

+

^

^

4-

+■

+

+

+

+

+

+

+

+

+
X

+-

^

X

+

■t-

+

+

X

X

X

X

X

X

X

X

X

X

X

X

X

X

X

X

X

X

X

X

X

X

X

X

50

50

50

50

1

48

50

50

49

50

50

1

1

14

2

1

9

I

1

50

I

2

50

50

2

50

50

1

50

2

50

50

4

50

5

1

16

4

50

2

47

50

12

2

50

1

7

3

2

1

1

50

5

50

4

1

6

1

50

50

1

2

50

4

41

1

5

87

Nitrofurantoin, NTP TR 341

TABLE A2.

INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF MALE RATS: LOW DOSE

(Continued)

WEEKS ON
STUDY

OOOOOOOOOOOOOllllllllllll
4677888999999000000000000
69091660 2 4679002 2 34444455

CARCASS
ID

2 '2^222 '2 2333222222222222 '2 2
67596 14800068 2 75728861913
1111 2 1L112332 12 2323452231

HEMATOPOIETIC SYSTEM

Blood
Leukemia mononuclear

Bone marrow
Leukemia mononuclear

Lymph node
Fibrosarcoma, metastatic, skin
Inguinal, leukemia mononuclear
Mediastinal, leukemia mononuclear
Pancreatic, leukemia mononuclear
Renal, leukemia mononuclear

Lymph node, mandibular
Leukemia mononuclear

Lymph node, mesenteric
Leukemia mononuclear

Spleen
Hemanposarcoma
Leukemia mononuclear
Mesothelioma malignant

Thymus

M 4 4 4 4 4 4

X XX XX

+ 4 + 4444444444444 + 444 4 + + 4 +

XXX XX X

+ + + + 44444444 + 4 + 4444 + 4 + + + +

X X

X XX
X X
X

+ + + + + + + + 44 + + + + 444 + + + + + + + +

XXX XX

+ + + + + 4 + + 44M4444444444444 +

X X XX

+ 4441-4444444 + 444444444 + + +

X

XXX XX X
X X

+ + 4 -^M4 + 4M4M4 +MMM4 4MMMM4 4 +

INTEGUMENTARY SYSTEM
Mammary gland

Fibroadenoma
Skin

Carcinoma

Keratoacanthoma

Keratoacanthoma. multiple

Papilloma squamous

Subcutaneous tissue, fibroma

Subcutaneous tissue, fibrosarcoma

Subcutaneous tissue, fibrosarcoma,
multiple

Subcutaneous tissue, schwannoma, NOS

Subcutaneous tissue, sebaceous gland.
adenoma

4444 + + + + 44444 + + + + + + + + + + + +

X X

+ + 4444 + + 4 + + + + 444444 + + + + 44

X

XX X

X X
X

X

MUSCULOSKELETAL SYSTEM
Bone

Femur, osteosarcoma
Skeletal muscle

Abdominal, mesothelioma malignant

4 + 4 + 44 + + + + + + + + + + + + + + + + + + +

X

4

X

NERVOUS SYSTEM
Brain

Leukemia mononuclear
Spinal cord

+ + + 444 + -»- + 44 + + + + + + 4 + + 44444

X

4 4

RESPIRATORY SYSTEM
Lung

Carcinoma, metastatic, skin

Fibrosarcoma. meUstatic. skin

Leukemia mononuclear

Bronchus, squamous cell carcinoma
Nose
Trachea

4444444444444444444444 + + +

X

XXX XX X

M+ + 4-t-44444 444444 + 44444 + + +
+ + 4444444444444444 + + + + + + +

SPECIAL SENSES SYSTEM

Ear

Eye

Hardenan gland

4 4 + + +

4 4 4 4
4 4

URINARY SYSTEM

Kidney
Fibrosarcoma, metastatic, skin
Leukemia mononuclear
Mesothelioma malignant
Renal tubule, adenoma

Unnary bladder
Leukemia mononuclear
Mesothelioma malignant

+ 444-»-4444444444444 + + + + + + +

XX X
X

+ + 4444444444 + + 4 + + + 4444444

X

X

Nitrofurantoin, NTPTR341

88

TABLE A2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF MALE RATS:

(Continued)

LOW DOSE

WEEKS ON
STUDY

1 1 I 1 I 1 1 1 1 1 1 I 1 I 1 1 I I 1 I I 1 1 L 1
OOOOOOOOUOOOOO 0 0000000000
5555555555555555555555555

TOTAL:
TISSUES
TUMORS

CAKCASS
ID

232222222222 'J 22222222 *i '2 2 3
9011222333344445556778990
3445345 2 345 2 3453454455455

HEMATOPOIETIC SYSTEM

Blood
Leukemia mononuclear

Bone marrow
Leukemia mononuclear

Lymph node
Fibrosarcoma, metastatic, skin
Inguinal, leukemia mononuclear
Mediastinal, leukemia mononuclear
Pancreatic, leukemia mononuclear
Renal, leukemia mononuclear

Lymph node, mandibular
Leukemia mononuclear

Lymph node, mesentenc
Leukemia mononuclear

Spleen
Hemangiosarcoma
Leukemia mononuclear
Mesothelioma malignant

Thymus

4 + 4 4 4 +4

XX X XX X

+ +-+-+- + + + + + + 4-+--t--t- + + + + + -t- + + + -t- +
X

+ -t-+-4--t--(--(- + + + + + +--t- + -t- + 4- + -|--*--t--»--+-4-

X

X

-»- + +- + + + + +■+• + + -»- + +■ + + + +■■*- + + + + ■♦--•-

X X

-t--t--t- + 4--t- + + 4-4- + +M + + + + -t- + -»- + + + + -t-
+ + 4-4- + + -F-t- + + + + + + +- + + + + + + + + -*- +

X X X X X X X

M + M + + -t- + f + + -t-+M + M+-t--t- + +- + + + -t--t-

13

11

50
7

50
1
2
4
2
1

50
7

48
4

50
1

13
2

36

INTEGUMENTARY SYSTEM
Mammary gland

Fibroadenoma
Skjn

Carcinoma

Keratoacanthoma

Keratoacanthoma, multiple

Papilloma squamous

Subcutaneous tissue, fibroma

Subcutaneous tissue, fibrosarcoma

Subcutaneous tissue, fibrosarcoma,
multiple

Subcutaneous tissue, schwannoma, NOS

Subcutaneous tissue, sebaceous gland,
adenoma

M + + -t- + 4--*-t-4- + + + + + + + + + + + -t- + + -t--f

X X

+ + + + + + ^■^ »• + ■(-+--»-■(--*--»- + ■(-■♦■+■ + + -*- + +

X X

X
X

XX X

X

X

49

4

50

1

5

1
1
5

1

1
1

1

MUSCULOSKELETAL SYSTEM
Bone

Femur, osteosarcoma
Skeletal muscle

Abdominal, mesothelioma malignant

+ + -^--^ + + + ^-+- + + + + + + + + + + + + + + + -♦-

50

1
1
1

NERVOUS SYSTEM
Brain

Leukemia mononuclear
Spinal cord

-(--(-■(- + + -f + -t- + + -(- + + -*--t- + -t- + + -t- + -t- + + -+-

50

1
2

RESPIRATORY SYSTEM

Lung

Carcinoma, metastatic, skin

Fibrosarcoma, metastatic, skin

Leukemia mononuclear

Bronchus, squamous cell carcinoma
Nose
Trachea

+ + -»- + -»- + +■-(-■(- + ■♦■ + + + + + + + + + •*- + -•- + +-

X XX X

X

-*- + ■*-■+- + + + -(- + + + + + +- + + + + -♦■■•- + + + + +

+- + + + + + +-4--t- + -l- + -t-4--t--t- + + + -f-t--t- + + +-

50

1

1

10

1

49

50

SPECIAL SENSES SYSTEM

Ear

Eye

Hardenan gland

+ + +

13
6
5

URINARY SYSTEM

Kidney
Fibrosarcoma, metastatic, skjn
Leukemia mononuclear
Mesothelioma malignant
Renal tubule, adenoma

Unnary bladder
Leukemia mononuclear
Mesothelioma malignant

-t--(--(- + + -f + + + + + + -+- + + + + + + -t-+--*- + + +
X

X

+ -(--(- + + + +--t- + + -(- + + + + + -t- + + -t- + + + -»--t-

50

1
3
1
1
50
1
1

89

Nitrofurantoin, NTP TR 341

TABLE A2.

INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF MALE RATS IN THE TWO-YEAR FEED
STUDY OF NITROFURANTOIN: HIGH DOSE

WEEKS ON

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1

1

1

1

I

I

1

I

1

1

STUDY

5

5

5

7

7

8

8

9

9

9

9

9

9

9

9

0

0

0

0

0

0

0

0

0

0

6

6

9

1

6

0

9

1

I

I

I

2

3

6

6

0

0

1

2

3

3

3

4

4

5

CARCASS

1

1

I

~r

"T

1

1

1

I

~2~

'i

1

1

1

T"

1

I

1

1

1

1

1

1

I

I

ID

7

1

8

4

8

1

3

7

2

0

0

5

4

8

0

5

5

I

9

8

1

3

1

6

2

1

1

1

1

2

2

1

2

1

1

2

1

2

3

3

2

3

3

1

4

4

2

5

1

2

ALIMENT ABY SYSTEM

Esophagus

■t-

+

+

+

+■

+■

■»-

+

+

+

+

-t-

4-

4-

4-

4-

4-

4-

4-

4-

4-

+

4-

4-

4-

Intestine large

■t-

+

+

+

+

A

+

+

+

+-

-t-

+

4-

4-

4-

A

4-

4-

4-

4-

4-

4-

4-

4-

4-

Intestine large, cecum

■(-

+

■*-

+

-*-

A

-(-

-t-

+

4-

+-

4-

4-

4-

4-

A

4-

f

4-

4-

4-

4-

4-

4-

4-

Intestine large, colon

f

+

■t-

+

-t-

A

+

+■

+

+

■f

4-

4-

4-

4-

A

4-

4-

4-

4-

4-

4-

4-

4-

4-

Musculans, leiomyosarcoma

X

Intestine large, rectum

■t-

M

+

+

+

A

■t-

-*■

+

+

■f

4-

4-

4-

4-

A

4-

1-

4-

4-

4-

4-

4-

4-

+

Intestine small

-t-

+

+-

+

■f

A

+

+

+

+

+

4-

4-

+

4-

4-

+

4-

4-

4-

4-

4-

4-

4-

4-

Intestine small, duodenum

+-

+

-f

+

-f

A

+-

+

+

-t-

■(■

4-

+

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

Intestine small, ileum

-t-

+

+

M

■f

A

+■

+

+

f

+

4-

+

4-

4-

A

4-

4-

4-

4-

4-

4-

4-

4-

4-

Intestine small, jejunum

+-

+

■f

f

f

A

+

+

+

-(-

+

f

4-

4-

4-

A

4-

4-

4-

-f

+

4-

+

4-

f

Liver

-t-

+

+

+

f

+

■♦-

+■

+

+

+

-t-

+

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

Leukemia mononuclear

X

X

X

X

X

X

X

X

X

X

X

Mesentery

+

4-

Pancreas

+

+

+

+

+

■t-

-4-

+■

+

+

+■

4-

4-

4-

4-

4-

f

4-

M

4-

4-

4-

4-

+■

4-

Leukemia mononuclear

X

Salivary glands
StomacD

+

+

+

+

+

+

+

+■

+

■»■

+■

4-

4-

+■

4-

4-

+

4-

+

f

4-

4-

4-

4-

f

-*-

+

+■

+

+

■*■

+

+

+

+

+

-♦-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

Stomach, forestomach

M

+

+

+

■f

+■

+

+

+

+

+

i-

+

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

Papilloma squamous

Stomach, glandular

+

+

+

+■

■f

-t-

■1-

■♦-

+

+

+

4-

4-

4-

4-

+■

4-

4-

4-

4-

4-

4-

+

4-

+

CARDIOVASCULAR SYSTEM

Heart

+

+

-t-

■(-

■f

-t-

+-

-t-

+

f

+■

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

Leukemia mononuclear

X

ENDOCRINE SYSTEM

Adrenal gland

■f

+

+

+

-f

+

+■

+

+

+

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

Adrenal gland, cortex

+

-f

-t-

+

■f

-)-

^■

+

+■

+

+

-(-

4-

4-

+-

4-

4-

4-

+

4-

4-

4-

4-

4-

4-

Adenoma

X

Adrenal gland, medulla

+

■f

+

+■

+

+■

+

+

+■

-t-

+

+•

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

Leukemia mononuclear

X

Pheochromocytoma malignant

Pheochromocytoma benign

X

X

X

X

X

X

X

X

X

X

Pheochromocytoma benign, multiple

Islets, pancreatic

+

+

+

+

■1-

+

+

+

+■

+

■f

4-

+

4-

4-

4-

4-

4-

M

+

+

+-

4-

4-

4-

Adenoma

X

Carcinoma

Parathyroid gland

+

+

■1-

+

+-

-♦-

+

-♦-

M

+

+

-t-

+

4-

4-

4-

4-

4-

4-

4-

4-

4-

+-

4-

4-

Pituitary gland

+

+

■f

+

+

-t-

+

+

-1-

+

■h

+

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

Leukemia mononuclear

X

X

Pars distaiis. adenoma

X

X

X

X

Pars distalis. carcinoma

X

X

X

Thyroid gland

+

■(-

■*-

+■

+

+

4-

+

4-

i-

+

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

C cell, adenoma

X

X

X

C cell, carcinoma

GENERAL BODY SYSTEM

None

GENTTAL SYSTEM

Epididymis

+-

-♦-

4-

+

+

4-

*■

■♦-

+

+■

■(-

4-

4-

4-

4-

4-

4-

4-

4-

4-

-1-

4-

+-

4-

4-

Preputial gland

M

M

M

+

*■

^-

f

-f

+

+

+-

■t-

4-

+

4-

+-

4-

4-

4-

4-

4-

4-

4-

4-

4-

Prostate

+

+

-»-

■(-

■»-

+-

+

■»-

4-

■t-

M

+■

4-

4-

4-

4-

4-

4-

+

4-

4-

4-

4-

4-

4-

Seminal vesicle

■t-

■»-

-f

■t-

+

-t-

+

+

+

-t-

f

4-

4-

4-

+

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

Testes

+

+

-*-

+

4-

■*-

4-

+

+

f

■*-

4-

-4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

Interstitial cell, adenoma

X

X

X

X

X

Interstitial cell, adenoma, multiple

X

X

Tunic, mesothelioma benign

X

X

Tunic, mesothelioma malignant

Tunic, sarcoma

X

HEMATOPOIETIC SYSTEM

Blood

M

■t-

f

4-

4-

4-

4-

Leukemia mononuclear

X

X

X

X

X

X

Bone marrow

+■

+

4-

+

+

+

■*-

■♦-

+

+

+

*■

4-

4-

4-

4-

4-

f

4-

4-

4-

4-

4-

4-

4-

Leukemia mononuclear

X

X

Lymph node

+

+

-f

+

-t-

+■

+

+

+

+

■1-

4-

4-

4-

4-

4-

4-

4-

4-

+

4-

»-

4-

4-

+

Bronchial, carcinoma, metastatic.

Zymbal gland

X

Deep cervical, leukemia mononuclear

X

Iliac, leukemia mononuclear

X

Inguinal, leukemia mononuclear

X

Mediastinal, leukemia mononuclear

X

Pancreatic, leukemia mononuclear

X

Renal, leukemia mononuclear

Lymph node, mandibular

-t-

+

-(-

+

+

+

+

+

+

+

M

4-

4-

4-

4-

4-

4-

4-

M

+

4-

4-

4-

4-

4-

Leukemia mononuclear

X

X

X

Lymph node, mesenteric

+

+

-♦-

+

+

+

+•

+

+

+

+

4-

4-

+

4-

M

4-

4-

4-

4-

4-

4-

4-

4-

4-

Leukemia mononuclear

X

X

X

Spleen

+■

+

■*-

+■

■»-

+

+

-f

+

+-

+

4-

4-

+

4-

4-

4"

4-

4-

4-

+

t-

4-

4-

4-

Leukemia mononuclear

X

X

X

X

X

X

X

X

X

Thymus

+

+

-t-

■f

■t-

-t-

M

+

M

M

+

4-

4-

M

4-

4-

M

4-

M

M

+

M

4-

4-

4-

Nitrofurantoin, NTP TR 341

90

TABLE A2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF MALE RATS:

(Continued)

HIGH DOSE

WEEKS On
STUDY

1 1 1 1 1 1 I 1 1 1 I 1 1 1 1 L I 1 1 I 1 1 1 1 I
0000000000000000000000000
5555555555555555555555555

TOTAL;

nssuES

TUMORS

CARCASS
ID

1 1 I I I 1 1 I I 1 1 1 I 1 1 1 1 1 1 1 1 I I ■-! '.i
2223334445566667778999900
3453453454523453455234545

ALIMENTARY SYSTEM
Esophagus
Intestine large
Intestine large, cecum
Intestine large, colon

Musculans. leiomyosarcoma
Intestine large, rectum
Intestine small
Intestine small, duodenum
Intestine small, ileum
Intestine small, jejunum
Liver

Leukemia mononuclear
Mesentery
Pancreas

Leukemia mononuclear
Salivary glands
Stomach
Stomach, forestomach

Papilloma squamous
Stomach, glandular

+ +++ + ++++++ ++++

+ + + + + +- + + + + + + + + +
+ -t- + + + + + -1- + + + + + + +

+ + + + ++ + + ■»■ + + + ++ + +
+ +++ + ++++++ ++++

50
48

48
48

1
47
49
49
47
48
50
15

6
49

1
50
50
49

L
50

CARDIOVASCULAR SYSTEM
Heart
Leukemia mononuclear

+, + +t-+- + + + +-+- + +-+-v-l- + -t--f + + + + -(- + -t-

50

1

ENDOCRINE SYSTEM
Adrenal gland
Adrenal gland, cortex

Adenoma
Adrenal gland, medulla

Leukemia mononuclear

Pheochromocytoma malignant

Pheochromocytoma benign

Pheochromocytoma benign, multiple
Islets, pancreatic

Adenoma

Carcinoma
Parathyroid gland
Pituitary gland

Leukemia mononuclear

Pars distahs, adenoma

Pars distahs, carcinoma
Thyroid gland

C-cell, adenoma

C cell, carcinoma

+ -t- + -(--t--h-t- + +--(- + + +- + + + + + + + + + -t- + +

+ + + -t- + -f+- + +-4--r + + + ■*- + + + + + ■♦- + -♦- + +
+ -t- + -F-»--t-+- + + -t- + -(- + + + + + + + + + -t-4- + -t-

X X

XX XX XX xxxxx

X X

+ + + + -»-■»- + + + + + + + + + + + + + + -♦- + +-■*- +

X X

+ + + -(- + + + + + + + + + + + + + + + + + + ■(- + +
+ + + -t- + + + + + 4- + + + + + -t--t--t-4--t- + + -(- + +

xxxxx XXX X

+ + -t--t--(--.- + -(--(--t--t- + + + -l--t- + -t- + + + + + -t--(-
XX X
X

50
50

1
50

1

2
21

2
49

1

2
49
50

2
13

3
50

6

1

GENERAL BODY SYSTEM
None

GENITAL SYSTEM

Epididymis

Preputial gland

Prostate

Seminal vesicle

Testes
Interstitial ceil, adenoma
Interstitial cell, adenoma, multiple
Tunic, mesothelioma benign
Tunic, mesothelioma malignant
Tunic, sarcoma

X + + -!- + +
X + + + + +

X + -t- + + +

X + + + + +

+ + + + +

t- + + + t

t + + + 1-
X + + + + +

+ + + + +
f + + + +

+ + + + +
+ + + + +

+ + + + +
X + + + + +

X + + +- + +

+ + + + +

X + + + + +

50

47

49

50

50

15

6

2

1

1

HEMATOPOIETIC SYSTEM

Blood
Leukemia mononuclear

Bone marrow
LeuJtemia mononuclear

Lymph node
Bronchial, carcinoma, metastatic.

Zymbal gland
Deep cervical, leukemia mononuclear
Iliac, leukemia mononuclear
Inffuinal. leukemia mononuclear
Mediastinal, leukemia mononuclear
Pancreatic, leukemia mononuclear
Renal, leukemia mononuclear

Lymph node, mandibular
Leukemia mononuclear

Lymph node, mesentenc
Leukemia mononuclear

Spleen
Leukemia mononuclear

Thymus

+- +■(-+-
X XXX

+- + + -(- + + + + + ■(- + + + + +■-»- + + + ■»-■♦- + + + +

X X

+ + 4-+- + -t- + + + + + + + + + + -t- + + + + + + + +

X

X

+ + + 4- + + +M + + + + + + + + + + + + + + + + +

+ + + + + + + + + + + + + + + + + -♦- + ■*--*- + + + +

+ + + + + -»- + + + + + ■*- + + -*- + + + + + -*- + + + +

X XXX
+ M + MM + M+ + + + + + + + MM+-1-M+-I- + M +

10
10
50
4
50

1

1

1

1

1

2

1

47

3

49

3

50

13

34

91

Nitrofurantoin, NTP TR 341

TABLE A2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF MALE RATS: HIGH DOSE

(Continued)

WEEKS On
STUDY

OOOOOOOOOOOOOOOllllllllll
5557788999999990000000000
6691609111123660012333445

CARCASS

ID

1111111112 2 1112 1111111111
7184813 720054805519813162
11112212 11212332331442512

INTEGUMENTARY SYSTEM

Mammary gland
Fibroadenoma

Skin
Basal cell cannnoma
Keratoacanthoma
Papilloma squamous
Subcutaoeous tissue, basosquamous

tumor benign
Subcutaneous tissue, fibroma
Subcutaneous tissue, fibrosarcoma
Subcutaneous tissue, hemangioma
Subcutaneous tissue, neoplasm, NOS

+ -(--(--F^-4-M-l-M + + +--t-M+-t- + -»- + -t-+- + -t- + +
X X

■l--*--(-l- + + +- + -t- + >--(--l- + f-l- + -(--l- + 4- + +- + +

X

X

X

X

X

X

X

MUSCULOSKELETAL SYSTEM
Bone

Femur, osteosarcoma

Vertebra, osteosarcoma

■l-+--4- + + + + + 4- + + -f-«--(-l- + -t--f-(--t- + -t- + + +

X
X

NERVOUS SYSTEM
Brain
Leukemia mononuclear

-f-f-t- + -»-i-+- + + + -t--i--i- + -t--t- + -t--t- + + + + > +

X

RESPrHATORY SYSTEM

Lung
Carcinoma, metastatic. Zymbal gland
LeuJtemia mononuciear
Osteosarcoma, metastatic, bone

Nose

Trachea

+ -hf + -t--(-4-*-+--t--t--f-*- + ^l--t-+--(-+f + -t-+-4-

X
X X XXX XXX
X

MM+- + +--i-+-T-+- + + + -t- + + -t- + + + -*-+' + + -t- +

4- + +- f + + + + + + + + -t- + + -t--t--t- + + + + + + +

SPECIAL SENSES SYSTEM
Ear
Eye

Hardenan gland
Zymbal gland
Carnnoma

+ *- + + + + +

+ 1- f 4- + +
+ + +

X

iraiNARY SYSTEM
Kidney

Carcinoma, metastatic. Zymbal gland

Leukemia mononuclear

Osteosarcoma, metastatic, bone

Pelvis, transitional epithelium,
cannnoma

Renal tubule, adenoma

Renal tubule, carcinoma
Ureter
Unnary bladder

Leukemia mononuclear

+ -t-1--t-+- + -f + + -f- + + -t- + + + + + i--l- + + + + 4-

X
X

X

X

+

+ + f + -t-4-f-f-t- + 4- + -(- + 4--f-f + + + f + + + +

X

Nitrofurantoin, NTP TR 341

92

TABLE A2.

INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF MALE RATS:

(Continued)

HIGH DOSE

WEEKS ON
STUDY

I I 1 1 1 1 1 1 I 1 I 1 1 1 1 L 1 I 1 1 1 1 I 1 1
0000000000000000000000000
5555555555555555555555555

TOTAL:
TISSUES
TUMORS

CARCASS
ID

1 1 1 1 1 1 1 1 1 I L I 1 1 1 1 1 1 I 1 L 1 1 -J -^
2223334445566667778999900
3453453454523453455234545

INTEGUMENTARY SYSTEM

Mammary gland
Fibroadenoma

Slun
Basal cell camnoma
Keratoacanthoma
Papilloma squamous
Subcutaneous tissue, basosquamous

tumor benipi
Subcutaneous tissue, fibroma
Subcutaneous tissue, fibrosarcoma
Subcutaneous tissue, hemangioma
Subcutaneous tissue, neoplasm, NOS

+ -*-M + + + + -»- + + +- + -*- + + + + '*- + "'" + "*"'*"'*" +

-t- + + -t--i--(--»- + + + -i- + + + -t- + + + + -t- + + + +- +

X X
XX X

X

X XX X

46
2

50
3
4
2

1
4

1
1
1

MUSCULOSKELETAL SYSTEM

Bone
Femur, osteosarcoma
Vertebra, osteosarcoma

+ + + + + + + + ■»-■*- + + + + + + + + -♦-+ ■*-■!- + + +

50

1
I

NERVOUS SYSTEM
Brain
Leukemia mononuclear

+ + + + -(--(--(-+■+- + + + + + + + ■*- + + + + + + + +

50

1

RESPnUTORY SYSTEM

Lung
Camnoma, metastatic, Zymbal gland
Leukemia mononuclear
Osteosarcoma, metastatic, bone

Nose

Trachea

+ -t- + + + -*--(- + -t- + + + -F-t- + +- + + -t- + +- + + + +

X X

+ ■(- + + + + + + + + + + + + + + + + + -*- + ■»- + + +

+ + + + + -t- + 4- + + + + + + + + + + -f + + -t--t- + +

50

I
10

I
48
50

SPECIAL SENSES SYSTEM
Ear
Eye

Hardenan gland
Zymbal gland
Carcinoma

-(- + + + + ■)-+ ■(- + + + + + + + + + + +
+ + + + + -(-+ + + + + ++ ■»- + +

+

26

22

4

1
1

URINARY SYSTEM
Kidney

Carcinoma, metastatic. Zymbal gland

Leukemia mononuclear

Osteosarcoma, metastatic, bone

Pelvis, transitional epithelium,
carcinoma

Renal tubuie. adenoma

Renal tubule, carcinoma
Ureter
Unnary bladder

Leukemia mononuclear

+ + f + +- + + -t- + + + + + + + + + + + + -t- + + + -)-

X

X X

+ 1- + +'+-4- + + + + -(- + + + + 4- + + -f + + -»--(--f +

50

I
1

1

1
2
1
1
50
1

93

Nitrofurantoin, NTP TR 341

TABLE A3.

ANALYSIS OF PRIMARY TUMORS IN MALE RATS IN THE TWO-YEAR FEED STUDY

OF NITROFURANTOIN

Control

1,300 ppm

2,500 ppm

Adrenal Gland: Pheochromocytoma

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

Cochran-Armitage Trend Test(d)

Fisher Exact Test (d)

Adrenal Gland: Malignant Pheochromocytoma

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

Cochran-Armitage Trend Test (d)

Fisher Exact Test (d)

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

Cochran-Armitage Trend Test (d)

Fisher Exact Test (d)

Preputial Gland: Adenoma

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates Ic)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

Cochran-Armitage Trend Test (d)

Fisher Exact Test (d)

Preputial Gland: Carcinoma

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

Cochran-Armitage Trend Test (d)

Fisher Exact Test (d)

Preputial Gland: Adenoma or Carcinoma

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

Cochran-Armitage Trend Test (d)

Fisher Exact Test (d)

23/50(46%)

20/50(40%)

23/50 (46%)

7L1%

63.3%

64.5%

15/24(63%)

16/27(59%)

14/26(54%)

591

548

559

P = 0.403N

P = 0.177N

P = 0.443N

P = 0.433N

P = 0.163N

P = 0.477N

P = 0.535N

P = 0.343N

P = 0.579N

3/50(6%)

1/50 (2%)

2/50(4%)

11.7%

3.1%

7.7%

2/24(8%)

0/27(0%)

2/26(8%)

729

722

730

P = 0.376N

P = 0.276N

P = 0.474N

P = 0.365N

P = 0.247N

P = 0.486N

P = 0.395N

P = 0.309N

P = 0.500N

It Pheochromocytoma

24/50(48%)

21/50(42%)

23/50 (46%)

74.3%

64.5%

64.5%

16/24(67%)

16/27(59%)

14/26(54%)

591

548

559

P = 0.330N

P = 0.174N

P = 0.368N

P = 0.345N

P = 0.152N

P = 0.386N

P = 0.456N

P = 0.344N

P = 0.500N

6/48(13%)

5/50(10%)

0/47(0%)

21.9%

15.2%

0.0%

4/22(18%)

3/27(11%)

0/26(0%)

533

630

P = 0.011N

P = 0.388N

P = 0.013N

P = 0.018N

P = 0.461N

P = 0.018N

P = 0.019N

P = 0.471N

P = 0.014N

6/48(13%)

6/50(12%)

0/47(0%)

18.3%

17.3%

0.0%

2/22(9%)

2/27(7%)

0/26(0%)

455

320

P = 0.019N

P = 0.522N

P = 0.015N

P = 0.038N

P = 0.603

P = 0.028N

P = 0.024N

P = 0.591N

P = 0.014N

12/48(25%)

11/50(22%)

0/47(0%)

37.6%

30.6%

0.0%

6/22(27%)

5/27(19%)

0/26(0%)

455

320

P<0.001N

P = 0.352N

P<0.001N

P = 0.001N

P = 0.494N

P<0.001N

P<0.001N

P = 0.455N

P<0.001N

Nitrofurantoin, NTP TR 341

94

TABLE A3.

ANALYSIS OF PRIMARY TUMORS IN MALE RATS IN THE TWO-YEAR FEED STUDY
OF NITROFURANTOIN (Continued)

Control

1,300 ppm

2,500 ppm

Pancreatic Islets: Adenoma or Carcinoma

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

Cochran-Armitage Trend Test (d)

Fisher Exact Test (d)

Kidney: Tubular Adenoma or Carcinoma

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d i

Cochran-Armitage Trend Test (d)

Fisher Exact Test (d)

Liver: Neoplastic Nodule

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

Cochran-Armitage Trend Test (d)

Fisher Exact Test (d)

Liver: Neoplastic Nodule or Hepatocellular Carcinoma

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table TesU(d)

Logistic Regression Tests (d)

Cochran-Armitage Trend Test (d)

Fisher Exact Test (d)

Mammary Gland: Fibroadenoma

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests Id)

Cochran-Armitage Trend Test (d) ,

Fisher Exact Test (d)

Mammary Gland: Adenoma or Fibroadenoma

Overall Rales (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

Cochran-Armitage Trend Test (d)

Fisher Exact Test (d)

2/50(4%)

2/50(4%)

3/49(6%)

8.3%

6.5%

10.9%

2/24(8%)

1/27(4%)

2/26(8%)

730

712

719

P = 0.443

P = 0.648N

P = 0.532

P = 0.441

P = 0.636N

P = 0.530

P = 0.404

P = 0.691

P = 0.490

0/50(0%)

1/50(2%)

3/50(6%)

0.0%

3.7%

10.9%

0/24(0%)

1/27(4%)

2/26(8%)

730

719

P = 0.068

P = 0.523

P = 0.134

P = 0.066

P = 0.523

P = 0.134

P = 0.063

P = 0.500

P = 0.121

1/50(2%)

3/50(6%)

0/50(0%)

4.2%

9.9%

0.0%

1/24(4%)

2/27(7%)

0/26(0%)

730

699

P = 0.366N

P = 0.360

P = 0.484N

P = 0.369N

P = 0.360

P = 0.484N

P = 0.399N

P = 0.309

P = 0.500N

;inoma

1/50(2%)

4/50(8%)

0/50(0%)

4.2%

13.5%

0.0%

1/24(4%)

3/27(11%)

0/26(0%)

730

699

P = 0.379N

P = 0.225

P = 0.484N

P = 0.380N

P = 0.228

P = 0.484N

P = 0.415N

P = 0.181

P = 0.500N

2/50(4%)

4/50(8%)

2/50(4%)

8.3%

12.4%

5.8%

2/24(8%)

2/27(7%)

0/26(0%)

730

644

635

P = 0.568N

P = 0.402

P = 0.663N

P = 0.584N

P = 0.390

P = 0.681N

P = 0.582

P = 0.339

P = 0.691 N

3/50(6%)

4/50(8%)

2/50(4%)

12.5%

12.4%

5.8%

3/24(13%)

2/27(7%)

0/26(0%)

730

644

635

P = 0.391N

P = 0.572

P = 0.463N

P = 0.403N

P = 0.569

P = 0.511N

P = 0.427N

P = 0.500

P = 0.500N

95

Nitrofurantoin, NTP TR 341

TABLE A3.

ANALYSIS OF PRIMARY TUMORS IN MALE RATS IN THE TWO-YEAR FEED STUDY
OF NITROFURANTOIN (Continued)

Control

1.300 ppm

2,500 ppm

Pituitary Gland/Pars Distalis: Adenoma

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

Cochran-Armitage Trend Test (d)

Fisher Exact Test (d)

Pituitary Gland/Pars Distalis: Carcinoma

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

Cochran-Armitage Trend Test (d)

Fisher Exact Test (d)

Pituitary Gland/Pars Distalis: Adenoma or Carcinoma

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

Cochran-Armitage Trend Test (d)

Fisher Exact Test (d)

Skin: Basal Cell Carcinoma

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

Cochran-Armitage Trend Test (d)

Fisher Exact Test (d)

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests <d)

Cochran-Armitage Trend Test (d)

Fisher Exact Test (d)

Skin: Keratoacanthoma

Overall Rates la I

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

Cochran-Armitage Trend Test (d)

Fisher Exact Test (d)

10/50(20%)

12/50(24%)

13/50(26%)

27.8%

32.1%

42.3%

3/24(13%)

5/27(19%)

9/26(35%)

506

489

646

P = 0.355

P = 0.520

P = 0.401

P = 0.282

P = 0.371

P = 0.333

P = 0.278

P = 0.405

P = 0.318

1/50(2%)

2/50(4%)

3/50(6%)

4.2%

6.0%

8.4%

1/24(4%)

1/27(4%)

1/26(4%)

730

644

387

P = 0.248

P = 0.548

P = 0.333

P = 0.223

P = 0.522

P = 0.303

P = 0.225

P = 0.500

P = 0.309

inoma

11/50(22%)

14/50(28%)

16/50(32%)

31.2%

36.7%

48.3%

4/24(17%)

6/27(22%)

10/26(38%)

506

489

387

P = 0.232

P = 0.453

P = 0.268

P = 0.157

P = 0.297

P = 0.188

P = 0.157

P = 0.322

P = 0.184

3/50(6%)

0/50(0%)

3/50(6%)

10.6%

0.0%

9.7%

2/24(8%)

0/27(0%)

2/26(8%)

621

559

P = 0.563N

P = 0.105N

P = 0.63IN

P = 0.588N

P = 0.113N

P = 0.660N

P = 0.589N

P = 0.121N

P = 0.661

ind Carcinoma, or Basal Cell Carcinoma

4/50(8%)

1/50(2%)

3/50(6%)

13.9%

3.7%

9.7%

2/24 ( 8% 1

1/27(4%)

2/26(8%)

621

730

559

P = 0.379N

P = 0.158N

P = 0.473N

P = 0.407N

P = 0.185N

P = 0.492N

P = 0.404N

P = 0.181N

P = 0.500N

4/50(8%)

6/50(12%)

4/50(8%)

1 1 .9%

19.1%

14.5%

1/24(4%)

4/27(15%)

3/26(12%)

455

644

716

P = 0.535N

P = 0.441

P = 0.614N

P = 0.569N

P = 0.367

P = 0.642N

P = 0.564

P = 0.370

P = 0.643N

Nitrofurantoin, NTP TR 341

96

TABLE A3. ANALYSIS OF PRIMARY TUMORS IN MALE RATS IN THE TWO-YEAR FEED STUDY

OF NITROFURANTOIN (Continued)

Control

1,300 ppm

2,500 ppm

Subcutaneous Tissue: Fibroma

Overall Rates (a)
Adjusted Rates (b)
Terminal Rates (c)
Day of First Observation
Life Table Tests (d)
Logistic Regression Tests ( d )
Cochran-Armitage Trend Test (d)
Fisher Exact Test (d)

Subcutaneous Tissue: Fibroma or Fibrosarcoma

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests Id)

Logistic Regression Tests Id)

Cochran-Armitage Trend Test (d)

Fisher Exact Test (d)

Testis: Interstitial Cell Adenoma

Overall Rates (a I

Adjusted Rates (b I

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

Cochran-Armitage Trend Test (d)

Fisher Exact Test (d )

Thyroid Gland: C-Cell Adenoma

Overall Rates (a)

Adjusted Rates (bl

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

Cochran-Armitage Trend Test (d)

Fisher Exact Test (d)

Thyroid Gland: C-Cell Carcinoma

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

Cochran-Armitage Trend Test (d)

Fisher Exact Test (d)

Thyroid Gland: C-Cell Adenoma or Carcinoma

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

Cochran-Armitage Trend Test (d)

Fisher Exact Test (d)

0/50(0%)

5/50(10%)

4/50 (8%)

0.0%

16.2%

15.4%

0/24 10%)

3/27(11%)

4/26(15%)

644

730

P = 0.077

P = 0.047

P = 0.071

P = 0.076

P = 0.042

P = 0.071

P = 0.067

P = 0.028

P = 0.059

1/50(2%)

7/50(14%)

5/50(10%)

2.7%

21.9%

17.5%

0/24(0%)

4/27(15%)

4/26(15%)

633

644

636

P = 0.127

P = 0.054

P = 0.128

P = 0.109

P = 0.039

P = 0.109

P = 0.101

P = 0.030

P = 0.102

47/50(94%)

45/50(90%)

21/50(42%)

100.0%

100.0%

61.8%

24/24(100%)

27/27(100%)

14/26(54%)

455

548

559

P<0.001N

P = 0.162N

P<0.001N

P<0.001N

P = 0.221N

P<0.001N

P<0.001N

P = 0.357N

P<0.001N

3/50(6%)

7/50(14%)

6/50(12%)

12.5%

22.6%

20.1%

3/24(13%)

5/27(19%)

4/26(15%)

730

561

639

P = 0.241

P = 0.207

P = 0.285

P = 0.227

P = 0.200

P = 0.278

P = 0.205

P = 0.159

P = 0.243

0/50(0%)

3/50(6%)

1/50(2%)

0.0%

8.4%

3.8%

0/24(0%)

1/27(4%)

1/26(4%)

656

730

P = 0.394

P = 0.159

P = 0.516

P = 0.373

P = 0.121

P = 0.516

P = 0.367

P = 0.121

P = 0.500

3/50(6%)

9/50(18%)

7/50(14%)

12.5%

26.4%

23.7%

3/24(13%)

5/27(19%)

5/26(19%)

730

561

639

P = 0.176

P = 0.100

P = 0.193

P = 0.155

P = 0.076

P = 0.189

P = 0.140

P = 0.061

P = 0.159

97

Nitrofurantoin, NTP TR 341

TABLE A3.

ANALYSIS OF PRIMARY TUMORS IN MALE RATS IN THE TWO-YEAR FEED STUDY
OF NITROFURANTOIN (Continued)

Control

1,300 ppm

2,500 ppm

Thyroid Gland: Follicular Cell Adenoma or Carcinoma

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

Cochran-Armitage Trend Test (d )

Fisher Exact Test(d)

Hematopoietic System: Mononuclear Leukemia

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

CochranArmitage Trend Test (d)

Fisher Exact Test (d)

All Sites: Malignant Mesothelioma

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d )

CochranArmitage Trend Test (d)

Fisher Exact Test (d)

All Sites: All Mesothelioma

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

CochranArmitage Trend Test (d)

Fisher Exact Test (d)

0/50(0%)

3/50(6%)

0/50(0%)

0.0%

9.7%

0.0%

0/24(0%)

2/27(7%)
656

0/26 (0%)

p = 0.644N

P = 0.150

(e)

P = 0.635

P = 0.133

(e)

P = 0.623

P = 0.121

(e)

23/50 (46%)

14/50(28%)

15/50(30%)

57.0%

39.2%

38.1%

8/24(33%)

7/27(26%)

4/26(15%)

554

561

413

P = 0.053N

P = 0.037N

P = 0.076N

P = 0.054N

P = 0.044N

P = 0.075N

P = 0.055N

P = 0.048N

P = 0.074N

3/50(6%)

5/50(10%)

1/50(2%)

9.3%

13.9%

3.8%

1/24(4%)

1/27(4%)

1/26(4%)

506

630

730

P = 0.247N

P = 0.437

P = 0.284N

P = 0.277N

P = 0.349

P = 0.307N

P = 0.278N

P = 0.357

P = 0.309N

3/50(6%)

6/50(12%)

3/50(6%)

9.3%

17.2%

8.6%

1/24(4%)

2/27(7%)

1/26(4%)

506

630

635

P = 0.538N

P = 0.320

P = 0.622N

P = 0.564

P = 0.242

P = 0.659

P = 0.564

P = 0.243

P = 0.661

(a I Number of tumor-bearing animals/number of animals examined at the site

(b) Kaplan-Meier estimated tumor incidences at the end of the study after adjusting for intercurrent mortality

(c) Observed tumor incidence at terminal kill

(d) Beneath the control incidence are the P values associated with the trend test. Beneath the dosed group incidence are the
P values corresponding to pairwise comparisons between that dosed group and the controls. The life table analysis regards tu-
mors in animals dying prior to terminal kill as being (directly or indirectly) the cause of death. The logistic regression test re-
gards these lesions as nonfatal. The CochranArmitage and Fisher exact tests compare directly the overall incidence rates.
A negative trend or lower incidence in a dosed group is indicated by (N).

(e) No P value is reported because no tumors were observed in the 2,500-ppm and control groups.

Nitrofurantoin, NTP TR 341

98

TABLE A4a. HISTORICAL INCIDENCE OF KIDNEY TUBULAR CELL TUMORS IN MALE F344/N RATS

RECEIVING NO TREATMENT (a)

Study Incidence of Adenomas or Adenocarcinomas in Controls

Historical Incidence at Southern Research Institute

HC Blue No. 2 0/50

C.I. Disperse Blue 1 0/49

Eugenol 0/40

Stannous chloride 0/60

D-Mannitol 0/50

Ziram 0/50

Propyl gallate (b)l/50

Zearalenone 0/50

HCBlueNo. 1 (c) 1/50

TOTAL 2/439(0.5%)

SD(d) 0.88%

Range (e)

High 1/50

Low 0/50

Overall Historical Incidence

TOTAL (f) 8/1,929 (0.4%)

SD(d) 0.94%

Range (e)

High 2/50

Low 0/50

(a) Data as of August?, 1986,for studies of at least 104 weeks

(b) Tubular cell adenoma

(c) Adenoma, NOS

(d) Standard deviation

(e) Range and SD are presented for groups of 35 or more animals.

