Background Lesions in Laboratory Animals

A Color Atlas
Background Lesions in
Laboratory Animals
A Color Atlas
Elizabeth F McInnes BVSc, PhD, FRCPath, MRCVS, FIAP
Department of Pathology, Huntingdon Life Sciences, Cambs,
United Kingdom

Foreword by
Peter Mann DVM, DACVP
Manager, EPL NorthWest, Seattle, WA, USA

Edinburgh  London  New York  Oxford  Philadelphia  St Louis  Sydney  Toronto  2012
© 2012 Elsevier Ltd. All rights reserved.

First published 2012

Reprinted 2012

No part of this publication may be reproduced or transmitted in any form or by any means, electronic
or mechanical, including photocopying, recording, or any information storage and retrieval system,
without permission in writing from the publisher. Details on how to seek permission, further
information about the Publisher’s permissions policies and our arrangements with organizations such as
the Copyright Clearance Center and the Copyright Licensing Agency, can be found at our website:
www.elsevier.com/permissions.

This book and the individual contributions contained in it are protected under copyright by the
Publisher (other than as may be noted herein).

ISBN 978 0 7020 3519 7

British Library Cataloguing in Publication Data

A catalogue record for this book is available from the British Library

Library of Congress Cataloging in Publication Data

A catalog record for this book is available from the Library of Congress

Notices
Knowledge and best practice in this field are constantly changing. As new research and experience
broaden our understanding, changes in research methods, professional practices, or medical treatment
may become necessary.

Practitioners and researchers must always rely on their own experience and knowledge in evaluating and
using any information, methods, compounds, or experiments described herein. In using such information
or methods they should be mindful of their own safety and the safety of others, including parties for
whom they have a professional responsibility.

With respect to any drug or pharmaceutical products identified, readers are advised to check the most
current information provided (i) on procedures featured or (ii) by the manufacturer of each product to
be administered, to verify the recommended dose or formula, the method and duration of
administration, and contraindications. It is the responsibility of practitioners, relying on their own
experience and knowledge of their patients, to make diagnoses, to determine dosages and the best
treatment for each individual patient, and to take all appropriate safety precautions.

To the fullest extent of the law, neither the Publisher nor the authors, contributors, or editors, assume
any liability for any injury and/or damage to persons or property as a matter of products liability,
negligence or otherwise, or from any use or operation of any methods, products, instructions, or ideas
contained in the material herein.

The
Working together to grow publisher’s
policy is to use
libraries in developing countries paper manufactured
from sustainable forests
www.elsevier.com | www.bookaid.org | www.sabre.org

Printed in China
FOREWORD
The volume you hold in your hands reveals the first secret we
should learn as toxicologic pathologists: untreated animals are not
animals without lesions; the normal wear and tear of life causes a
number of histologic changes in many organs. That Dr McInnes and
her colleagues—Drs Chamanza, Taylor and Bradley, and Professor
Scudamore—are all experienced pathologists is obvious from their
selection and description of these changes; it is clear that each of these
authors has struggled with these, often perplexing, changes at some
point in their career. Each lesion has been carefully cataloged, described
and illustrated with color images. For each of the major species of labo-
ratory animal commonly used in toxicology studies, there is a complete,
well organized description of the background changes expected for each
of the major organ systems. In addition, there is an invaluable and
exhaustive list of references for each species; this should prove very
useful when documenting these changes in pathology narratives or
when presenting material to regulatory agencies. In addition to the
chapters on individual species, there is an additional chapter covering
vi
artifacts, i.e. pseudo-changes that may mimic background changes
but which must be separated out so that they are not reported as
treatment-related findings. Finally, Dr Creasy has admirably described
the normal changes in both the male and female reproductive tracts
in a manner which should help us mere mortals to separate changes
arising from treatment from background changes in these very complex
systems.
The second great secret we learn as toxicologic pathologists is
that treatment-related changes can be exacerbations of normal back-
ground changes. With the aid of this book and an adequate set of
control animals, the working toxicologic pathologist is much better
equipped to wend their way through mountains of slides, confident that
they are truly describing only those changes which are the result of
treatment.
Peter Mann
2011