(f) Includes one adenoma, NOS, six tubular cell adenomas, and one tubular cell adenocarcinoma

99 Nitrofurantoin, NTP TR 341

TABLE A4b. HISTORICAL INCIDENCE OF SKELETAL SYSTEM TUMORS IN MALE F344/N RATS

RECEIVING NO TREATMENT (a)

Study Incidence of Osteosarcomas in Controls

Historical Incidence at Southern Research Institute

HC Blue No. 2 2/50

C.I. Disperse Blue 1 0/49

Eugenol 0/40

Stannous chloride 0/50

D-Mannitol 1/50

Ziram 0/50

Propyl gallate 0/50

Zearalenone 0/50

HC Blue No. 1 0/50

TOTAL 3/439 (0.7%)

SD(b) 1.41%

Range (c)

High 2/50

Low 0/50

Overall Historical Incidence

TOTAL 8/1,937(0.4%)

SD(b) 0.94%

Range (c)

High 2/50

Low 0/50

(a) Data asof August 7, 1986, for studies of at least 104 weeks; no osteomas have been observed.

(b) Standard deviation

(c) Range and SD are presented for groups of 35 or more animals.

Nitrofurantoin, NTP TR 341 100

TABLE A4c. HISTORICAL INCIDENCE OF SUBCUTANEOUS TISSUE TUMORS IN MALE F344/N RATS

RECEIVING NO TREATMENT (a)

Incidence in Controls

Study Fibroma Fibrosarcoma Fibroma or Fibrosarcoma

Historical Incidence at Southern Research Institute

HC Blue No. 2
C.I. Disperse Blue 1
Eugenol

Stannous chloride
D-Mannitol
Ziram

Propyl gallate
Zearalenone
HC Blue No. 1

TOTAL

SD(c)

Range (d)
High
Low

Overall Historical Incidence

TOTAL
SD(c)

Range (d)
High
Low

5/50

(b)2/50

(b)7/50

4/49

1/49

5/49

3/40

0/40

3/40

1/50

1/50

2/50

2/50

0/50

2/50

2/50

0/50

2/50

1/50

0/50

1/50

3/50

1/50

4/50

2/50

0/50

2/50

23/439(5.2%)

5/439(1.1%)

28/439(6.4%)

2.79%

1.46%

3.85%

5/50

2/50

7/50

1/50

0/50

1/50

(e) 107/1,937 (5.5%)

(f) 38/1,937 (2.0%)

(e,f) 144/1,937 (7.4%)

3.22%

2.69%

4.31%

6/50

7/50

12/50

0/50

0/50

0/49

(a) Data as of August?, 1986, for studies of at least 104 weeks

(b) Includes one sarcoma, NOS, and one neurofibrosarcoma

(c) Standard deviation

(d) Range and SD are presented for groups of 35 or more animals.

(e) Includes five neurofibromas

(f) Includes eight sarcomas, NOS, and six neurofibrosarcomas

101 Nitrofurantoin, NTP TR 341

TABLE A4d. HISTORICAL INCIDENCE OF TESTICULAR TUMORS IN MALE F344/N RATS RECEIVING

NO TREATMENT (a)

Study Incidence of Adenomas in Controls (b)

Historical Incidence at Southern Research Institute

HC Blue No. 2 45/50

C.I. Disperse Blue 1 44/49

Eugenol 38/40

Stannous chloride 34/50

D-Mannitol 45/50

Ziram 41/50

Propyl gallate 47/50

Zearalenone 45/50

HC Blue No. 1 45/50

TOTAL 384/439 (87.5%)

SD(c) 8.21%

Range (d)

High 38/40

Low 34/50

Overall Historical Incidence

TOTAL (6)1,681/1,909(88.1%)

SD(c) 7.32%

Range (d)

High 49/50

Low 34/50

(a) Data as of August 7, 1986, for studies of at least 104 weeks

(b) Diagnosed as interstitial cell tumors

(c) Standard deviation

(d) Range and SD are presented for groups of 35 or more animals.

(e) Includes one malignant mterstitial cell tumor

Nitrofurantoin, NTP TR 341 102

TABLE A4e. HISTORICAL INCIDENCE OF PREPUTIAL GLAND TUMORS IN MALE F344/N RATS

RECEIVING NO TREATMENT (a)

Incidence in Controls

Study Adenoma Carcinoma Adenoma or Carcinoma

Historical Incidence at Southern Research Institute

HC Blue No. 2
C.I. Disperse Blue 1
Eugenol

Stannous chloride
D-Mannitol
Ziram

Propyl gallate
Zearalenone
HCBlueNo. 1

TOTAL
SD(d)

Range (e)
High
Low

Overall Historical Incidence

TOTAL (f) 57/1,937 (2.9%) (g)66/l,937(3.4%) (f,g) 123/1,937 (6.4%)

SD(d) 4.02% 2.93% 4.77%

Range (e)

High 8/50 5/50 9/50

Low 0/50 0/50 0/50

(a) Data as of August 7, 1986, for studies of at least 104 weeks

(b) Adenocarcinoma, NOS

(c) Includes one squamous cell carcinoma

(d) Standard deviation

(e) Range and SD are presented for groupsof 35 or more animals.

(f) Includes one papillary adenoma and one cystadenoma, NOS

(g) Includes two squamous cell carcinomas, eight adenocarcmomas, NOS, and one sebaceous adenocarcinoma

0/50

1/50

1/50

0/49

4/49

4/49

0/40

2/40

2/40

0/50

0/50

0/50

0/50

(b)l/50

(b)l/50

3/50

4/50

7/60

0/50

1/50

1/50

1/50

(c) 2/50

(c)3/50

2/50

2/50

4/50

6/439(1.4%)

17/439(3.9%)

23/439(5.2%)

2.24%

2.79%

4.37%

3/50

4/49

7/50

0/50

0/50

0/50

103 Nitrofurantoin, NTPTR 341

TABLE A5.

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS IN THE
TWO-YEAR FEED STUDY OF NITROFURANTOIN

Untreated Control

Low Dose

High Dose

Animals initially in study

Animals removed

Animals exammed histopathologically

50
50
50

50
50
50

50
50
50

ALIMENTARY SYSTEM

Intestine large, cecum

(50)

(50)

(48)

Parasite metazoan

I

(2%)

2

(4%)

Submucosa, edema

1

(2%)

1

(2%)

3

(6%)

Intestine large, colon

(50)

(50)

(48)

Parasite metazoan

6

(12%)

5

(10%)

2

(4%)

Intestine large, rectum

(50)

(48)

(47)

Parasite metazoan

1

(2%)

Liver

(50)

(50)

(50)

Angiectasis

2

(4%)

2

(4%)

1

(2%)

Basophilic focus

1

(2%)

1

(2%)

Congestion

2

(4%)

Cytologic alterations, focal

5

(10%)

5

(10%)

4

(8%)

Cytologic alterations, multifocal

1

(2%)

2

(4%)

2

(4%)

Degeneration, cystic, focal

4

(8%)

2

(4%)

2

(4%)

Developmental malformation

1

(2%)

1

(2%)

Hematopoietic cell proliferation

1

(2%)

Hyperplasia, nodular

1

(2%)

1

(2%)

Infiltration cellular, lymphocytic, multifocal

1

(2%)

Inflammation, granulomatous, multifocal

3

(6%)

4

(8%)

Mixed cell focus

1

(2%)

Necrosis, coagulative, multifocal

3

(6%)

3

(6%)

Vacuolization cytoplasmic, diffuse

1

(2%)

1

(2%)

Vacuolization cytoplasmic, focal

3

(6%)

1

(2%)

1

(2%)

Vacuolization cytoplasmic, multifocal

2

(4%)

Bile duct, hyperplasia

10

(20%)

15

(30%)

21

(42%)

Bile duct, hyperplasia, multifocal

1

(2%)

Centrilobular, degeneration

1

(2%)

Portal, fibrosis

1

(2%l

1

(2%)

Serosa, fibrosis, focal

1

(2%)

Mesentery

(6)

(9)

(6)

Fibrosis, focal

1

(17%)

Inflammation, granulomatous

I

(17%)

Pigmentation, hematoidin, hemosiderin

1

(17%)

Artery, inflammation, chronic

1

(11%)

1

(17%)

Artery, mineralization

1

(17%)

Artery, thrombus

1

(17%)

Fat, edema

1

(17%)

Fat, necrosis, focal

1

(17%)

5

(56%)

3

(50%)

Vein, mineralization

1

(11%)

Pancreas

(50)

(50)

(49)

Hyperplasia, focal

1

(2%)

Pigmentation, hematoidin, hemosiderin

1

(2%)

Acinus, atrophy

9

(18%)

10

(20%)

15

(31%)

Adventitia, edema

1

(2%)

1

(2%)

Artery, inflammation, chronic

2

(4%)

1

(2%)

Salivary glands

(48)

(50)

(50)

Hyperplasia

1

(2%)

Stomach, forestomach

(49)

(50)

(49)

Edema

2

(4%)

2

(4%)

Inflammation, chronic

3

(6%)

1

(2%)

Inflammation, suppurative

1

(2%)

Mineralization

3

(6%)

3

(6%)

Necrosis

1

(2%)

Ulcer, focal

1

(2%)

2

(4%)

Stomach, glandular

(49)

(50)

(50)

Edema

1

(2%)

Mineralization

1

(2%)

8

(16%)

14

(28%)

Nitrofurantoin, NTP TR 341

104

TABLE A5. SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS IN THE
TWO-YEAR FEED STUDY OF NITROFURANTOIN (Continued)

Untreated Control

Low

Dose

High

Dose

CARDIOVASCULAR SYSTEM

Blood vessel

(1)

Aorta, mineralization

1

(100%)

Heart

(50)

(50)

(50)

Fibrosis, multifocal

44

(88%)

45

(90%)

39

(78%)

Inflammation, suppurative

1

(2%)

Mineralization, multifocal

1

(2%)

Atrium, thrombus

3

(6%)

3

(6%)

2

(4%)

Valve, bacterium

1

(2%)

ENDOCRINE SYSTEM

Adrenal gland

(50)

(50)

(50)

Fibrosis

1

(2%)

Adrenal gland, cortex

(50)

(50)

(50)

Angiectasis

1

(2%)

1

(2%)

Cyst

I

(2%)

Hyperplasia, focal

3

(6%)

2

(4%)

Hypertrophy, focal

2

(4%)

Necrosis, multifocal

1

(2%)

Vacuolization cytoplasmic, diffuse

1

(2%)

Vacuolization cytoplasmic, focal

11

(22%)

6

(12%)

15

(30%)

Vacuolization cytoplasmic, multifocal

1

(2%)

2

(4%)

Adrenal gland, medulla

(50)

(50)

(50)

Angiectasis

1

(2%)

Hemorrhage

1

(2%)

Hyperplasia, focal

10

(20%)

8

(16%)

7

(14%)

Hyperplasia, multifocal

1

(2%)

2

(4%)

Mineralization

1

(2%)

1

(2%)

Parathyroid gland

(49)

(47)

(49)

Hyperplasia

3

(6%)

18

(38%)

23

(47%)

Pituitary gland

(50)

(50)

(50)

Pars distalis, angiectasis

9

(18%)

7

(14%)

14

(28%)

Pars distalis, cyst

1

(2%)

2

(4%)

3

(6%)

Pars distalis, cyst, multiple

I

(2%)

2

(4%)

Pars distalis, hemorrhage

1

(2%)

Pars distalis, hyperplasia, focal

7

(14%)

6

(12%)

5

(10%)

Pars distalis, hyperplasia, multifocal

1

(2%)

Pars distalis, necrosis

1

(2%)

Pars nervosa, mfiltration cellular

1

(2%)

Thyroid gland

(50)

(50)

(50)

Ultimobranchial cyst

1

(2%)

2

(4%)

C-cell, hyperplasia, focal

6

(12%)

2

(4%)

5

(10%)

C-cell, hyperplasia, multifocal

1

(2%)

1

(2%)

Follicle, cyst

1

(2%)

1

(2%)

Follicle, cyst, multiple

1

(2%)

Follicle, hyperplasia, cystic

2

(4%)

1

(2%)

GENERAL BODY SYSTEM

None

GENITAL SYSTEM
Coagulating gland
Hyperplasia
Inflammation, chronic
Inflammation, suppurative

(1)

(1)

I (100%)

1 (100%)

1 (100%)

105

Nitrofurantoin, NTP TR 341

TABE A5. SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS IN THE
TWO-YEAR FEED STUDY OF NITROFURANTOIN (Continued)

Untreated Control

Low Dose

High Dose

GENITAL SYSTEM (Continued)
Epididymis

Atypical cells

Degeneration

Depletion

Fibrosis

Inflammation, granulomatous
Preputial gland

Atrophy

Fibrosis

Hyperplasia

Inflammation, suppurative

Duct, cyst
Prostate

Cyst

Cyst, multiple

Fibrosis

Inflammation

Inflammation, chronic

Inflammation, chronic, focal

Inflammation, suppurative

Mineralization, focal

Mineralization, multifocal

Epithelium, hyperplasia
Seminal vesicle

Atrophy

Dilatation

Fibrosis

Mineralization

Epithelium, hyperplasia
Testes

Aspermatogenesis

Atrophy

Degeneration

Hemorrhage

Mineralization

Arteriole, necrosis, fibrinoid

Interstitial cell, hyperplasia

Perivascular, infiltration cellular,
mononuclear cell

(50)

(50)

40

(80%)

(48)

1

(2%)

7

(15%)

1

(2%)

11

(23%)

9

(19%)

(50)

3 (6%)

1 (2%)

24 (48%)

1 (2%)

(50)

1 (2%)

(50)
38 (76%)
41 (82%)

1 (2%)

1 (2%)

1 (2%)

4 (8%)

3 (6%)

43
1

1
(50)

(86%)

(2%)

(2%)

3

1

6

13

(50)

2

(6%)
(2%)
(12%)
(26%)

(4%)

4

1
2

(8%)
(2%)
(4%)

19

(38%)

1

(50)

8

(2%)
(16%)

1
1

(2%)
(2%)

(50)
40
44

(80%)
(88%)

(50)

12

(24%)

1

(2%)

46

(92%)

(47)

5

(11%)

3

(6%)

(49)

2

(4%)

3

(6%)

8 (16%)
3 (6%)

9 (18%)

39 (80%)

1 (2%)
1 (2%)
(50)

8 (16%)
1 (2%)

4 (8%)

(50)

44 (88%)

48 (96%)

36 (72%)

15 (30%)
2 (4%)

19 (38%)

HEMATOPOIETIC SYSTEM
Blood

Anemia

Hypochromasia

Polychromasia
Bone marrow

Atrophy

Hyperplasia, reticulum cell

Myelofibrosis

Myeloid cell, hyperplasia
Lymph node

Inguinal, hyperplasia, lymphoid

Mediastinal, angiectasis

Mediastinal, congestion

Mediastinal, hyperplasia, lymphoid

Mediastinal, pigmentation, hemosiderin

Pancreatic, hyperplasia, lymphoid

Pancreatic, inflammation, granulomatous

Pancreatic, mineralization

Renal, angiectasis

Renal, hyperplasia, lymphoid

1

(6%)

1

(6%l

(501

2

(4%)

1

(2%)

1

(2%)

(50)

1

(2%)

1 (2%)

1 (2%)
1 (2%)

(13)
1 (8%)

1 (8%)
(50)

2 (4%)
(50)
2 (4%)

1 (2%)
1 (2%)

1 (2%)

1 (2%)

(10)

1 (10%)
(50)

1 (2%)

2 (4%)

(50)

1 (2%)

2 (4%)

2 (4%)

1 (2%)

1 (2%)

1 (2%)

4 (8%)

3 (6%)

Nitrofurantoin, NTP TR 341

106

TABLE A5. SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS IN THE
TWO-YEAR FEED STUDY OF NITROFURANTOIN (Continued)

Untreated Control

Low

Dose

High

Dose

HEMATOPOIETIC SYSTEM (Continued)

Lymph node, mandibular

(46)

(50)

(47)

Angiectasis

1

(2%)

1

(2%)

Cyst

1

(2%)

1

(2%)

1

(2%)

Hyperplasia, lymphoid

4

(9%)

2

(4%)

10

(21%)

Infiltration cellular, plasma cell

1

(2%)

Lymph node, mesenteric

(49)

(48)

(49)

Angiectasis

3

(6%)

3

(6%)

Ectasia

2

(4%)

Hyperplasia, lymphoid

3

(6%)

4

(8%)

Inflammation, chronic

1

(2%)

Mineralization

1

(2%)

Spleen

(50)

(50)

(50)

Congestion

4

(8%)

4

(8%)

Fibrosis

1

(2%)

Fibrosis, diffuse

1

(2%)

Fibrosis, focal

1

(2%)

2

(4%)

6

(12%)

Hematopoietic cell proliferation

3

(6%)

1

(2%)

2

(4%)

Hyperplasia, lymphoid

2

(4%)

Necrosis

1

(2%)

Pigmentation, hemosiderin

1

(2%)

Thymus

(40)

(36)

(34)

Artery, inflammation

1

(3%)

INTEGUMENTARY SYSTEM

Mammary gland

(46)

(49)

(46)

Duct, cyst

6

(13%)

11

(22%)

7

(15%)

Skin

(50)

(50)

(50)

Alopecia

2

(4%)

Cyst epithelial inclusion

2

(4%)

3

(6%)

Fibrosis, focal

3

(6%)

1

(2%)

Hyperkeratosis, focal

2

(4%)

2

(4%)

Hyperplasia, focal

I

(2%)

2

(4%)

Inflammation, chronic

2

(4%)

Ulcer, focal

1

(2%)

1

(2%)

Prepuce, ulcer

1

(2%)

Subcutaneous tissue, mineralization, focal

1

(2%)

Subcutaneous tissue, necrosis, focal

1

(2%)

MUSCULOSKELETAL SYSTEM

Bone

(50)

(50)

(50)

Fibrous osteodystrophy

5

(10%)

5

(10%)

NERVOUS SYSTEM

Brain

(50)

(50)

(50)

Compression

1

(2%)

2

(4%)

2

(4%)

Cerebellum, degeneration

1

(2%)

Cerebellum, hemorrhage, acute, multifocal

1

(2%)

Cerebellum, hemorrhage, focal

1

(2%)

1

(2%)

RESPIRATORY SYSTEM

Lung

(50)

(50)

(50)

Atelectasis

1

(2%)

2

(4%)

Congestion

2

(4%)

2

(4%)

3

(6%)

Hemorrhage, multifocal

1

(2%)

1

(2%)

Infiltration cellular, lymphocytic, focal

1

(2%)

Infiltration cellular, histiocytic
Inflammation, suppurative

3

(6%)

8

1

(16%)
(2%)

2

(4%)

107

Nitrofurantoin, NTP TR 341

TABLE A5. SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE RATS IN THE
TWO-YEAR FEED STUDY OF NITROFURANTOIN (Continued)

Untreated Control

Low Dose

High Dose

RESPIRATORY SYSTEM

Lung (Continued) (50)

Alveolar epithelium, hyperplasia, focal
Alveolar epithelium, hyperplasia, multifocal
Alveolus, edema

Alveolus, mineralization, multifocal

Arteriole, hypertrophy 1 (2%)

Bronchiole, foreign body
Interstitium, edema
Interstitium, infiltration cellular,

mononuclear cell
Interstitium, infiltration cellular,

histiocytic, diffuse 1 (2% I

Mediastinum, pigmentation, hemosiderin
Nose (50)

Lumen, foreign body

Lumen, fungus 2

Lumen, inflammation, suppurative 3

Nasolacrimal duct, inflammation, suppurative 1

(4%)
(6%)
(2%)

(50)

I

(2%)

1

(2%)

1

(2%)

1

(2%)

I

(2%)

1

(2%)

1 (2%)
(49)

1 (2%)

1 (2%)
4 (8%)

(50)

3 (6%)

1 (2%)

1 (2%)

(48)

1 (2%)

2 (4%)

SPECIAL SENSES SYSTEM

Eye (4)

Cataract 1 (25%)

Anterior chamber, hemorrhage

Anterior chamber, inflammation, suppurative

Cornea, infiltration cellular, lymphocytic

Cornea, inflammation, granulomatous, focal

Cornea, mineralization

Lens, mineralization

Retina, degeneration 2 (50%)

Harderian gland (I)

Atrophy

Ectasia

Fibrosis 1 (100%)

Lacrimal gland (1)

Atrophy 1 (100%)

Inflammation, chronic 1 (100%)

>)

(22)

14 (64%)

1 (5%)

1 (17%)

1 (5%)

1 (17%)

1 (5%)

1 (17%)

(5)

2 (40%)

3 (60%)

2

(9%)

17

(77%)

(4)

1

(25%)

4

(100%)

URINARY SYSTEM
Kidney

Embolus bacterial, multifocal

Infiltration cellular, lymphocytic, multifocal

Inflammation, chronic

Inflammation, suppurative

Mineralization

Nephropathy, chronic

Cortex, cyst

Cortex, cyst, multiple

Cortex, necrosis

Medulla, congestion

Pelvis, hydronephrosis

Pelvis, transitional epithelium, hyperplasia

Renal tubule, hyperplasia, focal

Renal tubule, pigmentation, hemosiderin
Ureter

Dilatation
Urinary bladder

Mucosa, hyperplasia

Mucosa, inflammation, chronic

(50)

50 (100%)
2 (4%)

1 (2%)

2 (4%)
1 (2%)

(50)

(50)

1

(2%)

1

(2%)

2

(4%)

48

(96%)

2

(4%)

3

(6%)

5

(10%)

5

(10%)

2

(4%)

1

(2%)

(50)

1

(2%)

1

(2%)

(50)
1

(2%)

1

(2%)

1

(2%)

3

(6%)

48

(96%)

2

(4%)

1

(2%)

1

(2%)

2

(4%)

2

(4%)

1

(2%)

(1)

1

(100%)

(50)

2

(4%)

Nitrofurantoin, NTP TR 341

108

APPENDIX B

SUMMARY OF LESIONS IN FEMALE RATS IN

THE TWO-YEAR FEED STUDY OF NITROFURANTOIN

PAGE

TABLE Bl

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE RATS IN THE
TWO-YEAR FEED STUDY OF NITROFURANTOIN

111

TABLE 82

INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE RATS IN THE TWO-YEAR
FEED STUDY OF NITROFURANTOIN

114

TABLE B3

ANALYSIS OF PRIMARY TUMORS IN FEMALE RATS IN THE TWO-YEAR FEED
STUDY OF NITROFURANTOIN

126

TABLE B4a

HISTORICAL INCIDENCE OF CLITORAL GLAND TUMORS IN FEMALE F344/N
RATS RECEIVING NO TREATMENT

129

TABLE B4b

HISTORICAL INCIDENCE OF MAMMARY GLAND TUMORS IN FEMALE F344/N
RATS RECEIVING NO TREATMENT

130

TABLE B5

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS
IN THE TWO-YEAR FEED STUDY OF NITROFURANTOIN

131

109

Nitrofurantoin, NTP TR 341

Nitrofurantoin, NTPTR 341 110

TABLE Bl.

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE RATS IN THE TWO-YEAR
FEED STUDY OF NITROFURANTOIN

Untreated Control

Low

Dose

High

Dose

Animals initially in study

50

50

50

Animals removed

50

50

50

Animals examined histopathologica

illy

50

50

50

ALIMENTARY SYSTEM

Liver

(50)

(50)

(50)

Leukemia mononuclear

12

(24%)

13

(26%)

9

(18%)

Neoplastic nodule

1

(2%)

Osteosarcoma, metastatic, bone

1

(2%)

Sarcoma stromal, metastatic,

uterus

1

(2%)

Mesentery

*(50)

♦(50)

*(50)

Leukemia mononuclear

I

(2%)

1

(2%)

Sarcoma stromal, metastatic,

uterus

I

(2%)

Pancreas

(50)

(48)

(49)

Leukemia mononuclear

1

(2%)

2

(4%)

Sarcoma stromal, metastatic,

uterus

1

(2%)

Acinus, adenoma

1

(2%)

Salivary glands

(49)

(49)

(50)

Adenocarcinoma

1

(2%)

CARDIOVASCULAR SYSTEM

Heart

(50)

(50)

(50)

Leukemia mononuclear

2

(4%)

2

(4%)

2

(4%)

ENDOCRINE SYSTEM

Adrenal gland, cortex

(50)

(49)

(50)

Adenoma

2

(4%)

1

(2%)

Leukemia mononuclear

4

(8%)

7

(14%)

4

(8%)

Adrenal gland, medulla

(48)

(48)

(50)

Leukemia mononuclear

4

(8%)

4

(8%)

4

(8%)

Pheochromocytoma malignant

1

(2%)

Pheochromocytoma benign

2

(4%)

1

(2%)

4

(8%)

Islets, pancreatic

(50)

(48)

(49)

Carcinoma

1

(2%)

Parathyroid gland

(49)

(50)

(47)

Adenoma

1

(2%)

Pituitary gland

(50)

(48)

(48)

Leukemia mononucli?ar

3

(6%)

3

(6%)

1

(2%)

Pars distalis, adenoma

23

(46%)

16

(33%)

21

(44%)

Pars distalis, carcinoma

3

(6%)

9

(19%)

2

(4%)

Thyroid gland

(50)

(50)

(50)

Leukemia mononuclear

1

(2%)

C-cell, adenoma

3

(6%)

2

(4%)

2

(4%)

C-cell, adenoma, multiple

1

(2%)

2

(4%)

C-cell, carcinoma

4

(8%)

2

(4%)

2

(4%)

GENERAL BODY SYSTEM
Tissue, NOS

Osteosarcoma, metastatic, bone

*(50)

♦(50)

♦(50)

I (2%)

111

Nitrofurantoin, NTP TR 341

TABLE Bl.

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE RATS IN THE TWO-YEAR
FEED STUDY OF NITROFURANTOIN (Continued)

Untreated Control

Low Dose

High Dose

GENITAL SYSTEM

Clitoral gland
Adenoma
Carcinoma

Ovary

Leukemia mononuclear

Uterus

Leiomyosarcoma
Leukemia mononuclear
Polyp stromal
Polyp stromal, multiple
Sarcoma stromal
Cervix, sarcoma stromal
Cervix, squamous cell carcmoma
Endometrium, adenoma
Endometrium, sarcoma stromal

144)

1 (2%)

4 (9%)

(50)

(50)

1 (2%)

9 (18%

1 (2%)

2 (4%)

>b)

(38)

(42)

7 (18%)

4

(10%)

3 (8%)

(50)

(50)

4 <8%)

1

(2%)

(50)

(50)

1

(2%)

1

(2%)

16 (32%)

10

(20%)

1

(2%)

1 (2%)

2

(4%)

1 (2%)

HEMATOPOIETIC SYSTEM

Blood

*(50)

♦(50)

•(50)

Leukemia mononuclear

7

(14%)

1

(2%)

6

(12%)

Bone marrow

(49)

(48)

(50)

Leukemia mononuclear

7

(14%)

4

(8%)

6

(12%)

Sarcoma

1

(2%)

Lymph node

(50)

(50)

(50)

Inguinal, leukemia mononuclear

1

(2%)

Mediastinal, leukemia mononuclear

2

(4%)

2

(4%)

Pancreatic, leukemia mononuclear

1

(2%)

1

(2%)

I

(2%)

Lymph node, mandibular

(48)

(47)

(50)

Leukemia mononuclear

2

(4%)

3

(6%)

5

(10%)

Lymph node, mesenteric

(50)

(48)

(49)

Leukemia mononuclear

3

(6%)

3

(6%)

3

(6%)

Spleen

(50)

(49)

(50)

Leukemia mononuclear

12

(24%)

13

(27%)

9

(18%)

Sarcoma stromal, metastatic.

uterus

1

(2%)

Thymus

(43)

(34)

(40)

Leukemia mononuclear

1

(2%)

1

(3%)

INTEGUMENTARY SYSTEM
Mammary gland
Adenocarcinoma
Adenocarcinoma, multiple
Fibroadenoma
Fibroadenoma, multiple
Fibrosarcoma
Skin

Basal cell carcinoma
Keratoacanthoma
Papilloma squamous
Foot, papilloma squamous
Subcutaneous tissue, fibroma
Subcutaneous tissue, fibrosarcoma
Subcutaneous tissue, hemangiosarcoma
Subcutaneous tissue, leukemia mononuclear
Subcutaneous tissue, sarcoma

(49)

6 (12%)

23 (47%)

5 (10%)

1 (2%)
(50)

1 (2%)

1 (2%)

1 (2%)

1 (2%)

(50)

4 (8%)

1 (2%)

16 (32%)

10 (20%)

(49)

1 (2%)

2 (4%)
1 (2%)

(50)

16 (32%)
15 (30%)

(50)

1 (2%)
1 (2%)
1 (2%)

MUSCULOSKELETAL SYSTEM

Skeletal muscle *(50)

Diaphragm, sarcoma stromal, metastatic, uterus 1

(2%)

•(50)

•(50)

Nitrofurantoin, NTP TR 341

112

TABLE Bl. SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE RATS IN THE TWO-YEAR

FEED STUDY OF NITROFURANTOIN (Continued)

Untreated Control

Low Dose

High Dose

NERVOUS SYSTEM
Brain

Astrocytoma malignant

Carcinoma, metastatic, pituitary gland

Leukemia mononuclear

(50)

(50)

1 (2%)
1 (2%)
3 (6%)

(50)

1 (2%)

RESPIRATORY SYSTEM
Lung

Alveolar/bronchiolar adenoma
Alveolar/bronchiolar carcinoma
Carcinoma, metastatic, thyroid gland
Leukemia mononuclear
Sarcoma stromal, metastatic, uterus

iO)

(50)

(50)

2 (4%)

2 (4%)

2 (4%)

1 (2%)

1 (2%)

12 (24%)

12 (24%)

8 (16%)

1 (2%)

SPECIAL SENSES SYSTEM
Zymbal gland
Carcinoma

•(50)

1 (2%)

•(50)

1 (2%)

•(50)

URINARY SYSTEM
Kidney

Leukemia mononuclear

Sarcoma stromal, metastatic, uterus

Renal tubule, adenoma
Urinary bladder

Leukemia mononuclear

(50)
3 (6%)
1 (2%)

(50)

(50)
4 (8%)

(50)

1 (2%)

(50)
2 (4%)

1 (2%)
(50)

1 (2%)

SYSTEMIC LESIONS
Multiple organs

Leukemia mononuclear
Hemangiosarcoma

(50)

•(50)

13 (26%)

13 (26%

1 (2%)

•(50)

9 (18%)

ANIMAL DISPOSITION SUMMARY
Animals initially in study
Moribund
Terminal sacrifice
Dead

50

22

25

3

50
12
26
12

50

17

31

2

TUMOR SUMMARY

Total animals with primary neoplasms ** 49

Total primary neoplasms 109

Total animals with benign neoplasms 43

Total benign neoplasms 70

Total animals with malignant neoplasms 3 1

Total malignant neoplasms 39

Total animals with secondary neoplasms *** 1

Total secondary neoplasms 7

48

116

41

79

28

37

1

1

46

100

42

82

17

18

3

4

' Number of animals receiving complete necropsy examinations; all gross lesions including masses examined microscopically.

'* Primary tumors: all tumors except secondary tumors

'*• Secondary tumors: metastatic tumors or tumors invasive into an adjacent organ

113

Nitrofurantoin, NTP TR 341

TABLE B2. INDIVIDUAL AmMAL TUMOR PATHOLOGY OF FEMALE RATS IN THE TWO-YEAR FEED

STUDY OF NITROFURANTOIN: UNTREATED CONTROL

WEEKS ON
STUDY

OOOOOOOOOOOOOOIlIlll lllll
6678889999999900000000000
2932691345667901 12 3344455

CARCASS
ID

33333^3334433333343433333
93427261700352243 07029719
1 I 11121 1212 21342333452423

ALIMENTARY SYSTEM
Esophagus
Intestine large
Intestine large, cecum
Intestine large, colon
Intestine large, rectum
Intestine small
Intestine small, duodenum
Intestine small, ileum
Intestine small, jejunum
Liver

Leukemia mononuclear

Sarcoma stromal, metastatic, uterus
Mesentery

Leukemia mononuclear

Sarcoma stromal, metastatic, uterus
Pancreas

Sarcoma stromal, metastatic, uterus
Salivary glands
Stomach

Stomach, forestomach
Stomach, glandular

+ + + + + + + + + + + + + + + + + + 4- + + + + + +
+ + + + + + 4- + + + + + + + + 4- + + + + + + + + +
+ + + + + + + + + + 4- + 4- + 4- + + + + + + + + + +
+ + + + + + + + + + + + + + + + + + + + + + + + +
+ + + + + + + + + + + + + + + + + + + + + + + + +
+ + + + + + + + + + + + + + + + + + + + + + + + +
+ + + + + + + + + + + + + + + + + + + + + 4-4- + +
+ +A-(-4- + + + + + + + + + + + + + + + + 4- + + 4-
+ + + + + + + + + + + + + + + + + + + 4- + + + + +
+ + + + + 4- + f + + + + + + + + + + + + + + 4- + +

XXX XX xxxxx

X

+ + + + +

X
X

+ + + + + + + + + + + + + + + + + + + + + + + + +
X

+ + -t- + + + + + +M + 4- + + + + + + + + + + 4- + +
+ + + + + + + + + + + + + + + + + + + + + 4- + + +
+ + + + + + + + + + + + + -I- + + + + + + + 4- + 4- +
4- + f + + + + + + + + + + + + + + + -f + 4- + + + 4-

CARDIOVASCULAil SYSTEM
Heart
Leukemia mononuclear

+ + + + + + + + + + + + + + + + + + + + + 4- + + +

X X

ENDOCRINE SYSTEM

Adrenal gland

Adrenal gland, cortex
Leukemia mononuclear

Adrenal gland, medulla
Leukemia mononuclear
Pheochromocytoma benign

Islets, pancreatic
Carcinoma

Parathyroid gland

Pituitary gland
Leukemia mononuclear
Pars distaiis. adenoma
Parsdistahs, carcinoma

Thyroid gland
C-cell. adenoma
C-cell, carcinoma

+ + + + + + + + + + + + + + + + + + + + + + + + 4-
+ + + + + + + + + + + + + + + + + + + + + + 4- + +

XX X

+ + +M4-M+4- + + + + + + + 4- + + + + + + + + +

XXX X

X

+ + + + + + + + + + + + + + + + + + + + 4- + + + +

+ + + +M4- + + + + + 4- + + -I- + + + + + + + + + 4-
+ + + + + + + + + + + + + + + + + + + + + + + + 4-

XXX

X xxxxx XXX X X ' X

X X

4--f--f+- + -t--f + + -«- + + + + +- + -t- + -t-+-4--t- + + +

X

X

GENERAL BODY SYSTEM
None

GENITAL SYSTEM
Clitoral gland

Adenoma

Carcinoma
Ovary
Uterus

Leukemia mononuclear

Polyp stromal

Polyp stromal, multiple

Sarcoma stromal

+ -t-MMM-t- + + +M+ + + +■■♦--(- + + + + 4- + + + +
X
X X
+ + + + + + + + + + + + + ^ + + + + ^-*--*- + + + +

+ + + + + + + -*-^ + 4--»-+'+- + + -f^-t- + + + + -f +

X
X XXX

X

fTEMATOPOIETIC SYSTEM
Blood

Leukemia mononuclear
Bone marrow

Leukemia mononuclear

Sarcoma
Lymph node

Mediastinal, leukemia mononuclear

Pancreatic, leukemia mononuclear
Lymph node, mandibular

Leukemia mononuclear
Lymph node, mesentenc

Leukemia mononuclear
Spleen

Leukemia mononuclear

Sarcoma stromal, metastatic, uterus
Thymus

Leukemia mononuclear

+ + + + + -*- + +
X X X X X X

+ +--t-4--f+-f + +- + -t->-f + + + + + +- + + -f + + +

X X X X XX

+ + ■*- + + + +- + + + + + -»- + ■»- + + + + + ■*- + + + +-
X X

X
M + + f + + f+-+M+ + +-+- + + + + 't- + + +- + + +

X X
+ f + + -t- + + +--t- + -t- + + + + +- + + -t- + -f-+- + + f
X XX

+ + + + + 4-+- + -f + -t- + -(- + + -t- + + -h+-+--f+- + -t-

XXX XX xxxxx

X

+ + + + + + + + +M-fMM + + M-t- + -t-+-M+ + + +
X

+ : Tissue examined microscopically
Not examined

— : Present but not examined microscopically
I: Insufficient tissue

M: Missing

A: Autolysis precludes examination

X: Incidence of listed morphology

Nitrofurantoin, NTP TR 341

114

TABLE B2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE RATS:

(Continued)

UNTREATED CONTROL

WEEKS ON

1

1

1

1

1

1

1

1

1

1

I

1

1

1

1

I

1

1

1

I

I

1

1

1

I

STUDY

0

0

n

n

0

n

0

0

0

n

0

U

0

0

U

U

U

0

0

0

U

0

0

0

U

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

b

b

5

5

b

5

TOTAL:

CARCASS

.1

.1

.1

.1

a

s

s

.1

.1

3

.1

S

11

;)

:t

a

a

a

a

a

a

a

a

i

TISSUES

ID

«

1

1

1

3

3

4

4

4

s

S

S

S

6

6

6

6

7

8

8

8

8

9

9

0

TUMORS

L

3

4

5

4

5

3

4

5

2

3

4

5

2

3

4

5

5

2

3

4

5

4

5

5

ALIMENTARY SYSTEM
Esophagus
Intestine large
Intestine large, cecum
Intestine large, colon
Intestine large, rectum
Intestine small
Intestine small, duodenum
Intestine small, ileum
Intestine small, jejunum
Liver

Leukemia mononuclear

Sarcoma stromal, metastatic, uterus
Mesentery

Leukemia mononuclear

Sarcoma stromal, metastatic, uterus
Pancreas

Sarcoma stromal, metastatic, uterus
Salivary glands
Stomach

Stomach, forestomach
Stomach, glandular

CARDIOVASCULAR SYSTEM
Heart

Leukemia mononuclear

ENDOCRINE SYSTEM
Adrenal gland
Adrenal gland, cortex

Leukemia mononuclear
Adrenal gland, medulla

Leukemia mononuclear

Pheochromocytoma benign
Islets, pancreatic

Carcinoma
Parathyroid gland
Pituitary gland

Leukemia mononuclear

Pars distahs. adenoma

Pars distahs. carcinoma
Thyroid gland

C cell, adenoma

C-cel!, carcinoma

GENERAL BODV SYSTEM "

None

GENITAL SYSTEM
Clitoral gland

Adenoma

Carcinoma
Ovary
Uterus

Leukemia mononuclear

Polyp stromal

Polyp stromal, multiple

Sarcoma stromal

HEMATOPOIETIC SYSTEM
Blood

Leukemia mononuclear
Bone marrow

Leukemia mononuclear

Sarcoma
Lymph node

Mediastinal, leukemia mononuclear

Pancreatic, leukemia mononuclear
Lymph node, mandibular

Leukemia mononuclear
Lymph node, mesentenc

Leukemia mononuclear
Spleen

Leukemia mononuclear

Sarcoma stromal, metastatic, uterus
Thymus

Leukemia mononuclear

+ +

+ + +

+ + +
+ + +
+ + -f

+ +

-*--»- + -♦- + +

+ + + + + +

XX XX

-t- + 4- + + +

X X

-t- +
+ +

■(- + +
+ + +

■t- +■
X

+ + M *

XX X X

X

+ +

+ +
X

■(- + +

+ + + + -*-
X

50

50

50

50

50

50

50

49

50

50

12

1

8

1

I

50

1

49

50

50

50

50
2

50

50

4

48
4
2

50
1

49

50
3

23
3

50
3
4

44
1
4
50
50
1
9
1
2

9

7
49

7

1
50

2

1
48

2
50

3
50
12

1
43

1

115

Nitrofurantoin, NTP TR 341

TABLE B2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE RATS: UNTREATED CONTROL

(Continued)

WEfiKS OK
STUDY

OOOOOOOOOOOOOOlllLlllllll
6678889999999900000000000
2932691345667901 123344455

CARCASS
ID

3333333334433333343433333
93427 2 61700352243070 2 9719
111112 1121221342333452423

INTEGUMENTARY SYSTEM
Mammary gland

Adenocarcinoma

Fibroadenoma

Fibroadenoma, multiple

Fibrosarcoma
Skin

Subcutaneous tissue, fibroma

Subcutaneous tissue, heraangiosarcoma

Subcutaneous tissue, leukemia
mononuclear

Subcutaneous tissue, sarcoma

+ + + +M-t- + + + + + + + + -t- + + -(- + + + + -(- + +

X XX
XX XXX X XXX XXX

X X
X
+ + + -t- + '+--»- + + + -t- + + + + + + + +-+-+- + -l--(- +

X

X

X

MUSCULOSKELETAI. SYSTEM
Bone

Skeletal muscle
Diaphragm, sarcoma stromal, metastatic,
uterus

+ + -t- + -t- + *-+- + + + + + + + + + + 4- + + + -»-4- +

NERVOUS SYSTEM

Brain

Spinal cord

+ + + ■*- + -•-+♦- + + •(- + + + + ■+- + ■*- + + + + + + +

+

RESPIRATORY SYSTEM
Lung

Alveolar/bronchiolar adenoma

Alveotar/bronchiolar carcinoma

Leukemia mononuclear

Sarcoma stromal, metastatic, uterus
Nose
Trachea

■t- + -t- + -«- + -t- + -l- + + + -t- + + -(- + + -*- + -(- + + + +

X

XXX XX xxxxx

X

+ + + + + + + + + -+- + + + + + + + + + + + + + ■♦■■•-

-t- + -t- + + + + + + + + f + + f4- + -(- + + 4- + + -(- +

iSPECIAL SENSES SYSTEM
Ear
Eye

Hardenan gland
Zymbal gland
Carcinoma

+ ++ + +■»- +

+

+
X

URINARY SYSTEM
Kidney

Leukemia mononuclear

Sarcoma stromal, metastatic, uterus
Unnary bladder

Leukemia mononuclear

+ + +■ + + -*- + +- + + + + + + + + + +■ + + + + ■♦-■*--»-
XX X
X

-t- + + + + -t--t- + -4--t--(- + + -»- + + -(- + + +-l--t- + + -t-

X

Nitrofurantoin, NTP TR 341

116

TABLE B2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE RATS: UNTREATED CONTROL

(Continued)

WEEKS ON
STUDY

CARCASS
ID

INTEGUMENTAHY SYSTEM
Mammary gland

Adenocartinoma

Fibroadenoma

Fibroadenoma, multiple

Fibrosarcoma
Skin

Subcutaneous tissue, fibroma

Subcutaneous tissue, hemangiosarcoma

Subcutaneous tissue, leukemia
mononuclear

Subcutaneous tissue, sarcoma

MUSCULOSKELETAL SYSTEM
Bone

Skeletal muscle
Diaphragm, sarcoma stromal, metastatic,
uterus

NERVOUS SYSTEM

Brain

Spinal cord

RESPIRATORY SYSTEM
Lung

Alveoiar'bronchiolar adenoma

Alveolar/bronchiolar carcinoma

Leukemia mononuclear

Sarcoma stromal, metastatic, uterus
Nose
Trachea

SPECIAL SENSES SYSTEM
Ear
Eye

Hardenan gland
Zymbal gland
Carcinoma

URINARY SYSTEM
Kidney

Leukemia mononuclear

Sarcoma stromal, metastatic, uterus
Unnary bladder

Leukemia mononuclear

1111
0 0 0 0
5 5 5 5

1 1 1
0 0 0

5 5 5

^ — 5 3-
8 1 1
1 3 4

T — 3 — 3 — 3~
4 4 4 5
3 4 5 2

1 3 3"

5 5 5
3 4 5

-3 3 3 3 3-

6 6 6 6 7
2 3 4 5 5

"T 3 3 3~

8 8 9 9
4 5 4 5

TOTAL:
TISSUES
TUMORS

+ +
X

X

+ 4- + + + -*- + + + + + +

X
X X XX XX X

X

+ + + +

+ +

■t- + +

+ + + + -(- + + 4- +

+ +
+ +

49
6

23
5
1

50
1
1

1

1

50
1

50

1

50
2
1

12
1

50

50

11
1
1
1
1

50
3
1

50

1

117

Nitrofurantoin, NTP TR 341

TABLE B2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE RATS IN THE TWO YEAR FEED