Evaluating the pathology of tissues involves not only the recognition of
lesions caused directly by treatment or disease but also the identifica­
tion of background lesions. An experienced pathologist should be fami­
liar with the spontaneous, incidental background lesions present in a
particular organ or tissue (Shackleford et al 2002). Toxicologic patholo­
gists need to be able to recognize background lesions in animals of
different ages so that these changes are not incorrectly attributed to
the test article. Toxicologists and regulators are understandably keen to
ensure that background findings are not incorrectly attributed to test
article effects. This book is for veterinary and toxicologic pathologists,
at all stages of their training or career, who want to know more about
background lesions in laboratory animals.
Background lesions are described as an array of individual variations
with an accepted reference range (Shackelford et al 2002); they are
generally congenital or hereditary findings, i.e. normal findings that are
unique to the histology of an animal species. Occasionally, background
lesions include the effects of trauma. Background lesions also include
normal aging changes as well as some degenerative conditions. In
rodents, background lesions include normal findings such as hematopoi­
esis in the spleen, incomplete growth-plate resorption in the long bones,
continuous eruption of incisor teeth, high cellularity of bone marrow,
and hepatocyte polyploidy, karyomegaly and binucleation in the liver.
Traumatic background lesions include fractures and gavage injuries, bite
wounds and foot lesions. Background findings, which also include condi­
tions such as thymic involution, ovarian and uterine atrophy, may be
related to normal physiological or hormonal status and often include
minor developmental anomalies such as cysts or ectopic tissue. Degen­
erative background lesions resulting from the aging process include
chronic nephropathy, amyloidosis, atrial thrombosis, cardiomyopathy,
polyarteritis, hepatic foci of alteration, and urologic syndrome.
A drift away from the normal range of background lesions may be
associated with factors such as changes in supplier and/or geographic
source, genetics, age at sexual maturity (dogs and monkeys), diet,
environment, the experience and training of the pathologist, subjective
threshold preferences for background changes, and familiarity with
previous studies using animals from the same source or a different
source. Genetic and geographic influence on background findings can
occur in cynomolgus monkeys (Drevon-Gaillot et al 2006, Stevison &
Kohn 2008), and disease susceptibility, disease serology status and
stress response can also affect the incidence of certain background
lesions (Menninger et al 2002, Drevon-Gaillot et al 2006). This atlas
aims to provide pathologists with clear descriptions and photographs of
various lesions to facilitate recognition of background lesions despite
diagnostic drift.
Some pathologists will use thresholds to filter out background
changes—which may lead to under-diagnosis of a particular lesion.
Pathologists may also choose to use broad terms to include a number
PREFACE
of lesions; for instance, a pathologist may use the term ‘cardiomyopa­
thy’ to refer to myocardial necrosis, myocardial fibrosis and myocardial
inflammatory cell infiltration. Grouping findings together in this way
may lead to masking of a treatment-related finding, particularly in long-
term studies. In general, experienced pathologists tend to record fewer
background findings compared to their inexperienced counterparts.
Although standardized terminology is encouraged, there is an infinite
variety of findings and very often several terms exist for the same lesion.
The use of different diagnostic terms to describe the same lesion makes
accurate retrieval of historical control data challenging. In this atlas,
every effort has been made to give the lesion its correct name, but
synonyms for the same lesion have also been included.
This atlas describes background lesions and illustrates most of them
with a color image. There are chapters on each of the major species of
laboratory animal commonly used in toxicology studies, with a com­
plete, well-organized description of the background changes expected
for each of the major organ systems. There is also a chapter on back­
ground lesions in the reproductive system of all laboratory animals;
the reproductive system is discussed separately from the other organ
systems because of the considerable number of background lesions in
the reproductive tissues encountered due to normal physiology, housing
conditions and the impact of age (i.e. before or after the onset of
puberty). Evaluation of the reproductive system in non-human pri­
mates can be difficult owing to the small sample sizes and the timing
around puberty which can result in a large number of background
lesions that may mimic test-article-related findings; the reproductive
chapter contains detailed descriptions of the hormonal changes recorded
in female non-human primates, dogs and rodents. Finally, there is also
a chapter on artifacts caused during the death process (agonal) and
during the processing of tissues. The book is fully referenced.
The authors trust that you will find the book both useful and
interesting.
References
Drevon-Gaillot, E., Perron-Lepage, M.F., Clément, C., et al., 2006. Review of
background findings in cynomolgus monkeys (Macaca fascicularis) from
three different geographical origins. Exp. Toxicol. Pathol. 58, 77–88.
Menninger, K., Wieczorek, G., Riesen, S., et al., 2002. The origin of
cynomolgus monkey affects the outcome of kidney allografts under neoral
immunosuppression. Transplant Proc. 34, 2887–2888.
Shackelford, C., Long, G., Wolf, J., et al., 2002. Qualitative and quantitative
analysis of non-neoplastic lesions in toxicology studies. Toxicol. Pathol. 30,
93–96.
Stevison, L.S., Kohn, M.H., 2008. Determining genetic background in captive
stocks of cynomolgus macaques (Macaca fascicularis). J. Med. Primatol. 37,
311–317.
vii
ACKNOWLEDGMENTS

The authors would like to thank the following people for the generous Blanco, Stuart Naylor (Charles River), Heinrich Ernst (Fraunhofer
gift of photographs: Carlos Lopez, Antonio de Molina, Nigel Young, Institute for Toxicology and Experimental Medicine) and Gloria del
David Bell (HLS), David J. Lewis, J. Bowles (GSK), Matt Jacobsen, Fiero (Cerberus). Where possible, the individual photographs have
Jayne Harris (Astrazeneca), Alys Bradley, Ronnie Chamanza, Ana been acknowledged; however, in some cases this was not possible.
Dedication
For Peter, Edward and Simon (EM).

viii
 CONTRIBUTORS

Alys Bradley Cheryl Scudamore

BSc, BVSc, MAnimSc, DipRCPath, FRIPH, MRCVS, FRCPath BVSc, Dip Mgmt, PhD, MRCVS, FRCPath
Director of Pathology, Preclinical Services, Professor of Toxicological Pathology, Royal Veterinary
Charles River Laboratory, Tranent, Edinburgh, UK College, Hatfield, UK

Ronnie Chamanza Ian Taylor

BVSc, MSc, MRCVS, FRCPath BSc, DIBT, CBiol, MSB
Principal Pathologist, Preclinical Services, Principal Pathologist, Department of Pathology
Charles River Laboratory, Tranent, Edinburgh, UK Huntingdon Life Sciences, Suffolk, UK

Dianne Creasy
PhD, DipRCPath (Tox), FRCPath
Senior Scientific Advisor, Huntingdon Life Sciences,
New Jersey, USA

ix

Non-human primates: cynomolgus (Macaca fascicularis) and rhesus (Macaca mulatta)
Introduction
Non-human primates are extensively used as a
non-rodent species of choice in preclinical toxicity
testing, primarily because of their phylogenetic
and physiological proximity to humans. The most
commonly used species is the cynomolgus monkey
also known as the crab-eating or long-tailed
macaque (Macaca fascicularis), rhesus macaque
(Macaca mulatta) and the common marmoset
(Callithrix jacchus). Cynomolgus and rhesus
macaques are Old World monkeys (from Africa
and Asia) of the subfamily cercopithicine and
genus Macaca, while marmosets are New Word
monkeys (from Central and South America) of
the genus Callithridae. Cynomolgus monkeys
have gradually replaced rhesus monkeys as the
most commonly used laboratory non-human
primate mainly because of costs and availability.
Marmosets are preferred because of their small
size which is ideal for testing products in limited
amounts. They are also easier to handle and less
expensive to house. All three species can now be
obtained from accredited suppliers as young
animals (one to five years old), purpose-bred for
laboratory use and free of specific disease agents.
Young macaques may be easily housed in captivity
once obtained from suppliers without major
disease outbreaks, but the captive management of
marmosets remains a huge challenge and can be
associated with problems related to stress and the
strict dietary requirements of marmosets.
In spite of the immense value of non-human
primates in predicting human responses, there
are important ethical and scientific considerations
regulating their use. Current regulations dictate
that non-human primates should only be used
when there is no other appropriate alternative.
It has to be scientifically demonstrated that no
other non-rodent species commonly used in safety
testing is appropriate for the purpose of the study.
However, with the advent of biotechnology-
derived products or biologics, such as humanized
monoclonal antibodies, which due to their high
species specificity require testing in a pharmaco-
logically relevant animal species, the demand for
utilizing species that closely match human phar-
macodynamic responses has increased. This has
also increased the need to understand the back-
ground pathology of non-human primates, includ-
ing the range of incidental pathology findings that
might be confused with treatment effects.
This chapter describes and presents photo-
graphic images of common incidental findings that
might be encountered in young cynomolgus and
rhesus macaques and marmosets used in toxicity
testing as control animals. The spectrum of find-
ings are reflective of the young age of the animals,
the strict barrier-conditions they are kept under,
and for marmosets, the problems associated with
diet and nutrition of captive-reared New World
monkeys. In general congenital and degenerative
macaques and the common marmoset (Callithrix jacchus)
findings are the most common findings and prolif-
erative lesions are rare, while inflammatory lesions
are largely limited to mononuclear cell infiltra-
tions in several tissues.
Cardiovascular system
Spontaneous findings of the heart are among the
most commonly encountered findings in non-
human primates used as controls in preclinical
safety studies. Of these, the most frequently
recorded incidental findings are idiopathic myo-
cardial inflammatory cell infiltrations and focal
myocarditis. Although various diagnostic terms
have been used to describe these two seemingly
separate inflammatory findings of the myocardium
(Drevon-Gaillot et al 2006, Keenan & Vidal 2006,
Lowenstine 2003, Qureshi 1979, Scott 1999,
Shimoi et al 1998), it is generally accepted that
they represent a single pathological entity that
differs only in the degree of severity (Chamanza
et al 2006). The pathology continuum starts
with minimal to mild, focal, lymphoplasmacytic
inflammatory cell infiltration of the myocardium,
with little or no evidence of cardiac myocyte
necrosis or degeneration (Fig. 1.1), and progresses
to focal myocarditis characterized by some evi-
dence of cardiac myocyte necrosis and associated
edema or fibrin deposition (Fig. 1.2). Both condi-
tions occur more commonly in the subendocardial
or subepicardial areas of the heart and may occur
simultaneously in the same heart. No clinical signs
or grossly visible lesions are known to be associated
with these findings, and no etiological agent has
been isolated or demonstrated (Chamanza et al
2006). The most likely cause is therefore consid-
ered to be stress and the release of catecholamines
associated with capture or captivity; hence, higher
incidences of the findings have been reported in
wild-caught animals (Qureshi 1979) compared
with laboratory non-human primates. A proposal
has been put forward to apply umbrella terms
such as ‘focal idiopathic myocardial inflammation’
FIGURE 1.1  Inflammatory cell infiltration of the myocar-
dium of the cynomolgus heart. ×200.
Non-human primates 1
CHAPTER 1 
Ronnie Chamanza
FIGURE 1.2  Focal myocarditis in a female cynomolgus
monkey. ×200.
for these lesions and to grade them according
to the presence or absence of degenerative
or inflammatory changes in the adjacent myocar-
dial tissues when evaluating non-human primate
hearts in routine toxicity studies (Chamanza et al
2006).
Myocardial degeneration with karyomegaly is
an idiopathic focal myofibre degeneration or car-
diomyopathy (Vidal et al 2010, Zabka et al 2009)
that has been described in cynomolgus macaques
from various sources and origins. Even though it
has occurred at a low frequency, high incidences
have been recorded in the few studies in which it
has occurred (Chamanza et al 2010). The finding
is characterized by minimal to moderate focal
myocardial degeneration associated with mild to
moderate karyomegaly, cardiac myofibre hyper-
trophy and vacuolation, with minimal inflamma-
tion or fibrosis (Fig. 1.3). In early lesions, only
lipid-depleted vacuolation of cardiac myofibres
and karyomegaly with increased nuclear basophilia,
are present, while more advanced lesions may be
associated with inflammation, hemorrhage, miner-
alization and extensive fibrosis. The most com-
monly affected areas of the heart are, in decreasing
order, the subepicardial areas of the apex, the
interventricular septum (just below the atrio-
ventricular valves), and the tip of the papillary
muscle and subendocardial areas of both ventri-
cles (Chamanza et al 2010).
2 BACKGROUND LESIONS IN LABORATORY ANIMALS
FIGURE 1.3  Myocardial degeneration and karyomegaly in
a male cynomolgus monkey. ×200.
Rarely eosinophilic intranuclear inclusion bodies,
believed to be caused by invagination of the
cytoplasm into the enlarged nuclei, and intracyto­
plasmic eosinophilic or hyaline granules (usually
located at the poles of the nucleus) within the
hypertrophic cardiac myofibres, may be observed
(Fig. 1.4). Intranuclear (pseudo)inclusion bodies
in hypertrophic cardiac myofibres resulting from
intranuclear invagination of cytoplasmic organelles,
including mitochondria, have been reported in man
(Engedal et al 1977), while hyaline, ceroid and
lipofuscin cytoplasmic granules located at the poles
of cardiac myofibre nuclei have been reported in
cynomolgus monkeys (Jasty et al 1984).
FIGURE 1.4  Eosinophilic intranuclear inclusions and
cytoplasmic granules in the myocardium. ×400.
Acute hemorrhagic necrosis (Fig. 1.5) and/or
fibrosis (Fig. 1.6a & b) of the papillary muscle or
subendocardial areas, resembling ischemic lesions
such as those associated with beta-agonist and
other cardio-active drugs (Greaves 2000), have
also been observed in occasional animals, including
marmosets (Fig. 1.6a & b).
FIGURE 1.5  Myocardial degeneration and hemorrhage in
a female cynomolgus monkey. ×100.
FIGURE 1.6a  Fibrosis in the subendocardial myocardium
of a marmoset. ×50.
FIGURE 1.6b  Fibrosis of the heart of a marmoset.
Masson’s trichrome stain. ×100.
The possible role of catecholamines in this
idiopathic myocardial degeneration has been
suggested. Findings similar to those described
above, such as cardiac myofibre hypertrophy,
karyomegaly and vacuolation, have been observed
in a rhesus macaque (Macaca mulatta) with an
active angiomatous phaeochromocytoma (Vogel &
Fritz 2003) and in animals directly injected with
catecholamines (Khullar et al 1989).
Ectopic cysts or glandular structures lined by
various types of epithelium, from squamous to
cuboidal, or columnar, have been described in
various species, including man (De Lacroix &
Hübner 1974, Thomas & van Wesep 1990), cattle
(Bundza & Dukes 1978) and mice (Elwell &
Mahler 1999). In non-human primates, three
main types of ectopic cyst have been described in
the cynomolgus and rhesus macaques and marmo-
sets (Chamanza et al 2006, Drevon-Gaillot et al
2006, Kaspareit et al 2003).
Keratinized or non-keratinized squamous cysts
(Fig. 1.7), squamous plaques with no central
lumen, and thyroid follicle-like epithelial cysts
filled with an eosinophilic fluid have been reported
at very low incidences in the three species. Squa-
mous keratinized cysts are lined with a complete
or incomplete thin layer of flattened epithelium
filled with concentric layers of keratin (Fig. 1.8)
and may be associated with a foreign-body inflam-
matory reaction where the epithelial wall has been
broken, or appears incomplete, and the keratin is
in direct contact with adjacent tissues (Chamanza
et al 2006). Squamous non-keratinized cysts are
composed of a thicker layer of stratified squamous
epithelium around a central lumen filled with a
few inflammatory cells, amorphous cellular debris
or eosinophilic colloid-like material. The base of
the epithelial wall of the non-keratinized squa-
mous cyst is usually surrounded by a layer of
fibrous connective tissue of varying thickness.
FIGURE 1.7  Squamous cell lining of cyst in the heart of
a monkey. ×100.
FIGURE 1.8  Epithelial squamous cyst in the heart of a
cynomolgus monkey. ×100.
Squamous plaques, which are incomplete cysts
with no central lumen, may be found lying adja-
cent to squamous cysts, in which case they are
thought to represent tangential sections of walls
of adjacent squamous cysts. However, some epi-
cardial or endocardial squamous plaques, unasso-
ciated with complete squamous cysts, have also
been observed. Keratinized or non-keratinized
squamous cysts and squamous plaques are usually
located in the subepicardial or subendocardial
regions between the apex and the base of the
heart, and are therefore easily identified as yellow-
white nodules on the heart surface or interven-
tricular septum at necropsy.