STUDY OF NITROFURANTOIN: LOW DOSE

WEEKS ON
STUDY

CARCASS
ID

ALIMENTARY SYSTEM
Esophapis
Intestine large
Intestine large, cecum
Intestine large, colon
Intestine large, rectum
Intestine small
Intestine small, duodenum
Intestine small, ileum
Intestine small, jejunum
Liver

Leukemia mononuclear

Neoplastic nodule
Mesentery
Pancreas

Leukemia mononuclear

Acinus, adenoma
Salivary glands
Stomach

Stomach, forestomach
Stomach, glandular
Tongue

OOOOOOOOOOOOOOOOOIlllllIl
5666778888899999900000000
6I36I20116914556923344445

■3 — 5 — 5 — B — 5"
8 2 10 9
I 1 1 L I

CARDIOVASCULAR SYSTEM
Heart
Leukemia mononuclear

ENDOCRINE SYSTEM
Adrenal gland
Adrenal gland, cortex

Adenoma

Leukemia mononuclear
Adrenal gland, medulla

Leukemia mononuclear

Pheochromocytoma benign
Islets, pancreatic
Parathyroid gland

Adenoma
Pituitary gland

Leukemia mononuclear

Pars distalis, adenoma

Pars distalis. carcinoma
Thyroid gland

Leukemia mononuclear

C-cell. adenoma

C-cell, adenoma, multiple

C eel!, carcinoma

GENERAL BODY SYSTEM
Tissue. NOS

GENTTAL SYSTEM
Clitoral gland

Adenoma

Carcinoma
Ovary

Leukemia mononuclear
Uterus

Polyp stromal

Cervix, squamous cell carcinoma

Endometnum. sarcoma stromal

HEMATOPOIETIC SYSTEM
Blood

Leukemia mononuclear
Bone marrow

Leukemia mononuclear
Lymph node

Pancreatic, leukemia mononuclear
Lymph node, mandibular

Leukemia mononuclear
Lymph node, mesentenc

Leukemia mononuclear
Spleen

Leukemia mononuclear
Thymus

"5 — 5 — 5 — 5 — S — 5 — 5 — 5 — 5 5 5 5 5 5 B 5 5 5 5 5~
67125669473898053392
112212 32121233222343

+ M M +

M + + +

-t- + + M

+ + + M

+ M + M

+ + + M

A

A
A
A

A
A
A

A

+ -t- +

+ ^- +

+ + +

+ + +

■»- + +
+ -I- +

-t- +
X

+

+

+

+

+

+

+

+

+

+

+

+

+

•»-

+

+

+

■1-

+

-t-

+

■t-

X

X

X

X

+

+

+

+

+

^

+
X

+

+

X

+

+

+

+

+

M

+

+

+

+

+

^

+

+

-f

+

-f

-t-

+

■f

-(■

+

+

+

M

+

+

+

M

+

+

+
X

+■

-f
X

+- + -(- + -f-t- + + +A + + + +
■t- + -(- + + -(- + + + A + + +- +

X

X X

+ 4- + + + + + + +A-t- + -f- +

X

+ + +

+ A + + + +
+ + + + + +

X

+ -♦- + + +

+ + + + + + -*- + +

X

X XXX

XXX X

+ -f+' + + -(- + + + + + +

X

M M M M

M +- -t- + M
X

+ 4-
X

X X

+ M M

X

M

A +

M M

Nitrofurantoin, NTP TR 341

118

TABLE B2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE RATS: LOW DOSE

(Continued)

WEEKS ON

I

1

1

1

I

1

L

l

1

I

1

1

1

I

1

1

1

I

I

I

1

1

I

1

l

STUDY

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

TOTAL;
TISSUES

CARCASS

5

~s

5

S

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

_g_

6

(5

ID

3

5

6

7

7

I

I

1

2

2

3

4

4

4

4

5

5

6

7

8

8

9

0

0

0 TUMORS 1

4

3

4

3

4

3

4

5

4

5

5

2

3

4

5

4

5

5

5

4

5

5

3

4

5

ALIMENTARY SYSTEM

Esophagus

+

+

+

•^■

+

-(-

+

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

+

4-

4-

50

iDtestine large

+

+

■t-

+

+

+

+

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

+

4-

4-

4-

4-

4-

4-

4-

48

Intestine large, cecum

■t-

+

+

+

+

+

+

4-

4-

4-

4-

4-

4-

4-

4-

4-

+

4-

4-

4-

+■

4-

4-

4-

+

48

Intestine large, colon

-t-

+

■♦-

-(-

+

+

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

■t-

4-

4-

4-

4-

4-

4-

4-

46

Intestine large, rectum

+

+

+

■t-

+

+

+

4-

4-

4-

4-

4-

4-

4-

4-

■t-

4-

4-

4-

4-

-(-

4-

+

4-

4-

47

Intestine small

+

-t-

+

+■

+

+

-t-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

47

Intestine small, duodenum

+

+

+

+■

+

+

+

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

+

4-

4-

4-

4-

4-

4-

4-

47

Intestine small, ileum

+

+

+

+-

4-

+

+

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

+

4-

4-

4-

4-

46

Intestine small, jejunum

+

+

+

+-

+

+

+

4-

f

+

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

+

4-

4-

4-

47

Liver

+

+

+

+

+

■h

+

+

+■

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

+

4-

50

Leukemia mononuclear

X

X

X

X

X

13

Neoplastic nodule

1

Mesentery

+

+■

+

4-

+

4-

4-

4-

9

Pancreas

+

+

+

+

-)■

■f

+

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

-t-

4-

+

+

4-

4-

4-

4-

48

Leukemia mononuclear

X

1

Acinus, adenoma

X

1

Salivary glands
Stomach

+

+

■(-

+-

+

+

+

4-

4-

4-

4-

4-

4-

4-

4-

4-

+

4-

4-

4-

4-

4-

4-

4-

4-

49

+

+■

+

+

+

■(-

+

4-

4-

4-

4-

+

4-

4-

4-

4-

M

4-

4-

4-

4-

4-

4-

4-

4-

48

Stomach, forestomach

+

+■

■t-

■(-

+

■1-

+

4-

4-

4-

4-

4-

4-

4-

4-

4-

M

+■

4-

4-

4-

4-

4-

4-

4-

48

Stomach, glandular

-(-

+

f

-f

+

+■

+

4-

4-

+

4-

4-

4-

4-

4-

4-

M

+

+

+

4-

+

4-

4-

4-

48

Tongue

1

CARDIOVASCULAR SYSTEM

Heart

+

+

+

f

+

+■

+

4-

4-

4-

4-

4-

4-

+

4-

4-

4-

4-

-(-

4-

4-

4-

4-

4-

4-

50

Leukemia mononuclear

2

ENDOCRINE SYSTEM

Adrenal gland

+

+

+

+■

+

+

4-

4-

4-

+

4-

4-

4-

4-

4-

4-

4-

4-

+

4-

4-

4-

4-

4-

4-

49

Adrenal gland, cortex

+

+

+

+

+

+■

+

4-

4-

4-

+.

4-

4-

4-

4-

4-

+

4-

4-

+

4-

4-

4-

4-

4-

49

Adenoma

X

2

Leukemia mononuclear

X

7

Adrenal gland, medulla

+

+

+

M

+

+

4-

4-

4-

+

4-

4-

4-

4-

4-

4-

4-

4-

4-

+

4-

4-

4-

4-

4-

48

Leukemia mononuclear

X

4

Pheochromocytoma benign

X

1

Islets, pancreatic

+

+

+

+

+

+

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

48

Parathyroid gland

+

+

+

+■

+

+

+

+■

4-

4-

4-

4-

+

4-

4-

4-

4-

4-

+

4-

+

+

4-

4-

4-

50

Adenoma

L

Pituitary gland

+

-f

+

■f

+

+

+

4-

+

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

+

4-

+

4-

4-

48

Leukemia mononuclear

X

3

Pars distalis. adenoma

X

X

X

X

X

X

X

X

X

X

16

Pars distalis, carcinoma

X

X

X

X

9

Thyroid gland

+

+

+

+

+

+

4-

4-

4-

4-

4-

4-

4-

4-

4-

+

4-

4-

4-

4-

4-

4-

4-

4-

4-

50

Leukemia mononuclear

X

1

C-cell, adenoma

2

C-cell. adenoma, multiple

1

C-cell. carcinoma

X

X

2

GENERAL BODY SYSTEM

Tissue. NOS

-t-

3

GENITAL SYSTEM

Chtoral gland

+

-t-

M

M

M

-1-

4-

1-

4-

4-

4-

4-

4-

4-

4-

4-

4-

M

4-

4-

4-

4-

4-

4-

4-

38

Adenoma

X

X

X

X

7

Carcinoma

X

X

X

3

Ovary

+

+

+

■t-

+

+-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

+

+

4-

+

4-

4-

4-

4-

50

Leukemia mononuclear

X

4

Uterus

+

+

+

+

+

+

4-

+

+

4-

4-

4-

4-

4-

4-

4-

4-

4-

+

4-

4-

4-

4-

4-

4-

50

Polyp stromal

X

X

X

X

X

X

X

X

X

16

Cervix, squamous cell carcinoma

X

1

Endometnum. sarcoma stromal

1

HEMATOPOIETIC SYSTEM

Blood

■*■

2

Leukemia mononuclear

1

Bone marrow

+

+

+

•1-

+

+

4-

4-

4-

4-

4-

4-

4-

4-

4-

+

4-

4-

4-

4-

4-

4-

4-

4-

4-

48

Leukemia mononuclear

X

X

4

Lymph node

+

+

+

+

+

f

4-

4-

+

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

+

4-

4-

4-

4-

4-

50

Pancreatic, leukemia mononuclear

1

Lymph node, mandibular

+

+■

+■

■t-

■t-

■»-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

4-

47

Leukemia mononuclear

3

Lymph node, mesentenc

+

-t-

+

-*-

-«-

+

4-

4-

4-

4-

4-

4-

4-

4-

M

+

+

4-

4-

4-

4-

4-

4-

4-

4-

48

Leukemia mononuclear

3

Spleen

+

-t-

+

+■

+

+■

4-

4-

4-

4-

4-

4-

+

+

4-

4-

4-

4-

4-

4-

f

4-

4-

4-

4-

49

Leukemia mononuclear

X

X

X

X

X

13

Thymus

+

+

+

¥

+

+

4-

M

M

M

+

+

M

4-

M

4-

M

M

+

4-

4-

4-

M

4-

4-

34

119

Nitrofurantoin, NTP TR 341

TABLE B2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE RATS: LOW DOSE

(Continued)

WEEKS ON

0

0

0

0

0

0

0

0

0

0

0

"0~

0

0

"0-

"6-

0

1

1

1

I

1

I

1

1

STUDY

5

6

6

6

7

7

8

8

8

8

8

9

9

9

9

9

9

0

0

0

0

0

0

0

0

6

1

3

6

1

2

0

1

1

6

9

I

4

5

5

6

9

2

3

3

4

4

4

4

5

CARCASS

b

«

s

"5

~^~

=)

"S~

~5~

"T~"

b

"3"

~T~

T"

~r~

5

5

6

5

5

5

5

5

ID

e

2

1

0

9

6

7

1

2

5

6

6

9

4

7

3

8

9

8

0

5

3

3

9

2

1

1

1

1

1

1

I

2

2

1

2

3

2

1

2

1

2

3

3

2

2

2

3

4

3

INTEGUMENTARY SYSTEM

Mammary gland

+

+

+

+

+

+

+

+

+

+

+

■t-

■*-

■f

■*-

+

+

-4-

■(-

+

+

+

+

+

+

Adenocamnoma

X

Adenocarcinoma, multiple

X

Fibroadenoma

X

X

X

X

X

X

X

X

X

X

Fibroadenoma, multiple

X

X

Skin

+

t-

+

M

f

+

+

+

+

+

+

+

■♦-

+

+

+

■(-

+-

+

+

+

+

+

+

+

Foot, papilloma squamous

Subcutaneous tissue, fibroma

X

Subcutaneous tissue, fibrosarcoma

MUSCULOSKELETAL SYSTEM

Bone

+

+-

+

+

+

+

+

+

+

+

+

+

+

+

-f

+

+

+-

+

■♦-

+

+

+-

+

+

NEHVOUS SYSTEM

Brain

-)■

■(-

-♦-

+

+

+

-f

+

+

+

+

+

+

+

+

+

+

+■

+

-(-

+

+

+

+

+

Astrocytoma malignant

X

Carcinoma, metastatic, pituitary gland

Leukemia mononuclear

X

X

X

Spinal cord

+■

RESPnUTORY SYSTEM

Lung

■t-

+

+

+-

+

+

+

+

+

+

+

+

+

+

+

+

■t-

+

+

■t-

+

+

+

+

+

AJveolar/bronchiolar adenoma

X

Leukemia mononuclear

X

X

X

X

X

X

X

X

Nose

M

■t-

-f

+

+

*-

■1-

f

+

■t-

+

+

+

-+-

+■

+

+

+■

+

+

+

4-

-(-

+

+

Trachea

-)-

+

-4-

+

+

+■

+

■1-

+

+

+

+

+

+

+

■1-

+

+

+

+

■(-

+

■t-

+

■1-

SPECIAL SENSES SYSTEM

Ear

+

Eye

+

+

+

+

+

+

+

Hardenan gland

+

+

+

-t-

+

Zymbal gland

+

Carcinoma

X

UTUNARY SYSTEM

Kidney

+

■f-

+

-*-

f

+

+■

+

+

+

+

+

+

+-

-(-

+■

+

+

+

+

+

+

+

+

+

Leukemia mononuclear

X

X

X

X

Unnary bladder

+

+

+

+

+■

+

+

+

+

+

+

+

+

+

+

+

+

-(-

+

+

+

+

■1-

+

+

Nitrofurantoin, NTP TR 341

120

TABLE B2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE RATS: LOW DOSE

(Continued)

WEEKS ON
STUDY

1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 I 1 1 1 1 1 1 1 1
OOOOOOOOOOOOOOOOOOOOOOOOO
5555555555555555555555555

TOTAL:
TISSUES
TUMORS

CARCASS
ID

5555555555555555555555Bee
3567711122344445567889000
4343434545523454555455345

INTEGUMENTARY SYSTEM
Mammary gland

Adenocarcinoma

Adenocaranoma, multiple

Fibroadenoma

Fibroadenoma, multiple
Skin

Foot, papilloma squamous

Subcutaneous tissue, fibroma

Subcutaneous tissue, fibrosarcoma

+ +- + -t- + + + + -*- + -l--f + 4- + + + + + + -+- + + -t- +

XX X

XX X X X X
XXX XX X X X

+- + + +--t- + -(-+--t- + -«- + + + 4- + + + + + + 4- + -*- +

X
X

X

50

4

1

16

10

49

1

2

1

MUSCULOSKELETAL SYSTEM
Bone

+ + -*--*--l-+- + ^- + + -t- + -t- + -*--t- + + + + + -l- + -(--t-

50

NERVOUS SYSTEM
Brain

Astrocytoma malignant

Carcinoma, metastatic, pituitary gland

Leukemia mononuclear
Spinal cord

+ + + + -t- + -t- + + l- + + + -t- + + + + -f+--t--(- + + +
X

50

1
1
3
1

RESPIRATORY SYSTEM

Lung
Alveolar/bronchiolar adenoma
Leukemia mononuclear

Nose

Trachea

+ + + + -t--f-4- + -*-*- + +--f + -l- + + -+--t- + + + 4-f +

X

X XX X

+ -t-+- + -t- + -t- + + *--f + -t- + 4- + -(- + + + + + + -l- +
+ + + + -»--+- + + -*- + + + 4- + -t- + -(- + + + -l- + + -(- +

50
2
12
49
50

SPECIAL SENSES SYSTEM
Ear
Eye

Hardenan gland
Zymbal gland
Carcinoma

+ + + ■+--»--*- *- + + + + + + + + + +
+ 4- + 4-4- -;■ + + + + ■*- +

+ + + +

18
19
9

1
1

URINAEY SYSTEM
Kidney

Leukemia mononuclear
Unnary bladder

+ -t- + -»- + + 4->->--(--t--t--(-+-4- + + + -»--t--(--l- + -t--t-
4--t- + + + + -(-4- + + + + + -f4- + + + + -f4- + + + +

50

4

50

121

Nitrofurantoin, NTP TR 341

TABLE B2.

INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE RATS IN THE TWO-YEAR FEED
STUDY OF NITROFURANTOIN: HIGH DOSE

WEEKS ON
STUDY

OOOOOOOOOOOOOOOllllIlllll
5 566778899999990000000000
7944140603777890001555555

CARCASS
ID

4444444454544445444444444
177935230205946026433411 1
112 11 L121222 2 11332 2 343234

ALIMENTARY SYSTEM
Esophagus
Intestine lar^
Intestine large, cecum
Intestine large, colon
Intestine large, rectum
Intestine small
Intestine small, ducxienum
Intestine small, ileum
Intestine small, jejunum
Liver

Leukemia mononuclear

Osteosarcoma, metastatic, bone
Mesentery

Leukemia mononuclear
Pancreas

Leukemia mononuclear
Pharynx
Salivary glands

Adenocarcinoma
Stomach

Stomach, forestomach
Stomach, glandular

+ + + + -t- + + -t- + + + -t- + + + + + + + -t--f-f + + +
+ -*- + ■»- + + + + + + +A + + + + +A + + -t--t- + -t- +
+ + -4- + + + + -t- + + +A + + + + + A + + + -t--»--+- +
+ -t--*- + -t-i- + + + -f-»-A + 4- + + -t-A ■♦- + -t- + -*- + 4-
+ + M + + ■t- + + + -f-t-A + +- + + -t-A + -*- + + + + -*-
+- + -*-+--(-4-i- + + + +A-t- + + 4 + A + + -t- + + + +
+ + -t- + 4- + + + + +-+A ■t- + + + + A ■(- + + + +■ + +
+ 4 + + + -i- + -t- + -t-+A+-+--t- + +A ■(-+-+■ + + + +
+ + 4-t- + f-(-4 + -t-4-A+->-(-+-+A +- + + -f + -t--t-

+ + -<--(- + + + -(- + + + -ff-t--(- + + + + + + + + + V

XX X XX X X

X

+ -f- -t- -(-■)-

X

+ -t--t- + + + + + -f--*--t-A4 + + + + -t- + + + f + +--)-
X X
+
+ + + -t- + + -f + + + + + + + + + + + + -t--l- + + + -t-
X

+ + -*- + 4--t- + + + + -(--(- + + + + + + + + + + + + -f

+ -»- + + +--t-f-t- + f + 4 + + -l--f + + + + + -f+ f-l-
+ -f-t- + + -(- + + -f + + + + -*- + + + -t- + + + + + + -(-

CAIIDIOVASCULAR SYSTEM
Heart
Leukemia mononuclear

+ + + + -t--f-»--*- + -(--»--t- + + + + + + -t- + + + + + +
X X

ENDOCRINE SYSTEM
Adrenal gland
Adrenal gland, cortex

Adenoma

Leukemia mononuclear
Adrenal gland, medulla

Leukemia mononuclear

Pheochroraocytoma malignant

Pheochromocytoma benign
Islets, pancreatic
Parathyroid gland
Pituitary gland

Leukemia mononuclear

Pars distalis, adenoma

Pars distalis. carcinoma
Thyroid gland

C-cell, adenoma

C cell, adenoma, multiple

C ceil, cannnoma

+ + + -t- + -t- + + + + + + + + + + + + + +- + -f-l-+- +
-*- + + +- + + -(-■*- + + + + + + + + + + + + + + -*- + -*-

X
X XXX
+ + -t- + -(- + + -(- + + -f-t--t- + -f + + -*- + + + +- + -f-t-
X XXX
X
X
+ + + + -»--*- + + -t- + +A + + + + + + + -»- + -t- + -*-+-
+ + -t-+M + -*- + -t--»-+- + + -(- + + + + + + + + + + +
+ M-t-+- + + -(- + + + + +--(- + -t- + + A + + + + + + ■+-

X
X XX XXX XX XX X
X

■(- + + + -t--^4--t- + + -t- + -t- + 4+- + + + -t- + + + + +

GENERAL SODY SYSTEM

Tissue. NOB
Osteosarcoma, metastatic, bone

X

GENITAL SYSTEM
Chtoral gland

Adenoma
Ovary

Leukemia mononuclear
Uterus

Leiomyosarcoma

Leukemia mononuclear

Polyp stromal

Cervix, sarcoma stromal

Endometnum, adenoma

MMMM-*--*- + + + + + + + + -t- + + +M4M-i--t- + -t-

X
+ + ■♦- ■t--f+--i--t--(- + + -t--f + + + +- + -(- + -f-f + + -*-

X
+ -(--t--*--t->-+--*-f + -t--(--f + -»- + + +- + + -<--»--t- + +

X

XX X X
X

FIEMATOPOIETTC SYSTEM
Blood

Leukemia mononuclear
Bone marrow

Leukemia mononuclear
Lymph node

Inguinal, leukemia mononuclear

Mediastinal, leukemia mononuclear

Pancreatic, leukemia mononuclear
Lymph node, mandibular

Leukemia mononuclear
Lymph node, mesentenc

Leukemia mononuclear
Spleen

Leukemia mononuclear
Thymus

Leukemia mononuclear

+ -(■*■ + +■*-■•-
XX XX X

+ + -t- + -t--»--l- + + + + + -(- + + + -t--»- + + + + -f + +

X X X X X
+ + + + + + + + + + +-4-(--t--f + >--+- + 4 + -f-f + +

X
X X
X
+ + +- + -f + -l- + + + + + + + + + + + + + + + + + +

X X XXX
+ + + + + + + + +■ + +! + + + + f + + + + f + -*- +
X X

+ -F-»--)--l-f + + -(- + + + + + + + 4- + -t--»- + + + -)--«-

XX X XX X X

+ + + -i- + -t- + + + + + -F + + + +MM-*--*--t-M't--t-M

X

Nitrofurantoin, NTP TR 341

122

TABLE B2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE RATS: HIGH DOSE

(Continued)

WEEKS ON
STUDY

CARCASS
ID

ALIMENTARY SYSTEM

Esopfaa^s
Intestine large
Intestine large, cecum
Intestine large, colon
Intestine large, rectum
Intestine small
Intestine small, duodenum
Intestine small, ileum
Intestine small, jejunum
Liver

Leukemia mononuclear

Osteosarcoma, metastatic, bone
Mesentery

Leukemia mononuclear
Pancreas

Leukemia mononuclear
Pharynx
Salivary glands

Adenocarcinoma
Stomach

Stomach, forestomach
Stomach, glandular

CARDIOVASCULAR SYSTEM

Heart
Leukemia mononuclear

ENDOCRINE SYSTEM

Adrenal gland
Adrenal gland, cortex

Adenoma

Leukemia mononuclear
Adrenal gland, medulla

Leukemia mononuclear

Pheochromocytoma malignant

Pheochromocytoma benign
Islets, pancreatic
Parathyroid gland
Pituitary gland

Leukemia mononuclear

Pars distalis. adenoma

Pars distalis, carcinoma
Thyroid gland

C-cell. adenoma

C ceil, adenoma, multiple

C-cell, carcinoma

GENERAL BODY SYSTEM

Tissue. NOS
Osteosarcoma, metastatic, bone

GENITAL SYSTEM

Clitorai gland

Adenoma
Ovary

Leukemia mononuclear
Uterus

Leiomyosarcoma

Leukemia mononuclear

Polyp stromal

Cervix, sarcoma stromal

Endometrium, adenoma

HEMATOPOIETIC SYSTEM

Blood

Leukemia mononuclear
Bone marrow

Leukemia mononuclear
Lymph node

Inguinal, leukemia mononuclear

Mediastinal, leukemia mononuclear

Pancreatic, leukemia mononuclear
Lymph node, mandibular

Leukemia mononuclear
Lymph node, mesenteric

Leukemia mononuclear
Spleen

Leukemia mononuclear
Thymus

Leukemia mononuclear

1

1

I

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

I

1

1

0

0

0

0

U

0

0

0

0

0

0

0

0

0

0

n

0

0

n

n

0

0

0

0

0

b

i>

b

b

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

TOTAL;

4

4

4

4

4

4

4

4

4

4

4

4

4

4

4

4

4

4

4

4

4

4

4

S

S

TISSUES

1

2

•i

3

4

4

5

5

5

6

6

6

7

7

7

8

8

8

8

8

9

9

9

0

0

TUMORS

5

4

5

5

4

5

3

4

5

3

4

5

3

4

5

1

2

3

4

5

3

4

5

4

5

■»-

+

+

^

^.

-t-

+

+

+

-*-

+

+

+

+

-(-

+

+

■t-

+

^

+

^

+

+

+

50

+-

+

+

-*-

■t-

+

+

+■

4-

-t-

-t-

■1-

+

+

-*-

+

+

■t-

+

4-

+

4-

4-

+

4-

48

+

+

+

+

^-

+

+

+

+-

+

+

+

+

■»■

+

+

+

■(-

4-

4-

+

+

+

+

+

48

+

+

+

+

+

+

-1-

+

-t-

+

+■

+

+

+

■t-

+

■f

4-

4-

+

+

+

+

+

+

48

+

+

+

+

-(-

-(-

+

+

+

-t-

+■

-1-

+

+

+

+

+

+

4-

+

+

+

+

+

+

47

■♦-

+

+

+

-(-

+-

+

+

+-

-t-

■¥

+

+

■»■

■»■

+

+

+-

-t-

+

4-

4-

4-

+

4-

48

+

■t-

■«-

+

-(-

■(-

-(-

+

+

+

+

+

+

+

+

+

-t-

+

+

+

+

+

■(-

+

+

48

+

-(-

+

+

■(-

+-

+

■t-

+

-t-

+

+■

-(-

+

+

4-

4-

4-

4-

+

+

4-

+

4-

4-

48

-*-

+

+

■f

+■

-t-

+

+

■»■

+

+

+■

+

+

+

+

■t-

+-

+

+

+

+

+

+

+

48

-t-

+

X

+

+

+

+

-»-

+

+

-t-

X

+

+

■t-

+

4-

4-

4-

+

+

+

4-

4-

4-

4-

50
9

■*-

+

+

+

4-

10

■»-

+

+

-t-

+

+

+

+

+

+■

+

■I-

+

+

+

+

4-

4-

+

+

+

+

-1-

4-

+

49
2

■»-

-1-

+

+

-t-

■1-

+

+

+

+

+

+

+

+

+

4-

4-

4-

4-

+

+

+

4-

+

+

1
50

+

-)-

+

+

+

-(-

■1-

+

4-

-t-

+

+

+

+

+

+

+

+

4-

+

+

+

+

+

-(-

50

+

+

f

f

f

¥

-(-

+

+

■f

+

+

+

+

+

+

4-

4-

4-

■t-

+

+

4-

+

+

50

+

+

f

-t-

-t-

+-

+

■»-

+

+

+

+

•¥

-1-

+

+

+

+

4-

-(-

-(-

4-

+

4-

4-

50

+ 4-
+ +

+ +

+ +

+ +

+ +

X

+ +

+ +
X

+ + +
X X

+ + +

4-

+

-*-

4-

+

M

4-

X

M

4-

4-

4-

4-

4-

X

4-

4-

4-

4-

-*-

+

4-

+
X

+

4-

4-

4-

4-

+

-^

+

+

4-

4-

-*■

4-

4-

4-

4-

4-

+

4-

4-

4-

4-

+

4-

4-

■t-

4-

4-

4-

4-

+

+

4-

+

4-

4-

4-

4-

+

4-

4-

+

+

X

4-

4-

4-

+

4-

4-

+

4-

4-

4-

4-

X

X

X

X

X

X

X

M

4-

4-

4-

4-

4-

4-

4-

+

4-

4-

+

4-

f

4-

4-

+

4-

4-

4-

4-

+

4-

4-

4-

+

4-

M

+

4-

4-

+ + + -*-

50

50

1

4

50

4

I

4

49

47

48

I

21

42
4

50
1

50
I
1

10
1

50
1
2
1

50
5

49
3

50
9

40
I

123

Nitrofurantoin, NTP TR 341

TABLE B2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE RATS: HIGH DOSE

(Continued)

WfiEKS OK
STUDY

OOOOOOOOOOOOOOOllllllllll
5566778899999990000000000
7944140603777890001555555

CARCASS
ID

4444444454544445444444444
1779352302059460 2 64334111
112 11112 12 2 22113322343234

INTEGUMENTARY SYSTEM
Mammary gland

Fibroadenoma

Fibroadenoma, multiple
Skin

Basal cell cannnoma

Keratoacanthoma

Papilloma squamous

+ + + + + + + + + + + + + -4- + + + -»- + -f + -t- + -f +

X XX XXXX XXX
X XXX

+ + + + + + + + -t- + + +- + + -t- + + -t--(- + + + + -t- +

X
X

MUSCULOSKELETAL SYSTEM
Bone

+ +- + + -(- + + + + f+--(-+--»--t-+- + + + -(-4--f + -(-+-

NERVOUS SYSTEM
Brain
Carcinoma, metastatic, pituitary gland

+ + -f + + + ■f + ■^--^-■♦--t-■^-(- + + + ■^- + ■t- + + + + +

X

RESPIRATORY SYSTEM

Lung
Alveolar/bronchiolar adenoma
Carcinoma, metastatic, thyroid gland
Leukemia mononuclear

Nose

Trachea

■t- + + + + + + -t-4-+- + + + -t- + -t--t--t- + 4--t-4- + -(- +

X

XX X XX X X

+-MMM+-t--t- + -*- + +- + -(-4- + + +--t--(-+- + + +-+- +

+ + -(- + + + + 4- + + + + -(- + + + + + + 4- + + -t--t- +

SPECIAL SENSES SYSTEM

Ear

Eye

+ 4-

+ -t- +

URINARY SYSTEM

Kidney
Leukemia mononuclear
Renal tubule, adenoma

Unnary bladder
Leukemia mononuclear

+ + 4--t- + + + + + + + + + + +- + + + + + + + -t- + +

X X

+ + + + + + + + + + + + + + -t--»- + -*--(- + + + + ^ +

X

Nitrofurantoin, NTP TR 341

124

TABLE B2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE RATS: HIGH DOSE

(Continued)

WEEKS ON
STUDY

lllllllllllllllltllllllll
0000000000000000000000000
5555555555555555555555555

TOTAL:
TISSUES
TUMORS

CARCASS
ID

4444444444444444444444455

1223445556667778888899900
5455453453453451234534545

INTEGUMENTARY SYSTEM
Mammary gland

Fibroadenoma

Fibroadenoma, multiple
Skin

Basal cellcamnoma

Keratoacanthoma

Papilloma squamous

+ -+- + +- + + ■♦- + ^ + + + + 4- + + + + + + -+- + + + +

X X X X X X

X X X X X X X XXX X

-t-4-+- + ++- + -t- + + + + + + + 4--f+- + + + + + + +

X

50
16
15
50

1
I
1

MUSCULOSKELETAL SYSTEM
Bone

+ +- + ♦- + + ■»- + + + + + -(- + ■♦- + ■»--(-■♦- + + + -+- + +

50

NERVOUS SYSTEM
Brain
Carcinoma, metastatic, pituitary gland

-*-■+--♦--»-+■ + + + + -(■ + + + + ■♦- + ■*--(- + + + + -+- + +

50

1

RESPIRATORY SYSTEM

Lung
Alveoiar/bronchiolar adenoma
Carcinoma, metastatic, thyroid gland
Leukemia mononuclear

Nose

Trachea

+ + + + + -t--(-4- + + + + + -t- + 4- + -t- + 4- + + + -t- +

X

X

X

-t-^-»- + + +-+--t- + + + + -t'-t- + + + + -t- + +- + + -t- +

4--+- + +- + 4- + + + + + + + 4--f+- + >- + + + + + + +

50

2

I

8

47

50

SPECIAL SENSES SYSTEM

Ear

Eye

+ + +

5
6

URDJARY SYSTEM

Kidney
Leukemia mononuclear
Renal tubule, adenoma

Unnary bladder
Leukemia mononuclear

+ + + + + 4- + + + + + -»- + 4- + + + + + + -t--t- + + +

X

+ + -t- + -t-+--+- + + -h + + + + -t- + + 4- + + + + + + +

50
2

1

50

1

125

Nitrofurantoin, NTP TR 341

TABLE B3.

ANALYSIS OF PRIMARY TUMORS IN FEMALE RATS IN THE TWO-YEAR FEED STUDY

OF NITROFURANTOIN

Control

600 ppm

1,300 ppm

Adrenal Gland: Pheochromocytoma

Overall Rates (a)
Adjusted Rates (c)
Terminal Rates (d)
Day of First Observation
Life Table Tests (e)
Logistic Regression Tests (e)
Cochran-Armitage Trend Test (e)
Fisher Exact Test (e)

Clitoral Gland: Adenoma

Overall Rates (a)
Adjusted Rates (c)
Terminal Rates (d)
Day of First Observation
Life Table Tests (e)
Logistic Regression Tests (e)
Cochran-Armitage Trend Test (e)
Fisher Exact Test (e)

Clitoral Gland: Carcinoma

Overall Rates (a)

Adjusted Rates (c)

Terminal Rates (d)

Day of First Observation

Life Table Tests (e)

Logistic Regression Tests (e)

Cochran-Armitage Trend Test (e)

FisherExactTest(e)

Clitoral Gland: Adenoma or Carcinoma

Overall Rates (a)
Adjusted Rates (c)
Terminal Rates (d)
Day of First Observation
Life Table Tests (e)
Logistic Regression Tests (e)
Cochran-Armitage Trend Test le)
Fisher Exact Test (e)

Lung: Alveolar/Bronchiolar Adenoma or Carcinoma

Overall Rates (a)
Adjusted Rates (c)
Terminal Rates (d)
Day of First Observation
Life Table Tests (e)
Logistic Regression Tests (e)
Cochran-Armitage Trend Test (e)
Fisher ExactTest(e)

Mammary Gland: Fibroadenoma

Overall Rates (a)
Adjusted Rates (c)
Terminal Rates (d)
Day of First Observation
Life Table Tests (e)
Logistic Regression Tests (e)
Cochran-Armitage Trend Test (e)
Fisher Exact Test(e)

2/48 < 4% 1

1/48(2%)

(b) 4/50 (8%)

7.6%

4.0%

11.6%

1/25(4%)

1/25(4%)

3/31(10%)

729

730

491

P = 0.297

P = 0.504N

P = 0.413

P = 0.235

P = 0.520N

P = 0.353

P = 0.244

P = 0.500N

P = 0.359

1/4412%)

7/38(18%)

4/42(10%)

2.4%

26.8%

13.3%

0/23(0%)

5/22(23%)

3/28(11%)

658

621

700

P = 0.270

P = 0.027

P = 0.212

P = 0.189

P = 0.018

P = 0.163

P = 0.191

P = 0.017

P = 0.166

4/44(9%)

3/38(8%)

0/42(0%)

14.1%

13.6%

0.0%

2/23(9%)

3/22(14%)

0/28 (0%)

633

730

P = 0.036N

P = 0.532N

P = 0.052N

P = 0.057N

P = 0.607N

P = 0.069N

P = 0.055N

P = 0.583N

P = 0.064N

5/44(11%)

10/38(26%)

4/42(10%)

16.1%

39.7%

13.3%

2/23(9%)

8/22(36%)

3/28(11%)

633

621

700

P = 0.315N

P = 0.107

P = 0.436N

p = 0.449N

P = 0.063

P = 0.532N

P = 0.443 N

P = 0.072

P = 0.530N

>ma

3/50(6%)

2/50(4%)

2/50(4%)

11.1%

6.4%

5.8%

2/25(8%)

1/26(4%)

1/31(3%)

727

665

686

P = 0.365N

P = 0.499N

P = 0.439N

P = 0.432N

P = 0.534N

P = 0.512N

P = 0.418N

P = 0.500N

P = 0.500N

28/50(56%)

26/50(52%)

31/50(62%)

70.4%

69.0%

79.2%

14/25(56%)

15/26(58%)

23/31(74%)

428

456

448

P = 0.503N

P = 0.449N

P = 0.517N

P = 0.230

P = 0.507N

P = 0.262

P = 0.297

P = 0.421N

P = 0.342

Nitrofurantoin, NTP TR 341

126

TABLE B3. ANALYSIS OF PRIMARY TUMORS IN FEMALE RATS IN THE TWO-YEAR FEED STUDY

OF NITROFURANTOIN (Continued)

Control

600 ppm

1,300 ppm

Mammary Gland: Adenocarcinoma

Overall Rates (a)

6/50(12%)

5/50(10%)

0/50(0%)

Adjusted Rates (c)

19.4%

17.2%

0.0%

Terminal Rates (d)

3/25(12%)

3/26(12%)

0/31(0%)

Day of First Observation

661

687

Life Table Tests (e)

P = 0.013N

P = 0.497 N

P = 0.015N

Logistic Regression Tests (e)

P = 0.018N

P = 0.553N

P = 0.019N

Cochran-Armitage Trend Test (e)

P = 0.016N

Fisher Exact Test (e)

P = 0.500N

P = 0.013N

Pituitary Gland/Pars Distalis: Adenoma

Overall Rates (a)

23/50(46%)

16/48(33%)

21/48(44%)

Adjusted Rates (c)

59.1%

50.4%

51.7%

Terminal Rates (d)

11/25(44%)

11/26(42%)

12/31(39%)

Day of First Observation

568

567

448

Life Table Tests (e)

P = 0.264N

P = 0.148N

P = 0.293N

Logistic Regression Tests (e)

P = 0.492N

P = 0.163N

P = 0.501N

Cochran-Armitage Trend Test (e)

P = 0.471N

Fisher Exact Test (e)

P = 0.141N

P = 0.492N

Pituitary Gland/Pars Distalis: Carcinoma

Overall Rates (a)

3/50 (6%)

9/48(19%)

2/48(4%)

Adjusted Rates (c)

9.7%

26.9%

5.6%

Terminal Rates (d)

1/25(4%)

4/26(15%)

1/31 (3%)

Day of First Observation

700

564

675

Life Table Tests (e)

P = 0.342N

P = 0.069

P = 0.462N

Logistic Regression Tests (e)

P = 0.426N

P = 0.048

P = 0.527N

Cochran-Armitage Trend Test (e)

P = 0.417N

Fisher Exact Test (e)

P = 0.052

P = 0.520N

Pituitary Gland/Pars Distalis: Adenoma or

Carcinoma

Overall Rates (a)

26/50(52%)

25/48(52%)

23/48(48%)

Adjusted Rates (c)

64.3%

68.6%

55.4%

Terminal Rates (d)

12/25(48%)

15/26(58%)

13/31(42%)

Day of First Observation

568

564

448

Life Table Tests (e>

P = 0.192N

P = 0.524N

P = 0.236N

Logistic Regression Tests (e)

P = 0.407N

P = 0.524

P = 0.435N

Cochran-Armitage Trend Test (e)

P = 0.381N

Fisher Exact Test (e)

P = 0.577

P = 0.420N

Skin: Fibroma or Fibrosarcoma

Overall Rates (a)

1/50(2%)

3/50(6%)

0/50(0%)

Adjusted Rates (c)

4.0%

10.1%

0.0%

Terminal Rates (d)

1/25(4%)

2/26(8%)

0/31 (0%)

Day of First Observation

730

653

Life Table Tests (e)

P = 0.313N

P = 0.307

P = 0.457N

Logistic Regression Tests (e)

P = 0.368N

P = 0.280

P = 0.457N

Cochran-Armitage Trend Test (e) ,

P = 0.357N

Fisher Exact Test (e)

P = 0.301

P = 0.500N

Skin: Fibroma, Sarcoma, or Fibrosarcoma

Overall Rates (a)

2/50(4%)

3/50(6%)

0/50 (0%)

Adjusted Rates (c)

6.9%

10.1%

0.0%

Terminal Rates (d)

1/25(4%)

2/26(8%)

0/31(0%)

Day of First Observation

709

653

Life Table Tests (e)

P = 0.169N

P = 0.502

P = 0.223N

Logistic Regression Tests (e)

P = 0.202N

P = 0.469

P = 0.243N

Cochran-Armitage Trend Test (e)

P = 0.192N

Fisher Exact Test (e)

P = 0.500

P = 0.247N

127

Nitrofurantoin, NTP TR 341

TABLE B3. ANALYSIS OF PRIMARY TUMORS IN FEMALE RATS IN THE TWO-YEAR FEED STUDY

OF NITROFURANTOIN (Continued)

Control

600 ppm

1,300 ppm

Thyroid Gland: C-Cell Adenoma

Overall Rates (a)

3/50(6%)

3/50(6%)

4/50(8%)

Adjusted Rates (c)

n.4%

10.2%

12.9%

Terminal Rates (d)

2/25(8%)

0/26(0%)

4/31(13%)

Day of First Observation

729

726

730

Life Table Tests (e)

P = 0.514

P = 0.656N

P = 0.611

Logistic Regression Tests (e)

P = 0.434

P = 0.656

P = 0.547

Cochran-Armitage Trend Test (e)

P = 0.423

Fisher Exact Test (e)

P = 0.661N

P = 0.500

Thyroid Gland: C-Cell Carcinoma

Overall Rates (a)

4/50(8%)

2/50(4%)

2/50(4%)

Adjusted Rates (c)

13.8%

7.7%

6.5%

Terminal Rates (d I

3/25(12%)

2/26(8%)

2/31(6%)

Day of First Observation

481

730

730

Life Table Tests (e)

P = 0.199N

P = 0.326N

P = 0.262N

Logistic Regression Tests { e )

P = 0.276N

P = 0.344N

P = 0.338N

Cochran-Armitage Trend Test (e)

P = 0.265N

Fisher Exact Test (e)

P = 0.339N

P = 0.339N

Thyroid Gland: C-Cell Adenoma or Carcinoma

Overall Rates (a)

7/50(14%)

5/50(10%)

6/50(12%)

Adjusted Rates (c)

24.5%

17.1%

19.4%

Terminal Rates (d)

5/25(20%)

2/26(8%)

6/31(19%)

Day of First Observation

481

726

730

Life Table Tests (e)

P = 0.329N

P = 0.372N

P = 0.356N

Logistic Regression Tests (e)

P = 0.476N

P = 0.425N

P = 0.527N

Cochran-Armitage Trend Test (e)

P = 0.453N

Fisher Exact Test(e)

P = 0.380N

P = 0.500N

Uterus: Endometrial Stromal Polyp

Overall Rates (a)

10/50(20%)

16/50(32%)

10/50(20%)

Adjusted Rates (c)

32.8%

47.2%

30.2%

Terminal Rates (dl

6/25(24%)

9/26(35%)

8/31(26%)

Day of First Observation

633

601

700

Life Table Tests (e)

P = 0.377N

P = 0.142

P = 0.456N

Logistic Regression Tests (e)

P = 0.543

P = 0.081

P = 0.574

Cochran-Armitage Trend Test (e)

P = 0.522N

Fisher Exact Test(e)

P = 0.127

P = 0.598N

Hematopoietic System: Mononuclear Leukemia

Overall Rates (a)

13/50(26%)

13/50(26%)

9/50(18%)

Adjusted Rates (c)

32.6%

33.7%

21.9%

Terminal Rates (d)

2/25(8%)

5/26(19%)

2/31(6%)

Day of First Observation

481

390

513

Life Table Tests (e)

P = 0.201N

P = 0.545

P = 0.243N

Logistic Regression Tests (e)

P = 0.147N

P = 0.447N

P = 0.19lN

Cochran-Armitage Trend Test (e)

P = 0.200N

Fisher Exact Test (e)

P = 0.590N

P = 0.235N

(a) Number of tumor-bearing animals/number of animals examined at the site

(b) A malignant pheochromocytoma was observed in an animal with a benign pheochromocytoma.