Ectopic thyroid tissue or thyroid follicle-like
structures have been observed mainly in the sub-
epicardial areas at the base of the heart, including
atrial appendages and walls of the great vessels,
and are not readily identified at necropsy. Squa-
mous cysts and squamous plaques may be of
foregut origin while ectopic thyroid tissue could
be of thyroglossal duct origin (Elwell & Mahler
1999, Kaspareit et al 2003).
Various types of vascular lesions have been
reported to occur spontaneously, albeit at very low
frequencies in laboratory non-human primates
(Chamanza et al 2006, 2010). With the exception
of continuous intravenous infusion studies, in
which local injection sites and systemic vascular
lesions are common background findings associ-
ated with the test item administration procedure
(Lilbert & Burnett 2003), inflammatory lesions of
blood vessels are uncommon in laboratory non-
human primates.
Two isolated cases of polyarteritis nodosa, a sys-
temic necrotizing vasculitis of small to middle-
sized arteries, have been described in cynomolgus
macaques (Albassam et al 1993, Porter et al
2003). The most frequently encountered inflam-
matory vascular lesions are minimal to moderate,
localized vasculitis or perivasculitis characterized
by a lymphoplasmacytic infiltration of the vascular
wall without extensive necrosis of the tunica
media or substantial fibrin deposition. The most
commonly affected organs include the large intes-
tinal wall, lungs, meninges (brain and spinal cord),
heart (Fig. 1.9), urinary bladder and sciatic nerve.
FIGURE 1.9  Perivasculitis in the heart. ×200.
Naturally occurring, degenerative lesions of
arteries, such as atherosclerosis, do not occur
commonly in non-human primates, but intimal
thickening and the formation of arteriosclerotic
lesions (fibrous plaques) in the coronary artery
and aorta, sometimes leading to occlusion, are
encountered infrequently (Chamanza et al 2010,
Scott 1999). The intima is infiltrated by smooth
muscle cells, mucopolysaccharides and fibrous
tissue with little or no foam cells or extracellular
lipids seen in diet-induced, atherosclerotic plaques
(Fig. 1.10). Intimal proliferation or fibrocellular
intimal thickening may also be encountered in the
lung of normal control cynomolgus macaques fol-
lowing organization of small pulmonary emboli
derived from infusion sites.
FIGURE 1.10  Intimal thickening and atherosclerosis in a
blood vessel. ×200.
Another closely-related, degenerative lesion of
arteries seen in young macaques is the accumula-
tion of mucopolysaccharides, or mucification, in
the subendothelial areas of the aorta and the
heart. Mucin accumulation in the tunica intima
and, to a lesser extent, tunica media, of the aorta,
with proliferation of intimal cells and splitting of
elastic fibres of the tunica media (Fig. 1.11a), has
been observed at low frequency in cynomolgus
macaques used in toxicity studies (Chamanza et al
2010). Mucin or mucopolysaccharide accumula-
tion in the tunica intima of the aorta and sub­
endocardial tissues of the heart chambers and
valves (Fig. 1.11b) has also been described in
humans and other non-human primates (Lindsay
& Chaikoff 1966, Scott 1999). The mucopolysac-
charide content of the walls (particularly the
intima) of arteries of non-human primates is
generally more abundant than it is in humans
(Scott 1999). This aortic degenerative change in
macaques results in a massive accumulation of
mucopolysaccharides in the subintimal areas,
causing localized swelling or expansion of the
intima with disruption of the elastic fibres of the
intima media (Fig. 1.11a).
FIGURE 1.11a  Intimal degeneration and mucopolysac-
charide accumulation of the aorta in a cynomolgus
monkey. ×200.
Non-human primates 3
FIGURE 1.11b  Mucinous degeneration of the atrium.
×100.
Hemolymphoreticular system
Extramedullary hematopoiesis (EMH) occurs fre-
quently in the adrenal gland (Fig. 1.53), liver,
kidney and other tissues in marmosets. The sever-
ity and incidence of this finding is usually associ-
ated with the frequency of blood sampling, with
marmosets being very sensitive to frequent bleeds
(Tucker 1984). This spontaneous finding should
be differentiated from inflammation, inflammatory-
cell infiltrates or treatment-related EMH. Sponta-
neous EMH in macaques is extremely rare, but
has sporadically been observed in lymph nodes of
healthy control cynomolgus macaques (Chamanza
et al 2010).
Multinucleate lymphocytic syncytia, resem-
bling the Warthin–Finkeldey bodies associated
with measles (Fig. 1.12), have been observed with
some frequency in the bronchial-associated lym-
phoid tissue (BALT) of the larynx and the gut-
associated lymphoid tissue (GALT) of the large
intestines in normal healthy cynomolgus macaques
(Chamanza et al 2010). It is not known whether
these represent subclinical measles infection, but
no necrosis of lymphocytes is observed within the
lymphoid nodules, nor are viral inclusions present
within the cells. Natural measles infections which
may originate from animal handlers, are known to
occur in both marmosets and macaques (Scott
1999).
FIGURE 1.12  Warthin–Finkeldey bodies in the gastroin-
testinal lymphoid tissue (GALT) of a non-human primate.
×100.
The presence of prominent lymphoid follicles
within the bone marrow is a common background
finding in macaques (Fig. 1.13). This lesion,
together with lymphoid follicular hyperplasia and
prominent germinal centres in other sites, such as
the spleen (Fig. 1.14), lymph nodes and salivary
4 BACKGROUND LESIONS IN LABORATORY ANIMALS
glands, have been associated with subclinical
type D retroviral infection (Guzman et al 1999,
Lowenstine 1993). However, most non-human
primates used in preclinical studies are routinely
screened for, and are known to be free from,
simian retroviruses (type D). Thus hyperplastic
lymphoid follicles in the spleen and other tissues
are considered to indicate heightened non-specific
immunosurveillance in animals which have been
reared in a relatively disease-free environment and
are known to be free from major pathogens. Peri-
odontal disease, glossitis and tonsillitis are other
common minor inflammatory lesions known to be
associated with lymphoid follicular hyperplasia
and mononuclear cell infiltrations in other organs.
FIGURE 1.13  Germinal centres within the bone marrow
in a cynomolgus monkey. ×100.
FIGURE 1.14  Prominent lymph follicle with hyalinization
in the lymph node in a cynomolgus monkey. ×100.
The accumulation of brightly eosinophilic
amorphous material (hyalinization) in the centre
of lymphoid follicles in non-human primate
spleens is a common observation of no known
clinical significance. The eosinophilic substance is
thought to be proteinaceous in content and com-
posed of antigen–antibody complexes. Russell
bodies can occasionally be observed in the vicinity
of hyalinized germinal centres, giving more cred-
ibility to the theory of immunoglobin composition
of the amorphous material.
Respiratory system
Pneumonia in laboratory macaques is not common,
but focal interstitial inflammation or fibrosis of
minimal to slight grades of severity, does occur
with some frequency. The lesions occur in subp-
leural areas or at the tips of lung lobes. There may
be proliferation of bronchiolo-alveolar epithelium,
fibrosis and accumulation of alveolar macrophages
in alveolar spaces (Fig. 1.15). Other associated
lesions commonly encountered in the lungs of
macaques include focal pleuritis and pleural or
subpleural fibrosis, with or without lung adhesions
to adjacent lobes or to the parietal pleura. These
are usually associated with necropsy findings of
lung adhesions or pale white foci of the pleura.
FIGURE 1.15  Interstitial pneumonia in a subpleural loca-
tion in a cynomolgus monkey. ×200.
Other inflammatory lesions of the lung include
focal, foreign-body granulomas with minimal to
mild interstitial inflammation. The lesions occur
as a result of inhalation of plant, food or other
small particles (Fig. 1.16) and should be distin-
guished from hair emboli, which usually occur as
a result of the inadvertent injection of hair or skin
fragments during intravenous injection of test sub-
stances. Hair emboli or fragments of skin lodged
in macaque lungs may appear as focal, foreign-
body granulomas in the lung or within arterial
thrombi with a perivascular reaction (Kast 1994).
FIGURE 1.16  Foreign body pneumonia in the lung in a
cynomolgus monkey. ×200.
Pulmonary vascular disease in laboratory
primates is rare, but arteritis/periarteritis, throm-
boembolism, perivascular edema and arterioscle-
rosis may be encountered in control animals. Focal
thickening of the intima, due to cellular prolifera-
tion in earlier lesions and later replacement by
fibrosis (with or without smooth muscle hyperpla-
sia) of the media can be encountered. Intimal
lesions can lead to complete occlusion of the
artery concerned (Fig. 1.17), but changes in adja-
cent tissues are usually absent. However, occlusive
intimal proliferation and thrombosis associated
with continuous intravenous infusion with saline
may be associated with bronchopneumonia, which
is usually confined to the distal parts of lung lobes,
due to the dual blood supply of the lungs. Diffuse
interstitial inflammation, periarteritis and peri­
vascular edema are some of the more common
findings observed in saline-injected control animals
on continuous intravenous infusion studies.
FIGURE 1.17  Thrombi and intimal proliferation in a blood
vessel of the lung in a cynomolgus monkey. ×200.
Yellowish brown to dark brown pigment in
alveolar macrophages is often found distributed
perivascularly or peribronchiolarly within the
lungs of macaques (Fig. 1.18). Although iron-
positive pigment associated with the lung mite
Pneumonyssus simincola is occasionally encoun-
tered, especially in wild-caught macaques, the
majority of the brown pigment present in the
lungs of purpose-bred macaques is usually not
associated with any parasite sections or pulmonary
parenchymal pathological changes, and is there-
fore believed to be anthracosis. Anthracosis, is
caused by the inhalation of atmospheric carbon
particles by laboratory non-human primates
housed near or within urban areas. Similar pigment
can be encountered within macrophages in the
bronchial or mediastinal nodes and should be dif-
ferentiated from tattoo pigment.
FIGURE 1.18  Brown pigment in the lungs in a cynomol-
gus monkey. ×200.
Vascular or parenchymal mineralization in the
lungs (Fig. 1.19) of marmosets occurs commonly
in control animals. This occurs as part of meta-
static mineralization in this species and may
involve other major organs such as the kidneys,
the heart (mostly the atria), the aorta and adrenal
glands. Excessive calcium absorption as a result of
hypervitaminosis D, or high calcium levels in the
diet, are believed to be the main causes of meta-
static calcification in marmosets (Kaspareit 2006,
Scott 1999). The kidneys and the urinary tract
are the most commonly affected areas. Minerali-
zation of other organs, such as the lungs, can also
be observed in marmosets with severe renal
disease and nephrocalcinosis (Fig. 1.20) which can