(c) Kaplan-Meier estimated tumor incidences at the end of the study after adjusting for intercurrent mortality

(d) Observed tumor incidence at terminal kill

(e) Beneath the control incidence are the P values associated with the trend test. Beneath the dosed group incidence are the
P values corresponding to pairwise comparisons between that dosed group and the controls. The life table analysis regards tu-
mors in animals dying prior to terminal kill as being (directly or indirectly) the cause of death. The logistic regression test re-
gards these lesions as nonfatal. The Cochran-Armitage and Fisher exact tests compare directly the overall incidence rates.
A negative trend or lower incidence in a dosed group is indicated by ( N ).

Nitrofurantoin, NTP TR 341

128

TABLE B4a. HISTORICAL INCIDENCE OF CLITORAL GLAND TUMORS IN FEMALE F344/N RATS

RECEIVING NO TREATMENT (a)

Study

Adenoma

Incidence in Controls

Carcinoma

Adenoma or Carcinoma

Historical Incidence at Southern Research Institute

HC Blue No. 2
C.I. Disperse Blue 1
Eugenol

Stannous chloride
D-Mannitol
Ziram

Propyl gallate
Zearalenone
HC Blue No. 1

TOTAL

SD(b)

Range (c)
High
Low

0/50

0/50

0/50

1/49

2/49

3/49

0/40

1/40

1/40

0/50

0/50

0/50

1/50

0/50

1/50

2/50

3/50

5/50

2/50

0/50

2/50

0/50

1/50

1/50

1/50

3/50

4/50

7/439(1.6%)

10/439(2.3%)

17/439(3.9%)

1.67%

2.55%

3.51%

2/50

3/50

5/50

0/50

0/50

0/50

Overall Historical Incidence

TOTAL
SD(b)

(d)39/l,984(2.0%)
2.31%

(6)57/1,984(2.9%)
2.95%

(d,e) 96/1 ,984 (4.8%)
3.40%

Range (c)
High
Low

5/49
0/50

6/49
0/50

6/49
0/50

(a) Data as of August 7, 1986, for studies of at least 104 weeks

(b ) Standard deviation

(c) Range and SD are presented for groups of 35 or more animals.

(d) Includes one cystadenoma, NOS

(e) Includes five squamous cell carcinomas and five adenocarcinomas, NOS

129

Nitrofurantoin, NTP TR 341

TABLE B4b.

HISTORICAL INCIDENCE OF MAMMARY GLAND TUMORS IN FEMALE F344/N RATS

RECEIVING NO TREATMENT (a)

Incidence in

Controls

Study

Adenoma

Adenocarcinoma

Adenoma or Adenocarcinoma

Historical Inc

idence at Southern

Research Institute

HC Blue No. 2

1/50

1/50

2/50

C.I. Disperse Bl

ue 1

0/49

0/49

0/49

Eugenol

0/40

0/40

0/40

Stannous chloride

1/50

3/50

4/50

D-Mannitol

0/50

3/50

3/50

Ziram

0/50

3/50

3/50

Propyl gallate

0/50

1/50

1/50

Zearalenone

0/50

1/50

1/50

HC Blue No. 1

0/50

3/50

3/50

TOTAL

2/43910.5%)

15/439(3.4%)

17/439(3.9%)

SD(b)

0.88%

2.65%

2.91%

Range (c)

High

1/50

3/50

4/50

Low

0/50

0/49

0/49

Overall Historical

Incidence

TOTAL

13/1,984(0.7%)

(d) 51/1 ,984 (2.6%)

(d) 64/1,984 (3.2%)

SD(b)

1.32%

2.32%

2.81%

Range (c)

High

3/49

4/49

7/49

Low

0/50

0/50

0/50

(a) Data as of August 7, 1986, for studies of at least 104 weeks

(b) Standard deviation

(c) Range and SD are presented for groups of 35 or more animals.

(d) Includes one squamous cell carcinoma, six papillary adenocarcinomas, and two papillary cystadenocarcinomas, NOS

Nitrofurantoin, NTP TR 341 130

TABLE B5. SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS IN THE

TWO-YEAR FEED STUDY OF NITROFURANTOIN

Untreated Control

Low Dose

High Dose

Animals initially in study

Animals removed

Animals examined histopathologically

50
50
50

50
50
50

50
50
50

ALIMENTARY SYSTEM
Intestine large, cecum

Parasite metazoan

Submucosa, edema, diffuse

Submucosa, hemorrhage, multifocal

Submucosa, inflammation, diffuse
Intestine large, colon

Parasite metazoan

Mucosa, mineralization
Intestine large, rectum

Parasite metazoan
Liver

Angiectasis, focal

Basophilic focus, multiple

Clear cell focus

Cytologic alterations, focal

Developmental malformation

Focal cellular change

Hematopoietic cell proliferation

Hyperplasia, focal

Infiltration cellular, lymphocytic, multifocal

Inflammation, granulomatous, focal

Inflammation, granulomatous, multifocal

Necrosis, coagulative, diffuse

Necrosis, coagulative, focal

Vacuolization cytoplasmic, diffuse

Vacuolization cytoplasmic, focal

Vacuolization cytoplasmic, multifocal

Bile duct, hyperplasia

Centrilobular, congestion

Kupffer cell, pigmentation, bile
Mesentery

Hemorrhage, focal

Infiltration cellular, lymphocytic

Inflammation, granulomatous, focal

Fat, fibrosis, focal

Fat, necrosis, focal
Pancreas

Acinus, atrophy

Acinus, atrophy, focal

Acinus, atrophy, multifocal

Acinus, hyperplasia, focal

Duct, cyst
Pharynx

Hyperplasia

Infiammation, suppurative
Stomach

Inflammation, chronic
Stomach, forestomach

Cyst

Edema

Hyperkeratosis

Hyperplasia

Inflammation, chronic

Mineralization

Ulcer, focal

Epithelium, hyperplasia

(50)

1 (2%)
1 (2%)
1 (2%)

(501

6 (12%)

(50)

(50)

1

(2%)

10

(20%

1

(2%)

1

(2%)

2

(4%)

1

(2%)

2

(4%)

2

(4%)

2

(4%)

2 (4%)

3 (6%)
2 (4%)
1 (2%)

(8)

6 (75%)
(50)

5 (10%)
1 (2%)

1 (2%)

(50)

1 (2%)
(50)

3 (6%)

3 (6%)

(48)

1 (2%)

(46)

3 (7%)
1 (2%)

(47)

(50)

8 (16%)

4 (8%)
1 (2%)

5

(10%)

1

(2%)

5

(10%)

(2%)

(4%)

(14%)

(2%)

(2%)

(9)

(11%)

(11%)

7 (78%)

(48)

5 (10%)

2 (4%)

(48)

1 (2%)
(48)

1 (2%)
3 (6%)

2 (4%)

2 (4%)
1 (2%)
1 (2%)

3 (6%)

(48)

1 (2%)

(48)
2 (4%)

(47)

1 (2%)
(50)

10 (20%)

1 (2%)

1 (2%)

1 (2%)

1 (2%)

1 (2%)

2 (4%)
7 (14%)

2 (4%)
2 (4%)
1 (2%)
1 (2%)
4 (8%)

(10)

1 (10%)

1 (10%)
8 (80%)
(49)

1 (2%)
8 (16%)

1 (2%)

(1)
1 (100%)

1 (100%)
(50)

(50)

2 (4%)
1 (2%)

1 (2%)

1 (2%)
1 (2%)

131

Nitrofurantoin, NTP TR 341

TABLE B5. SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS IN THE
TWO-YEAR FEED STUDY OF NITROFURANTOIN (Continued)

Untreated Control

Low

Dose

High

Dose

ALIMENTARY SYSTEM (Continued)

Stomach, glandular

(50)

(48)

(50)

Dysplasia, focal

1

(2%)

Erosion

1

(2%)

2

(4%)

Mineralization

2

(4%)

3

(6%)

Necrosis, focal

1

(2%)

Ulcer

1

(2%)

Tongue

(1)

Inflammation

1

(100%)

CARDIOVASCULAR SYSTEM

Heart

(50)

(50)

(50)

Fibrosis, multifocal

33

(66%)

34

(68%)

25

(50%)

Inflammation, multifocal

1

(2%)

1

(2%)

1

(2%)

ENDOCRINE SYSTEM

Adrenal gland, cortex

(50)

(49)

(50)

Angiectasis

6

(12%)

5

(10%)

1

(2%)

Congestion

1

(2%)

Cyst

1

(2%)

Degeneration, fatty, focal

2

(4%)

5

(10%)

7

(14%)

Hemorrhage, diffuse

1

(2%)

Hemorrhage, focal

1

(2%)

Hyperplasia, focal

3

(6%)

3

(6%)

6

(12%)

Hypertrophy, focal

1

(2%)

Necrosis

2

(4%)

Vacuolization cytoplasmic, diffuse

1

(2%)

Vacuolization cytoplasmic, focal

4

(8%)

2

(4%)

Extra adrenal tissue, accessory adrenal

cortical nodule

1

(2%)

Adrenal gland, medulla

(48)

(48)

(50)

Angiectasis

1

(2%)

1

(2%)

1

(2%)

Cyst, multiple

1

(2%)

Hyperplasia, focal

2

(4%)

1

(2%)

Infiltration cellular, lymphocytic, multifocal

I

(2%)

Parathyroid gland

(49)

(50)

(47)

Hyperplasia

1

(2%)

Pituitary gland

(50)

(48)

(48)

Parsdistalis, angiectasis

26

(52%)

24

(50%)

26

(54%)

Pars distalis, cyst

8

(16%)

8

(17%)

4

(8%)

Pars distalis, cyst, multiple

5

(10%)

I

(2%)

3

(6%)

Parsdistalis, hemorrhage

1

(2%)

1

(2%)

Pars distalis, hyperplasia, focal

2

(4%)

7

(15%)

5

(10%)

Pars distalis, necrosis

1

(2%)

Pars distalis, pigmentation

1

(2%)

Thyroid gland

(50)

(50)

(50)

Ultimobranchial cyst

1

(2%)

3

(6%)

C-cell, hyperplasia, focal

5

(10%)

9

(18%)

4

(8%)

C-cell, hyperplasia, multifocal

3

(6%)

5

(10%)

2

(4%)

Follicle, cyst

1

(2%)

Follicle, hyperplasia, cystic

1

(2%)

GENERAL BODY SYSTEM

Tissue, NOS

(3)

(1)

Hemorrhage

1

(33%)

Nitrofurantoin, NTP TR 341

132

TABLE B5. SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS IN THE
TWO-YEAR FEED STUDY OF NITROFURANTOIN (Continued)

Untreated Control

Low Dose

High Dose

GENITAL SYSTEM
Clitoral gland

Inflammation, suppurative

Duct, cyst
Ovary

Cyst
Uterus

Hemorrhage, chronic, focal

Hydrometria

Cervix, abscess

Cervix, cyst

Cervix, cyst, multiple

Cervix, inflammation, suppurative

Endometrium, cyst

Endometrium, hyperplasia, cystic

Endometrium, intlammation, suppurative

Endometrium, necrosis, focal

Lumen, inflammation, suppurative

Muscularis, cyst

(44)

(38)

(42)

2 (5%)

5

(13%)

1

(2%)

12 (27%)

13

(34%)

7

(17%)

(50)

(50)

(50)

2 (4%)

1

(2%)

(50)

(50)

(50)

1 (2%)

2

(4%)

2

(4%)

4

(8%)

2

(4%)

2 (4%)

5

1

(10%)
(2%)

1

(2%)

1 (2%)

4

(8%)

3

(6%)

1 (2%)

5 (10%)

10

(20%)

10

(20%)

2

(4%)

1

(2%)

1

(2%)

1

(2%)

1 (2%)

HEMATOPOIETIC SYSTEM

Blood

(9)

Anemia

1

(11%

Anisocytosis

Bone marrow

(49)

Hyperplasia, reticulum cell

1

(2%)

Myelofibrosis

Myeloid cell, hyperplasia

Lymph node

(50)

Axillary, hyperplasia, lymphoid

Axillary, infiltration cellular, plasma cell

Axillary, infiltration cellular, histiocytic

Iliac, hyperplasia, lymphoid

Inguinal, hyperplasia, lymphoid

1

(2%)

Mediastinal, congestion

1

(2%)

Mediastinal, erythrophagocytosis

Mediastinal, hyperplasia, lymphoid

1

(2%)

Renal, sinus, ectasia

Lymph node, mandibular

(48)

Erythrophagocytosis

Hyperplasia, lymphoid

2

(4%)

Infiltration cellular, plasma cell

Inflammation, suppurative

1

(2%)

Lymph node, mesenteric

(50)

Angieclasis

1

(2%)

Erythrophagocytosis

Hyperplasia, lymphoid

1

(2%)

Infiltration cellular, histiocytic

Spleen

(50)

Atrophy

Congestion

1

(2%)

Fibrosis

Hematopoietic cell proliferation

4

(8%)

Hemorrhage

Hyperplasia, lymphoid, focal

Necrosis, coagulative

Pigmentation, hemosiderin

1

(2%)

Thymus

(43)

Congestion

Hyperplasia, lymphoid

(2)

(48)

2

2

(50)
1

(4%)
(4%)

(2%)

1

(2%)

1

(2%)

(8)

1 (13%)
(50)

2 (4%)
1 (2%)
4 (8%)

(50)

1 (2%)

1 (2%)

1 (2%)

1 (2%)

(47)

(50)

1

(2%)

1 (2%)

1

(2%)

2

(4%)

(48)

(49)

1 (2%)

1

(2%)

2 (4%)

1 (2%)

1

(2%)

(49)

(50)

1 (2%)

4 (8%)

2

(4%)

1 (2%)

6 (12%)

8

(16%

1

(2%)

2 (4%)

1

(2%)

1 (2%)

1

(2%)

(34)

(40)

1 (3%)
1 (3%)

133

Nitrofurantoin, NTP TR 341

TABLE B5 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS IN THE
TWO-YEAR FEED STUDY OF NITROFURANTOIN (Continued)

Untreated Control

Low Dose

High Dose

INTEGUMENTARY SYSTEM
Mammary gland
Hyperplasia, cystic
Hyperplasia, lobular
Duct, cyst
Skin

Cyst epithelial mclusion

Hyperkeratosis, focal

Hyperplasia, focal

Subcutaneous tissue, cyst

Subcutaneous tissue, inflammation, suppurative

(49)

(50)

(50)

5 (10%)

19

(38%)

11

(22%)

5 (10%)

5

(10%)

3

(6%)

38 (78%)

33

(66%)

39

(78%)

(50)

(49)

(50)

1 (2%)

1

(2%)

2

(4%)

1

(2%)

1

(2%)

1

(2%)

2

(4%)

(50)

(50)

(50)

1 (2%)

4 (8%)

4

(8%)

5

(10%)

MUSCULOSKELETAL SYSTEM
Bone

Osteopetrosis
Cranium, hyperostosis

NERVOUS SYSTEM
Brain

Compression
Degeneration, multifocal
Hemorrhage, focal
Hemorrhage, multifocal
Hydrocephalus

(50)

(50)

6 (12%)

2 (4%)

1 (2%)

1 (2%)

1 (2%)

1 (2%)

1 (2%)

(50)

1 (2%)
3 (6%)

1 (2%)

RESPIRATORY SYSTEM
Lung

Angiectasis

Congestion

Edema

Infiltration cellular, lymphocytic, multifocal

Infiltration cellular, histiocytic, focal

Inflammation, chronic

Alveolar epithelium, hyperplasia, focal

Alveolar epithelium, hyperplasia, multifocal

Bronchiole, inflammation, chronic active

Bronchiole, inflammation, suppurative

Interstitium, edema
Nose

Lumen, foreign body

Lumen, fungus

Lumen, inflammation, suppurative

Nasolacrimal duct, inflammation, suppurative
Trachea

Infiltration cellular, lymphocytic

(50)

1 (2%)

(4%)
(8%)

(50)

1
2
4

(2%)

(4%)
(8%)

(50)

1

(2%)

(11)
1

(1)

(9%)

1

(100%)

(50)

(50)

2

(4%)

4

(8%)

4

(8%)

1

(2%)

2

(4%)

3

(6%)

5

(10%)

2

(4%)

1

(2%)

1

(2%)

2

(4%)

1

(2%)

1

(2%)

1

(2%)

2

(4%)

(49)

(47)

1

(2%)

1

(2%)

1

(2%)

1

(2%)

3

(6%)

5

(10%)

4

(9%)

(50)

(50)

SPECIAL SENSES SYSTEM
Ear

Middle ear, inflammation, suppurative
Eye

Cataract

Inflammation, suppurative

Synechia

Cornea, edema

Lens, mineralization

Retina, degeneration
Hardenan gland

Fibrosis

Infiltration cellular, lymphocytic, multifocal

(1)

1 (100%)

(18)

19)
16

1

(84%)
(5%)

2
17
(9)

8

I

(11%)
(89%)

(89%)
(11%)

(5)

(6)

2 (33%)

1 (17%)

2 (33%)

3 (50%)

Nitrofurantoin, NTP TR 341

134

TABLE B5. SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE RATS IN THE
TWO-YEAR FEED STUDY OF NITROFURANTOIN (Continued)

Untreated Control

Low

Dose

High

Dose

URINARY SYSTEM

Kidney

(50)

(50)

(50)

Mineralization, multifocal

1

(2%)

1

(2%)

Nephropathy, chronic

44

(88%)

40

(80%)

48

(96%)

Pigmentation, hemosiderm

1

(2%)

Cortex, fibrosis, focal

1

(2%)

Pelvis, hydronephrosis

1

(2%)

Pelvis, infiltration cellular, lym

phocytic

I

(2%)

Renal tubule, degeneration

I

(2%)

2

(4%)

Renal tubule, hyperplasia, focal

I

1

(2%)

1

(2%)

Renal tubule, pigmentation, hemosiderin

1

(2%)

2

(4%)

2

(4%)

Urinary bladder

(50)

(50)

(50)

Mucosa, hyperplasia, diffuse

1

(2%)

135 Nitrofurantoin, NTP TR 341

Nitrofurantoin, NTP TR 341 136

APPENDIX C

SUMMARY OF LESIONS IN MALE MICE IN THE

TWO-YEAR FEED STUDY OF NITROFURANTOIN

PAGE

TABLE CI SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE IN THE

TWO-YEAR FEED STUDY OF NITROFURANTOIN

TABLE C2 INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF MALE MICE IN THE TWO-YEAR

FEED STUDY OF NITROFURANTOIN

TABLE C3 ANALYSIS OF PRIMARY TUMORS IN MALE MICE IN THE TWO-YEAR FEED

STUDY OF NITROFURANTOIN

TABLE C4 HISTORICAL INCIDENCE OF ADRENAL MEDULLARY PHEOCHROMOCYTOMAS

IN MALE B6C3Fi MICE RECEIVING NO TREATMENT

TABLE C5 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE MICE

IN THE TWO-YEAR FEED STUDY OF NITROFURANTOIN

139

142

154

157

158

137

Nitrofurantoin, NTPTR 341

Nitrofurantoin, NTPTR 341 138

TABLE CI. SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE IN THE TWO-YEAR

FEED STUDY OF NITROFURANTOIN

Untreated Control

Low Dose

High Dose

Animals initially in study

Animals removed

Animals examined histopathologically

50
50
50

50
50
50

50
50
50

ALIMENTARY SYSTEM
Esophagus

Lymphoma malignant lymphocytic
Intestine small, ileum

Lymphoma malignant mixed
Intestine small, jejunum

Lymphoma malignant lymphocytic
Liver

Hemangiosarcoma

Hemangiosarcoma, multiple

Hepatocellular carcinoma

Hepatocellular carcinoma, multiple

Hepatocellular adenoma

Hepatocellular adenoma, multiple

Lymphoma malignant histiocytic

Lymphoma malignant lymphocytic

Lymphoma malignant mixed
Mesentery

Alveolar/bronchiolar carcinoma, metastatic,
lung

Lymphoma malignant histiocytic
Pancreas

Alveolar/bronchiolar carcinoma, metastatic,
lung
Stomach, forestomach

Fibrosarcoma

Squamous cell carcinoma

(49)

(49)

(50)

1 (2%)

(47)

(43)

(47)

1

(2%)

(49)

(45)

1

(2%)

(49)

(50)

(50)

1

(2%)

(50)

1 (2%)

8 (16%)

6

(12%)

7

(14%)

1 (2%)

1

(2%)

1 (2%)

4

(8%)

3

(6%)

1 (2%)

1

(2%)

(2%)

2

(4%)

(2%)

3 (6%)

(2%)

(50)

•(50)

•(50)

(2%)

1

(2%)

(2%)

(50)

(49)

(50)

(2%)

(49)

(48)

(49)

1 (2%)
1 (2%)

CARDIOVASCULAR SYSTEM
Heart

Lymphoma malignant lymphocytic

(50)

1 (2%)

(50)

(50)

ENDOCRINE SYSTEM
Adrenal gland, cortex

Adenoma

Spindle cell, carcinoma
Adrenal gland, medulla

Pheochromocytoma malignant

Pheochromocytoma benign
Islets, pancreatic

Adenoma
Thyroid gland

Follicular cell, adenoma

(50)

(49)
2 (4%)
4 (8%)

(50)

(48)

2 (4%)

(49)

1 (2%)
1 (2%)

(48)

(49)

1 (2%)

(47)

(50)

(49)

1 (2%)
(50)

(48)

GENERAL BODY SYSTEM
None

139

Nitrofurantoin, NTP TR 341

TABLE CI. SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE IN THE TWO-YEAR

FEED STUDY OF NITROFURANTOIN (Continued)

Untreated Control

Low Dose

High Dose

GENITAL SYSTEM
Epididymis

Lymphoma malignant lymphocytic
Preputial gland

Hemangiosarcoma
Testes

Interstitial cell, adenoma, multiple

(50)

•(50)

1 (2%)
(49)

1 (2%)

(49)

•(50)
(49)

(50)

1 (2%)
♦(50)

(50)

HEMATOPOIETIC SYSTEM

Blood •(50)

Leukemia 1 (2%)

Bone marrow (50)

Hemangiosarcoma 1 (2%)

Lymph node (50)

Lymphoma malignant lymphocytic 1 (2%)

Bronchial, lymphoma malignant lymphocytic 1 (2%)

Bronchial, mediastinal, fibrosarcoma,
metastatic, stomach 1 (2%)

Pancreatic, lymphoma malignant mixed

Renal, lymphoma malignant lymphocytic
Lymph node, mandibular (45)

Lymphoma malignant lymphocytic 1 (2%)

Bronchial, mediastinal, alveolar/bronchiolar
carcinoma, metastatic, lung
Lymph node, mesenteric (47)

Lymphoma malignant histiocytic

Lymphoma malignant lymphocytic 3 (6%)

Lymphoma malignant mixed
Spleen (50)

Hemangiosarcoma 1 (2%)

Lymphoma malignant histiocytic

Lymphoma malignant lymphocytic 4 (8%)

Lymphoma malignant mixed
Thymus (44)

Lymphoma malignant lymphocytic 2 (5%)

Mediastinum, fibrosarcoma, metastatic, skin

♦(50)
(50)
(49)

1 (2%)
(41)

(46)

1

(2%)

(49)

2
1

(4%)
(2%)

(43)

♦(50)
(50)
(49)

1 (2%)
(46)

1 (2%)
(44)

1 (2%)

2 (5%)
(50)

1 (2%)

1 (2%)

2 (4%)
(41)

1 (2%)

INTEGUMENTARY SYSTEM

Skin (50)

Melanoma benign
Squamous cell carcinoma

Subcutaneous tissue, fibroma 1 (2%)

Subcutaneous tissue, fibroma, multiple

Subcutaneous tissue, fibrosarcoma 7 (14%)

Subcutaneous tissue, fibrosarcoma, multiple 2 (4%)

Subcutaneous tissue, schwannoma malignant 2 (4%)

Subcutaneous tissue, schwannoma malignant,
multiple 1 (2%)

(50)

1 (2%)

1 (2%)

1 (2%)

1 (2%)

5 (10%)

(50)

3 (6%)

11 (22%)
1 (2%)

MUSCULOSKELETAL SYSTEM
Skeletal muscle

Fibrosarcoma, metastatic, stomach
Diaphragm, intercostal, alveolar/bronchiolar
carcinoma, metastatic, lung

•(50)

I (2%)

•(50)

•(50)

1 (2%)

NERVOUS SYSTEM
Brain

Lymphoma malignant lymphocytic

(50)

1 (2%)

(50)

(50)

Nitrofurantoin, NTP TR 341

140

TABLE CI. SUMMARY OF THE INCIDENCE OF NEOPLASMS IN MALE MICE IN THE TWO-YEAR

FEED STUDY OF NITROFURANTOIN (Continued)

Untreated Control

Low Dose

High Dose

RESPIRATORY SYSTEM

Lung (50)

Alveolar/bronchiolar adenoma 5 (10%)

Alveolar/bronchiolar carcinoma 1 (2%)

Alveolar/bronchiolar carcinoma, multiple

Fibrosarcoma, metastatic, stomach 1 (2%)

Hepatocellular carcinoma, metastatic, liver 1 (2%)

Lymphoma malignant histiocytic

Lymphoma malignant lymphocytic 3 (6%)

(50)
2 (4%)
2 (4%)

2 (4%)

(50)
4 (8%)
2 (4%)
I (2%)

1 (2%)
1 (2%)

SPECIAL SENSES SYSTEM
Harderian gland
Adenoma

•(50)
2 (4

•(50)

3 (6%)

•(50)

2 (4%)

URINARY SYSTEM
Kidney

Fibrosarcoma, metastatic, stomach
Lymphoma malignant lymphocytic
Lymphoma malignant mixed

(50)

I (2%)
1 (2%)

(50)

(50)

1 (2%)

SYSTEMIC LESIONS
Multiple organs

Lymphoma malignant lymphocytic

Hemangiosarcoma

Leukemia

Lymphoma malignant histiocytic

Lymphoma malignant mixed

(50)

*(50)

5 (10%)

1 (2%)

3 (6%)

1 (2%)

I (2%)

2 (4%)

(50)

1 (2%)

1 (2%)
4 (8%)

ANIMAL DISPOSITION SUMMARY
Animals initially in study
Dead

Terminal sacrifice
Moribund

50

7

28

15

50

12

29

9

50

2

33

15

TUMOR SUMMARY

Total animals with primary neoplasms •* 31

Total primary neoplasms 52

Total animals with benign neoplasms 12

Total benign neoplasms 17

Total animals with malignant neoplasms 28

Total malignant neoplasms 35

Total animals with secondary neoplasms *** 1

Total secondary neoplasms 5

27
35
11
15
20
20
2
2

33
42
14
14
24
28
3
6

• Number of animals receiving complete necropsy examinations; all gross lesions including masses examined microscopically.

*• Primary tumors: all tumors except secondary tumors

•*• Secondary tumors: metastatic tumors or tumors invasive into an adjacent organ

141

Nitrofurantoin, NTP TR 341

TABLE C2.

INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF MALE MICE IN THE TWO-YEAR FEED
STUDY OF NITROFURANTOIN: UNTREATED CONTROL

WEEKS ON
STUDY

CARCASS
ID

ALIMENTARY SYSTEM
Esophagus

Lvmpnoma malignant lymphocytic
Gallbladder
Intestine large
Intestine large, cecum
Intestine large, colon
Intestine large, rectum
Intestine small
Intestine small, duodenum
Intestine small, ileum
Intestine small, jejunum
Liver

Hemangiosarcoma. multiple

Hepatocellular carcinoma

Hepatocellular carcinoma, multiple

Hepatocellular adenoma

Hepatocellular adenoma, multiple

Lymphoma malignant lymphocytic
Mesentery
Pancreas
Salivary glands
Stomacn
Stomach, forestomach

Fibrosarcoma

Squamous cell carcinoma
Stomach, glandular
Tooth

CARDIOVASCULAR SYSTEM

Heart
Lymphoma malignant lymphocytic

ENDOCRINE SYSTEM

Adrenal gland
Adrenal gland, cortex
Adrenal gland, medulla

Pheochromocytoma malignant

Pheochromocytoma benign
Islets, pancreatic
Parathyroid gland
Pituitary gland
Thyroid grand

Follicular cell, adenoma

GENERAL BODY SYSTEM
None

GENITAL SYSTEM

Coagulating gland
Epididymis
Penis
Preputial gland

Hemangiosarcoma
Prostate
Seminal vesicle
Testes

Interstitial cell, adenoma, multiple

0

0

0

0

II

U

0

0

()

II

0

0

0

0

1)

1)

fl

II

1

1

0

0

u

1

1

3

5

6

6

6

B

7

H

9

9

9

9

9

n

n

n

9

9

■i

9

0

9

5

6

6

8

7

8

1

3

5

9

9

0

2

0

1

1

0

0

0

0

0

0

0

0

1

1

n

1

0

0

0

(1

n

1

0

0

6

4

5

4

3

3

7

8

0

0

3

0

4

2

2

R

9

1

1

2

1

1

1

2

1

2

1

2

3

4

3

5

3

I

2

2

1

~D — (T
4 9
4 2

u — or

1 I

2 3

X

M

+ ■»-■»- +
-*- + ■♦- +

+ + + +

+ + + +

+ + + +

-(- + + + -*--♦- + + -*- + +

+ + + + + -*- + + + ■*- +

■»- + + + + ■*- + + + + +

+ + + ■(- + + + + + + ■♦-

+ + + + + + + -f- + + +-

■t- + -»-+M+ + -*--t- + +

f -f + + + +
X

+ *- + + +
■♦- + + + ■*-
M + + + -t-

+ -(• + -»-+'■>-

M M M + + M

+ M + + + +

+ M + -t- + M

+ + f + +
+ +- + + +
+ + ■*- + +

-(- + + -*- + ♦--(- + + + +

-t--»-M-t- + -»-M + M + +

-(-■»--*- + -t- + + + -i--*- +

■t- + -t--t- + 4--(- + + -»- +

X

-(- + ■♦-■(-■*-
■•■ + + +-■*-

+ +
X

+ : Tissue examined microscopically

Not examined
— : Present but not examined microscopically
I: Insufficient tissue

M: Missing

A: Autolysis precludes examination

X: Incidence of listed morphology

Nitrofurantoin, NTP TR 341

142

TABLE C2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF MALE MICE: UNTREATED CONTROL

(Continued)

WEEKS ON
STUDY

CARCASS
ID

ALIMENTARY SYSTEM
Esophagus

Lymphoma malignant lymphocytic
Gallbladder
Intestine large
Intestine large, cecum
Intestine large, colon
Intestine large, rectum
Intestine small
Intestine small, duodenum
Intestine small, ileum
Intestine small, jejunum
Liver

Hemangiosarcoma. multiple

Hepatocellular carcinoma

Hepatocellular carcinoma, multiple

Hepatocellular adenoma

Hepatocellular adenoma, multiple

Lymphoma malignant lymphocytic
Mesentery
Pancreas
Salivary glands
Storaacn
Stomach, forestomach

Fibrosarcoma

Squamous cell carcinoma
Stomach, glandular
Tooth

CARDIOVASCULAR SYSTEM

Heart
Lymphoma malignant lymphocytic

ENDOCRINE SYSTEM

Adrenal gland
Adrenal gland, cortex
Adrenal gland, medulla

Pheochromocytoma malignant

Pheochromocytoma benign
Islets, pancreatic
Parathyroid gland
Pituitary gland
Thyroid Bland

Folhcular cell, adenoma

None

BODY SYSTEM

GENITAL SYSTEM

Coagulating gland
Epididymis
Penis
Preputial gland

Hemangiosarcoma
Prostate
Seminal vesicle
Testes

Interstitial cell, adenoma, multiple

1

1

1

1

1

1

I

1

1

1

1

1

0

0

0

0

0

0

0

0

0

0

n

n

5

5

5

5

5

5

5

5

5

5

5

5

U

0

i)

0

0

I)

(1

0

I)

(1

0

(1

1

1

■I

2

3

3

4

5

5

5

5

6

4

5

4

5

4

5

5

•I

3

4

5

3

1 1 1 1 1 I 1 1

00000000
55555555

11111
0 0 0 0 0
5 5 5 5 5

"D — D — n — n — 0 — 0 — 0 — s — d — d — d — o w

6677778888999
45 2 3451345345

TOTAL:
TISSUES
TUMORS

M +

+ +

-♦- +
+ +
+ +

+ + -*- + +

+-■*- + ■»- +

+ M + 4- 4-

+ -K + + +

+ + + +- +

X X
X

-I- +
+ +

-t- +-

X

+ + -(-
+ + +
+ + ■(-

+ +
+ +

M
+
+
X

+ •!■ + +

+ + -(- +

X

49

1

44

50

50

50

48

50

49

47

49

50

1

8

1

1

1

3

3

50

50

50

49

1

1

50

10

50

1

50

50

49

2

4

50

41

46

48

2

50

2
9
1

49
6

49
1

143

Nitrofurantoin, NTP TR 341

TABLE C2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF MALE MICE: UNTREATED CONTROL

(Continued)

WEEKS ON
STUDY

CARCASS
ID

HEMATOPOIETIC SYSTEM
Blood

Leukemia
Bone marrow

Hemanposarcoma
Lymph node

Lymphoma malignant lymphocytic

Bronchial, lymphoma malignant
lymphocytic

Bronchial, mediastinal, fibrosarcoma,
metastatic, stomach
Lymph node, mandibular

Lymphoma malignant lymphocytic
Lymph node, mesenteric

Lymphoma malignant lymphocytic
Spleen

Hemangiosarcoma

Lymphoma malignant lymphocytic
Thymus

Lymphoma malignant lymphocytic

INTEGUMENTARY SYSTEM
Mammary gland
Skin
Subcutaneous tissue, fibroma
Subcutaneous tissue, fibrosarcoma
Subcutaneous tissue, fibrosarcoma,

multiple
Subcutaneous tissue, schwannoma

malignant
Subcutaneous tissue, schwannoma
malignant, multiple

MUSCULOSKELETAL SYSTEM
Bone

Skeletal muscle
Fibrosarcoma, metastatic, stomach

NERVOUS SYSTEM

Brain
Lymphoma malignant lymphocytic

RESPIRATORY SYSTEM

Lung
Alveolar/bronchiolar adenoma
Alveolar/bronchiolar carcinoma
Fibrosarcoma, metastatic, stomach
Hepatocellular carcinoma, metastatic.

liver
Lymphoma malignant lymphocytic

Nose

Trachea

5PECTAL SENSES SYSTEM
Hardenan gland
Adenoma

URINARY SYSTEM

Kidney
Fibrosarcoma, metastatic, stomach
Lymphoma malignant lymphocytic

Ureter
Urethra
Unnary bladder

0

0

n

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

1

1

I

1

1

1

1

0

1)

0

1

1

;t

5

B

B

B

B

7

8

9

9

9

9

9

0

0

0

0

U

0

0

8

9

9

2

9

0

9

5

6

6

8

7

8

1

3

5

9

9

0

2

2

2

5

5

5

n

1

1

0

n

(1

(1

II

(1

(1

(1

1

1

11

1

II

II

II

II

II

(1

(1

(1

11

11

1

0

0

6

4

5

4

3

3

7

8

0

0

3

0

4

2

2

6

9

4

9

2

I

1

1

1

2

1

1

1

2

1

2

1

2

3

4

3

5

3

1

2

2

L

4

2

3

2

3

+

■f

-t-

^-

+

+

+

+

-t-

+

+-

-(-

X

+

+

+

+

+

+

4-

-t-

+

+

+

■t-

+

+

-t-

+

+

■♦-

-♦■

+

+

■K

■♦-

-t-

+

+

+

-t-

+

+

+

+

+

+

4-

+

4-

■♦- M -t- -t- -t-

+ -*-■*- + +

-t- H- + -f -t-

f -H + + +

-»-

4-

+

4-

+

-t-

X

+

+

M

+■

+

+■

+

+

M

■f

X

■(-

+

+

+

+

+

+

+

4-

X

X

+-

+

-*-

+

+

M

+
X

+

+
X

MMMMMMM +

+ + + + + + + +

MMMMMMMMMMMMMMM

+ + +

+- + + 4-

MMMMMMMM

+-

+

+

+

-t-
X

X

+

+

X

X

X

X

+

■t-

+■

4-

+

+

t-

+

+

■(-

+■

+

+

+■

+

-♦-

+ + -(-

Nitrofurantoin, NTPTR 341

144

TABLE C2. INDIVIDUAL A>aMAL TUMOR PATHOLOGY OF MALE MICE;

(Continued)

UNTREATED CONTROL

WEEKS ONf
STUDY

1 1 1 1 1 1 1 1 1 L 1 1 1 1 1 1 1 I 1 1 1 1 1 I 1
0000000000000000000000000
5555555555555555555555555

TOTAL:

CARCASS
ID

0000000000000000000000000
1122334555566677778888999
4545455234534523451345345

TISSUES
TUMORS

HEMATOPOIETIC SYSTEM
Blood

Leukemia
Bone marrow

Hemangnosarcoma
Lymph node

Lymphoma mahgnant lymphocytic

Bronchial, lymphoma malignant
lymphocytic

Bronchial, mediastinal, fibrosarcoma,
metastatic, stomach
Lymph node, mandibular

Lymphoma malignant lymphocytic
Lymph node, mesentenc

Lymphoma malignant lymphocytic
Spleen

Heraangiosarcoma

Lymphoma malignant lymphocytic
Thymus

Lymphoma malignant lymphocytic

+ +

X

+ + + + + + + 4--t--(- + + + + + -t- + +--t- + + -t- + + +
+ + + + 4-^- + -+-+- + + + + + + + + + + + + + + + +

X

+ + + + 4- + + + 4-MM-t- + -t- + -t--*--t-+- + M + + + +

-t--f + -f+M + + -t- + + + + + + + -»--»- + + + + + + +
X X

+- + + + -t--*- + + + + + 4- + + + + + + + + + + + + +

XX X

+ + -t- + + + + + + + + +M + + + + -t- + -*-+MMM +

2

1
50

1
50

1

1

1
45

1
47

3
50

1

4
44

2

INTEGUMKNTAilY SYSTEM
Mammary gland
Skin
Subcutaneous tissue, fibroma
Subcutaneous tissue, fibrosarcoma
Subcutaneous tissue, fibrosarcoma.

multiple
Subcutaneous tissue, schwannoma

malignant
Subcutaneous tissue, schwannoma
malignant, multiple

MMM + MMMM + +MMMMMMMMMMMMMM +

+ -t- + + + +- + + + -t-+'+- + + -f + + + + + + + + + +

X X

X

X

6
50

1
7

2

2

1

MUSCULOSKELETAL SYSTEM
Bone

Skeletal muscle
Fibrosarcoma, metastatic, stomach

+ + 4- + + +--t- + -(- + -f + + -f + -t--+- + + + + + -f-(--f-

50

1
1

NERVOUS SYSTEM
Brain

Lymphoma malignant lymphocytic

-(- + + + + + -(->--*--(--(--t- + + + + + + + + + + + + +

50
1

RESPIRATORY SYSTEM

Lung
Alveolar/bronchiolar adenoma
Alveolar/bronchiolar carcinoma
Fibrosarcoma, metastatic, stomach
Hepatocellular carcinoma, metastatic,

liver
Lymphoma malignant lymphocytic

Nose

Trachea

^- + + + ■t- + ■*■■*--^^ + + + + + + * + + + + + + + +

X X
X

X

+ -^--(--^-^^-■(-■t-+-+- + ^-^--(■^- + -^ + -f + -(-■t--^- + +
+ + + 4■-t- + + + + + -*■-t-^^ + *-■*■ + ^■-*■■*■^■*■■^ +

50
5
1

1

1

3

42

49

SPECIAL SENSES SYSTEM
Hardenan gland
Adenoma

4- +

X X

2
2

URINARY SYSTEM

Kidney
Fibrosarcoma, metastatic, stomach
Lymphoma malignant lymphocytic

Ureter

Urethra

Unnary bladder

+ + 4--(-4 + + -t- + +--(- + -t--t- + + -*- + + + + + + -t- +

X

+ -(--(-+- + -t--t--f4-4 + -t- + -t-+- + + +- + + + -t-4- + +

50
1
I
1
2

50

145

Nitrofurantoin, NTP TR 341

TABLE C2.

INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF MALE MICE IN THE TWO-YEAR FEED
STUDY OF NITROFURANTOIN: LOW DOSE

WEEKS ON
STUDY

CARCASS
ID

ALIMENTARY SYSTEM
Esopha^s
Gallbladder
Intestine large
Intestine large, cecum
Intestine large, colon
Intestine large, rectum
Intestine small
Intestine small, duodenum
Intestine small, ileum
Intestine small, jejunum

Lymphoma mahgTiant lymphocytic
Liver

Hemangiosarcoma

Hepatocellular carcinoma

Hepatocellular carcinoma, multiple

Hepatocellular adenoma

Hepatocellular adenoma, multiple

Lymphoma malignant histiocytic
Mesentery

Lymphoma malignant histiocytic
Pancreas
Salivary glands
Stomacn

Stomach, forestomach
Stomach, glandular
Tooth

CARDIOVASCULAR SYSTEM

Heart

ENDOCRINE SYSTEM

Adrenal gland
Adrenal gland, cortex

Adenoma

Spindle cell, carnnoma
Adrenal gland, medulla
Islets, pancreatic

Adenoma
Parathyroid giand
Pituitary gland
Thyroid gland

GENERAL BODY SYSTEM

None

GENITAL SYSTEM

Epididymis
Penis

Preputial gland
Prostate
Seminal vesicle
Testes

0

(1

0

(»

0

0

I)

0

0

0

0

0

0

)l

0

(1

0

1

1

1

1

1

1

1

1

0

2

2

3

4

f>

7

7

8

8

8

8

8

8

9

9

9

0

0

0

0

0

0

0

0

B

3

'^

4

8

4

1

6

2

3

6

7

8

9

1

5

7

0

2

4

4

5

5

5

5

'2

'2

'1

'I

'2

'2

'2

'2

'2

'2

'2

'2

M

'2

'2

'2

M

'2

'2

'2

t\

'2

'2

'2

2

7

2

6

2

5

6

4

3

5

3

4

8

0

9

3

6

0

3

6

9

0

3

1

I

I

1

1

1

2

1

2

1

1

2

2

2

1

1

1

3

3

2

4

4

2

3

5

I

2

3

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+■

+

+

+

+

+

+

+

+

+

+

M

M

+

+

+

+

+

M

+

+

+

+

+

+

+■

+

+

+

+

+

+

+

+

+

+

A

A

-(-

+-

+

+

+

+

+

+

+

+

+

+

+■

+

+

+

+

+

+

+

+

+

+■

A

A

+

+

M

+

M

+

+

+

+

+

+

+

+

+

+

+

+

+

M

+

+

+

+

A

A

M

+

+

+

M

+

+

+

-t-

+

+

+

-f

-t-

+

+

+

+

+

+

■H

+

+

A

A

M

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

-t-

+

+

M

+

+

+

A

A

+

A

+

+

+

+

+

+

-f

+

+

■t-

+

+

+

+

+

+

+

M

+

+

+

A

A

+

A

+

-*-

+

+

+

+

+

+

-(-

-(-

M

+

-t-

+

+

+

A

M

¥

+

-♦-

A

A

+

A

+

A

+

+

+

+

+

+

+

-t-

■♦-

+

-t-

+

+

+

-t-

M

■(■

+

+

A

A

+

A

+

A

+

+

■t-

+

+

+

-t-

-t-

+

+

4-

+

■f

■(-

+ f

M
M

M

Nitrofurantoin, NTP TR 341

146

TABLE C2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF MALE MICE: LOW DOSE

(Continued)

WEEKS ON
STUDY

CARCASS

ID

ALIMENTARY SYSTEM
Esophagus
Gallbladder
Intestine large
Intestine large, cecum
Intestine large, colon
Intestine large, rectum
Intestine small
Intestine small, duodenum
Intestine small, ileum
Intestine small, jejunum

Lymphoma malignant lymphocytic
Liver

Hemangiosarcoma

Hepatocellular carcinoma

Hepatocellular carcinoma, multiple

Hepatocellular adenoma

Hepatocellular adenoma, multiple

Lymphoma malignant histiocytic
Mesentery

Lymphoma malignant histiocytic
Pancreas
Salivary glands
Stomach

Stomach, forestomach
Stomach, glandular
Tooth

CARDIOVASCULAR SYSTEM

Heart

ENDOCRINE SYSTEM

Adrenal gland
Adrenal gland, cortex

Adenoma

Spindle cell, carcinoma
Adrenal gland, medulla
Islets, pancreatic

Adenoma
Parathyroid gland
Pituitary gland
Thyroid gfand

GENERAL BODY SYSTEM

None

GENITAL SYSTEM

Epididymis
Penis

Preputial gland
Prostate
Seminal vesicle
Testes

1 1
0 0
5 5

I 1 1 1 1
0 0 0 0 0
5 5 5 5 5

1 1 1
0 0 0
5 5 5

1 1 1
0 0 0
5 5 5

'2

•i

2

2

2

1

2

2

2

2

2

2

2

2

2

2

2

2

2

2

2

2

2

.1

3

1

1

2

2

2

4

4

4

5

5

5

6

7

7

7

7

8

8

8

8

9

9

9

0

0

4

5

3

4

5

3

4

5

3

4

5

5

2

3

4

5

2

3

4

5

3

4

5

4

5

M
M

+ + +
+ + +

■f +

4- + + + + +

+ ■*- + -*-

■*-}- +
+ -t- +

TOTAL;
TISSUES
TUMORS

49

45

48

46

45

47

46

44

43

45

1

50

1

6

1

4

1

2

1

1

49

50

48

48

48

12

49

49

1

1

48

49

1

41

48

47

50

4

49

147

Nitrofurantoin, NTP TR 341

TABLE C2. INDIVIDUAL ANIMAL

TUMOR PATHOLOGY OF MALE MICE: LOW DOSE

(Continued)

WEEKS On

0

~0"

~0"

0

"0-

0

0

0

0

0

0

0

0

0

"6"

^0"

0

I

1

1

1

1

1

I

1

STUDY

0

2

2

3

4

5

7

7

8

8

8

8

8

8

9

9

9

0

0

0

0

0

0

0

0

8

3

7

4

8

4

1

6

2

3

6

7

8

9

1

5

7

0

2

4

4

5

5

5

5

CARCASS

ii

~T~

~T~

T"

'^

'2

'2

'i

""2~

'2

a

"2~

~T~

~T~

a

'2

~T~

""3"

'2

•2

'2

'2

ID

7

2

6

2

5

6

4

3

5

3

4

8

0

9

3

6

0

3

6

9

0

3

1

1

1

1

1

1

2

1

■2

I

1

2

2

2

1

1

I

3

3

2

4

4

2

3

5

I

2

3

HEMATOPOIETIC SYSTEM

Bone marrow

+

+

+

4-

-(-

+

+

+

+

+■

+

■♦-

+

+

-(-

•t-

+

+

+

+

+

+■

+

-t-

■K

Lymph node

+

+

-*-

■t-

+

M

+

+

+

+

+

+

+

+

-t-

+

+

+

+

+

+

+■

+■

+

4-

Renal, lymphoma malignant lymphocytic

Lymph node, mandibular

+

+

+

4-

+

M

+

+

+

M

+

+

+

M

■f

+

+

+

+

M

+

4-

+

4-

M

Lymph node, mesenteric

+

+

+

■t-

+

M

+

M

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

Lymphoma malignant lymphocytic

Spleen

+

+

+

+■

+

M

+

+

+

+

+

+

+

+

+

■*-

+

+

+

+

+■

-f

+■

+

+

Lymphoma malignant histiocytic

X

X

Lymphoma malignant lymphocytic

Thymus

+

+

+

+

■•"

M

+

+

M

+

+

+

+

+

+

+

+

+

+

+

+

M

*-

+

+

INTEGUMENTARY SYSTEM

Mammary gland

M

+

M

M

M

M

M

M

+

M

+

M

M

M

M

M

M

M

M

M

M

M

+

M

■h

Skin

+

■t-

+

-t-

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

Melanoma benign

Squamous cell carcinoma

Subcutaneous tissue, fibroma

Subcutaneous tissue, fibroma, multiple

Subcutaneous tissue, fibrosarcoma

X

X

X

X

X

MUSCULOSKELETAL SYSTEM

Bone

+

+

+

4-

+

•*■

+

-(-

+

+

+

+

+•

+■

+■

+

+

+

+

+

+■

+

+

+

+

NERVOUS SYSTEM

Brain

+

-t-

+

+

+

+

+

+

+

■1-

+

4-

+

+

+

+

+

+

+

+

+■

+

-*-

^-

+■

RESPIRATORY SYSTEM

Lung

+

+

+

+

+

+

+

f

+

-(-

-t-

+

+

+

+

+

+

+

+

+

+-

+

+

+

■1-

Aiveolar/bronchiolar adenoma

X

Alveolar/bronchiolar carcinoma

Hepatocellular carcinoma, metastatic.

liver

X

X

Nose

M

M

M

M

M

M

+

+

+

+

+

+

-t-

+■

+

+

+

+

-f

+

+

+

+

+

■f

Trachea

+

+

+

-f

+

+

+

+

+

■h

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

SPECIAL SENSES SYSTEM

Hardenan gland

+

Adenoma

X

URINARY SYSTEM

Kidney

+

4-

+

+

-♦-

+

+

+

+

-f

■t-

+

-t-

+

+

+

+

■(-

+

+

+

■t-

*■

+■

4-

Unnary bladder

+

+

+

+-

"*"

+

■1-

+

■1-

■t-

+

+

-t-

+

+

+

+

+

+

+

■¥■

+

+

■f

+

Nitrofurantoin, NTP TR 341

148

TABLE C2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF MALE MICE: LOW DOSE

(Continued)

WEEKS ON
STUDY

CARCASS
ID

HEMATOPOIETIC SYSTEM
Bone marrow
Lymph node

Renal, lymphoma mahg. lymphocytic
Lymph node, mandibular
Lymph node, mesentenc

Lymphoma malignant lymphocytic
Spleen

Lymphoma malignant histiocytic

Lymphoma malignant lymphocytic
Thymus

INTEGUMENTARY SYSTEM

Mammary gland

Skin
Melanoma benign
Squamous cell carcinoma
Subcutaneous tissue, fibroma
Subcutaneous tissue, fibroma, multiple
Subcutaneous tissue, fibrosarcoma

MUSCULOSKELETAL SYSTEM

Bone

NERVOUS SYSTEM
Brain

RESPIRATORY SYSTEM

Lung
Alveolar/bronchiolar adenoma
Alveolar/bronchiolar carcinoma
Hepatocellular carcinoma, metastatic,
liver

Nose

Trachea

SPECIAL SENSES SYSTEM
Hardenan gland
Adenoma

URINARY SYSTEM

Kidney

Unnary bladder

11111
0 0 0 0 0
5 5 5 5 5

11111
0 0 0 0 0
5 5 5 5 5

1 1
0 0
5 5

■^ — 2 — 2 — T
112 2
4 5 3 4

"2 — T

4 4
4 5

"2 2 — T
5 5 5
3 4 5

"2 T

6 7
5 2

"2 2 T

7 7 7
3 4 5

"2 — 2"

"2 — 2 — 2 — 2 — T
8 8 9 9 9
4 5 3 4 5

1 — r

0 0
4 5

+ +

+ +

+ M + M + M M

+ M

+ +

■(- +
-t- +

X

M M +

M M M M

M M

-t- +
X

MMMMMMMMMMMMMM

M M M

+ + +

X

X

TOTAL:
TISSUES
TUMORS

+

-t-

X

X

+

+

+

+

+

-(-

+■

+■

+

+

+

+

+

^

^

+

+

+

+

+

+

-t-

+

+

+

-t-

+

■^

+

-*-

^

+

M

+

4-

+

+■

+

+

+

+

+

+

-f

■(-

"•"

+■

+

+

+

50
49

1
41
46

1
49

2

1
43

5
50

50
2

2
44
49

50
49

149

Nitrofurantoin, NTP TR 341

TABLE C2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF MALE MICE IN THE TWO-YEAR FEED

STUDY OF NITROFURANTOIN: HIGH DOSE

wfiEKS On

STUDY

OOOOOOOOOOOOlllIlllI lllll
1788889999990000000000000
0633692566890055555555555

CARCASS
ID

I 1 2 1 '2 1 I 1 1 I 1 1 1 1 1 1 I I 1 I I 1 I 1 I
540901472I5675381I1222333

II 11212112212311345234234

ALIMENTASY SYSTEM
Esophagus
Gallbladder
Intestine largo
Intestine large, cecum
Intestine large, colon
Intestine large, rectum
Intestine small
Intestine small, duodenum
Intestine small, ileum

Lymphoma malignant mixed
Intestine smalt, jejunum
Liver

Hepatocellular camnoma

Hepatocellular adenoma

Hepatocellular adenoma, multiple

Lymphoma malignant histiocytic

Lymphoma malignant mixed
Mesentery

Alveoiar/bronchiolar carcinoma,
metastatic, lung

Lymphoma malignant histiocytic
Pancreas

Aiveolar/bronchiolar carcinoma,
metastatic, lung
Salivary glands
Stomach

Stomach, forestomach
Stomach, glandular
Tooth

4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4--«-4-4-4-4-4--»-
4-M4-4-4-4-4-4-M4-4-4-4-4- + 4-4-M-«-4-M-»-+M +

4- + 4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4- + 4-4-4-
4-4- + 4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4- + 4-4-4-4-
4-M4-4-4-4-4-4-4--)-4-4-4-4-4-4-4-4-4-4- f4-4-4-4-
4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4- + 4-4-4-4-4-
4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4- + 4- + 4-4-4-4-
4-4-4-4-4--f4-M4-4-4-4-4-4-4-4-4-4-4-4-+- + 4-4-4-
4-M4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4- + 4-

X

+-A + + 4-4-4-4-4-4- + 4-4-4-4-4-4-4- + 4-44-4-4-4-
4- + 4-4-4-4-4-4-4--t-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-

X XXX X

X

X

X

4- 4-

X

X

■(-■l- + 4-4- + 4-4-4-4- + 4-4-4-4-4-4-4-4--«-4- + 4--t- +

X

4-4-4- + 4- + -(-4-4--(-4-4-4--f4--*-4-4-4-4- + 4--(--f4-
4-4- + 4-4-4-4-4-4-4-4- + + 4-4-4-4-4-4-4-4- + 4-4- +
4-4- + 4-4-4-4-4-4-4- + 4-4--l-4-4--(-4-4-H- + -t-4-4-4-
4-4- + 4-4-4-4-4-4-4-4-4- + 4-4-4--t-4--f4-4--t-4- ■t-4-

-)- -t-

CARDIOVASCULAR SYSTEM
Heart

4"4-4-4-4--l--(-4-4-4-4- + 4-4-4-4-4- + 4-4-4-4- + 4- +

ENDOCRINE SYSTEM
Adrenal gland
Adrenal gland, cortei
Adrenal gland, medulla

Pheochromocytoma benign
Islets, pancreatic
Parathyroid gland
Pituitary gland
Thyroid gland

4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4- + 4-4-4-4--^4--t-4-
4-4--h4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4- + f4- ■t-4-4-
4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4--f-(-4-4-

X

4--I-4- + 4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-
M-t- + 4-4-4-4-4-4-4-4-4-4- + 4-4--*-M-l- + + 4-4-4- +
4-M-t--t-4- + 4-4-4-4-4-4-4-4-4-4--t-4-4-4-4-M4-4-4-
M-t--t-4--»-4-4-4-4-4-4-4- + + 4-4--t-4-4-4--t-4-4-4-4-

GENERAL BODY SYSTEM
None

GENITAL SYSTEM
Epididymis

Lymphoma malignant lymphocytic
Pen is

Preputial gland
Prostate
Seminal vesicle
Testes

4-4-4-4-4--t-4- + 4-4--l- + 4-4-4-4-4-4--)- + -t--f4--»-4-

4--f-f4-4--f4-4-4-4--»-4- + 4- + 4--(-4-4-4-i- + + -l- +

+
4--(-4--(- + 4-4-4--(-4- + 4-4-4-4-4--l-4--l-^--(--l-4--f4-

Nitrofurantoin, NTP TR 341

150

TABLE C2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF MALE MICE:

(Continued)

HIGH DOSE

WEEKS ON

1

1

1

1

I

1

1

I

1

1

1

I

I

I

1

1

I

1

I

1

1

1

I

1

1

STUDY

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

TOTAL:
TISSUES

CARCASS

1

~~I

~~I

1

1

I

I

1

1

1

1

I

1

I

I

I

1

~I

1

1

1

'2

y

2

1

ID

3

4

4

4

5

5

6

6

6

6

7

7

7

8

8

8

8

9

9

9

9

0

0

0

2

TUMORS

5

3

4

5

4

5

2

3

4

5

3

4

5

2

3

4

5

2

3

4

5

3

4

5

5

ALIMENTARY SYSTEM

Esophagus
Galtbladder

+

+

+

-^

-t-

+

-t-

+

+

4-

+

+

+

+

+

+

+

4-

+

+

+

4-

+

+

4-

50

+

+

+

-f

M

+

M

+

4-

M

+

M

+

-t-

M

+

+

4-

+

M

M

+

+■

M

4-

37

Intestine large

+

+

+

^-

+

+

+

+

+

4-

4-

4-

4-

+

4-

+

+

4-

4-

+

+

+

+

4-

4-

50

Intestine large, cecum

+

+

+

+

+

+

+

+

4-

4-

4-

4-

+

4-

4-

+

+

4-

+

+

+

4-

+

+

4-

50

Intestine large, colon

+■

■t-

-f

+■

+

+

+

+

+

4-

+

+

+

4-

4-

+

4-

4-

+

+

+

4-

4-

+

+

49

Intestine large, rectum

+

+

+

+

+

+

+

4-

+

4-

+

+

+

+

+

+

+

+

+

+

+

+

+

+

4-

50

Intestine small

+

+

+

+

+

+

4-

+

4-

4-

4-

4-

4-

4-

4-

+

4-

4-

4-

+

4-

4-

4-

•f-

4-

50

Intestine small, duodenum

+

+

+

+

^>

+

M

■*-

+

+

+

+

4-

+

+

+

+

+

+

+

+

4-

M

+

4-

47

Intestine small, ileum

+

+

-t-

M

+

+

+

■f

4-

4-

+

M

4-

4-

4-

+

4-

4-

4-

+

4-

4-

+

+

4-

47

Lymphoma malignant muced

1

Intestine small, jejunum

+

+

+

4-

-*-

-t-

+

+

4-

4-

4-

+

4-

+

4-

+

4-

4-

4-

4-

4-

4-

4-

4-

4-

49

Liver

+

+

+

■t-

+

+

4-

■•-

4-

4-

-t-

+

+

+

+

+

+

+

-*-

■*-

+

+

+

+

4-

50

Hepatocellular carcinoma

X

X

7

Hepatocellular adenoma

X

X

3

Hepatocellular adenoma, multiple

X

1

Lymphoma malignant histiocytic

1

Lymphoma malignant mixed

1

Mesentery

2

Alveolar/bronchioiar carcinoma.

metastatic, lung

1

Lymphoma malignant histiocytic

1

Pancreas

+

+

+

■f-

+

+

+

+■

+

+-

+

+

+

+

+

+

+

+

+

+

+

+

+

4-

+

50

Alveolar/bronchioiar carcinoma.

metastatic, lung

I

Sahvarv glands
Stomacti

+

+

+

+

+

+

+

+

+

+

+

4-

4-

+

4-

4-

+

4-

4-

4-

4-

+

4-

+

+

50

+

+

+

M

+

+

+

f

+

+

+

+

-t-

+

+

+

-(-

4-

4-

4-

4-

4-

4-

4-

4-

49

Stomach, forestoroach

+

+

+

M

+

+

+■

4-

4-

4-

4-

4-

4-

+

4-

4-

■t-

+

4-

4-

4-

4-

■(-

+

4-

49

Stomach, glandular

+

+

4-

M

+

■f

4-

4-

+

+

4-

+

+

4-

+

4-

4-

+

4-

4-

4-

4-

4-

4-

49

Tooth

4-

+■

4

CARDIOVASCULAR SYSTEM

Heart

+

+

+

^

+

+

+

4-

+

+

+

4-

-*"

+

+

+

+

4-

4-

+

+

+

4-

4-

4-

50

ENDOCRINE SYSTEM

Adrenal gland

+

+

+

-t-

+

+

+

+

4-

4-

+

4-

+

+

+

4-

+

4-

4-

4-

4-

4-

+

+

4-

50

Adrenal gland, cortex

+

+

+

+

-(-

+■

-F

+

4-

4-

+

+

+

+

+

+

+

+

+

+

■f

+

+

+

4-

50

Adrenal gland, medulla

+

+

+

+.

+

+

4-

+

+

4-

+

M

+

+

+

+

+

4-

+

+

4-

4-

+

+

4-

49

Pheochromocytoma benign

I

Islets, pancreatic

+

+

+

-t-

-t-

-f

-f

4-

+

4-

+

4-

+

+

-f

+

+

+

+

+

4-

+

■1-

+

4-

50

Parathyroid gland

+

+

+

■(-

-f

+-

4-

M

+

4-

4-

4-

4-

-t-

4-

+

M

+

+

-f

■1-

4-

+

+

+

46

Pituitary gland
Thyroid gland

+

+

+

+

4-

M

+

f

4-

M

+

M

M

-t-

4-

+

+

+

+

+

4-

M

4-

4-

4-

43

+

+

+

+

■t-

+

4-

A-

+

4-

4-

4-

4-

4-

4-

+

M

+

4-

4-

+

+

■»-

+

-*-

48

GENERAL BODY SYSTEM

None

GENITAL SYSTEM

Epididymis

+

+

+

+■

+

+

+

■»-

+

4-

■1-

+

+

4-

+

+

+

4-

4-

+

*■

-t-

+

+-

4-

50

Lymphoma malignant lymphocytic

X

1

Penis

+

I

Preputial gland

+

■ +

+

4

Prostate

+

+

+

+

+■

+

+

+

4-

4-

+

4-

4-

4-

4-

4-

-f

4-

4-

4-

4-

-t-

+

4-

4-

50

Seminal vesicle

1

Testes

+

+

+

+

+

■»■

4-

+

4-

4-

4-

4-

4-

4-

4-

4-

+

■*■

■*■

4-

+

+

■*■

-*-

4-

50

151

Nitrofurantoin, NTP TR 341

TABLE C2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF MALE MICE: HIGH DOSE

(Continued)

WEEKS ON
STUDY

CARCASS
ID

HEMATOPOIETIC SYSTEM
Bone marrow
Lymph node

Pancreatic, lymphoma malignant mixed
Lymph node, mandibular

Bronchial, mediastinal,
alveolar/bronchiolar carcinoma,
metastatic, lung
Lymph node, mesenteric

Lymphoma malignant histiocytic

Lymphoma malignant mixed
Spleen

Lymphoma malignant histiocytic

Lymphoma malignant lymphocytic

Lymphoma malignant mixed
Thymus

Mediastinum, fibrosarcoma, metastatic,
skin

INTEGUMENTARY SYSTEM

Mammary gland

Skin
Subcutaneous tissue, fibroma
Subcutaneous tissue, fibrosarcoma
Subcutaneous tissue, fibrosarcoma,
multiple

MUSCULOSKELETAL SYSTEM
Bone

Skeletal muscle
Diaphragm, intercostal.
alveolar/broncbiolar carcinoma,
metastatic, lung

NERVOUS SYSTEM

Brain

RESPT!UTORY SYSTEM

Lung
Alveolar/bronchiolar adenoma
Alveolar/bronchiolar carcinoma
Alveolar/bronchiolar carcinoma.

multiple
Hepatocellular carnnoma. metastatic.

liver
Lymphoma malignant histiocytic

Nose

Trachea

SPECIAL SENSES SYSTEM

Ear

Hardenan gland
Adenoma

URINARY SYSTEM

Kidney

Lymphoma malignant mixed
Unnary bladder

1 I I L 1
0 0 0 0 0
0 0 5 5 5

1 I I
0 0 0
5 5 5

1

\

2

1

2

1

1

1

1

I

1

1

1 1

1

1

1

1

1

1

1

1

5

4

0

9

0

4

7

2

1

5

6

7

S

3

a I

I

1

2

2

2

3

3

3

1

1

1

1

2

2

1

1

2

2

1

2

3

1

1 3

4

5

2

3

4

2

3

4

X

•I- +

M M M M M

M

+

M

M

M

M

+

+

+

+-

+

+

X

X

X

+ + + + ■*- + +
+• + + + + ■*- +

M M M M

+ + + + + + -h + + + + + +
XXX X

X

-t- -t- + + ■<■
+ >■■»- + ^-

Nitrofurantoin, NTP TR 341

152

TABLE C2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF MALE MICE: HIGH DOSE

(Continued)

WEEKS ON

1

1

1

I

1

1

1

1

I

1

1

1

L

1

1

1

1

1

1

L

1

1

1

1

1

STUDY

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

TOTAL:

CARCASS

1

I

~~I

~~I

I

I

1

1

1

I

I

"T

~T

I

1

1

I

1

1

1

1

'2

ir~

2

1

TISSUES

ID

3

4

4

4

5

5

6

6

6

6

7

7

7

8

8

8

8

9

9

9

9

0

0

0

2

TUMORS

5

3

4

5

4

5

2

3

4

5

3

4

5

2

3

4

5

2

3

4

5

3

4

5

5

HEMATOPOIETIC SYSTEM

Bone marrow

+

+

+

+

+

+

+

+■

+■

+

+

-t-

+

+

+

+

-(-

+

+

+

+

■f

+■

+

+

50

Lymph node

+

+

+

+

■*-

-f

+

+

+

+

+

■t-

+

4-

■(-

■(-

+■

+

+

+

+

+

+

+

+

49

Pancreatic, lymphoma malignant muced

1

Lymph node, mandibular
Bronchial, mediastinal.

+

■t-

+

+

+■

+

+

+■

+■

+

M

+

M

+

-(-

■t-

M

■♦-

+

+

4-

■t-

+

+

+

46

alveolar/bronchiolar carcinoma.

metastatic, lung

1

Lymph node, mesentenc

+

+

+

+

+

+

+

+

+■

+

+■

+

+

M

M

■(-

■f

M

+

+

M

+

+•

-t-

M

44

Lymphoma malignant histiocytic

1

Lymphoma malignant mixed

2

Spleen

+

+

+

+

+-

-t-

-(-

■t-

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

■t-

■I-

50

Lymphoma malignant histiocytic

1

Lymphoma malignant lymphocytic

X

I

Lymphoma malignant mixed

X

2

Thymus

+

+

+

+

+

+

+

+

M

+

+

+

+

+

+

M

+

+■

+

+

M

■t-

-♦-

+

+

41

Mediastinum, fibrosarcoma, metastatic.

skin

X

1

INTEGUMENTARY SYSTEM

Mammary gland

M

M

M

M

M

M

M

M

M

M

+

M

M

M

M

M

M

M

M

M

M

M

M

M

M

8

Skin

f

+■

+-

+

+

-t-

+

+

+

+

+

•1-

+

+

+

+

+

■f

■t-

+

f

+

+

+

-(-

50

Subcutaneous tissue, fibroma

X

X

3

Subcutaneous tissue, fibrosarcoma

X

X

X

U

Subcutaneous tissue, fibrosarcoma.

multiple

I

MUSCULOSKELETAL SYSTEM

Bone

+

+

+

-(-

+

+

+

+

+

+

+

+

+

+

•*-

■»-

+

+

+

+

+

+

+

+

+

50

Skeletal muscle

1

Diaphragm, intercostal,

alveolar/bronchiolar carcinoma.

metastatic, lung

1

NERVOUS SYSTEM

Brain

+

+

+

+-

+

+

+

+

+

+

-t-

+

+

+

+

+

+

+■

+

+

+

■)-

+

+

+

50

RESPIRATORY SYSTEM

Lung

+

+

+

+

+

+

+

+

+

+

-f

+

+

+

+

+■

•(-

+

+

+

+

+

+

+

+

50

Alveolar/bronchiolar adenoma

4

Alveolar/bronchiolar carcinoma

2

Alveolar/bronchiolar carcinoma.

multiple

1

Hepatocellular carcinoma, metastatic.

liver

X

1

Lymphoma malignant histiocytic

1

Nose

-t-

+

+

+

■I-

+

+

+

+

+

+

+

+

+

+

4-

+

+

+

+

+

+

+

+

M

48

Trachea

+

+

+

-*-

+

+

+

+

+

+

+

+

+

+

-t-

■♦-

M

f

+

+

+

■(-

+

+■

+

49

SPECIAL SENSES SYSTEM

Ear

+

1

Hardenan gland

-f

2

Adenoma

X

2

URINARY SYSTEM

Kidney

+

-f

+

+

■f

+

+

+

+

+

-♦-

+

+

+

+

+

+

+

-f

+

+

+

+

+■

+

50

Lymphoma malignant mixed

1

Unnary bladder

+

-*-

*-

+

-(-

f

+■

"*"

+

+

■f

■»•

+

+

+-

+

+

+

+

+

^

4-

+

¥

+

50

153

Nitrofurantoin, NTP TR 341

TABLE C3. ANALYSIS OF PRIMARY TUMORS IN MALE MICE IN THE TWO-YEAR FEED STUDY

OF NITROFURANTOIN

Control

1,300 ppm

2,500 ppm

Adrenal Gland: Pheochroraocytoma

Overall Rates (a)

Adjusted Rates ( b )

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

Cochran-Armitage Trend Test (d)

Fisher Exact Test (d)

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table TesU(d)

Logistic Regression Tests (d)

Cochran-Armitage Trend Test (d)

Fisher Exact Test (d)

Harderian Gland: Adenoma

Overall Rates (a)

Adjusted Rates (b I

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

Cochran-Armitage Trend Test (d)

Fisher Exact Test (d)

Liver: Hepatocellular Adenoma

Overall Rates (a I

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

Cochran-Armitage Trend Test (d)

Fisher Exact Test (d )

Liver: Hepatocellular Carcinoma

Overall Rates (a)

Adjusted Rates cb)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

Cochran-Armitage Trend Test (d)

Fisher Exact Test (d)

Liver: Hepatocellular Adenoma or Carcinoma

Overall Rates (a)

Adjusted Rates ( b )

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

Cochran-Armitage Trend Test(d)

Fisher Exact Test (d )

4/49(8%)

0/48(0%)

1/49(2%)

13.3%

0.0%

3.0%

3/28(11%)

0/28(0%)

1/33(3%)

688

730

P = 0.062N

P = 0.065N

P = 0.138N

P = 0.059N

P = 0.061N

P = 0.132N

P = 0.079N

P = 0.061N

P = 0.181N

it Pheochromocytoma

6/49(12%)

0/48(0%)

1/49(2%)

19.7%

0.0%

3.0%

4/28(14%)

0/28(0%)

1/33(3%)

688

730

P = 0.010N

P = 0.018N

P = 0.038N

P = 0.009N

P = 0.016N

P = 0.034N

P = 0.015N

P = 0.014N

P = 0.056N

2/50(4%)

3/50(6%)

2/50(4%)

7.1%

9.9%

5.5%

2/28(7%)

2/29(7%)

1/34(3%)

730

723

694

P = 0.515N

P = 0.522

P = 0.624N

P = 0.518N

P = 0.521

P = 0.626N

P = 0.592

P = 0.500

P = 0.691

2/50(4%)

5/50(10%)

4/50(8%)

7.1%

17.2%

11.4%

2/28(7%)

5/29(17%)

3/34(9%)

730

730

729

P = 0.380

P = 0.226

P = 0.434

P = 0.400

P = 0.225

P = 0.448

P = 0,278

P = 0.218

P = 0.339

9/50(18%)

7/50(14%)

7/50(14%)

26.6%

20.2%

17.3%

5/28(18%)

4/29(14%)

3/34(9%)

611

496

602

P = 0.220N

P = 0.380N

P = 0.263N

P = 0.260N

P = 0.362N

P = 0.297N

P = 0.338N

P = 0.393N

P = 0.393N

10/50(20%)

12/50(24%)

11/50(22%)

29.8%

36.2%

27.4%

6/28(21%)

9/29(31%)

6/34(18%)

611

496

602

P = 0.449N

P = 0.433

P = 0.503N

P = 0.499N

P = 0.445

P = 0.551N

P = 0.450

P = 0.405

P = 0.500

Nitrofurantoin, NTP TR 341

154

TABLE C3. ANALYSIS OF PRIMARY TUMORS IN MALE MICE IN THE TWO-YEAR FEED STUDY

OF NITROFURANTOIN (Continued)

Control

1.300 ppm

2,300 ppm

Lung: Alveolar/Bronchiolar Adenoma

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

Cochran-Armitage Trend Test (d)

Fisher Exact Test (d)

Lung: Alveolar/Bronchiolar Carcinoma

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

Cochran-Armitage Trend Test (d)

Fisher Exact Test (d)

Lung: Alveolar/Bronchiolar Adenoma or Carcinoma

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Tab le Tests ( d )

Logistic Regression Tests ( d )

Cochran-Armitage Trend Test (d)

Fisher Exact Test (d)

Skin: Fibroma

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

Cochran-Armitage Trend Test (d)

Fisher Exact Test (d)

Skin: Fibrosarcoma

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

Cochran-Armitage Trend Test (d)~

Fisher Exact Test (d)

Skin: Fibroma or Fibrosarcoma

Overall Rates (a 1
Adjusted Rates (b)
Terminal Rates (c)
Day of First Observation
Life Table Tests (d)
Logistic Regression Tests (d)
Cochran-Armitage Trend Test(d)
Fisher Exact Test (d)

5/50(10%)

2/50(4%)

4/50(8%)

15.0%

6.4%

11.8%

2/28(7%)

1/29(3%)

4/34(12%)

611

699

730

P = 0.328N

P = 0.214N

P = 0.394N

P = 0.325N

P = 0.200N

P = 0.397N

P = 0.417N

P = 0.218N

P = 0.500N

1/50(2%)

2/50(4%)

3/50(6%)

3.6%

6.9%

7.7%

1/28(4%)

2/29(7%)

1/34(3%)

730

730

602

P = 0.296

P = 0.512

P = 0.384

P = 0.280

P = 0.513

P = 0.352

P = 0.225

P = 0.500

P = 0.309

}ma

6/50(12%)

4/50(8%)

7/50(14%)

18.3%

13.1%

18.9%

3/28(11%)

3/29(10%)

5/34(15%)

611

699

602

P = 0.549N

P = 0.356N

P = 0.592N

P = 0.560N

P = 0.343N

P = 0.618N

P = 0.448

P = 0.370N

P = 0.500

1/50(2%)

2/50(4%)

3/50(6%)

2.9%

6.9%

8.4%

0/28(0%)

2/29(7%)

2/34(6%)

688

730

695

P = 0.285

P = 0.505

P = 0.366

P = 0.287

P = 0.513

P = 0.367

P = 0.225

P = 0.500

P = 0.309

9/50(18%)

5/50(10%)

12/50(24%)

25.8%

12.8%

27.0%

4/28(14%)

0/29(0%)

4/34(12%)

408

335

575

P = 0.429

P = 0.192N

P = 0.495

P = 0.269

P = 0.190N

P = 0.336

P = 0.269

P = 0.194N

P = 0.312

10/50(20%)

7/50(14%)

14/50(28%)

28.0%

18.8%

31.9%

4/28(14%)

2/29(7%)

6/34(18%)

408

335

575

P = 0.372

P = 0.289N

P = 0.433

P = 0.229

P = 0.285N

P = 0.282

P = 0.204

P = 0.298N

P = 0.241

155

Nitrofurantoin, NTP TR 341

TABLE C3.

ANALYSIS OF PRIMARY TUMORS IN MALE MICE IN THE TWO-YEAR FEED STUDY
OF NITROFURANTOIN (Continued)

Control

1,300 ppm

2,500 ppm

Skin: Malignant Schwannoma

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

CochranArmitage Trend Test(d)

Fisher Exact Test(d)

Circulatory System: Hemangiosarcoma

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table TesU(d)

Logistic Regression Tests (d)

CochranArmitage Trend Test (d)

Fisher Exact Test (d)

Hematopoietic System: Lymphoma, All Malignant

Overall Rates(a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

CochranArmitage Trend Test(d)

Fisher Exact Test (d)

3/50(6%)

0/50(0%)

0/50(0%)

8.8%

0.0%

0.0%

1/28(4%)

0/29(0%)

0/34(0%)

533

P = 0.029N

P = 0.119N

P = 0.096N

P = 0.033N

P = 0.117N

P = 0.114N

P = 0.036N

P = 0.12IN

P = 0.121N

3/50(6%)

1/50(2%)

0/50(0%)

9.6%

2.3%

0.0%

2/28(7%)

0/29(0%)

0/34(0%)

611

530

P = 0.047N

P = 0.293N

P = 0.092N

P = 0.057N

P = 0.298N

P = 0.099N

P = 0.061N

P = 0.309N

P = 0.121N

It

5/50(10%)

3/50(6%)

6/50(12%)

16.4%

9.0%

15.3%

3/28(11%)

1/29(3%)

3/34(9%)

697

605

527

P = 0.567

P = 0.336N

P = 0.614N

P = 0.535

P = 0.333N

P = 0.601

P = 0.444

P = 0.357N

P = 0.500

(a ) Number of tumor-bearing animals/number of animals examined at the site

(b) Kaplan- Meier estimated tumor incidences at the end of the study after adjusting for intercurrent mortality

(c) Observed tumor incidence at terminal kill

(d) Beneath the control incidence are the P values associated with the trend test. Beneath the dosed group incidence are the
P values corresponding to pairwise comparisons between that dosed group and the controls. The life table analysis regards tu-
mors in animals dying prior to terminal kill as being (directly or indirectly) the cause of death. The logistic regression test re-
gards these lesions as nonfatal. The CochranArmitage and Fisher exact tests compare directly the overall incidence rates.
A negative trend or lower incidence in a dosed group is indicated by (N).

Nitrofurantoin, NTP TR 341

156

TABLE C4. HISTORICAL INCIDENCE OF ADRENAL MEDULLARY PHEOCHROMOCYTOMAS IN MALE

B6C3Fi MICE RECEIVING NO TREATMENT (a)

Study Incidence in Controls

Historical Incidence at Southern Research Institute

HC Blue No. 2 0/50

C.I. Disperse Blue 1 1/49

D-Mannitol 0/49

Ziram 0/49

Eugenol 0/43

Propyl gallate 1/49

Zearalenone 0/50

HC Blue No. 1 2/49

Stannous chloride 0/49

TOTAL 4/437(0.9%)

SD(b) 1.48%

Range (c)

High 2/49

Low 0/50

Overall Historical Incidence

TOTAL (d) 25/1,962 (1.3%)

SD(b) 1.78%

Range (c)

High 3/49

Low 0/50

(a) Data as of August 7, 1986, for studies of at least 104 weeks

(b) Standard deviation

(c) Range and SD are presented for groups of 35 or more animals.

(d) Includes one malignant pheochromocytoma

157 Nitrofurantoin, NTP TR 341

TABLE C5. SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE MICE IN THE

TWO-YEAR FEED STUDY OF NITROFURANTOIN

Untreated Control

Low

Dose

High Dose

Animals initially in study

50

50

50

Animals removed

50

50

50

Animals examined histopathologically

50

50

50

ALIMENTARY SYSTEM

Gallbladder

(441

(45)

(37)

Hyperplasia

1

(3%)

Intestine large, rectum

(481

(47)

(50)

Cyst

2

(4%)

1

(2%)

Hyperplasia, focal

1

(2%)

Inflammation, chronic, focal

1

(2%)

Inflammation, subacute

2

(4?o)

1

(2%)

Perforation

1

(2%)

Prolapse

3

(6%)

1

(2%)

2

(4%)

Intestine small, jejunum

(49)

(45)

(49)

Cyst

1

(2%)

Liver

(50)

(50)

(50)

Basophilic focus

I

(2%)

Cyst

1

(2%)

Fibrosis, focal

1

(2%)

Granuloma, multiple

1

(2%)

Hematopoietic cell proliferation

1

(2%)

Mixed cell focus

2

(4%)

Necrosis, multifocal

1

(2%)

1

(2%)

1

(2%)

Thrombus

1

(2%)

Mesentery

(3)

(1)

(2)

Inflammation, subacute, focal

1

(33%)

Inflammation, suppurative, acute, multifocal

2

(67%)

Thrombus

1

(33%)

Stomach, forestomach

(49)

(48)

(49)

Cyst

1

(2%)

1

(2%)

Hyperkeratosis

1

(2%)

Hyperplasia

2

(4%)

1

(2%)

Inflammation, chronic, focal

1

(2%)

Inflammation, suppurative, acute, focal

1

(2%)

Mineralization

1

(2%)

Stomach, glandular

(50)

(48)

(49)

Mineralization

1

(2%)

Tooth

(10)

(12)

(4)

Dysplasia

10

(100%)

12

(100%)

4

(100%)

CARDIOVASCULAR SYSTEM

Heart

(50)

(50)

(50)

Inflammation, acute, multifocal

1

(2%)

Inflammation, subacute, multifocal

2

(4%)

Thrombus

1

(2%)

ENDOCRINE SYSTEM

Adrenal gland, cortex
Hyperplasia, focal
Spindle cell, hyperplasia
Spindle cell, hyperplasia, focal

Adrenal gland, medulla
Hyperplasia
Hyperplasia, focal

Pituitary gland

Pars distalis, cyst

(50)

1 (2%)
5 (10%)

(49)
3 (6%)
1 (2%)

(46)

(49)

3 (6%)

(48)

6 (13%)

(48)

(50)

2

(4%)

3

(6%)

I

(2%)

(49)

5

(10%)

(43)

1

(2%)

Nitrofurantoin, NTP TR 341

158

TABLE C5. SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE MICE IN THE
TWO-YEAR FEED STUDY OF NITROFURANTOIN (Continued)

Untreated Control

Low Dose

High Dose

ENDOCRINE SYSTEM (Continued)
Thyroid gland

Hyperplasia, cystic
Follicle, cyst
Follicle, degeneration
Follicle, hyperplasia, cystic

(48)

1 (2%)

1 (2%)

(47)

1 (2%)

1 (2%)

2 (4%)

(48)

3 (6%)

1 (2%)

2 (4%)

GENERAL BODY SYSTEM
None

GENITAL SYSTEM

Epididymis

(50)

(49)

(50)

Atypical cells

26

(52%)

Depletion

1

(2%)

1

(2%)

15

(30%)

Granuloma sperm

3

(6%)

Penis

(2)

(8)

(1)

Developmental malformation

2

(100%)

8

(100%)

1

(100%)

Preputial gland

(9)

(2)

(4)

Inflammation, subacute

5

(56%)

Inflammation, suppurative, acute

1

(11%)

Duct, cyst

6

(67%)

2

(100%)

4

(100%)

Prostate

(49)

(50)

(50)

Inflammation, chronic

1

(2%)

1

(2%)

Inflammation, suppurative, acute

4

(8%)

4

(8%)

Seminal vesicle

(6)

(4)

(1)

Dilatation

3

(50%)

4

(100%)

1

(100%)

Inflammation, chronic

1

(17%)

1

(25%)

Inflammation, suppurative, acute

2

(33%)

Testes

(49)

(49)

(50)

Aspermatogenesis

1

(2%)

1

(2%)

16

(32%)

Atrophy

1

(2%)

Mineralization

1

(2%)

1

(2%)

Germinal epithelium, degeneration

3

(6%)

23

(46%)

Seminiferous tubule, dilatation, focal

1

(2%)

HEMATOPOIETIC SYSTEM

Blood

(2)

Anemia

1

(50%)

Lymph node

(50)

(49)

(49)

Axillary, hyperplasia

1

(2%)

Deep cervical, hyperplasia

1

(2%)

Iliac, hyperplasia

1

(2%)

1

(2%)

Inguinal, hyperplasia

1

(2%)

Lymph node, mesenteric

(47)

(46)

(44)

Angiectasis

6

(13%)

Ectasia

1

(2%)

Hyperplasia

1

(2%)

Thrombus

1

(2%)

Spleen

(50)

(49)

(50)

Angiectasis

1

(2%)

1

(2%)

Atrophy

3

(6%)

1

(2%)

Hematopoietic cell proliferation

10

(20%)

U

(22%)

4

(8%)

Necrosis, focal

1

(2%)

Thymus

(44)

(43)

(41)

Cyst

I

(2%)

1

(2%)

159

Nitrofurantoin, NTP TR 341

TABLE C5. SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE MICE IN THE
TWO-YEAR FEED STUDY OF NITROFURANTOIN (Continued)

Untreated Control

Low Dose

High Dose

INTEGUMENTARY SYSTEM
Skin

Alopecia

Alopecia, multifocal

Fibrosis

Fungus

Hyperplasia, focal

Inflammation, chronic

Inflammation, granulomatous, focal

Inflammation, subacute, focal

Inflammation, suppurative, acute

Mineralization, focal

Ulcer

(50)
4
3
7

(8%)
(6%)
(14%)

1 (2%)

7 (14%)
2 (4%)
1 (2%)

(50)

5 (10%)

5

(10%)

1

(2%)

1

(2%)

2

(4%)

12

(24%)

2

(4%)

1

(2%)

4

(8%)

(50)
3 (6%)

12 (24%)

1 (2%)

2 (4%)

1 (2%)

9 (18%)

1 (2%)

2 (4%)
1 (2%)

MUSCULOSKELETAL SYSTEM
Bone

Cranium, hyperostosis

(50)

1 (2%)

(50)

(50)

NERVOUS SYSTEM
None

RESPIRATORY SYSTEM

Lung

Congestion

Fibrosis, multifocal

Infiltration cellular, histiocytic, multifocal

Inflammation, subacute, multifocal

Alveolar epithelium, hyperplasia

Nose

Hemorrhage

(50)
3

32 (64%)
1 (2%)
(42)

(50)

(50)

1 (2%)

1 (2%)

34 (68%)

30 (60%)

2 (4%)

14)

(48)

1 (2%)

SPECIAL SENSES SYSTEM
None

URINARY SYSTEM
Kidney

Calculus gross observation

Hydronephrosis

Inflammation, suppurative, acute, multifocal

Metaplasia, osseous, focal

Nephropathy

Cortex, infarct

Cortex, infarct, focal

Medulla, mineralization

Papilla, necrosis

Renal tubule, degeneration, multifocal

Renal tubule, dilatation

Renal tubule, hyperplasia

Renal tubule, hyperplasia, focal

Renal tubule, necrosis, multifocal

Renal tubule, nephropathy
Ureter

Inflammation, suppurative, acute

(50)

1

(2%)

1

(2%)

1

(2%)

1

(2%)

3

(6%)

1

(2%)

1

(2%)

1 (2%)
(1)
1 (100%)

(50)

2 (4%)

1 (2%)

1 (2%)