complicate the etiological diagnosis. Metastatic
calcification in macaques is very rare, and almost
all forms of mineral deposition seen spontaneously
are dystrophic in nature.
FIGURE 1.19  Mineralization of the lung of a marmoset.
×200.
FIGURE 1.20  Mineralization of the kidney of a marmoset.
×200.
Gastrointestinal system
Superficial inflammation, erosion or ulceration of
the tongue and esophageal mucosa (Fig. 1.21),
associated with Candida albicans infection, is
occasionally encountered in stressed and deb­
ilitated marmosets (Chalmers et al 1983) or
macaques on long-term antibiotic treatment. The
lesion is characterized grossly by the presence of
pale white pseudomembranes on the oral or
esophageal mucous membranes, with superficial
ulceration beneath the pseudomembrane. Upon
histopathological examination, erosion and ulcera-
tion that rarely extend below the basement mem-
brane are observed in association with minimal,
mixed inflammatory cell infiltrates and fungi in
the superficial epithelium. The organisms consist
of septate pseudohyphae and budding blastospores
and are easier to visualize with PAS or Grocott
Gomori’s methenamine silver stains.
FIGURE 1.21  Mycotic oesophagitis in a marmoset. ×200.
The normal histology of the stomach of the
cynomolgus macaque is of particular importance
because some pathological changes affecting the
mucosa are associated with an increase or decrease
in some cell populations that make up the gastric
mucosa. The stomach of the cynomolgus macaque
can be divided into four anatomical regions: the
cardia, fundus, body and antrum. It is the histo-
logical appearance of the fundic region of the
stomach in the non-human primate that differs
slightly from that of humans and dogs. Unlike
humans, dogs and pigs, the fundic region and the
body of the stomach of macaques are histologi-
cally different, with the fundic mucosa having few
or no parietal cells and the transition between the
two zones being marked by the abrupt appearance
of parietal cells (Vidal et al 2008). The histologi-
cal appearance of the antral mucosa does not
differ greatly from that of humans and dogs and
has numerous, deep gastric pits, no parietal cells
and a few chief cells. Lymphoid follicles, or foci
of lymphoplasmacytic cells, can be observed
throughout the gastric mucosa in normal stomach
sections, but these can be increased in gastritis.
Gastritis, especially chronic gastritis of the
antral region, is very common in laboratory non-
human primates and may affect the majority of
animals in a toxicity study (McKeag & McInnes
2012). Chronic gastritis is characterized by
minimal to marked infiltration of the mucosa and
submucosa with lymphocytes and plasma cells,
an increase in lymphoid follicles and mucosal
intestinalization of the antral mucosa typified by
atrophy of gastric glands with an accompanying
increase in the lamina propria area (Fig. 1.22).
While chronic gastritis is observed mainly in the
antral mucosa, acute or chronic-active gastritis
associated with hemorrhage, erosions or glandular
micro-abscesses is more common in the fundic/
body of the stomach (Fig. 1.23).
Non-human primates 5
FIGURE 1.22  Chronic gastritis in the pylorus in a
cynomolgus monkey. ×100.
FIGURE 1.23  Gastric erosion in the fundus of the stomach
in a cynomolgus monkey. ×100.
Helicobacter pylori and Helicobacter heilmannii-
like organisms (also known as Gastrospirillum
hominus) are the most commonly identified path-
ogens in the stomach of non-human primates used
in toxicity studies and are believed to be associ-
ated with subclinical gastritis (Dubois et al 1994,
Reindel et al 1999). Helicobacter pylori is a small
(2–4 µm long) bacterium, shaped like a comma, a
loose spiral or a seagull’s wing, that occurs largely
in the gastric antral mucosa. Lesions associated
with H. pylori include a diffuse, mononuclear cell
infiltration of the mucosal lamina propria with
mucosal thickening, lamina propria expansion
with separation of glands, epithelial hyperplasia,
prominent lymphoid follicles or mucosal erosion.
The lymphoplasmacytic infiltrates can be made
up of a large percentage of plasma cells with
Russell bodies but occasionally, and in the more
severe lesions, neutrophilic infiltrates in the glan-
dular lumen (glandular micro-abscesses) may be
observed.
Helicobacter heilmannii-like organisms
(HHLOs) are larger than H. pylori (approximately
7–10 µm) and have a tightly wound spiral shape
with approximately 4 to 12 coils. They typically
colonize the fundic/body mucosa and can be
observed within parietal cells. Although HHLOs
are more prevalent than H. pylori in the stomach
of both cynomolgus and rhesus monkeys, evidence
shows that HHLOs are relatively non-pathogenic,
although subtle damage to infected parietal cells
has been reported (Dubois et al 1991).
Gastric infarction is an uncommon, but clini-
cally important, finding in young adult cynomol-
gus macaques. It may be the cause of death or
poor health in study animals or may be observed
6 BACKGROUND LESIONS IN LABORATORY ANIMALS

as an incidental finding in clinically normal animals.
The cause of gastric infarction is not always appar-
ent but in most cases a severe systemic insult that
predisposes to disseminated intravascular coagula-
tion, such as trauma, hernia, intussusception or
some other natural disease, is usually present
(Chamanza et al 2010, Fikes et al 1996). The
lesion is characterized by extensive necrosis,
hemorrhage and edema of the muscularis and
submucosa of the fundic or pyloric region of
the stomach (Fig. 1.24) associated with micro-
thrombi, particularly in the venous system.
non-human primates used in toxicity studies.
Outbreaks of Campylobacter or Shigella enteritis
occur occasionally even in the most well main-
tained modern facilities. Campylobacter displays
a histological picture of villus atrophy and crypt
micro-abscesses in the colon, while acute shigel-
losis manifests as ulceration, erosion, crypt micro-
abscesses and exudation of neutrophils leading
to accumulation of fibrinopurulent or fibrinon-
ecrotic exudate within the lumen or areas overly-
ing the damaged enteric mucosa (Fig. 1.26).
Hemorrhage is usually present at the periphery of
the lesion while the lamina propria is filled with a
predominantly neutrophilic cellular infiltrate with
variable numbers of mononuclear cells. FIGURE 1.27  Balantidium coli associated with colonic
mucosal glandular herniation. ×100.