1 (2%)

1 (2%)

(50)

1 (2%)

1 (2%)

17 (34%)

1 (2%)

14 (28%)

1 (2%)

Nitrofurantoin, NTP TR 341

160

TABLE C5. SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN MALE MICE IN THE
TWO-YEAR FEED STUDY OF NITROFURANTOIN (Continued)

Untreated Control Low Dose High Dose

URINARY SYSTEM (Continued)
Urethra

Inflammation, suppurative, subacute

Bulbourethral gland, dilatation
Urinary bladder (50) (49) (50)

Calculus gross observation

Calculus micro observation only

Inflammation, chronic 1 (2%)

Inflammation, suppurative, acute

Transitional epithelium, hyperplasia

(2)

1

(50%)

I

(50%)

(50)

1

(2%)

1

(2%)

2

(4%)

2

(4%)

161 Nitrofurantoin, NTP TR 341

Nitrofurantoin, NTP TR 341 162

APPENDIX D

SUMMARY OF LESIONS IN FEMALE MICE IN

THE TWO-YEAR FEED STUDY OF NITROFURANTOIN

PAGE

TABLE Dl SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICE IN THE

TWO-YEAR FEED STUDY OF NITROFURANTOIN 165

TABLE D2 INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE MICE IN THE TWO-YEAR

FEED STUDY OF NITROFURANTOIN 170

TABLE D3 ANALYSIS OF PRIMARY TUMORS IN FEMALE MICE IN THE TWO-YEAR FEED

STUDY OF NITROFURANTOIN 182

TABLE D4a HISTORICAL INCIDENCE OF OVARIAN TUMORS IN FEMALE B6C3Fi MICE

RECEIVING NO TREATMENT 185

TABLE D4b HISTORICAL INCIDENCE OF HEMATOPOIETIC SYSTEM TUMORS IN FEMALE

B6C3Fi MICE RECEIVING NO TREATMENT 186

TABLE D4c HISTORICAL INCIDENCE OF UTERINE GLANDULAR TUMORS IN FEMALE B6C3Fi

MICE RECEIVING NO TREATMENT 187

TABLE D4d HISTORICAL INCIDENCE OF HEPATOCELLULAR TUMORS IN FEMALE B6C3Fi

MICE RECEIVING NO TREATMENT 188

TABLE D4e HISTORICAL INCIDENCE OF LIVER ITO CELL TUMORS IN FEMALE B6C3Fi MICE

RECEIVING NO TREATMENT 188

TABLE D5 SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE

IN THE TWO YEAR FEED STUDY OF NITROFURANTOIN 189

163

Nitrofurantoin, NTP TR 341

Nitrofurantoin, NTP TR 341 1 64

TABLE Dl. SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICE IN THE TWO-YEAR

FEED STUDY OF NITROFURANTOIN

Untreated Control

Low Dose

High Dose

Animals initially in study

Animals removed

Animals examined histopathologically

50
50
50

50
50
50

50
50
50

(44)

(2%)

(2%)

(50)

(2%)

(2%)

(2%)

(2%)

1

i2%i

5

(10%)

ALIMENTARY SYSTEM

Gallbladder (41)

Lymphoma malignant histiocytic
Intestine large, cecum (43)

Lymphoma malignant histiocytic
Intestme small, ileum

Lymphoma malignant lymphocytic
Intestine small, jejunum

Lymphoma malignant lymphocytic
Liver

Hemangioma

Hemangiosarcoma

Hepatocellular carcinoma

Hepatocellular adenoma

Itocell tumor, NOS

Lymphoma malignant histiocytic

Lymphoma malignant lymphocytic

Lymphoma malignant mixed
Mesentery '(50)

Fibrosarcoma

Lymphoma malignant histiocytic

Lymphoma malignant lymphocytic 3

Sarcoma
Pancreas (47)

Fibrosarcoma, metastatic, mesentery

Lymphoma malignant histiocytic

Lymphoma malignant lymphocytic

Sarcoma
Salivary glands (49)

Lymphoma malignant histiocytic

Lymphoma malignant lymphocytic 1

Stomach (49)

Squamous cell carcinoma 1

Stomach, forestomach (49)

Lymphoma malignant lymphocytic

Papilloma squamous 1 (2%)

Sarcoma

Glandular, fibrosarcoma, metastatic, mesentery
Stomach, glandular l49)

Sarcoma
Tongue *(50)

Lymphoma malignant lymphocytic 1

(6%)

(2%)
(2%)

(39)
(49)
(47)
(48)
(50)

1 (2%)

1 (2%)

1 (2%)

4 (8%)

3 (6%)

3 (6%)
•(50)

1 (2%)
(50)

1 (2%)
1 (2%)
(50)

(49)
(49)

1 (2%)

(49)

1 (2%)
♦(50)

(42)

1

(2%)

(50)

1

(2%)

(50)

(50)

(50)

2

(4%)

7

(14%)

1

(2%)

6

(12%)

4

(8%)

■(50)

1

(2%)

1

(2%)

3

(6%)

1

(2%)

(50)

1

(2%)

2

(4%)

1

(2%)

(50)

1

(2%)

(50)

(50)
1
1

1
(49)

♦(50)

(2%)
(2%)

(2%)

(2%)

CARDIOVASCULAR SYSTEM
Heart

Hemangiosarcoma

Lymphoma malignant lymphocytic

Sarcoma

(50)

1 (2%)
1 (2%)

(50)

1 (2%)

(50)

165

Nitrofurantoin, NTP TR 341

TABLE Dl. SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICE IN THE TWO YEAR

FEED STUDY OF NITROFURANTOIN (Continued)

Untreated Control

Low Dose

High Dose

ENDOCRINE SYSTEM
Adrenal gland

Fibrosarcoma, metastatic, mesentery
Adrenal gland, cortex

Adenoma

Osteosarcoma, metastatic, bone

Spindle cell, adenoma
Adrenal gland, medulla

Hepatocellular carcinoma, metastatic, liver

Pheochromocytoma malignant

Pheochromocytoma benign
Islets, pancreatic

Carcinoma
Pituitary gland

Pars distalis, adenoma
Thyroid gland

Lymphoma malignant lymphocytic

Follicular cell, adenoma

Follicular cell, carcmoma

(50)
(50)

(48)

1

(2%)

1

(2%)

1

(2%)

(47)

(48)

3

(6%)

(48)

1

(2%)

3

(6%)

1

(2%)

(50)

(50)

1 (2%)
1 (2%)
(49)

(50)

(44)

1 (2%)
(49)

1 (2%)

(50)

1 (2%)
(50)

1 (2%)

(49)

(49)

1 (2%)
(49)

2 (4%)
(50)

GENERAL BODY SYSTEM
None

GENITAL SYSTEM
Ovary

Adenoma, tubular

Cystadenoma

Cystadenoma, papillary

Granulosa cell tumor malignant

Granulosa cell tumor benign

Hemangioma

Lymphoma malignant histiocytic

Lymphoma malignant lymphocytic

Mixed tumor benign

Neoplasm, NOS
Uterus

Adenocarcinoma

Lymphoma malignant histiocytic

Lymphoma malignant lymphocytic

Lymphoma malignant mixed

Polyp stromal

Sarcoma stromal

Cervix, lymphoma malignant mixed

(50)

2 (4%)

(50)

1 (2%)

2 (4%)

(50)

1 (2%)

3 (6%)

1 (2%)

1 (2%)

(50)

1 (2%)

2 (4%)

1 (2%)

2 (4%)

1 (2%)

(50)

5

(10%)

1

(2%)

I

(2%)

1

(2%)

1

(2%)

3

(6%)

5

(10%)

4

(8%)

1

(2%)

(50)

1

(2%)

2

(4%)

1

(2%)

2

(4%)

HEMATOPOIETIC SYSTEM

Bone marrow (50)

Lymphoma malignant lymphocytic 3 (6%)

Lymph node (50)

Axillary, lymphoma malignant histiocytic
Bronchial, lymphoma malignant histiocytic
Bronchial, lymphoma malignant lymphocytic 1 (2%)

Bronchial, lymphoma malignant mixed
Deep cervical, lymphoma malignant lymphocytic 1 (2%)
Iliac, lymphoma malignant lymphocytic

(50)

(50)

1

(2%)

1
1

(2%)
(2%)

(50)

(49)

1 (2%)
1 (2%)
3 (6%)

1 (2%)

Nitrofurantoin, NTP TR 341

166

TABLE Dl. SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICE IN THE TWO-YEAR

FEED STUDY OF NITROFURANTOIN (Continued)

Untreated Control

Low Dose

High Dose

HEMATOPOIETIC SYSTEM

Lymph node (Continued)

Iliac, lymphoma malignant mixed
Inguinal, lymphoma malignant histiocytic
Inguinal, lymphoma malignant lymphocytic
Inguinal, lymphoma malignant mixed
Lumbar, lymphoma malignant lymphocytic
Mediastinal, lymphoma malignant histiocytic
Mediastinal, lymphoma malignant lymphocytic
Mediastinal, lymphoma malignant mixed
Pancreatic, lymphoma malignant lymphocytic
Pancreatic, lymphoma malignant mixed
Renal, lymphoma malignant histiocytic
Renal, lymphoma malignant lymphocytic
Renal, lymphoma malignant mixed

Lymph node, mandibular

Lymphoma malignant histiocytic
Lymphoma malignant lymphocytic
Lymphoma malignant mixed

Lymph node, mesenteric

Fibrosarcoma, metastatic, mesentery
Lymphoma malignant histiocytic
Lymphoma malignant lymphocytic
Lymphoma malignant mixed

Spleen

Fibrosarcoma, metastatic, mesentery

Hemangiosarcoma

Lymphoma malignant histiocytic

Lymphoma malignant lymphocytic

Lymphoma malignant mixed

Thymus

Lymphoma malignant histiocytic

Lymphoma malignant lymphocytic

Lymphoma malignant mixed

Mediastinum, lymphoma malignant lymphocytic

(50)

(50)
2

(4%)

(49)

1

1
1
1

(2%)

(2%)
(2%)
(2%)

1

(2%)

1
4

(2%)
(8%)

1

(2%)

1

(2%)

2 (4%)

2

1

(4%)
(2%)

I (2%)

1 (2%)

1
1

(2%)
(2%)

(50)

(50)

(45)

1 (2%)

2

(4%)

3

(7%)

7 (14%)

6

(12%)

12

(27%)

2

(4%)

I

(2%)

(45)

(45)

(48)

1

(2%)

2

(4%)

4

(8%)

5 (11%)

2

(4%)

10

(21%)

1

(2%)

1

(2%)

(49)

(50)

(50)

1

(2%)

1 (2%)

1 (2%)

3

(6%)

4

(8%)

9 (18%)

10

(20%)

16

(32%)

3

(6%)

1

(2%)

(471

(48)

(48)

1 (2%)

2

(4%)

8 (17%)

2
2

(4%)
(4%)

2

(4%)

1 (2%)

INTEGUMENTARY SYSTEM
Mammary gland
Adenoacanthoma
Adenocarcinoma
Adenocarcinoma, multiple
Skin

Basal cell carcinoma

Lymphoma malignant lymphocytic

Squamous cell carcinoma

Sebaceous gland, adenoma

Subcutaneous tissue, fibrosarcoma

Subcutaneous tissue, hemangiosarcoma

Subcutaneous tissue, schwannoma benign

(50)

(48)

5 (10%)

3 (6%)

(50)

(50)

1 (2%)

1 (2%)

1 (2%)

1 (2%)

1 (2%)

(48)

1 (2%)

1 (2%)
(50)

1 (2%)

1 (2%)
1 (2%)

MUSCULOSKELETAL SYSTEM

Bone

Osteosarcoma

Skeletal muscle

Fibrosarcoma, metastatic, mesentery
Lymphoma malignant histiocytic
Lymphoma malignant lymphocytic
Sarcoma

(50)
•(50)

1 (2%)

(50)

1 (2%)
♦(50)

1 (2%)

(50)

•(50)

1 (2%)
1 (2%)

167

Nitrofurantoin, NTP TR 341

TABLE Dl. SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICE IN THE TWO-YEAR

FEED STUDY OF NITROFURANTOIN (Continued)

Untreated Control

Low Dose

High Dose

NERVOUS SYSTEM
Brain

Lymphoma malignant lymphocytic
Meningioma benign

(49)

1 (2%)

(50)

1 (2%)

(50)

1 (2%)
1 (2%)

RESPIRATORY SYSTEM
Lung

Adenocarcinoma, metastatic, mammary gland

Alveolar/bronchiolar adenoma

Alveolar/bronchiolar carcinoma

Basal cell carcinoma, metastatic, skin

Hepatocellular carcmoma, metastatic, liver

Lymphoma malignant histiocytic

Lymphoma malignant lymphocytic

Lymphoma malignant mixed

Osteosarcoma, metastatic, bone

Sarcoma

Mediastinum, lymphoma malignant lymphocytic

Mediastinum, sarcoma

(50)

2
1

1
1
3

(4%)
(2%)

(2%)
(2%)
(6%)

3 (6%)

(50)

1 (2%)

2 (4%)

2 (4%)
4 (8%)
1 (2%)
1 (2%)
1 (2%)

1 (2%)

(50)

1 (2%)
1 (2%)

5 (10%)

6 (12%)

SPECIAL SENSES SYSTEM
Harderian gland
Adenocarcinoma
Adenoma

•(50)

1 (2%)
1 (2%)

•(50)

1 (2%)

2 (4%)

•(50)

1 (2%)

URINARY SYSTEM
Kidney

Lymphoma m&lignant histiocytic
Lymphoma malignant lymphocytic
Lymphoma malignant mixed
Sarcoma

Renal tubule, carcinoma
Urinary bladder

Lymphoma malignant lymphocytic

(50)

3 (6% I

(50)

1 (2%)

(50)

(50)

2

(4%)

5 (10%

5

(10%)

5 (10%

1

(2%)

1 (2%)

1

(2%)

1 (2%)

(50)

(50)

3 (6%)

SYSTEMIC LESIONS
Multiple organs
Hemangioma

Lymphoma malignant lymphocytic
Lymphoma malignant histiocytic
Hemangiosarcoma
Lymphoma malignant mixed

'(50)

*(50)

•(50)

1 (2%)

1 (2%)

1 (2%)

11 (22%)

10 (20%)

17 (34%)

1 (2%)

5 (10%)

6 (12%)

2 (4%)

4 (8%)

1 (2%)

ANIMAL DISPOSITION SUMMARY
Animals initially in study
Dead
Moribund
Terminal sacrifice

50
19
12
19

50
5
9

36

50

2

11

37

Nitrofurantoin, NTP TR 341

168

TABLE Dl.

SUMMARY OF THE INCIDENCE OF NEOPLASMS IN FEMALE MICE IN THE TWO-YEAR
FEED STUDY OF NITROFURANTOIN (Continued)

Untreated Control

Low Dose

High Dose

TUMOR SUMMARY

Total animals with primary neoplasms ** 30

Total primary neoplasms 42

Total animals with benign neoplasms 15

Total benign neoplasms 17

Total animals with malignant neoplasms 21

Total malignant neoplasms 25

Total animals with secondary neoplasms *** I

Total secondary neoplasms 2
Total animals with neoplasms-
uncertain benign or malignant

Total uncertain neoplasms

35
55
15
17
27
37
2
3

1

1

41
67
22
27
31
38
3
8

2
2

* Number of animals receiving complete necropsy examination; all gross lesions including masses examined microscopically.

*• Primary tumors: all tumors except secondary tumors

*•* Secondary tumors: metastatic tumors or tumors invasive into an adjacent organ

169

Nitrofurantoin, NTP TR 34!

TABLE D2.

INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE .MICE IN THE TWO YEAR FEED
STUDY OF NITROFURANTOIN: UNTREATED CONTROL

WEEKS ON
STUDY

CARCASS

ID

ALIMENTARY SYSTEM

Esophagus

Gallbladder
Intestine large
Intestine large, cecum
Intestine large, colon
Intestine large, rectum
Intestine small
Intestine small, duodenum
Intestine small, ileum

Lymphoma malignant lymphocytic
Intestine small, jejunum

Lymphoma malignant lymphocytic
Liver

Hemangioma

Hemangiosarcoma

Hepatocellular carcinoma

Hepatocellular adenoma

Lymphoma malignant histiocytic

Lymphoma malignant lymphocytic
Mesentery

Lymphoma malignant lymphocytic
Pancreas
Salivary glands

Lymphoma malignant lymphocytic
Stomach

Squamous cell carcinoma
Stomach, forestomach

Papilloma squamous
Stomach, glandular
Tongue

Lymphoma malignant lymphocytic

CARDIOVASCULAR SYSTEM

Heart

Hemanposarcoma

Lymphoma malignant lymphocytic

ENDOCRINE SYSTEM

Adrenal gland
Adrenal gland, corten
Adrenal gland, medulla

Hepatocellular carcinoma, metastatic,
liver

Pheochromocytoma malignant

Pheochromocytoma benign
Islets, pancreatic
Parathyroid gland
Pituitary gland

Pars distalis. adenoma
Thyroid gland

Lymphoma malignant lymphocytic

Follicular cell, adenoma

Follicular cell, carcinoma

GENERAL BODY SYSTEM
None

GENITAL SYSTEM
Clitoral gland
Ovary

Cystadenoma. papillary
Uterus

Lymphoma malignant lymphocytic

Polyp stromal

+ ; Tissue examined microscopically

Not examined
- : Present but not examined microscopically
I: Insufficient tissue

0 0
2 6
2 4

0 0 0
9 9 9

1 3 4

0 1 1.
9 0 0
9 0 2

"3 T

3 0

1 1

"T — T"
8 0
2 2

T T

7 7
I 2

~i 5 3 3 3~
0 9 6 9 9
3 112 3

"3 — 3 — 3 — 3 — 3~
4 5 5 17
12 3 15

"3 — 3 — T-
9 1 2
4 2 2

M +
M M

M M

A +

A +

A +

A +

A +

A -1-

A +

M

+
M
-♦-
M

+ + + +

+ + -F + +
+ + + <- +

+ + + 4- +
-I- + + -t- +

+ +

+ + + +

+ + + +

+ + + +

+ + + +

+ f •*-+■ + + +

A A +

M + M

+ + + -1- + + +A
M + + +MM + +

+ + + +- + + >• +

+ + + + + + + +-

+ +f-f + +- + +-

+ 4- + -t-+--f- + -t-

+ +--t- + + -(- + +

X

M, Missing

A; Autolysis precludes examination

X: Incidence of listed morphology

Nitrofurantoin, NTP TR 341

170

TABLE D2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE MICE: UNTREATED CONTROL

(Continued)

WEEKS ON
STUDY

CARCASS
ID

ALIMENTARY SYSTEM

Esopbaeus
Gallbladder
Intestine large
Intestine large, cecum
Intestine large, colon
Intestine large, rectum
Intestine small
Intestine small, duodenum
Intestine small, ileum

Lymphoma malignant lymphocytic
Intestine small, jejunum

Lymphoma malignant lymphocytic
Liver

Hemangioma

Heraangiosarcoma

Hepatocellular carcinoma

Hepatocellular adenoma

Lymphoma malignant histiocytic

Lymphoma malignant lymphocytic
Mesentery

Lymphoma malignant lymphocytic
Pancreas
Salivary glands

Lymphoma malignant lymphocytic
Stomach

Squamous cell carcinoma
Stomach, forestomach

Papilloma squamous
Stomach, glandular
Tongue

Lymphoma malignant lymphocytic

C.JiRDIOVASCULAR SYSTEM

Heart
Hemangiosarcoma

Lymphoma malignant lymphocytic

ENDOCRINE SYSTEM

Adrenal gland
Adrenal gland, cortex
Adrenal gland, medulla

Hepatocellular carcinoma, metastatic,
liver

Pheochromocytoma malignant

Pheochromocytoma benign
Islets, pancreatic
Parathyroid gland
Pituitary gland

Pars distahs, adenoma
Thyroid gland

Lymphoma malignant lymphocytic

Follicular cell, adenoma

Follicular cell, carcinoma

GENERAL BODY SYSTEM

None

GENITAL SYSTEM
Clitoral gland
Ovary

Cystadenoma, papillary
Uterus

Lymphoma malignant lymphocytic

Polyp stromal

1 1
0 0
5 5

1 1
0 0
5 5

1 1 1 1 1 I 1 1 1 1
0000000000
5555555555

M +

+■ +

+ +
+ +

+ +

X

+
X

+
+

M

M
M

+
M

M
+
X

3

3

3

3

3

3

3

T"

3

3

3

~3~

~T-

3

3

IT

3

3

"T"

3

"TT

~T-

T-

4

4

it

1

5

8

S

1

1

2

2

2

3

3

3

4

4

4

S

6

6

6

6

8

0

0

■i

b

3

4

3

4

4

5

3

4

5

3

4

5

3

4

5

5

2

3

4

5

5

4

5

TOTAL:
TISSUES
TUMORS

49

41

48

43

46

46

47

47

44

1

45

1

50

1

1

1

1

1

5

21

3

47

49

I

49

1

49

1

49

1

1

50
1
1

50
50
48

1

1

1

47

43

48

3

48

1

3

I

1
50

2
50

1

2

171

Nitrofurantoin, NTP TR 341

TABLE D2.

INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE MICE: UNTREATED CONTROL

(Continued)

WEEKS ON
STUDY

CARCASS
ID

HEMATOPOIETIC SYSTEM
Bone marrow

Lymphoma malignant lymphocytic
Lympn node

Bronchial, lymphoma malignant
lymphocytic

Deep cervical, lymphoma malignant
lymphocytic

Pancreatic, lymphoma malignant
lymphocytic

Renal, lymphoma malignant histiocytic

Renal, lymphoma malignant lymphocytic
Lymph node, mandibular

Lymphoma malignant histiocytic

Lymphoma malignant lymphocytic
Lymph node, mesentenc

Lymphoma malignant lymphocytic
Spleen

Hemangiosarcoma

Lymphoma malignant histiocytic

Lymphoma malignant lymphocytic
Thymus

Lymphoma malignant histiocytic

Lymphoma malignant lymphocytic

INTEGUMENTARY SYSTEM

Mammary gland
Adenocarcinoma

Skin
Lymphoma malignant lymphocytic
Subcutaneous tissue, hemangiosarcoma

MUSCULOSKELETAL SYSTEM

Bone

Skeletal muscle
Lymphoma malignant lymphocytic

NERVOUS SYSTEM

Brain
Lymphoma malignant lymphocytic

RESPIRATORY SYSTEM

Lung
Alveolar/bronchiolar adenoma
Alveolar/bronchiolar carcinoma
Hepatocellular carcinoma, metastatic,

liver
Lymphoma malignant histiocytic
Lymphoma malignant lymphocytic
Mediastinum, lymphoma malignant
lymphocytic

Nose

Trachea

SPECIAL SENSES SYSTEM
Hardenan gland

Adenocarcinoma

Adenoma

URINARY SYSTEM

Kidney

Lymphoma malignant lymphocytic
Unnary bladder

Lymphoma malignant lymphocytic

0

0

0

0

0

0

Q

0

0

0

0

0

0

0

0

0

0

n

n

n

0

n

n

1

1

■i

B

6

6

6

7

7

7

7

H

H

H

8

8

8

8

9

9

9

9

9

9

9

0

n

■i

4

5

6

8

2

5

5

7

1

5

5

6

7

7

7

1

1

3

4

6

8

9

0

2

a

4

a

3

4

a

a

4

a

a

a

a

a

a

a

a

a

a

a

a

a

a

a

a

a

3

0

8

8

0

7

7

0

9

B

9

9

7

3

5

7

2

4

5

5

1

7

9

1

2

I

1

1

2

■I

1

■2

3

1

I

2

3

3

2

I

4

1

1

2

3

1

5

4

2

2

+

+

+

+

+

+

+

+

+

4-

+

+

+

+

+

+

X

+

+

+

*

+

+

+

+
X

-t-

+

4-

+

+

+

+

+

+

^■

+

■f

+

+

+

+

+

-t-

+

-t-

-t-

+

■t-

+

+

■(■ + -(- ^-

X

+ +

f + t- -t-

+ +--I--I- + MM + +

X

+ + + + + + + + ■*-

-t- + M + -t-+M + + + + +

X

+ -f
X

+ -*--•- + +

+ + + + + + + + + +

X
+ + + ■(-■(--(-+■ + ■»--*-

M +

+ +
+ -t-

X

X

-(-

■(-

■t-

-t-

-t-

■*-

+ + + +
+ + + +

Nitrofurantoin, NTP TR 341

172

TABLE D2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE MICE:

(Continued)

UNTREATED CONTROL

WEEKS ON
STUDY

I 1 I I 1 1 1 1 1 L 1 I 1 I 1 1 1 1 1 1 1 1 1 1 L
0000000000000000000000000
22344455?>5555555555555555

TOTAL,
TISSUES
TUMORS

CARCASS
ID

3333333^33333333333333344
491588112 2 2 33344456666800
2534344534534534552345545

HEMATOPOIETIC SYSTEM
Bone marrow

Lymphoma malignant lymphocytic
Lymph node

Bronchial, lymphoma malignant
lymphocytic

Deep cervical, lymphoma malignant
lymphocytic

Pancreatic, lymphoma malignant
lymphocytic

Renal, lymphoma malignant histiocytic

Renal, lymphoma malig. lymphocytic
Lymph node, mandibular

Lymphoma malignant histiocytic

Lymphoma malignant lymphocytic
Lymph node, mesentenc

Lymphoma malignant lymphocytic
Spleen

Hemangiosarcoma

Lymphoma malignant histiocytic

Lymphoma malignant Wmphocvtic
Thymus

Lymphoma malignant histiocytic

Lymphoma malignant lymphocytic

4-4-4-4-4- + 4-4--)-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-

X

4-4--h4-4-4-4-4--«-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-4-

X

X X

+-'*-f-t- + + + -t- + + + + -»--)--t- + + + + + + + + + -F

X X X X X

+ + + -♦- + + + -(- + + + + + -»- + + ■»- + + + + + + + +

XXX
+- + f-*- + -f + + + + + -t- + + + + + + 4--t- + + + M-f
X

X X X X X X X

+ f + + + + 4- + + + + + + f-l- + -(-f-f-f+- + -(- + +

X X X X X X

50

3

50

1

1

2

1
1

50
1
7

45
5

49
I
1
9

47
1
8

INTEGUMENTARY SYSTEM

Mammary gland
Adenocarcinoma

Skin
Lymphoma malignant lymphocytic
Subcutaneous tissue, hemangiosarcoma

+ + + +- + + + + + + + + -t--(--t--t- + f + -t- + + -(- + +
X X

+ + + + -l- + -t- + + r + + 4--(--t--f4--(- + -t- + + + + -f

X
X

50

5

50

1
1

MUSCULOSKELETAL SYSTEM
Bone

Skeletal muscle
Lymphoma malignant lymphocytic

■t- + + + + + + + + + + + + + -(--(- + +-+--t- + + -t- + -t-
+-
X

50

L

NERVOUS SYSTEM
Brain
Lymphoma malignant lymphocytic

+ -t- + + + + + + + + M + 4- + ++- + + + + 4- + + + +

X

49

1

RESPIRATORY SYSTEM

Lung
AlveolarA>ronchiolar adenoma
Alveolar/bronchiolar carcinoma
Hepatocellular carcinoma, metastatic.

hver
Lymphoma malignant histiocytic
Lymphoma malignant lymphocytic
Mediastinum. lymphoma malignant
lymphocytic

Nose

Trachea

+ + + -*- + + + + + + + + + + ■*--»- + + + + + + + + +

X

X

X

X

+ >- + + -t--t--t- + + -»- + +'+ + + + + + + + + + -t- + -f

+ +- + + + 4--t- + + + + + + + + -t- + -t- + + + + + + +

50
2

1

1

1
3

3
48
49

SPECIAL SENSES SYSTEM
Hardenao gland

Adenocannnoma

Adenoma

+
X

2

1

1

URINARY SYSTEM
Kidney

Lymphoma malignant lymphocytic
Unnary bladder

Lymphoma malignant lymphocytic

+ + +- + -f^ + + + + -(--*--*- + + + + + + -t-+-+--t--»--t-
X X X

+ -(- + -t- + f + + + + + + -f + + -l- + + + + -t-4- + + +-

X

50

3

50

I

173

Nitrofurantoin, NTP TR 341

TABLE D2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE MICE IN THE TWO-YEAR FEED

STUDY OF NITROFURANTOIN: LOW DOSE

WEEKS ON
STUDY

CARCASS
ID

ALIMENTARY SYSTEM
Esophagus
Gallbladder
Intestine large
Intestine large, cecum
Intestine large, colon
Intestine large, rectum
Intestine small
Intestine small, duodenum
Intestine small, ileum
Intestine small, jejunum
Liver

Hepatocellular carcinoma

Hepatocellular adenoma

Ito cell tumor, NOS

Lymphoma malignant histiocytic

Lymphoma malignant lymphocytic

Lymphoma malignant mixed
Mesentery

Sarcoma
Pancreas

Lymphoma malignant lymphocytic

Sarcoma
Salivary glands
Stomach
Stomach, forestomach

Sarcoma
Stomach, glandular

Sarcoma

CARDIOVASCULAR SYSTEM

Heart
Sarcoma

ENDOCRINE SYSTEM
Adrenal gland
Adrenal gland, cortex

Osteosarcoma, metastatic, bone

Spindle cell, adenoma
Adrenal gland, medulla
Islets, pancreatic
Parathyroid gland
Pituitary gland

Parsdistalis. adenoma
Thyroid gland

Follicular cell, adenoma

GENERAL BODY SYSTEM
None

GENITAL SYSTEM
Ovary

Cystadenoma, papillary

Granulosa ceil tumor benign

Hemangioma

Lymphoma malignant histiocytic
Litems

Adenocarcinoma

Lymphoma malignant histiocytic

Lymphoma malignant mixed

Polyp stromal

Cervix, lymphoma malignant mixed

e

5

5

e

5

5

8

5

5

5

5

5

S

S

s

s

s

S

s

S

!>

S

0

9

8

0

6

7

4

0

a

B

3

3

5

7

7

1

1

1

1

1

2

2

2

2

2

1

1

1

2

1

1

1

3

2

2

1

2

I

2

3

1

2

3

4

5

1

2

3

4

5

A

-h

+■

+

+

M

-t-

+

-(-

+

A

■t-

-(-

-t-

+

A

+

+

+

+

+

4-

■»-

+
X

+
X

X

X

X

+ M M

+ + + + + ■)- +

+ + -t- + f

X

-(- +

+ + + + -t- + + + + + + -i--t- + + +

Nitrofurantoin, NTP TR 341

174

TABLE D2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE MICE:

(Continued)

LOW DOSE

WEEKS ON
STUDY

CARCASS

ID

ALIMENTARY SYSTEM

Esophaffus
Gallbladder
Intestine large
Intestine large, cecum
Intestine large, colon
Intestine large, rectum
Intestine small
Intestine small, duodenum
Intestine small, ileum
Intestine small, jejunum
Liver

Hepatocellular carcinoma

Hepatocellular adenoma

Ito cell tumor. NOS

Lymphoma malignant histiocytic

Lymphoma malignant lymphocytic

Lymphoma malignant mixed
Mesentery

Sarcoma
Pancreas

Lymphoma malignant lymphocytic

Sarcoma
Salivary glands
Stomach
Stomach, forestomach

Sarcoma
Stomach, glandular

Sarcoma

CAIIDIOVASCULAR SYSTEM

Heart
Sarcoma

ENDOCRINE SYSTEM

Adrenal gland
Adrenal gland, cortex

Osteosarcoma, metastatic, bone

Spindle cell, adenoma
Adrenal gland, medulla
Islets, pancreatic
Parathyroid gland
Pituitary gland

Parsdistalis, adenoma
Thyroid gland

Follicular cell, adenoma

GENERAL BODY SYSTEM ~"

Nona

GENITAL SYSTEM

Ovary

Cystadenoma. papillary

Granulosa cell tumor benign

Hemanpoma

Lymphoma malignant histiocytic
Uterus

Adenocarcinoma

Lymphoma malignant histiocytic

Lymphoma malignant miited

Polyp stromal

Cervix, lymphoma malignant muied

1 I 1
0 0 0
5 5 5

1 1
0 0
5 5

1 1 1 I 1
0 0 0 0 0
5 5 5 5 5

1 1
0 0
5 5

5

5

5

5

5

5

5

s

s

S

5

s

5

.1

.■>

S

S

5

5

5

5

e

6

3

3

3

4

4

4

4

5

5

5

5

6

6

6

7

7

8

8

8

9

9

9

9

0

0

3

4

5

2

3

4

5

2

3

4

5

3

4

5

4

5

3

4

5

2

3

4

5

4

5

TOTAL:
TISSUES
TUMORS

+ ■*--*■

+ + -t-

+ +- +

+ -t- +

M
M

4- +
+ +

+ + +
+ + +

M + +

M

+

M

M M

4- +

X

■(- + + + ■*-
X

+ + +

-(- + +
X

50

39

50

49

49

49

49

48

47

48

50

1

1

1

4

3

3

2

1

50

I

I

50

49

49

1

49

1

50

L

50
50

1

1
49
50
44
44

1
49

1

50
1
3
1
1

50
1
2
1

175

Nitrofurantoin, NTP TR 341

TABLE D2.

INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE MICE: LOW DOSE

(Continued)

WEEKS ON

STUDY

OOOOOOOllllllllllllllllll
6888899000000000000000000
73679690012 2 2555555555555

CARCASS
ID

5555555655555555555555555
0980674086335771 1 1 1 12222 2
11121 11322121231 2 ;i 4 5 12345

HEMATOPOIETIC SYSTEM
Bone marrow
Lymph node

Bronchial, lymphoma malignant
histiocytic

Bronchial, lymphoma malignant mixed

Deep cervical, lymphoma malignant
lymphocytic

Inguinal, lymphoma malignant
histiocytic

Mediastinal, lymphoma malignant
histiocytic

Mediastinal, lymphoma malignant mixed
Lymph node, mandibular

Lymphoma malignant histiocytic

Lymphoma malignant lymphocytic

Lymphoma malignant mixed
Lymph node, mesentenc

Lymphoma malignant histiocytic

Lymphoma malignant lymphocytic

Lymphoma malignant mixed
Spleen

Lymphoma malignant histiocytic

Lymphoma malignant lymphocytic

Lymphoma malignant mixed
Thymus

Lymphoma malignant lymphocytic

Lymphoma malignant mixed

+ + + + + + ■*- + + + + + + + + + + -*- + -(--*- + + + +
+ + + -i- + + + + -f + + + -f-t--(- + + -t- + -t- + + + +- +

X

X
X X

X

+ + + + + +- + + 4--t- + + + + + + -f-t- + f + -»--*- + +
X X

X XXX
X
■*- + -t-+-+- + + -+--t- + + + +Mt- + + -t- + + -«- + -H + -f
X X

X X
X

■l--t--t- + -t- + + + -)- + -t--t- + + f + 4-f + f-t- + -(-4- +

XX X

X XXX X
X X

+ + + + + -l--)- + -(--t--t- + 4 + -t-M+-t- + 'l"-^^--t'-*- +

X

INTEGUMENTARY SYSTEM

Mammary gland
Adenocarcinoma

Skm
Squamous cell carcinoma
Subcutaneous tissue, fibrosarcoma
Subcutaneous tissue, schwannoma benign

X X

+ + + -t- + -t-f-t- + + + -f-t--t- + + + + + + + + + -t--t-

X

X

MUSCULOSKELETAL SYSTEM
Bone
Osteosarcoma

Skeletal muscle
Sarcoma

+ + + -t- + + + -t--»- + + + + + + + +f + + + + -»- + +

X
+
X

NERVOUS SYSTEM
Brain
Meningioma benign

+ 4- + -t- + + -(- + + + + + + -(- + + + 4- + +--f-t-+- + -*-
X

RESPIRATORY SYSTEM

Lung
Adenocarcinoma, metastatic, mammary

gland
Alveolar/bronchiolar adenoma
Lymphoma malignant histiocytic
Lymphoma malignant lymphocytic
Lymphoma malignant mixed
Osteosarcoma, metastatic, bone
Sarcoma
Mediastinum, sarcoma

Nose

Trachea

+ + + + + + + + + + ■»- + + + -*--(- + -*- + -*-■»- + -*- + -*-

X

X
X X

X X
X

X
X
X
+ + -t- + + + + + + + + -t-f + -t- + + + -f + + f-*--f+-
+ + + + + + + + -(- + + -t--t-+- + -(- + + + + + -(--(- + 4-

SPECIAL SENSES SYSTEM
Hardenan gland

Adenocarcinoma

Adenoma

+ +

X

URINARY SYSTEM
Kidney

Lymphoma malignant histiocytic
Lymphoma malignant lymphocytic
Lymphoma malignant mixed
Sarcoma
Unnary bladder
Lymphoma malignant lymphocytic

+ -t-+- + + + +- + 4- + + + + + + + + -t- + -t- + + -f-f-t-
X X

X XX
X
X
+ + + + + + + + + + + + + + + + + + + -*- + +- + -»- +

X X

Nitrofurantoin, NTP TR 341

176

TABLE D2.

INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE MICE:

(Continued)

LOW DOSE

WEEKS ON
STUDY

I I 1 1 I 1 I 1 1 1 1 1 I I 1 1 1 1 1 I I 1 I 1 1
0000000000000000000000000
5555555555555555555555555

TOTAL:
TISSUES
TUMORS

CARCASS

ID

5555555555555555555555566
3334444555566677888999900
3452345234534545345234545

HEMATOPOIETIC SYSTEM
Bone marrow
Lymph node

Bronchial, lymphoma malignant
histiocytic

Bronchial, lymphoma malignant mixed

Deep cervical, lymphoma malignant
lymphocytic

Inguinal, lymphoma malignant
histiocytic

Mediastinal, lymphoma malignant
histiocytic

Mediastinal, lymphoma raahg. mixed
Lymph node, mandibular

Lymphoma malignant histiocytic

Lymphoma malignant lymphocytic

Lymphoma malignant mixed
Lymph node, mesenteric

Lymphoma malignant histiocytic

Lymphoma malignant lymphocytic

Lymphoma malignant mixed
Spleen

Lymphoma malignant histiocytic

Lymphoma malignant lymphocytic

Lymphoma malignant mixed
Thymus

Lymphoma malignant lymphocytic

Lymphoma malignant mixed

+ 4-444-4 + 4-4-44-4-44444 + + 44 + 4-44-
44444444-4-4-4-4444444 + 444444

X

X

44-444444-4-44-4-44-4-4-4-4-444 + 4-44

X X

X

444M4444M444444M44444444M
44444444 + 444444 + 44-4444444

XXX X X

X

4 4-44 + +M4 + + + + + + + 44 + + + + -*--t- + +

X

X

50
50

1
1

1

2

1
1

50
2
6
2

45
2
2
1

50
3

10
3

48
2

INTEGUMENT.-^Y SYSTEM
Mammary gland

Adenocaranoma
Skin

Squamous cell carcinoma

Subcutaneous tissue, fibrosarcoma

Subcut. tissue, schwannoma benign

4 + + 444- + + + 4 + 44444 + 444M4^ + 4

X

+ + + 4 + + + + + + + + 444 + + + + -*-44^-44

X

48

3

50

1
1

1

MUSCULOSKELETAL SYSTEM

Bone

Osteosarcoma
Skeletal muscle

Sarcoma

44-4444 + 444 + 444 + + + + + + 4444 +

50

1
1
1

NERVOUS SYSTEM
Brain
Meningioma benign

444444444 + + + + + + + + + + + + + 44 +

50

1

RESPIRATORY SYSTEM

Lung
Adenocarcinoma, metastatic, mammary

eland
Alveolar/bronchiolar adenoma
Lymphoma malignant histiocytic
Lymphoma malignant lymphocytic
Lymphoma malignant mixed
Osteosarcoma, metastatic, bone
Sarcoma
Mediastinum, sarcoma

Nose

Trachea

4444444 + + + + + + + 44 + + + 444 + 44

X
X X

+ + + + + 44 + 444 + + + + + + + 44 + + + + +
+ 4- + 44444 + + 4 + + + 44 + 444-44444

50

1
2
2
4
1
1
1
1
50
50

SPECIAL SENSES SYSTEM
Hardenan gland

Adenocarcinoma

Adenoma

+ + +

X X

5

1
2

URINARY SYSTEM

Kidney
Lymphoma malignant histiocytic
Lymphoma malignant lymphocytic
Lymphoma malignant mixed
Sarcoma

Unnary bladder
Lymphoma malignant lymphocytic

+ + -t- + + + + + + + + + + + + -»- + + + + + "*- + -^ +
X X

+ -t-+- + + + + + + + + + -t--f + + -t--t--*- + +-t--t--t- +

X

50
2
5
I
1

50
3

177

Nitrofurantoin, NTP TR 341

TABLE D2.

INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE MICE IN THE TWO-YEAR FEED
STUDY OF NITROFURANTOIN: HIGH DOSE

WEEKS ON
STUDY

CARCASS

ID

ALIMENTARY SYSTEM

Esophagus
Gallbladder

Lymphoma cnahgnant histiocytic
Intestine large
Intestine large, cecum

Lymphoma malignant histiocytic
Intestine large, colon
Intestine large, rectum
Intestine small
Intestine small, duodenum
Intestine small, ileum
Intestine small, jejunum
Liver

Hepatocellular carcinoma

Hepatocellular adenoma

Ito cell tumor. NOS

Lymphoma malignant histiocytic

Lymphoma malignant lymphocytic
Mesentery

Fibrosarcoma

Lymphoma malignant histiocytic

Lymphoma malignant lymphocytic

Sarcoma
Pancreas

Fibrosarcoma, metastatic, mesentery

Lymphoma malignant histiocytic

Lymphoma malignant lymphocytic
Salivary glands

Lyrapnoma malignant histiocytic
Stomach
Stomach, forestomach

Lymphoma malignant lymphocytic

Papilloma squamous

Glandular, fibrosarcoma, metastatic,
mesentery
Stomach, glandular

CARDIOVASCULAR SYSTEM

Heart

ENDOCRINE SYSTEM

Adrenal gland

Fibrosarcoma, metastatic, mesentery
Adrenal gland, cortex

Adenoma
Adrenal gland, medulla
Islets, pancreatic

Carcinoma
Parathyroid gland
Pituitary gland

Pars distalis, adenoma
Thyroid gland

GENERAL BODY SYSTEM

None

GENITAL SYSTEM

Ovary

Adenoma, tubular

Cystadenoma

Granulosa cell tumor malignant

Granulosa cell tumor benign

Hemangioma

Lymphoma malignant histiocytic

Lymphoma malignant lymphocytic

Mixed tumor benign

Neoplasm, NOS
Uterus

Adenocarcinoma

Lymphoma malignant histiocytic

Polyp stromal

Sarcoma stromal

I 1 1
0 0 0
5 5 5

1 — 3 — 5 — 3 — 5~
4 9 3 6 9
11112

~i — 3~

1 2
1 1

T — T"

7 5
1 L

"3 — T"

5 7
2 2

T — 3 — 3 — T"

1111

2 3 4 5

"3 — 3 — T"
2 3 3
5 2 3

+ M M + M +

+ + + -*-
■t- + + M

+ + + ^-■^ + -f +

+ + + ■♦- + + + -*-
X

+ + + + + + + +

+ + + + + + + +

+ + + + ■!- + + +

+ + + + + + + ■«■

+ + + -(- + + + +

+ + + + + + + +

+ + + + + + + +

-♦- + + +

+ + + +

+■ + + +

-)- + + +

+ + + +

■I- + + +

-»- + + +

X X X X

-t- ■(-
X

+ + +

+ + +
+ + +

■t- +

-I- ■»■
+ +

+ ■(- + +

X X

X

+ 4- +

X X

X

f +

X XX

+ + + + + + + +

X

■I- -»■

X X

Nitrofurantoin, NTP TR 341

178

TABLE D2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE MICE:

(Continued)

HIGH DOSE

WEEKS ON

1

1

1

1

1

1

1

1

1

1

1

1

I

1

1

1

1

I

1

I

1

1

1

1

1

STUDY

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

0

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

TOTAL:

nssuES

CARCASS

4

4

4

4

4

4

4

4

4

4

4

4

4

T-

-4—

T-

4

4

4

4

T~

h

t>

S

4

ID

4

4

4

4

5

5

6

6

6

6

7

7

7

8

3

8

8

8

9

9

9

0

0

0

5

rUMORS

2

3

4

5

3

4

2

3

4

5

3

4

5

1

2

3

4

5

3

4

5

3

4

5

5

ALIMENTARY SYSTEM

Esophagus
Gallbladder

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

-4-

+

+

+

•♦-

+

+

4-

50

+

+

M

+

+

+

+

+

4-

+

+

+

+

+

+

+

+

+

f

+

+

+

M

+

4-

42

Lymphoma malignant histiocytic

1

Intestine large

+

+

+

+

+

+■

+

+

+

-♦-

+

+

+

+

+

+

+

+

+

+

+■

+

+

+

4-

50

Intestine large, cecum

+

+

+

+

+

-t-

+

+

+

+

+

+

+

+

+

+

+

+

+

+

■*-

■(-

+

+

4-

50

Lymphoma malignant histiocytic
Intestine large, colon

1

+

+

+

+

+

-f

+

+

+

+

+

+

+

+

+

+

-f

+

+-

+

+

+

+

+

4-

50

Intestine large, rectum

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

-t-

+

+

+

+

+

■t-

4-

4-

50

Intestine small

+

+

+

+.

+

+

+

+

+

+

+

+

+

+

+

-t-

+

+

+

+

+

+

+■

+

4-

50

Intestine small, duodenum

+

+

+

+

+

-1-

+

+

+■

+

+

+

+

+

+

-♦-

+

+

+

+

+-

M

+

+

4-

49

Intestine small, ileum

+

+

4-

+

+

-(-

-*-

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

4-

50

Intestine small, jejunum

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

■f

+

■(-

+

4-

50

Liver

-1-

+

+

+

+

•1-

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

f

+

+

50

Hepatocellular carcinoma

X

2

Hepatocellular adenoma

X

X

X

X

X

7

Ito eel! tumor. NOS

X

1

Lymphoma malignant histiocytic

6

Lymphoma malignant lymphocytic

X

4

Mesentery

+

+

8

Fibrosarcoma

L

Lymphoma malignant histiocytic

I

Lymphoma malignant lymphocytic

X

3

Sarcoma

I

Pancreas

+

+

+

+

+

+

■t-

+

+

+

+

+

+

+

+

+

+

f

■t-

+

+

+

+

-t-

4-

50

Fibrosarcoma, metastatic, mesentery

i

Lymphoma malignant histiocytic

2

Lymphoma malignant lymphocytic

L

Salivarv glands

+

+

+

+

+

+

+

+

+

-:■

+

+

+

■H

+

-*-

+

f

■f

+

+

+

+

+

4-

50

Lymphoma malignant histiocytic

1

Stomach

+

+

+

-t-

+

-»-

+

-(-

+

+

+

+

+

+

+

+

+

■*-

+

^

+

■t-

+

••-

4-

50

Stomach, forestomach

+

+

+

+■

+

+-

+

+

+

+

+

+

+

+

+

-t-

+

+

+

t-

+

+

+

+

4-

50

Lymphoma malignant lymphocytic
Papilloma squamous

I

X

1

Glandular, fibrosarcoma, metastatic.

mesentery

I

Stomach, glandular

+

+

+

■t-

+

-t-

+■

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

-t-

+

4-

49

CARDIOVASCULAR SYSTEM

Heart

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

"*"

+

+■

-f

+

+

+

+

+

4-

50

ENDOCRINE SYSTEM

Adrenal giand

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

4-

+

-t-

+

-t-

+

4-

4-

50

Fibrosarcoma, metastatic, mesentery

1

Adrenal gland, cortex

+

+

+

+■

-1-

-♦-

+

+

■1-

+

+

+

+

+

+

+

+

+

+

-1-

+

■(-

+

■1-

4-

50

Adenoma

1

Adrenal gland, medulla

+

+

+

+

+

+

+

+

-f

+■

+

■1-

+

+

+

+

-t-

+

+

+

+

■t-

+

4-

4-

49

Islets, pancreatic

+

+

+

+

+

+

+

+■

+

+

+

+

+

+

+

+

+

+

+

■1-

+

+

-1-

4-

4-

49

Carcinoma

I

Parathyroid gland

+

+

+

M

+

■I-

+

■(-

+

+

+

+

+

+

+

+

+

M

+

+

M

-(-

+

4-

4-

45

Pituitary gland

-1-

+

+

+

+

-t-

+

+

+

+

+

+

+

■I-

+

+

+

+

-f

+

+

-(-

+

4-

4-

49

Pars distalis. adenoma

2

Thyroid gland

+

+

+

-t-

+

■f

+

"•"

^"

+

+

+

+

+

+

+

+■

+

+

'•'

+

-»-

■1-

4-

4-

50

GENERAL BODY SYSTEM

None

GENITAL SYSTEM

Ovary

-(-

+

+

■+-

+

+

+

+

+

+

+

+

+

+

+

+

+

-t-

->-

+

+

+

+

4-

4-

50

Adenoma, tubular

X

X

5

Cystadenoma

I

Granulosa cell tumor malignant

1

Granulosa cell tumor benign

X

1

Hemangioma

X

1

Lymphoma malignant histiocytic

3

Lymphoma malignant lymphocytic

X

X

5

Mixed tumor benign

X

X

4

Neoplasm, NOS

i

Uterus

■f

+

+

+

+

+

+

+

+

+

+

+•

+

+

-t-

+

+

+

+

■1-

■(-

■1-

+

+

4-

50

Adenocarcinoma

i

Lymphoma malignant histiocytic

■■

2

Polyp stromal

1

Sarcoma stromal

2

179

Nitrofurantoin, NTP TR 341

TABLE D2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE MICE: HIGH DOSE

(Continued)

WEEKS ON
STUDY

CARCASS
ID

HEMATOPOIETIC SYSTEM

Bone marrow
Lymph node
Axillary, lymphoma malignant

histiocytic
Bronchial, lymphoma malignant

histiocytic
Bronchial, lymphoma malignant

lymphocytic
Iliac, lymphoma malignant lymphocytic
Iliac, lymphoma malignant mixed
Ingumai. lyfTiphoma malignant

lymphocytic
Inguinal, lymphoma malignant mixed
Lumbar, lymphoma malignant lymphocyt.K
Mediastinal, lymphoma malignant

histiocytic
Mediastinal, lymphoma malignant

lymphocytic
Mediastinal, lymphoma malignant mixed
Pancreatic, lymphoma malignant

lymphocytic
Pancreatic, lymphoma malignant mixed
Renal, lymphoma malignant lymphocytic
Renal, lymphoma malignant mixed
Lymph node, mandibular
Lymphoma malignant histiocytic
Lymphoma malignant lymphocytic
Lymphoma malignant mixed
Lymph node, mesentenc
Fibrosarcoma, metastatic, mesentery
Lymphoma malignant histiocytic
Lymphoma malignant lymphocytic
Lymphoma malignant mixed
Spleen
Fibrosarcoma, metastatic, mesentery
Lymphoma malignant histiocytic
Lymphoma malignant lymphocytic
Lymphoma malignant mixed
Thymus
Lymphoma malignant histiocytic
Lymphoma malignant lymphocytic
Mediastinum, lymphoma malignant

lymphocytic

INTEGUMENTARY SYSTEM

Mammary gland

Adenoacanthoma

Adenocaranoma. multiple
Skin

Basal cell carcinoma

Sebaceous gland, adenoma

Subcutaneous tissue, fibrosarcoma

MUSCULOSKELETAL SYSTEM
Bone

Skeletal muscle
Fibrosarcoma, metastatic, mesentery

Lymphoma malignant histiocytic

NERVOUS SYSTEM
Brain

Lymphoma malignant lymphocytic

Meningioma benign

RESPIRATORY SYSTEM

Lung
Basal cell carcinoma, metastatic, skin
Hepatocellular carcinoma, metastatic,

liver
Lymphoma malignant histiocytic
Lymphoma malignant lymphocytic

Nose

Trachea

SPECUL SENSES SYSTEM

Hardenan gland
Adenoma

UklNAHY SYSTEM
Kidney

Lymphoma malignant histiocytic

Lymphoma malignant lymphocytic

Lymphoma malignant mixed

Renal tubule, carcinoma
Unnary bladder

0

0

0

0

0

0

0

0

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

1

H

9

9

9

9

9

9

9

0

0

0

0

0

0

0

0

0

0

0

0

n

0

0

0

0

2

0

0

2

3

4

5

7

0

2

5

5

5

5

5

5

5

5

5

5

5

5

5

5

5

i

4

4

4

4

4

4

4

4

h

4

4

s

4

4

4

4

4

4

4

4

4

4

4

4

4

9

3

6

9

1

2

7

5

0

5

7

0

1

1

1

1

2

2

■>

2

3

3

3

3

1

1

1

1

2

1

1

1

I

1

2

2

2

2

3

4

5

2

3

4

5

2

3

4

5

+

+

+

+

+

+

+

+

_j.

+

+

+

+

^

+

4-

+-

+

+

^

+

+

4-

+

■(-

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

+

4-

+

+

+

+

+

+

X X

X

+

M

-t-
X

+
X

+

+

+

+
X

+

+
X

+
X

+

+

+

■t-

+

+
X

+

+

+

+

M

+

X

■t-

X

M

+
X

+

X

+
X

+

+

+

+
X

+
X

+
X

-t-
X

*

+

+

"

■t-

+■
X

+

f

X

+

-t-

X

X

■(-

+

+

+
X

X

+

+

+

+
X

X

+
X

X

+
X

+

+

+

+

+■
X

■»■

+

X

"

■*-

X

+
X

+

M

+

+

-f

+

+

+

+

+

+

+

+

M

+

-t-

+

+■

+

■1-

+

+

+

+

■t-

+

X X

+ M + +

+ + + +

+ + + M

+ + +

+ -(- + + -f +

^-+^--f+++^--^-+++■t-+
+ +

X

+ *■ + *■

+ + + + f + + -t- + + -t-+- + -t- +
X

X

+ 4- >- f

X X X X

+ + + + 4- +

+ + + + + +

+ + +

+ + +

+ ■!■ + + -(-

+--(- + +

X X X X

Nitrofurantoin, NTP TR 341

180

TABLE D2. INDIVIDUAL ANIMAL TUMOR PATHOLOGY OF FEMALE MICE: HIGH DOSE

(Continued)

WEEKS ON

1 1 1 1 1 1 1 1 1 I I 1 1 1 I 1 1 1 1 1 1 1 I 1 1

STUDY

0000000000000000000000000
555555555555555555 5 555555

TOTAL:

nssuEs

CARCASS

4444444444444444444445554

ID

4444556666777888889990005 TUMORS

2345342345345123453453455

HEMATOPOIETIC SYSTEM

Bone marrow

+ -»-+- + + + -f + + -t- + -t- + + + + + + + + + + + + +

50

Lymph node

+ + + + + + + -f + + + + + + + + + +-+M+ + + + +

49

Axillary, lymphoma malignant

histiocytic

1

Bronchial, lymphoma malignant

histiocytic

1

Bronchial, lymphoma malignant

lymphocytic
Iliac. lymphoma malignant lymphocytic

X

3

1

Iliac, lymphoma malignant mixed

X

1

Inguinal, lymphoma malignant

lymphocytic

1

Inguinal, lymphoma malignant mixed

X

1

Lumbar, lymphoma malig. lymphocytic
Mediastinal, lymphoma malignant

1

histiocytic

1

Mediastinal, lymphoma malignant

lymphocytic

K

4

Mediastinal, lymphoma mahg. mixed
Pancreatic, lymphoma malignant

X

1

lymphocytic

2

Pancreatic, lymphoma malignant mixed

X

1

Renal, lymphoma maiig. lymphocytic

X

1

Renai, lymphoma malignant mixed

X

1

Lymph node, mandibular

+ ^ + + +- + -t- + + + + + + 4--t- + +M^-Mt- + + + +

45

Lymphoma malignant histiocytic

3

Lymphoma malignant lymphocytic

X X X X X X X

12

Lymphoma malignant mixed

X

1

Lymph node, mesentenc

+ -(- + -t- + + + + -t--h + + + + -t-M+- + +M + -t- + -*- +

48

Fibrosarcoma, metastatic, mesentery

1

Lymphoma malignant histiocytic

4

Lymphoma malignant lymphocytic

XX X X

10

Lymphoma malignant mixed

X

1

Spleen

+ -f + -t- + + + + + + + + + -»- + + + 4- + + -t- + + +- +

50

Fibrosarcoma, metastatic, mesentery

1

Lymphoma malignant histiocytic

4

Lymphoma malignant lymphocytic

XX XXX X XX XX

16

Lymphoma malignant mixed

X

1

Thymus

+ -*- + + -(- + + + -(- + + + + + + + + +■■*- + ■*- + + -*- +

48

Lymphoma malignant histiocytic

2

Lymphoma malignant lymphocytic

2

Mediastinum, lymphoma malignant

lymphocytic

1

INTEGUMENTAIiV SYSTEM

Mammary gland

+ + + + + + + + + -(- + -i- + + -f + -t- + + + -t--t- + v +

48

Adenoacanthoma

1

Adenocarcinoma, multiple

X

1

Skin

+ + + + + + 4- + + + + -f + -»- + + -»--t- + + -t- + + -*--t-

50

Basal cell carcinoma

X

1

Sebaceous gland, adenoma

1

Subcutaneous tissue, fibrosarcoma

X

1

MUSCULOSKELETAL SYSTEM

Bone

+ -f + + + -*--t--t- + + + + + + -*- + -*-t- + ++- + + + +

50

Skeletal muscle

2

Fibrosarcoma, metastatic, mesentery

1

Lymphoma malignant histiocytic

1

NERVOUS SYSTEM

Brain

+ + + ■»- + + + ■»- + + + + + + + + +- + + + + + +»■ +

50

Lymphoma malignant lymphocytic

1
1

Meningioma benign

RESPIRATORY SYSTEM

Lung

+ -!- + + +-+- + -(- + + + + + + +- + + + + + + '*- + -♦-■•■

50

Basal cell carcinoma, metastatic, skin

X

1

Hepatocellular carcinoma, metastatic.

liver

X

1

Lymphoma malignant histiocytic

5

Lymphoma malignant lymphocytic

X X

6

Nose

+ + +- + ■(-+- + + + + + + + + + + + ■(- + + + ■(- + + +

50

Trachea

-f + + + + -(-+--(- + + + -f-t--»--l- + -f-t-f + -t- + + +-4-

50

SPECIAL SENSES SYSTEM

Hardenan gland

+-

1

Adenoma

X

1

URINARY SYSTEM

Kidney

+ + + + + + ^- + -*- + -*-'*-^ + + + + + + + + + + + +

50

Lymphoma malignant histiocytic

5

Lymphoma malignant lymphocytic

X X

5

Lymphoma malignant mixed

X

1

Renal tubule, carcinoma

X

1

Unnary bladder

+ + + + -l--t- + + + -t--t- + -f + 4--t- + + -(- + + + + -+- +

50

181

Nitrofurantoin, NTP TR 341

TABLE D3. ANALYSIS OF PRIMARY TUMORS IN FEMALE MICE IN THE TWO-YEAR FEED STUDY

OF NITROFURANTOIN

Control

1,300 ppm

2,500 ppm

Harderian Gland: Adenoma or Adenocarcinoma

Overall Rates (a)

2/50(4%)

3/50(6%)

1/50(2%)

Adjusted Rates (b)

8.5%

8.1%

2.7%

Terminal Rates (c)

1/19(5%)

3/37(8%)

1/37(3%)

Day of First Observation

686

730

730

Life Table Tests (d)

P = 0.I98N

P = 0.596N

P = 0.308N

Logistic Regression Tests (d)

P = 0.259N

P = 0.673

P = 0.384N

Cochran-Armitage Trend Test (d)

P = 0.411N

Fisher Exact Test <d)

P = 0.500

P = 0.500N

Liver: Hepatocellular Adenoma

Overall Rates (a)

1/50(2%)

1/50(2%)

7/50(14%)

Adjusted Rates (b)

2.7%

2.7%

18.1%

Terminal Rates (cl

0/19(0%)

1/37(3%)

6/37(16%)

Day of First Observation

603

730

660

Life Table TesU(d)

P = 0.042

P = 0.658N

P = 0.147

Logistic Regression Tests (d)

P = 0.016

P = 0.758

P = 0.054

Cochran-Armitage Trend Test (d)

P = 0.012

Fisher Exact Test (d)

P = 0.753N

P = 0.030

Liver: Hepatocellular Adenoma or Carcinoma

Overall Rates (a)

2/50(4%)

2/50(4%)

8/50(16%)

Adjusted Rates <b)

6.6%

4.9%

20.0%

Terminal Rates (c)

0/19(0%)

1/37(3%)

6/37(16%)

Day of First Observation

603

670

660

Life Table Tests (d)

P = 0.093

P = 0.531N

P = 0.217

Logistic Regression Tests (d)

P = 0.029

P = 0.704N

P = 0.079

Cochran-Armitage Trend Test (d)

P = 0.023

Fisher Exact Test (d)

P = 0.691N

P = 0.046

Lung: Alveolar/Bronchiolar Adenoma or Carcinoma

Overall Rates (a)

3/50(6%)

2/50(4%)

0/50(0%)

Adjusted Rates (b)

10.1%

5.4%

0.0%

Terminal Rates (c)

1/19(5%)

2/37(5%)

0/37(0%)

Day of First Observation

561

730

Life Table Tests (d)

P = 0.030N

P = 0.292N

P = 0.065N

Logistic Regression Tests (d)

P = 0.099N

P = 0.526N

P = 0.161N

Cochran-Armitage Trend Test (d)

P = 0.085N

Fisher Exact Test (d)

P = 0.500N

P = 0.121N

Mammary Gland: Adenocarcinoma

Overall Rates (a)

5/50(10%)

3/50(6%)

1/50(2%)

Adjusted Rates (b)

19.6%

7.2%

2.7%

Terminal Rates (c)

2/19(11%)

1/37(3%)

1/37(3%)

Day of First Observation

635

596

730

Life Table Tests (dl

P = 0.016N

P = 0.144N

P = 0.026N

Logistic Regression Tests (d)

P = 0.053N

P = 0.301N

P = 0.056N

Cochran-Armitage Trend Test (d)

P = 0.071N

Fisher Exact Test (d)

P = 0.357N

P = 0.I02N

Ovary: Tubular Adenoma

Overall Rates (a)

0/50(0%)

0/50(0%)

5/50(10%)

Adjusted Rates (bl

0.0%

0.0%

13.0%

Terminal Rates (c)

0/19(0%)

0/37(0%)

4/37(11%)

Day of First Observation

729

Life Table Tests (d)

P = 0,019

(e)

P = 0.127

Logistic Regression Tests (d)

P = 0.018

(e)

P = 0.112

Cochran-Armitage Trend Test (d)

P = 0.007

Fisher Exact Test (d)

(e)

P = 0.028

Nitrofurantoin, NTP TR 341

182

TABLE D3. ANALYSIS OF PRIMARY TUMORS IN FEMALE MICE IN THE TWO-YEAR FEED STUDY

OF NITROFURANTOIN (Continued)

Control

1,300 ppm

2,500 ppm

Ovary: Granulosa Cell Tumor, Benign

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c )

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d )

Cochran-Armitage Trend Test (d)

Fisher Exact Test (d)

Ovary: Granulosa Cell Tumor, Benign or Malignant

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

Cochran-Armitage Trend Test (d)

Fisher Exact Test (d)

Ovary: Mixed Tumor, Benign

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d )

Cochran-Armitage Trend Test (d)

Fisher Exact Test (d)

Ovary: Tubular Adenoma or Mixed Tumor, Benign

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests ( d )

Cochran-Armitage Trend Test ( d )

Fisher Exact Test (d)

Pituitary Gland/Pars Distalis: Adenoma

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

Cochran-Armitage Trend Testjd)

Fisher Exact Test (d)

Thyroid Gland: Follicular Cell Adenoma

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table TesU(d)

Logistic Regression Tests (d )

Cochran-Armitage Trend Test (d )

Fisher Exact Test (d)

0/50(0%)

3/50(6%)

1/50(2%)

0.0%

8.1%

2.7%

0/19(0%)

3/37(8%)

1/37(3%)

730

730

P = 0.564

P = 0.260

P = 0.633

P = 0.564

P = 0.260

P = 0.633

P = 0.367

P = 0.121

P = 0.500

ant

0/50(0%)

3/50(6%)

2/50 (4%)

0.0%

8.1%

5.1%

0/19(0%)

3/37(8%)

1/37(3%)

730

729

P = 0.375

P = 0.260

P = 0.404

P = 0.362

P = 0.260

P = 0.374

P = 0.197

P = 0.121

P = 0.247

0/50(0%)

0/50(0%)

4/50(8%)

0.0%

0.0%

10.0%

0/19(0%)

0/37(0%)

3/37(8%)
630

P = 0.035

(e)

P = 0.162

P = 0.018

(e)

P = 0.084

P = 0.017

(e)

P = 0.059

gn

0/50(0%)

0/50(0%)

9/50(18%)

0.0%

0.0%

22.6%

0/19(0%)

0/37(0%)

7/37(19%)
630

P = 0.001

(e)

P = 0.028

P<0.001

(e)

P = 0.010

P<0.001

(e)

P = 0.001

3/48(6%)

1/44(2%)

2/49(4%)

16.7%

2.9%

4.7%

3/18(17%)

1/35(3%)

0/36(0%)

730

730

656

P = 0.178N

P = 0.107N

P = 0.229N

P = 0.256N

P = 0.107N

P = 0.330N

P = 0.390N

P = 0.342N

P = 0.490N

3/48(6%)

1/49(2%)

0/50(0%)

12.3%

2.8%

0.0%

1/19(5%)

1/36(3%)

0/37(0%)

635

730

P = 0.018N

P = 0.146N

P = 0.047N

P = 0.039N

P = 0.233N

P = 0.087N

P = 0.057N

P = 0.301N

P = 0.114N

183

Nitrofurantoin, NTP TR 341

TABLE D3.

ANALYSIS OF PRIMARY TUMORS IN FEMALE MICE IN THE TWO-YEAR FEED STUDY
OF NITROFURANTOIN (Continued)

Control

1,300 ppm

2,500 ppm

Thyroid Gland: Follicular Cell Adenoma or Carcinoma

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

Cochran-Armitage Trend Test (d)

Fisher Exact Test (d)

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

Cochran-Armitage Trend Test (d)

Fisher Exact Test(d)

Hematopoietic System: Lymphoma,

Overall Rates (a)

Adjusted Rates (b)

Terminal Rates (c)

Day of First Observation

Life Table Tests (d)

Logistic Regression Tests (d)

Cochran-Armitage Trend Test (d)

Fisher Exact Test(d)

4/48(8%)

1/49(2%)

0/50(0%)

17.2%

2.8%

0.0%

2/19(11%)

1/36(3%)

0/37(0%)

635

730

P = 0.005N

P = 0.058N

P = 0.016N

P = 0.012N

P = 0.109N

P = 0.033N

P = 0.023N

P = 0.175N

P = 0.054N

Hemangiosarcoma

3/50(6%)

1/50(2%)

1/50(2%)

14.8%

2.7%

2.7%

2/19(11%)

1/37(3%)

1/37(3%)

728

730

730

P = 0.067N

P = 0.116N

P = 0.112N

P = 0.072N

P = 0.134N

P = 0.120N

P = 0.201N

P = 0.309N

P = 0.309N

.11 Malignant

12/50(24%)

19/50(38%)

24/50(48%)

50.2%

43.4%

52.7%

8/19(42%)

13/37(35%)

16/37(43%)

631

596

568

P = 0.352

p = 0.447N

P = 0.449

P = 0.038

P = 0.295

P = 0.076

P = 0.008

P = 0.097

P = 0.011

(a) Number of tumor-bearing animals/number of animals examined at the site

(b) Kaplan- Meier estimated tumor incidences at the end of the study after adjusting for intercurrent mortality

(c) Observed tumor incidence at terminal kill

(d) Beneath the control incidence are the P values associated with the trend test. Beneath the dosed group incidence are the
P values corresponding to pairwise comparisons between that dosed group and the controls. The life table analysis regards tu-
mors in animals dying prior to terminal kill as being (directly or indirectly) the cause of death. The logistic regression test re-
gards these lesions as nonfatal. The Cochran-Armitage and Fisher exact tests compare directly the overall incidence rates.
A negative trend or lower incidence in a dosed group is indicated by ( N ).

(e) No P value is reported because no tumors were observed in the 1,300-ppm and control groups.

Nitrofurantoin, NTP TR 341

184

TABLE D4a. HISTORICAL INCIDENCE OF OVARIAN TUMORS IN FEMALE B6C3Fi MICE RECEIVING

NO TREATMENT (a)

Study

No. Examined

No. of Tumors

Diagnosis

Historical Incidence at Southern Research Institute

HC Blue No. 2 49

C.I. Disperse Blue 1
All others

Overall Historical Incidence

49
321

1.858

TOTAL

Granulosa cell tumor
Adenocarcinoma, NOS
Granulosa cell tumor

1

Adenoma, NOS

2

Papillary adenoma

3

Cystadenoma, NOS

8

Papillary cystadenoma, NOS

I

Luteoma

4

Tubular adenoma

1

Mixed tumor, benign

6

Granulosa cell tumor

1

Carcinoma, NOS

1

Adenocarcmoma, NOS

1

Cystadenocarcmoma, NOS

1

Papillary cystadenocarcinoma, NOS

1

Mucinous adenocarcinoma

1

Granulosa cell carcinoma

8(0.4%)

Stromal (granulosa cell or luteoma)

15(0.8%)

Epithelial (adenoma or carcinoma)

4(0.2%)

Glandular (adenocarcinoma)

1(0.1%)

Mixed tumor

4(0.2%)

Tubular

(a) Data as of August?, 1986,for studies of at least 104 weeks

185

Nitrofurantoin, NTP TR 341

TABLE D4b. HISTORICAL INCIDENCE OF HEMATOPOIETIC SYSTEM TUMORS IN FEMALE B6C3F,

MICE RECEIVING NO TREATMENT (a)

Study

Incidence in Controls

Lymphoma

Lymphoma or Leukemia

Historical Incidence at Southern Research Institute

HC Blue No. 2

C.I. Disperse Blue 1

D-Mannitol

Ziram

Eugenol

Propyl gallate

Zearalenone

HCBlueNo. 1

Stannous chloride

TOTAL
SD(b)

Elange (c)
High
Low

Overall Historical Incidence

TOTAL
SD(b)

Range (c)
High
Low

12/50

12/50

17/50

17/50

14/48

14/48

6/50

11/50

12/50

13/50

8/50

9/50

15/50

15/50

6/50

7/50

5/50

6/50

95/448(21.2%)

104/448(23.2%)

8.96%

7.46%

17/50

17/50

5/50

6/50

590/2,041(28.9%)

616/2,041 (30.2%)

12.56%

12.24%

37/50

38/50

5/50

6/50

(a) Data asof August 7, 1986,for studies of at least 104 weeks

(b) Standard deviation

(c) Range and SD are presented for groups of 35 or more animals.

Nitrofurantoin, NTP TR 341

186

TABLE D4c. HISTORICAL INCIDENCE OF UTERINE GLANDULAR TUMORS IN FEMALE B6C3Fi MICE

RECEIVING NO TREATMENT (a)

Study Incidence of Adenomas. Adenocarcinomas, or Carcinomas in Controls

Historical Incidence at Southern Research Institute

HC Blue No. 2 0/50

C.I. Disperse Blue 1 0/50

D-Mannitol 0/47

Ziram 0/50

Eugenol 0/50

Propyl gallate 0/50

Zearalenone 0/50

HCBlueNo. 1 (b)l/50

Stannous chloride 0/49

TOTAL 1/446(0.2%)

SD(c) 0.67%

Range (d)

High 1/50

Low 0/50

Overall Historical Incidence

TOTAL (6)7/2,010(0.3%)

SD(c) 0.78%

Range (d)

High 1/47

Low 0/50

(a) Data as of August 7, 1986, for studies of at least 104 weeks

(b) Squamous cell carcinoma

(c) Standard deviation

(d) Range and SD are presented for groups of 35 or more animals.

(e) Includes one adenoma, NOS, one squamous cell carcmoma, and five adenocarcinomas, NOS

187 Nitrofurantoin, NTP TR 341

TABLE D4d. HISTORICAL INCIDENCE OF HEPATOCELLL LAR TUMORS IN FEMALE BGCSF, MICE

RECEIVING NO TREATMENT (a)

Study

Adenoma

Incidence in Controls

Carcinoma Adenoma or Carcinoma

Historical Incidence at Southern Research Institute

HC Blue No. 2

C.I. Disperse Blue 1

D-Mannitol

Ziram

Eugenol

Propyl gallate

Zearalenone

HC Blue No. 1

Stannous chloride

TOTAL
SD(b)

Range (cl
High
Low

3/50

4/50

7/50

2/50

1/50

3/50

0/48

3/48

3/48

7/50

2/50

9/50

0/50

2/50

2/50

0/50

3/50

3/50

0/50

3/50

3/50

2/50

1/50

3/50

3/49

0/49

3/49

17/447(3.8%)

19/447(4.3%)

36/447(8.1%)

4.64%

2.56%

4.67%

7/50

4/50

9/50

0/50

0/49

2/50

Overall Historical Incidence

TOTAL
SD(b)

Range (c)
High
Low

97/2,033(4.8%)

(d) 83/2,033 (4.1%)

177/2,033(8.7%)

4.14%

2.61%

4.75%

9/49

5/50

10/49

0/50

0/50

0/50

(a) Data as of August 7, 1986, for studies of at least 104 weeks
(b) Standard deviation

(c) Range and SD are presented for groups of 35 or more animals.

(d) One hepatoblastoma was also observed.

TABLE D4e. HISTORICAL INCIDENCE OF LIVER ITO CELL TUMORS IN FEMALE B6C3F, MICE

RECEIVING NO TREATMENT (a)

Diagnosis Incidence in Controls

Historical Incidence at Southern Research Institute
Overall Historical Incidence

Ito cell tumors
Lipomas

0/447

0/2,033

1/2,033 (0.05% I

(a) Data as of August 7, 1986, for studies of at least 104 weeks

Nitrofurantoin, NTP TR 341

188

TABLE D5. SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE IN THE

TWO-YEAR FEED STUDY OF NITROFURANTOIN

Untreated Control

Low-

Dose

High

Dose

Animals initially in study

50

so

50

Animals removed

50

50

50

Animals examined histopathologically

60

50

50

ALIMENTARY SYSTEM

Gallbladder

(41)

(39)

(42)

Dilatation

(5%)

Liver

(50)

(50)

(50)

Basophilic focus

1

(2%)

Hematopoietic cell proliferation

(2%)

Necrosis, multifocal

(2%)

2

(4%)

2

(4%)

Vacuolization cytoplasmic, focal

(2%)

Kupffer cell, pigmentation

(2%)

Sinusoid, infiltration cellular.

polymorphonuclear

17

(34%)

Mesentery

(21)

(2)

(8)

Cyst

(5%)

Hemorrhage, focal

(5%)

Inflammation, subacute, focal

(5%)

Inflammation, suppurative, acute.

multifocal 14

(67%)

Fat, necrosis, focal

(19%)

1

(50%)

3

(38%)

Pancreas

(47)

(50)

(50)

Abscess

(2%)

Atrophy

(6%)

1

(2%)

2

(4%)

Inflammation, subacute

(2%)

Duct, cyst

(2%)

2

(4%)

Stomach, forestomach

(49)

(49)

(50)

Hyperkeratosis

(2%)

Hyperplasia

(2%)

1

(2%)

Inflammation, suppurative, acute.

focal 1

(2%)

Ulcer

(2%)

1

(2%)

Stomach, glandular

(49)

(49)

149)

Edema

1

(2%)

Mineralization

1

(2%)

Ulcer

1

i2%i

CARDIOVASCULAR SYSTEM
None

ENDOCRINE SYSTEM
Adrenal gland, cortex

Accessory adrenal cortical nodule

Congestion

Cyst

Degeneration, fatty

Spindle cell, hyperplasia
Adrenal gland, medulla

Hyperplasia
Pituitary gland

Pars distalis, angiectasis

Pars distalis, hyperplasia
Thyroid gland

Follicle, cyst

Follicle, degeneration, cystic

Follicle, hyperplasia, cystic

(50)

1

(2%i

1

(2%)

1

(2%)

3

(6%)

(48)

1

(2%)

(48)

1

(2%)

(48)

2

(4%)

3

(6%)

(50)

(50)

1 (2%)

4 (8%)

41

(82%)

45 (90%

(49)

(49)

2

(4%)

2 (4%)

(44)

(49)

1

(2%)

2 (4%)

3

(7%)

2 (4%)

(49)

(50)

2 (4%)

1 (2%)

189

Nitrofurantoin, NTP TR 341

TABLE D5. SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE IN THE
TWO-YEAR FEED STUDY OF NITROFURANTOIN (Continued)

Untreated Control

Low Dose

High Dose

GENERAL BODY SYSTEM
None

GENITAL SYSTEM
Clitoral gland

Cyst
Ovary

Abscess

Atrophy

Cyst

Hemorrhage

Inflammation, chronic

Mineralization
Uterus

Hydrometria

Hyperplasia, cystic

Inflammation, suppurative, acute

(1)

1 (100%)
(50)

18 (36%)

14 (28%)

1 (2%)

1 (2%)

(50)

2 (4%)
49 (98%)
11 (22%)

(50)

48 (96%)
10 (20%)

1 (2%)

2 (4%)
(50)

49 (987o)

(50)

49

(98%)

15

(30%)

3

(6%)

3

(6%)

50)

1

(2%)

49

(98%)

HEMATOPOIETIC SYSTEM

Lymph node (50)

Deep cervical, inflammation, suppurative, acute,
focal

Iliac, hyperplasia

Mediastinal, congestion

Mediastinal, hyperplasia

Renal, congestion

Renal, hyperplasia
Lymph node, mandibular

Congestion

Hyperplasia

Inflammation, suppurative, acute, focal
Lymph node, mesenteric

Angiectasis

Hyperplasia
Spleen

Angiectasis

Hematopoietic cell proliferation

Hemorrhage
Thymus (47)

Cyst

Hemorrhage 1

Mediastinum, inflammation, suppurative, acute 1

2

(4%)

2

(4%)

1

(2%)

1

(2%)

6

(12%)

(50)

1

(2%)

1

(2%)

1

(2%)

(45)

3

(7%)

2

(4%)

(49)

1

(2%)

21

(43%)

(50)

1 (2%)
(50)

(45)

(50)

4 (8%)
1 (2%)
(48)

1 (2%)

(49)

(45)

(48)

(50)

1 (2%)

2 (4%)

(48)

■2 (4%)

(2%)
(2%)

INTEGUMENTARY SYSTEM
Mammary gland

Duct, cyst
Skin

Alopecia

Fibrosis

Hemorrhage, focal

Inflammation, granulomatous, focal

Inflammation, suppurative, acute

(50)
3

(50)
2

(6%)
(4%)

1

(2%)

1

(2%)

(48)
(50)

I (2%)

(48)

3 (6%)

(50)

1 (2%)
1 (2%)

Nitrofurantoin, NTP TR 341

190

TABLE D5.

SUMMARY OF THE INCIDENCE OF NONNEOPLASTIC LESIONS IN FEMALE MICE IN THE
TWO-YEAR FEED STUDY OF NITROFURANTOIN (Continued)

L'ntreated Control

Low Dose

High Dose

MUSCULOSKELETAL SYSTEM
Skeletal muscle

Inflammation, suppurative, acute

(2)

1 150%)

(1)

(2)

NERVOUS SYSTEM
Brain

Compression

Hydrocephalus

Inflammation, suppurative, acute, multifocal

Necrosis

(49)

(50)

1 (2%)
1 (2%)
1 (2%)
1 (2%)

(50)

1 (2%)

RESPIRATORY SYSTEM
Lung

Congestion

Hemorrhage, focal

Infiltration cellular, histiocytic, multifocal

Inflammation, subacute

Inflammation, subacute, multifocal

Inflammation, suppurative, acute

Alveolar epithelium, hyperplasia

Mediastinum, inflammation, suppurative, acute
Nose

Hemorrhage

Inflammation, granulomatous

Inflammation, suppurative, acute

(50)

(50)

(50)

1 (2%)

1

(2%)

1 (2%)

1

(2%)

1 (2%)

1 (2%)

25 (50%)

19 (38%)

19

(38%)

I (2%)

2 (4%)

3

(6%)

2 (4%)

(48)

(50)

(50)

1

(2%)

1 (2%l

1 (2%)

1

(2%)

SPECIAL SENSES SYSTEM
Harderian gland
Hemorrhage

(2)

(5)
1 (20%)

(1)

URINARY SYSTEM
Kidney

Cyst

Fibrosis, focal

Hydronephrosis

Metaplasia, osseous, focal

Nephropathy

Cortex, infarct

Cortex, mineralization, diffuse

Medulla, mineralization

Renal tubule, degeneration, multifocal

Renal tubule, dilatation

Renal tubule, necrosis, multifocal
Urinary bladder

Inflammation, chronic

(50)

1 (2%)

1 (2%)
(50)
3 (6%)

(50)

1

(2%)

1

(2%)

1

(2%)

1

(2%)

3

(6%)

1

(2%)

1 (2%)

(50)

2 (4%)

(50)

1 (2%)

2 (4%)

1 (2%)

7 (14%)

1 (2%)

1 (2%)

(50)

191

Nitrofurantoin, NTP TR 341

Nitrofurantoin, NTP TR 341 192

APPENDIX E

GENETIC TOXICOLOGY OF

NITROFURANTOIN

TABLE El MUTAGENICITY OF NITROFURANTOIN IN SALMONELLA TYPHIMURWM

TABLE E2 MUTAGENICITY OF NITROFURANTOIN IN MOUSE L5178Y LYMPHOMA CELLS

TABLE E3 INDUCTION OF SISTERCHROMATID EXCHANGES IN CHINESE HAMSTER OVARY

CELLS BY NITROFURANTOIN

TABLE E4 INDUCTION OF CHROMOSOMAL ABERRATIONS IN CHINESE HAMSTER OVARY

CELLS BY NITROFURANTOIN

TABLE E5 INDUCTION OF SEX-LINKED RECESSIVE LETHAL MUTATIONS IN DROSOPHILA

BY NITROFURANTOIN

PAGE
194
195

196

197

198

193

Nitrofurantoin, NTPTR 341

TABLE El. MUTAGENICITY OF NITROFURANTOIN IN SALMONELLA TYPHIMURIUM (a)

Dose

Revertan

ts/Plate (b)

Strain (iig/plate)

-S9

^ S9 (hamster)

+ S9 (rat)

TA1535 0

21 ±

3.7

10 ±

1.7

7

+

2.5

0.1

17 ±

1.3

5 ±

3.4

14

+

0.6

0.3

22 ±

4.8

13 ±

3.5

9

+

2.0

1

21 ±

4.2

11 ±

1.7

13

+

0.3

3

17 +

5.7

15 ±

1.5

12

+

1.7

10

Toxic

13 ±

2.6

17

+

1.2

Trial summary

Negative

Negative

Nf

'gative

Positive control (c)

384 ±

17.9

429 ±

31.8

255

+

18.4

TA1537 0

8 ±

0.6

7 ±

3.0

13

±

2.9

0.1

10 ±

2.9

13 ±

3.2

11

±

2.2

0.3

9±

2.8

9 ±

1.5

14

+

1.9

1

13 ±

1.9

10 ±

3.3

8

+

0.9

3

7 +

1.2

17 ±

1.9

17

+

2.0

10

Toxic

18 ±

4.7

23

+

2.4

Trial summary

Negative

Negative

Negative

Positive controKc)

99 ±

3.5

479 ±

6.1

238

+

4.4

Revertants/Plate (b)

-S9

I-S9 (hamster)

+ S9

(rat)

Trial

1 1

T

rial 2

T

ria

1 1

Tri

iai 2

T

ria

il 1

Trial

2

TAIOO 0

145

+

1.5

120

+

4.7

160

t

11,8

126 ±

9.1

154

+

16.5

117 ±

7.8

0.03

136

+

13.2

156 ±

17.0

139 ±

12.5

0.1

337

+

7.5

228

+

16.7

288

+

166

214 ±

23.1

202

+

7.5

158 ±

7.3

0.3

860

+

17.9

528

+

38.2

501

±

24.2

440 ±

42.1

297

+

2.6

216 ±

14.2

1

1,471

+

31.2

1,216

+

34.0

990

+

30.6

824 ±

64.1

585

+

25.5

463 ±

20.4

3 (

dl469

+

183.2

404

+

24.0

931

+

43.9

670 +

87.9

1,131

+

53.2

870 ±

38.9

10

Tc

)xic

475

+

1202

(d)420

+

84.4

Trial summary

Positive

Posit

ive

F

'osl

tive

Positive

Posit

ive

Positive

Positive

control (c)

321

+

11.7

424

+

16.2

2,113

+

4.8

1,895 ±

83.7

1,055

+

61.4

900 ±

15 3

TA98 0

25

±

0.9

22

+

2.7

24

+

3.7

28 +

4.6

38

+

6.7

24 ±

2.6

0.03

18

+

2.6

29 +

0.7

-

34 ±

7.1

0.1

31

+

3.5

24

+

2.3

36

±

2.8

31 ±

2.2

29

+

3.8

24 ±

3.8

0.3

39

+

4.3

53

+

3.2

39

+

1.9

39 ±

0.9

33

+

3.5

29 ±

6.4

1

75

+

4.1

89

±

10.6

50

+

0.6

41 ±

2.9

40

+

4.4

38 ±

5.8

3

41

+

5.5

24

±

6.8

90

+

3.2

60 ±

1.5

59

+

4.6

56 ±

4.8

10

(d)0

+

0.0

33

+

3.7

65

+

6.1

Trial summary

Posr

tive

Po;

sitive

Positive

Positive

Positive

Posit:

ive

Positive

control (c)

830

+

33.4

760

+

8.0

1,779

±

45.1

1,761 ±

147.7

613

+

12.5

640 ±

8.7

(a) Study performed at SRI International The detailed protocol is presented in Haworth et al. (1983). Cells and study com-
pound or solvent (dimethy [sulfoxide I were incubated in the absence of exogenous metabolic activation ( -S9) or with Aroclor
1254-induced S9 from male Syrian hamster liver or male Sprague Dawley rat liver. High dose was limited by toxicity or
solubility but did not exceed 10 mg/plate;() pg/plate dose is the solvent control.

(b) Revertantsare presented as mean ± standard error from three plates.

(c) Positive control; 2-aminoanthracene was used on all strains in the presence of S9. In the absence of metabolic activation,
4-nitro-ophenylenediamine was used with TA98, sodium azide was used with TAIOO and TA1535, and 9-aminoacridine was
used with TA1537.

(d)Slight toxicity

Nitrofurantoin, NTP TR 341

194

TABLE E2. MUTAGENICITY OF NITROFURANTOIN IN MOUSE L5178Y LYMPHOMA CELLS (a.b)

Cloning Relative Mutant Mutant

Compound Concentration Efficiency Total Growth Count Fraction (c)

(Hg/ml) (percent) (percent)

Trial 1

Dimethyl sulfoxide (d) 108.0 ± 00 100.0 116.0 114.5 ± 4.5 35.5 ± 1.5

Nitrofurantoin 50 90.0 ± 46 65.0 ± 3.2 145.0 ± 5.6 53.7 ± 1.8

100 81.3 ± 5.9 44.7 ± 3.5 245.3+ 19.6 (e) 101.0 ± 3.6

150 79.3 ± 11.5 26.7+ 1.2 472.7+ 97.1 (e)196.3± 13.7

200 70.5 ± 4.5 22.5 ± 3.5 417.0 ± 181.0 (e) 194.0 ± 74.0

(f)300 67.3 ± 5.2 15.0 ± 1,5 586.7+ 44.6 (e)292.0± 12.1

500 54.3 1 3.9 9.0 1 0,6 485.7 1 41.2 (e)297.7+ 13,9

Methyl methanesulfonate 5 80.3 1 5.2 56.3 1 2.3 684.0 1 29.0 (e)285.3+ 6.1

Trial 2

Dimethyl sulfoxide (g) 84.0 1 17 100.0 1 6.1 63.5 1 8.1 25.3+ 3.0

Nitrofurantoin 50 84.7+ 5.6 65.0 1 2,6 102.7+ 26.5 (e)40.0+ 8.7

100 75.0+ 1,2 42,3+ 3,5 137.3 1 33.2 (e)60.7i 14.2

150 61.0 1 26 27.7 1 0.9 142.3 1 26.7 (el78.3 1 14.7

200 59.7 110,7 15.0 1 2.0 245.3 1 40.6 (e) 144.0 1 31.9

lf)300 37.3 1 5.6 6.3 1 1.2 297.0 1 38.2 (e)270.7 1 29.2

500 19.3 1 2.8 2.0 1 0.0 189.3 1 61.1 (ei374.0 1 141.5

Methyl methanesulfonate 5 56,7 1 6.1 44.0+ 3,5 247.7 1 20.2 (ei 149.0 1 20.0

(a I Study performed at Litton Bionetics, Inc. The experimental protocol is presented in detail by Myhr et al. (1985) and follows
the basic format of Clive et al. (1979). The highest dose of study compound is determined by solubility or toxicity and may not
exceed 5 mg/ml. All doses are tested in triplicate; the average for the three tests is presented in the table. Cells (6 X lOWml)
were treated for 4 hours at 37° C in medium, washed, resuspended in medium, and incubated for 48 hours at 37° C. After ex-
pression, 3 X 106 cells were plated in medium and soft agar supplemented with trifluorothymidine for selection of cells that
were mutant at the thymidine kinase (TK) locus, and 600 cells were plated in nonselective medium and soft agar to determine
the cloning efficiency. All trials were conducted in the absence of S9.

(b)Mean 1 standard error from replicate trials of approximately 1 x lO^cellseach, unlessotherwise noted. All data are evalu-
ated statistically for both trend and peak response ( P< 0.05 for at least one of the three highest dose sets). Both responses must
be significantly (P<0.05) positive for a chemical to be considered mutagenic. If only one of these responses is significant, the
call is "equivocal"; the absence of both trend and peak response results in a "negative" call.

(c) Mutant fraction (frequency) is a ratio of the mutant count to the cloning efficiency, divided by 3 (to arrive at MF per 1 X 10^
cells treated); MF = mutant fraction.

(d) Data presented are the average of two tests.

le) Significant positive response; occurs when the relative mutant fraction (average MF of treated culture/average MF of sol-
ventcontrol) isgreater than or equal to 1.6.

(f) Precipitate formed at this and all higher concentrations.

(g) Data presented are the average of four tests

195 Nitrofurantoin, NTFTR 341

TABLE E3. INDUCTION OF SISTER CHROMATID EXCHANGES IN CHINESE HAMSTER OVARY CELLS

BY NITROFURANTOIN (a)

Compound

No. of

SCEs/

Relative

Total

Chromo-

No. of

Chromo-

SCEs/

Hours

SCEs/Cell

Dose

Cells

somes

SCEs

some

Cell

In BrdU

(percent)

(Hg/mh

(b)

-S9 (c)

Trial I -Summary: Equivocal
Dimethyl sulfoxide
Nitrofurantoin

Mitomycin C

Trial 2-Summary: Positive
Dimethyl sulfoxide
Nitrofurantoin

Mitomycin C

10
25
33
50

0.001
0.01

-HS9 (e)

Trial 1 Summary: Weakly positive
Dimethyl sulfoxide
Nitrofurantoin

Cyclophosphamide

Trial 2--Summary: Weakly positive
Dimethyl sulfoxide
Nitrofurantoin

Cyclophosphamide

50

50

50

50

50

0

50
5

50

50

1 ,023

1,012

1 ,000
1,006
1,020

1,013
101

1,018

1 ,020

493

0.48

9.9

26.0

3.3

50

1,007

487

0.48

9.7

(d)32.0

98.0

10

50

1,026

489

0.48

9.8

(d)32.0

99.0

33.3

50

1,011

583

0.58

11.7

(dl32.0

118.2

100

0

26.0

0.001

50

1 ,027

,597

0,58

11.9

26.0

120.2

0.01

5

103

144

1.40

28.8

26.0

290.9

455

532
623

774

602
153

483

0.45

9.1

26.0

0.53

10.6

(d)31.5

116.5

0.62

12.5

(d)31.5

137.4

0.76

15.5

(d)31.5
26.0

170.3

0,59

12.0

26.0

131.9

1 51

30.6

26.0

336.3

0.47

9.7

26.0

33

50

1.006

483

0.48

9.7

(d)32.0

lOO.O

100

50

1,001

528

0.53

10.6

(d)32.0

109.3

333

50

1,016

602

0.59

12.0

(d)32.0

123.7

,000

0

26.0

0.4

50

1,033

673

0.65

13.5

26.0

139.2

2

5

104

178

1.71

35.6

26.0

367.0

457

0.45

9.1

26.0

330

50

1,028

498

0.48

10.0

(d)31.5

109.9

500

50

1,017

526

0.52

10.5

(d)31.5

115.4

750

50

1,001

634

0.63

12.7

(d)31.5

139.6

0.4

50

1,029

649

0.63

13.0

26,0

142.9

2

5

101

150

1.49

30.0

26.0

329,7

Nitrofurantoin, NTP TR 341

196

TABLE E3. INDUCTION OF SISTER CHROMATID EXCHANGES IN CHINESE HAMSTER OVARY CELLS

BY NITROFURANTOIN (Continued)

(a) Study performed at Litton Bionetics, Inc. SCE = sister chromatid exchange; BrdU = bromodeoxyuridine. A detailed de-
scription of the SCE protocol ispresentedby Galloway etal. (1985). Briefly, Chinese hamster ovary cells were incubated with
study compound or solvent (dimethylsulfoxide) as described in Ic) or (e) below and cultured for sufTicient time to reach second
metaphase division. Cells were then collected by mitotic shake-off, fixed, air-dried, and stained.
(b)SCEs/cell in treated culture expressed as a percent of the SCEs/cell in the control culture

(c) In the absence of 89, Chinese hamster ovary cells were incubated with study compound or solvent for 2 hours at 37° C. Then
BrdU was added, and incubation was continued for 24 hours. Cells were washed, fresh medium containing BrdU and colcemid
was added, and incubation was continued for 2-3 hours.

(d) Because some chemicals induce a delay in the cell division cycle, harvest times are occasionally extended to maximize the
proportion of second division cells available for analysis.

(e) In the presence of S9, cells were incubated with study compound or solvent for 2 hours at 37° C. Then cells were washed, and
medium containing BrdU was added. Cells were incubated for a further 26 hours, with colcemid present for the final 2-3 hours.
S9 was from the liver of Aroclor 1254-induced maleSprague Dawley rats.

197 Nitrofurantoin, NTPTR 341

TABLE E4.

INDUCTION OF CHROMOSOMAL ABERRATIONS IN CHINESE HAMSTER OVARY CELLS

BY NITROFURANTOIN (a)

Trial 1

Dose

Total

No. of

Abs/

Percent

((ig/ml)

Cells

Abs

Cell

Cells
with Abs

Trial 2

Dose

Total

No. of

Abs/

Percent

(|ig/ml)

Cells

Abs

Cell

Cells
with Abs

-S9(b)

Dimethyl sulfoxide

200

200

Nitrofurantoin

10

200

25

200

50

200

75

0

0.01

0.00

0.02
0.03
0,05

1.0
0.0

2.0
3.0
5.0

Summary: Weakly positive

Mitomycm C

0.05

200

44

0.22

19,0

0.08

25

25

1.00

60,0

+ S9 (c)

Dimethyl sulfoxide

Dimethyl sulfoxide

200

4

0.02

2,0

100

0

0.00

0.0

200

1

0.01

1.0

100

2

0.02

2.0

Nitrofurantoin

Nitrofurantom

250

200

4

0.02

2.0

747 100

10

0.10

8.0

500

200

2

0.01

1.0

900 100

23

0.23

17,0

750

200

17

0.09

6.0

950 100

33

0.33

16.0

1,000

0

Summary:

Weal<ly positi

ive

Summary:

Positive

Cyclophosphamide

Cyclophosphamide

6.25

200

10

0.05

4.0

6.25 100

15

0.15

13,0

12.5

25

7

0.28

28.0

12.5 25

12

0.48

40,0

(a) Study performed at Litton Bionetics, Inc, Abs = aberrations. Harvest time--20.0 hours. A detailed presentation of the tech-
nique for detecting chromosomal aberrations is presented by Galloway et al. ( 1985). Briefly, Chinese hamster ovary cells were
incubated with study compound or solvent (dimethyl sulfoxide) as indicated in (bl or (c). Cells were arrested in first metaphase
by addition of colcemid and harvested by mitotic shake-off, fixed, and stained in 6% Giemsa.

(b) In the absence of S9, Chinese hamster ovary cells were incubated with study compound or solvent (dimethyl sulfoxide! for 8-
10 hours at 37° C. Cells were then washed, and fresh medium containing colcemid was added for an additional 2-3 hours fol-
lowed by harvest.

(c) In the presence of S9, cells were incubated with study compound or solvent (dimethyl sulfoxide) for 2 hours at 37°C. Cells
were then washed, medium was added, and incubation was continued for 8- 10 hours. Colcemid was added for the last 2-3 hours
of incubation before harvest. S9 was from the liver of Aroclor 1254- induced male Sprague Daw ley rats.

Nitrofurantoin, NTP TR 341

198

TABLE E5.

INDUCTION OF SEX-LINKED RECESSIVE LETHAL MUTATIONS IN DROSOPHILA BY

NITROFURANTOIN (a)

Route of
Exposure

Dose

(ppm)

Incidence of

Deaths

(percent)

Incidence of
Sterility
(percent)

No. of Lethal
Mating 1

s/No,

. of X Chromosomes Tested Overall
Mating 2 Mating 3 Total

(b)

Feeding
Injection

2,000
0

10,000
0

52
13

25
1

5/5,063
5/4,271

1/2,822
2/1,964

4/3,279
3/2,942

1/2,856
1/1,919

3/3,400 12/11,742(0.10%)
2/2,396 10/9,609(0.10%)

1/2,591 3/8.269(0.04%)
1/1.882 4/5,765(0.07%)

(a) Study performed at Bowling Green State University. A detailed protocol of the se.x-linked recessive lethal assay is presented
in Zimmering et al. (1985). I Exposure by feeding was done by allowing 24- hour-old Canton-S males to feed for 3 days on a solu-
tion of the study chemical dissolved in 5% sucrose. In the mjection experiments, 24-hour-old Canton-S males were treated with
a solution of the chemical dissolved in 0.7% saline and allowed 24 hours to recover. I Exposed males were mated to three Base fe-
males for 3 days and given fresh females at 2-day intervals to produce three broods of 3, 2, and 2 days; sample sperm from suc-
cessive matings were treated as spermatozoa (mating 1 ), spermatids (mating 2), and spermatocytes (mating 3 1. F[ heterozygous
females were crossed to their siblings and placed ui individual vials. Fj daughters from the same parental male were kept to-
gether to identify clusters; no clusters were found. After 17 days, presumptive lethal mutations were identified as vials con-
taining no wild- type males; these were re tested. Results were notsignificant at the 5% level (Margolin et al., 1983).

(b) Combined total of number of lethal mutations/number of X chromosomes tested for three mating trials

199

Nitrofurantoin, NTP TR 341

Nitrofurantoin, NTP TR 341 200

APPENDIX F

SENTINEL ANIMAL PROGRAM

PAGE

TABLE Fl MURINE VIRUS ANTIBODY DETERMINATIONS FOR RATS AND MICE IN THE

TWO-YEAR FEED STUDIES OF NITROFURANTOIN 203

201 Nitrofurantoin, NTP TR 341

APPENDIX F. SENTINEL ANIMAL PROGRAM

Methods

Rodents used in the Carcinogenesis Program of the National Toxicology Program are produced in op-
timally clean facilities to eliminate potential pathogens that may affect study results. The Sentinel
Animal Program is part of the periodic monitoring of animal health that occurs during the toxicologic
evaluation of chemical compounds. Under this program, the disease state of the rodents is monitored
via viral serology on sera from extra (sentinel) animals in the study rooms. These animals are un-
treated, and these animals and the study animals are both subject to identical environmental condi-
tions. The sentinel animals come from the same production source and weanling groups as the ani-
mals used for the studies of chemical compounds.

Fifteen B6C3Fi mice and 15 F344/N rats of each sex were selected at the time of randomization and
allocation of the animals to the various study groups. Five animals of each designated sentinel group
were killed at 6, 12, and 18 months on study. Data from animals surviving 24 months were collected
from 5/50 randomly selected control animals of each sex and species. The blood from each animal was
collected and clotted, and the serum was separated. The serum was cooled on ice and shipped to
Microbiological Associates' Comprehensive Animal Diagnostic Service for determination of the viral
antibody titers. The following tests were performed:

Hemagglutination Complement

Inhibition Fixation ELISA

Mice PVM (pneumonia virus of mice) M. Ad. (mouse adenovirus) MHV (mouse hepatitis

Reo3(reovirus type 3) LCM (lymphocytic chorio- virus) (12, 18, 24 mo)

GDVII (Theiler's encephalo- meningitis virus)

myelitis virus) MHV (6 mo)

Poly (polyoma virus) Sendai(18mo)

MVM (minute virus of mice)
Ectro (infectious ectromelia)
Sendai(6, 12,24mo)

Rats PVM RCV(ratcoronavirus)

KRV(Kilham rat virus) SendaidSmo)

H-KToolan'sH-l virus)
Sendai (6, 12, 24 mo)

Results

Results are presented in Table Fl.

Nitrofurantoin, NTP TR 341 202

TABLE Fl.

MURINE VIRUS ANTIBODY DETERMINATIONS FOR RATS AND MICE IN THE TWO-YEAR
FEED STUDIES OF NITROFURANTOIN (a)

Interval (months)

Number of
Animals

Positive Serologic
Reaction for

RATS

2/10
10/10

RCV
Sendai

12

1/10
9/10

RCV
Sendai

18
24

4/9

3/10
2/10

Sendai

Sendai
KRV

MICE

6
12
18
24

10/10

4/10

9/9

5/10

Sendai
Sendai
Sendai
Sendai

(a) Blood samples were taken from sentinel animals at 6, 12, and 18 months after the start of dosing and from the control
animals just before they were killed; samples were sent to Microbiological Associates (Bethesda, MDjfor the Animal Disease
Screening Program.

203

Nitrofurantoin, NTP TR 341

Nitrofurantoin, NTP TR 341 204

APPENDIX G

FEED AND COMPOUND CONSUMPTION

BY RATS AND MICE IN THE TWO-YEAR FEED STUDIES

OF NITROFURANTOIN

TABLE Gl FEED AND COMPOUND CONSUMPTION BY MALE RATS IN THE TWO YEAR

FEED STUDY OF NITROFURANTOIN

TABLE G2 FEED AND COMPOUND CONSUMPTION BY FEMALE RATS IN THE TWO-YEAR

FEED STUDY OF NITROFURANTOIN

TABLE G3 FEED AND COMPOUND CONSUMPTION BY MALE MICE IN THE TWO-YEAR

FEED STUDY OF NITROFURANTOIN

TABLE G4 FEED AND COMPOUND CONSUMPTION BY FEMALE MICE IN THE TWO-YEAR

FEED STUDY OF NITROFURANTOIN

PAGE
206
207
208
209

205

Nitrofurantoin, NTP TR 341

TABLE Gl

FEED AND COMPOUND CONSUMPTION BY MALE RATS IN THE TWO-YEAR FEED STUDY

OF NITROFURANTOIN

Control

1,300 ppm

2,500 ppm

Grams

Body

Grams

Body

Dose/

Grams

Body

Dose/

Feed/

Weight

Feed/

Weight

Day (b)

Feed/

Weight

Day (b)

Week

Day (a)

(grams)

Day (a)

(grams)

Day (a)

(grams)

2

17

190

17

192

115

16

177

226

6

17

264

17

268

82

16

240

167

U

17

321

17

327

68

16

300

133

17

19

363

20

365

71

19

340

140

21

18

381

17

381

58

17

361

118

25

18

393

18

391

60

16

374

107

29

19

404

17

406

54

15

385

97

34

18

416

17

416

53

17

402

106

38

18

422

19

426

58

18

414

109

44

19

430

19

427

58

17

420

101

48

19

443

19

448

55

17

438

97

52

18

449

18

452

52

17

446

95

55

17

445

17

450

49

16

439

91

59

19

452

19

452

55

18

448

100

64

17

455

17

451

49

17

456

93

69

17

454

17

445

50

17

453

94

74

18

442

18

439

53

17

444

96

78

18

446

17

440

50

16

442

90

82

16

441

17

434

51

16

437

92

86

18

440

17

428

52

17

435

, 98

90

17

440

18

432

54

16

430

93

94

16

436

17

426

52

17

425

100

98

17

435

17

422

52

17

413

103

104

18

412

17

401

55

18

391

115

Mean

17.7

407

17.6

405

59

16.8

396

111

SD(c)

0.9

0.9

14

0.9

30

CV(d)

5.1

5.1

23.7

5.4

27.0

(a) Average grams of feed removed from feeder per animal per day. Not corrected for scatter.

(b) Estimated milligrams of nitrofurantoin consumed per day per kilogram of body weight

(c) Standard deviation

(d)CoefTicient of variation = (standard deviation/mean) X 100

Nitrofurantoin, NTP TR 341

206

TABLE

G2. FEED AND COMPOUND CONSUMPTION BY FEMALE

RATS IN THE TWO-YEAR FEED

STUDY OF NITROFURANTOIN

Control

600 ppm

1,300 ppm

Grams

Body

Grams

Body

Dose/

Grams

Body

Dose/

Feed/

Weight

Feed/

Weight

Day (b)

Feed/

Weight

Day (b)

Week

Day (a)

(grams)

Day (a)

(grams)

Day (a)

(grams)

2

13

135

13

138

57

13

136

124

6

12

170

14

174

48

12

167

93

11

11

193

11

198

33

11

191

75

17

10

210

12

216

33

11

206

69

21

12

214

11

223

30

11

212

67

25

11

225

11

231

29

11

217

66

29

11

228

11

239

28

11

223

64

34

11

237

U

248

27

11

233

61

38

12

245

12

256

28

11

237

60

44

13

256

13

271

29

11

246

58

48

U

267

11

279

24

10

256

51

52

13

283

13

294

27

12

270

58

55

12

288

13

300

26

13

277

61

59

13

303

13

310

25

13

291

58

64

13

316

12

325

22

12

302

52

69

13

328

13

339

23

12

314

50

74

13

332

13

343

23

13

318

53

78

14

340

13

347

22

13

324

52

82

13

343

14

355

24

13

331

51

86

13

345

14

359

23

14

339

54

90

13

352

14

367

23

14

344

53

94

14

353

14

368

23

14

346

53

98

14

361

14

375

22

13

352

48

104

14

352

13

359

22

14

351

52

Mean

12.5

278

12.6

288

28

12.2

270

62

SD(c)

1.1

1.1

8

1.2

17

CV(d)

8.8

8.7

28.6

9.8

27.4

(a) Average grams of feed removed from feeder per animal per day. Not corrected for scatter.

(b) Estimated milligrams of nitrofurantoin consumed per day per kilogram of body weight
(c)Standard deviation

(dlCoefTicient of variation = (standard deviation/mean) X 100

207

Nitrofurantoin, NTP TR 341

TABLE

G3. FEED AND COMPOUND CONSUMPTION BY MALE MICE IN THE TWO-YEAR FEED STUDY

OF NITROFURANTOIN

Control

1,300 ppm

2,500 ppm

Grams

Body

Grams

Body

Dose/

Grams

Body

Dose/

Feed;

Weight

Feed/

Weight

Day (b)

Feed/

Weight

Day (b)

Week

Day (a)

(grams)

Day (a)

(grams)

Day (a)

(grams)

4

6

27.6

6

27.6

283

6

25.3

593

8

7

30.9

8

29.7

350

7

28.5

614

13

7

33.4

8

31.7

328

8

31.0

645

18

7

34.9

7

33.9

268

7

32.0

547

22

8

36.0

8

35.0

297

8

33.1

604

26

8

37.2

7

35.3

258

8

34.1

587

30

7

37.4

8

36.4

286

7

34.0

515

35

10

38.0

9

36.8

318

9

34.9

645

40

8

38.1

8

36.2

287

8

35.4

565

45

9

39.1

8

37.9

274

8

36.0

556

49

8

40.0

7

37.6

242

7

36.6

478

53

8

41.4

7

39.0

233

7

37.6

465

57

9

41.5

8

39.6

263

8

37.8

529

61

9

41.3

9

39.1

299

8

37.7

531

65

9

41.4

8

40.2

259

8

37.9

528

69

9

41.1

8

39.6

263

8

37.5

533

74

9

40.7

9

38.0

308

8

36.4

549

78

9

40.4

9

38.2

306

8

36.3

551

82

9

41.0

9

38.6

303

8

37.0

541

86

10

39.6

10

37.2

349

9

36.3

620

90

9

40.0

11

38.6

370

9

36.3

620

94

10

40.1

10

38.5

338

9

36.0

625

98

9

39.9

12

38.9

401

11

36.3

758

104

5

39.6

6

39.1

199

6

36.5

411

Mean

8.3

38.4

8.3

36.8

295

7.9

35.0

567

SD(c)

1.3

1.4

46

11

71

CV(d)

15.7

16.9

15.6

13.9

12.5

(a) Average grams of feed removed from feeder per animal per day. Not corrected for scatter.

(b) Estimated milligrams of nitrofurantoin consumed per day per kilogram of body weight

(c ) Standard deviation

(djCoefTicientof variation = (standard deviation/mean) X 100

Nitrofurantoin, NTP TR 341

208

TABLE G4. FEED AND COMPOUND CONSUMPTION BY FEMALE MICE IN THE TWO-YEAR FEED

STUDY OF NITROFURANTOIN

Control

1,300 ppm

2,500 ppm

Grams

Body

Grams

Body

Dose/

Grams

Body

Dose/

Feedy

Weight

Feed/

Weight

Day (b)

Feed/

Weight

Day (b)

Week

Day (a)

(grams)

Day (a)

(grams)

Day (a)

(grams)

4

6

21.0

6

21.4

364

6

21.1

711

8

7

23.5

8

23.9

435

7

23.2

754

13

7

25.4

7

25.0

364

8

24.3

823

18

5

26.6

6

26.2

298

6

25.3

593

22

6

28.4

7

27.3

333

7

26.1

670

26

6

29.2

6

28.2

277

7

27.0

648

30

6

29.6

6

28.8

271

6

26.8

560

35

8

30.8

7

29.6

307

8

27.7

722

40

7

32.1

7

29.8

305

6

28.4

528

45

7

32.7

7

31.0

294

7

28.6

612

49

7

34.3

6

32.4

241

7

29.6

591

53

7

35.4

7

33.8

269

7

31.0

565

57

7

35.2

7

34.6

263

7

31.3

559

61

8

36.1

7

35.1

259

7

32.1

545

65

8

37.0

7

36.5

249

7

34.2

512

69

8

37.1

7

37.1

245

7

34.7

504

74

8

36.4

7

36.7

248

7

34.4

509

78

7

36.8

7

36.9

247

6

34.9

430

82

8

38.1

7

38.7

235

7

36.6

478

86

9

37.7

8

38.7

269

7

35.7

490

90

9

39.0

8

40.2

259

7

36.4

481

94

10

39.8

7

40.5

225

8

36.6

546

98

10

40.5

7

41.0

222

9

37.0

608

104

5

41.7

5

40.5

160

6

37.0

405

Mean

7.3

33.5

6.8

33.1

277

7.0

30.8

577

SD(c)

1.3

0.7

56

0.8

103

CV(d)

17.8

10.3

20.2

U.4

17.9

(a) Average grams of feed removed from feeder per anmial per day. Not corrected for scatter.

(b) Estimated milligramsofnitrofurantom consumed per day per kilogram of body weight
(c)Standard deviation

(d) Coefficient of variation = (standard deviation/mean) x 100

209

Nitrofurantoin, NTP TR 341

Nitrofurantoin, NTPTR 341 210

APPENDIX H

INGREDIENTS, NUTRIENT COMPOSITION, AND

CONTAMINANT LEVELS IN

NIH 07 RAT AND MOUSE RATION

Meal Diet: December 1980 to January 1983

(Manufactured by Zeigler Bros., Inc., Gardners, PA)

TABLE HI INGREDIENTS OF NIH 07 RAT AND MOUSE RATION

TABLE H2 VITAMINS AND MINERALS IN NIH 07 RAT AND MOUSE RATION

TABLE H3 NUTRIENT COMPOSITION OF NIH 07 RAT AND MOUSE RATION

TABLE H4 CONTAMINANT LEVELS IN NIH 07 RAT AND MOUSE RATION

PAGE
212

212
213
214

211

Nitrofurantoin, NTP TR 341

TABLE HI. INGREDIENTS OF NIH 07 RAT AND MOUSE RATION (a)

Ingredients (b)

Percent by Weight

Ground #2 yellow shelled corn

Ground hard winter wheat

Soybean meal (49% protein)

Fish meal (60% protein)

Wheat middlings

Dried skim milk

Alfalfa meal (dehydrated, 17% protein)

Corn gluten meal (60% protein)

Soy oil

Dried brewer's yeast

Dry molasses

Dicalcium phosphate

Ground limestone

Salt

Premixes (vitamin and mineral)

24.50

23.00

12.00

10.00

10.00

5.00

4.00

3.00

2.50

2.00

1.50

1.25

0.50

0.50

0.25

(a) NCI, 1976; NIH, 1978

(b) Ingredients ground to pass through a U.S. Standard Screen No. 16 before being mixed

TABLE H2. VITAMINS AND MINERALS IN NIH 07 RAT AND MOUSE RATION (a)

Amount

Source

Vitamins

A

5,500.000 lU

D3

4,600,000 lU

K3

2.8 g

d-Q-Tocopheryl acetate

20,000 lU

Choline

560.0 g

Folic acid

2.2 g

Niacin

30.0 g

d-Pantothenic acid

18.0 g

Riboflavin

3.4 g

Thiamine

lO.Og

Br2

4,000 ng

Pyridoxine

l-7g

Biotin

140.0 mg

Minerals

Iron

120.0 g

Manganese

60.0 g

Zinc

16.0 g

Copper

4.0 g

Iodine

1.4g

Cobalt

0.4 g

Stabilized vitamin A palmitate or acetate

D-activated animal sterol

Menadione

Choline chloride

d-Calcium pantothenate

Thiamine mononitrate

Pyridoxine hydrochloride
d-Biotin

Iron sulfate
Manganous oxide
Zinc oxide
Copper sulfate
Calcium iodate
Cobalt carbonate

(a) Perton (2,000 lb)offinished product

Nitrofurantoin, NTP TR 341

212

TABLE H3. NUTRIENT COMPOSITION OF NIH 07 RAT AND MOUSE RATION (a)

Mean ± Standard

Nutrients Deviation Range Number of Samples

Crude protein (percent by weight) 24.25 11.04 22.6-26.3 24

Crude fat (percent by weight) 5.10 10.44 4.4-6.0 24

Crude nber(percent by weight) 3.38 10.38 2.4-4.2 24

Ash (percent by weight) 6.59 + 0.34 5.97-7.42 24

Amino Acids (percent of total diet)

Arginine 1.260 1.21-1.31 2

Cystine 0.395 0.39-0.40 2

Glycine 1.175 1.15-1.20 2

Histidine 0.553 0.530-0.576 2

Isoleucine 0.908 0.881-0.934 2

Leucine 1.905 1.85-1.96 2

Lysine 1.250 1.20-1.30 2

Methionine 0.310 0.306-0.314 2

Phenylalanine 0.967 0.960-0.974 2

Threonine 0.834 0.827-0.840 2

Tryptophan 0.175 0.171-0.178 2

Tyrosine 0.587 0.566-0.607 2

Valine 1.085 1.05-1.12 2

Essential Fatty Acids (percent of total diet)

Linoleic 2.37 1

Linolenic 0.308 I

Arachidonic 0.008 1

Vitamins

VitammA(IU/kg) 11,188 11,239 8,900-14,000 24

Vitamin DdU/kg) 6,300 1

Q-TocopheroKppm) 37.6 31.1-44.0 2

Thiamine (ppm) 19.0 13.02 14.0-26.0 (b)23

Riboflavin (ppm) 6.9 6.1-7.4 2

Niacin (ppm) 75 65-85 2

Pantothenic acid (ppm) 30.2 . 29.8-30.5 2

Pyridoxine(ppm) 7.2 ' 5.6-8.8 2

Folic acid (ppm) 2.1 1.8-2.4 2

Biotin(ppm) 0.24 0.21-0.27 2

Vitamin Bi2(ppb) 12.8 10.6-15.0 2

Choline (ppm) 3,315 3,200-3,430 2

Minerals

Calcium (percent) 1.23 10.12 1.10 1.53 24

Phosphorus (percent) 0.97 + 0.06 0.84 1.10 24

Potassium (percent) 0.809 0.7720.846 2

Chloride (percent) 0.557 0.479-0.635 2

Sodium (percent! - 0.304 0.258-0.349 2

Magnesium (percent) 0.172 0.166-0.177 2

Sulfur (percent) 0.278 0.270-0.285 2

Iron (ppm) 418 409-426 2

Manganese (ppm) 90.8 86.0-95.5 2

Zinc(ppm) 55.1 54.2-56.0 2

Copper(ppm) 12.68 9.65-15.70 2

lodine(ppm) 2.58 1.52-3.64 2

Chromium (ppm) 1.86 1.79 1.93 2

Cobalt(ppm) 0.57 0.49-0.65 2

(a) One or two batches of feed analyzed for nutrients reported in this table were manufactured in January and/or April 1983.

(b) One batch (7/22/81 ) not analyzed for thiamine

213 Nitrofurantoin, NTP TR 341

TABLE H4. CONTAMINANT LEVELS IN NIH 07 RAT AND MOUSE RATION

Contaminants

Mean ± Standard
Deviation

Range

Number of Samples

Arsenic (ppm)
Cadmium (ppm) (al
Lead (ppm)
Mercury (ppm) (a)
Selenium (ppm)

Aflatoxins(ppb) (a,b)

Nitrate nitrogen (ppm) (c)
Nitrite nitrogen (ppm)(c)
BHA(ppm)(d,e)
BHT(ppm)(d)

Aerobic plate count (CFU/g)
Coliform(MPN/g)(f)
£. co/i(MPN/g)(f.g)
£.co/!(MPN/g){hl

Total nitrosamines(ppb) li.j)
Total nitrosamines(ppb) li.k)
N-Nitrosodimethylamine Ippb) (i,l)
A/-Nitrosodimethylamine Ippb) (i,m)
/V-Nitrosopyrrolidine (ppb)

Pesticides (ppm)

a BHC(a,n)

P BHC(a)

Y BHC-Lindane(a)

8BHC(a)

Heptachlor(a)

Aldrin(a)

Heptachlor epoxide (a)

DDE(o)

DDD(a)

DDT (a)

HCB(a)

Mirex(a)

Methoxychlor (p)

Dieldrin(a)

Endrin(a)

Telodrin(a)

Chlordane(a)

Toxaphene(a)

Estimated PCBs (a)

Ronnel(a)

Ethion(a)

Trithion(a)

Diazinon(o)

Methyl parathion (a)

Ethyl parathion (a)

Malathion(q)

Endosulfan I (a)

Endosulfan II (al

Endosulfan sulfate (a)

0.44 ± 0.14

<0.1
1.03 ± 0.75

<0.05
0.27 ± 0.05

<10

9.35 + 4.35

1.97 ± 1.28

5.83 ± 5.12

3.42 ± 2.57

105,438 ± 75.797

1,046 ± 973

8.0 ± 7.91

13.92 ± 30.0

5.13 ± 4.47

13.11 ± 27.39

3.82 ± 4.29

11.71 ± 27.03

1.21 ± 0.66

<0.01
<0.02
<0.01
<0.01
<0.01
<0.01
<0.01
<0.01
<0.01
<0.01
<0.01
<0.01
<0.05
<0.01
<0.01
<0.01
<0.05
<0.1
<0.2
<0.01
<0.02
<0.05
<0.1
<0.02
<0.02
0.08 ± 0.05
<0.01
<0.01
<0.03

<0.21-0.93

0.27-2.93

0.16-0.40

<5.0 10.0

0.6-18.0

0.4-5.3

0.4-20.0

<1.0-13.0

7,000-300,000

< 3-2,400
<3-23

<3-150

< 1.2-18.8
<1.2-101.6

0.6-16.8

0.6-99

<0.3-2.4

0.05(7/14/81)

0.13 (8/25/81 ); 0.06 (6/29/82)

<0.05-0.25

24
24
24
24
24

24

24
24
24
24

24
24
23
24

22
24
22
24
24

24
24
24
24
24
24
24
24
24
24
24
24
24
24
24
24
24
24
24
24
24
24
24
24
24
24
24
24
24

Nitrofurantoin, NTP TR 341

214

TABLE H4. CONTAMINANT LEVELS IN NIH 07 RAT AND MOUSE RATION (Continued)

(a) All values were less than the detection limit, given in the table as the mean.

(b) The detection limit was reduced from 10 ppb to 5 ppb after 7/81.

(c) Source of contamination: alfalfa, grains, and fish meal

(d) Source of contamination: soy oil and fish meal

(e)One batch contained less than 0.5 ppm. The value was < 0.04, and the batch was produced on 4/27/81.

(f)MPN = most probable number

(g) Mean, standard deviation, and range exclude one value of 150 ppm obtained for the batch produced on 8/26/82.

(h) Mean, standard deviation, and range include the high value listed in footnote (g).

(i) All values were corrected for percent recovery.

(j) Mean, standard deviation, and range exclude two very high values of 101.6 and 100.3 ppb obtained for batches produced on

1/26/81 and 4/27/81.

(k) Mean, standard deviation, and range include the high values listed in footnote (j).

(I) Mean, standard deviation, and range exclude two very high values in the range of 97.9 and 99 ppb obtained for batches

produced on 1/26/81 and 4/27/81 .

(m) Mean, standard deviation, and range include the very high values given in footnote (1).

(n)BHC = hexachlorocyclohexane or benzene hexachloride.

(o) There was one observation above the detection limit; the value and date it was obtained are given under the range.

(p) There were two observations above the detection hmit; the values and dates they were obtained are given under the range.

(q) Eleven batches contained more than 0.05 ppm.

215 Nitrofurantoin, NTP TR 341

Nitrofurantoin, NTPTR 341 216

APPENDIX I

AUDIT SUMMARY

217 Nitrofurantoin, NTPTR 341

APPENDIX I. AUDIT SUMMARY

The experimental data, documents, and pathology specimens for the 2-year toxicology and carcino-
genesis studies of nitrofurantoin in rats and mice were audited for accuracy, consistency, complete-
ness, and compliance with Good Laboratory Practice regulations of the Food and Drug Administra-
tion (fully implemented by the National Toxicology Program (NTP) beginning October 1, 1981). The
animal studies were conducted for the NTP by Southern Research Institute (Birmingham, AL) under
a subcontract with Tracor Jitco, Inc., until May 31, 1982, and then under contract with the National
Institute of Environmental Health Sciences (NIEHS). Animal exposure to the chemical in feed began
in February 1981. The retrospective audit was conducted for the NIEHS at the NTP Archives in June
1986 by Dynamac Corporation,. J. C. Bhandari, D.V.M., Ph.D., Principal Investigator. The other indi-
viduals who conducted the audit are listed in the full audit report, which is on file at the NIEHS. The
audit included a review of:

1. AH records concerning animal receipt, quarantine, randomization, and disposition prior to
study start.

2. Body weight, feed consumption, and clinical observation data for a random 10% sample of the
study animals.

3. All inlife records, including protocol, correspondence, environmental conditions, palpable
masses, mortality, animal identification, and correlation of final inlife observation of masses,
date of death, and disposition with necropsy records.

4. All chemistry records.

5. All postmortem records for individual animals concerning identification, disposition codes,
condition codes, correlations between gross observations and microscopic diagnoses, and tis-
sue accountability.

6. All wet tissue bags for inventory and wet tissues from a random 10% sample of the study ani-
mals, plus other relevant cases, to verify animal identification and examine for untrimmed
potential lesions

7. Blocks and slides of tissues from control and high dose groups of study animals to examine for
proper match and inventory.

8. Comparison of individual animal data tables.

9. Draft (4/87) of the NTP Technical Report on the 2-year studies of nitrofurantoin.

Inlife procedures and events were documented adequately by the archival records with the exception
of analytical data for chemical/vehicle samples in the animal room and bulk chemical reanalysis
within 30 days prior to initiation of the study. The relatively few audit findings from review of the in-
life records were miscellaneous and minor; for example, four mice (one control male, two control fe-
males, and one high dose female) were coded as moribund kill in clinical records and were coded as
found dead in necropsy records. No corresponding necropsy descriptions of masses were found for five
rats; these discrepancies were resolved by examination of the wet tissues. Animal disposition for one
control male rat (animal no. 10) and one high dose male mouse (animal no. 7) was found to be recorded
in error in EISRPTOl . Because the resulting changes in the statistical analyses did not infiuence in-
terpretation of the studies, these changes were not incorporated into the Technical Report.

Inspection of wet tissues for individual animal identifiers showed that 42/48 rats and 31/34 mice were
identified correctly by their residual tissues. All animals that were cross-checked were correctly
identified, with the exception of one control male rat with a torn ear that prevented further verifica-
tion. The audit identified five untrimmed potential lesions in the wet tissues of 48 rats examined and
three untrimmed potential lesions in 34 mice examined. These lesions were in nontarget tissues and
were examined by NTP pathologists. The correlation between gross observation and microscopic di-
agnoses was very good (only one noncorrelation in rats and three in mice). Full details of these and
other audit findings are presented in the audit report. In conclusion, the study records at the NTP
Archives support the data and results presented in the NTP Technical Report.

Nitrofurantoin, NTP TR 341 218

NATIONAL TOXICOLOGY PROGRAM TECHNICAL REPORTS
PRINTED AS OF JULY 1989

TR No. CHEMICAL

201 2,3,7 ,8Tetrachlorodibenzo-p-dioxin (Dermal)

206 Dibromochloropropane

207 Cytembena

208 FD&CYellowNo.6

209 2,3,7,8-Tetrachlorodibenzo-p-dioxin (Gavage)

210 1,2-Dibromoethane (Inhalation)

211 C.I. Acid Orange 10

212 Di(2-ethylhexyl)adipate

213 Butylbenzyl Phthalate

214 Caprolactam

215 BisphenolA

216 11-Aminoundecanoic Acid

217 Di(2-ethylhexyl)phthalate

219 2,6-Dichloro-p-phenylenediamine

220 C.I. Acid Red 14

221 Locust Bean Gum

222 C.I. Disperse Yellow 3

223 Eugenol

224 Tara Gum

225 D & C Red No. 9

226 C.l. Solvent Yellow 14

227 Gum Arabic

229 Guar Gum

230 Agar

231 Stannous Chloride

233 2-Biphenylaniine Hydrochloride

234 AUyl Isothiocyanate

235 Zearalenone

236 D-Mannilol

238 Ziram

239 Bis(2-chloro-l-methylethyl)ether

240 Propyl Gallate

242 Diallyl Phthalate (Mice)

244 Polybrominated Biphenyl Mixture

245 Melamine

247 L-Ascorbic Acid

248 4,4'-Methylenedianiline Dihydrochloride

249 Amosite Asbestos

250 Benzyl Acetate

251 Toluene Diisocyanate

252 Geranyl Acetate

253 Allyl Isovalerate
255 1,2-Dichlorobenzene

257 Diglycidyl Resorcinol Ether

259 Ethyl Acrylate

261 Chlorobenzene

263 1,2-Dichloropropane

266 Monuron

267 Propylene Oxide
269 Telone II®

271 HC Blue No. 1

272 Propylene

273 Trichloroethylene( Four strains of rats)

274 Tris(2-ethylhexyl)phosphate

275 2-Chloroethanol

276 8-Hydroxyquinoline

280 Crocidolite Asbestos

281 HC Red No. 3

282 Chlorodibromomethane

TR No

,. CHEMICAL

284

Diallylphthalate(Rats)

285

C.I. Basic Red 9 Monohydrochloride

287

Dimethyl Hydrogen Phosphite

288

1,3-Butadiene

289

Benzene

291

Isophorone

293

HC Blue No. 2

294

Chlorinated Trisodium Phosphate

295

Chrysotile Asbestos (Rats)

296

Tetrakis(hydroxymethyl)phosphonium Sulfate and

Tetrakis(hydrQxymethyl)phosphonium Chloride

298

Dimethyl Morpholinophosphorainidate

299

C.I. Disperse Blue 1

300

3-Chloro-2methylpropene

301

o-Phenylphenol

303

4-Vinylcyclohexene

304

Chlorendic Acid

305

Chlorinated Paraffins (C23, 43% chlorine)

306

Dichloromethane

307

Ephedrine Sulfate

308

Chlorinated Paraffins (C12, 60% chlorine)

309

Decabromodiphenyl Oxide

310

Marine Diesel Fuel and JP-5 Navy Fuel

311

Tetrachloroethylene (Inhalation)

312

n-Butyl Chloride

314

Methyl Methacrylate

315

Oxytetracycline Hydrochloride

316

1 -Chloro-2-methylpropene

317

Chlorpheniramine Maleate

318

Ampicillin Trihydrate

319

1,4-Dichlorobenzene

320

Rotenone

321

Bromodichloromethane

322

Phenylephrine Hydrochloride

323

Dimethyl Methylphosphonate

324

Boric Acid

325

Pentachloronitrobenzene

326

Ethylene Oxide

327

Xylenes (Mixed)

328

Methyl Carbamate

329

1,2-Epoxybutane

330

4'Hexylresorcinol

331

Malonaldehyde, Sodium Salt

332

Mercaptobenzothiazole

333

iV-Phenyl-2-naphthylamine

334

2-Amino-5-nitrophenol

335

C.I. Acid Orange 3

336

Penicillin VK

337

Nitrofurazone

338

Erythromycin Stearate

339

2-Amino-4-nitrophenol

343

Benzyl Alcohol

345

Roxarsone

348

a-Methyldopa Sesquihydrate

349

Pentachlorophenol

350

Tribromomethane

353

2,4-Dichlorophenol

356

Furosemide

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