Idiopathic chronic colitis, with or without

FIGURE 1.24  Extensive necrosis in the submucosa, due
to microthrombi, in the stomach in a cynomolgus monkey.
×100.
The presence of pigment-laden macrophages
within the lamina propria of the small intestinal
villi tips is a very common finding in laboratory
macaques. The finding has been reported at high
incidences by several authors (Drevon-Gaillot
et al 2006, Lowenstine 2003, Scott 1999), and
various theories have been suggested regarding the
nature of the pigment. The yellowish to light
brown, granular pigment is usually associated with
apoptotic bodies within the lamina propria
without any associated damage to the enterocytes.
The ileum is the most affected small intestinal
segment (Fig. 1.25). Iron (Scott 1999), bile
pigment, ceroid-lipofuscin and necrotic cellular
debris (Drevon-Gaillot et al 2006, Ito et al 1992)
have been suggested to be part of the constituents
of this pigment. The significance of this finding
has not been determined and it may represent
attempts by lamina propria phagocytes to engulf
foreign material.
FIGURE 1.25  Pigmented macrophages in the lamina
propria of the small intestine in a cynomolgus monkey.
×200.
Minimal to mild intestinal inflammation
associated with episodes of diarrhea is not an
uncommon background finding in purpose-bred,
FIGURE 1.26  Colitis associated with Shigella sp. in the
large intestine in a cynomolgus monkey. ×100.
Shigellosis should be differentiated from infec-
tions by Salmonella, Campylobacter and Yersinia
species. Disease caused by Campylobacter is less
debilitating, while infections with Salmonella and
Yersinia enterocolitica, unlike shigellosis, usually
become septicemic (Bennett et al 1998). Animals
with shigellosis die from dehydration if left
untreated. Spirochetosis is often present in the
crypts of the large intestine of non-human pri-
mates. Organisms can be found from the cecum
to the rectum, and species include Brachyspira.
The protozoan Balantidium coli may also be
present with this lesion. A high incidence of spi-
rochetes in the large intestine of immunocompro-
mised monkeys has been reported (Losco et al
2002).
Micro-herniation of colonic mucosal glands
through the muscularis mucosae occurs com-
monly in the non-human primate colon. The her-
niated mucosal glands are observed lying deep
within submucosal lymphoid patches of the gut-
associated lymphoid tissue (GALT) of the colon.
Herniation is thought to result from a defect in
the overlying muscularis mucosa, causing the
glands to protrude through the gap into the under-
lying GALT in the submucosa. An increase in the
intraluminal pressure, as part of normal peristalsis,
is thought to contribute to the herniation (Scott
1999). Micro-herniations play an important role
in the extension of pathogens through the wall of
the colon, so protozoal species such as Balantid­
ium (Fig. 1.27), and bacteria, can pass the mucosal
barrier and gain access to the deeper layers (Scott
1982, 1999). Parasites and fecal matter trapped
within these herniations or diverticula can incite
a chronic inflammatory reaction with foreign-body
giant cells.
chronic intermittent diarrhea, occurs commonly
in young adult macaques used in toxicity studies
(Adler et al 1993). Distension of the cecum and
colon with fluid, or thickening of the mucosa, may
be the sole necropsy finding, but occasionally
reddening of the colonic mucosa or the iliocecal
junction or multifocal ulceration or erosions are
observed in colitis. The most consistent histologi-
cal picture is that of a diffuse, lymphoplasmacytic
cellular infiltration of the lamina propria and sub-
mucosa, crypt atrophy, and micro-abscesses (Fig.
1.28), mucosal epithelial hyperplasia and glandu-
lar micro-herniation into GALT. Other findings
include goblet cell depletion, karyomegaly of
hyperplastic mucosal cells, micro-ulcers and
erosions caused by the rupture of crypt micro-
abscesses, and a few scattered macrophages.
Associated lesions in other organs such as hyper-
plasia of the lymphocytes in the mesenteric lymph
nodes, thymic atrophy, chronic inflammation in
the stomach, liver and gall bladder may also be
present (Bennett et al 1998).
FIGURE 1.28  Chronic colitis in a cynomolgus monkey.
×100.
A similarly common and less well-understood
chronic colitis also occurs in captive-bred marmo-
sets (David et al 2009). The condition is associ-
ated with diarrhea and weight loss and may form
part of wasting marmoset syndrome (WMS).
Gross and histopathology presentations are essen-
tially similar to those of chronic colitis of macaques,
with crypt micro-abscesses, herniation of glands,
micro ulcerations, atrophy and loss of crypts as
well as lymphoplasmacytic inflammatory cell infil-
tration into the lamina propria as the most common
microscopic findings (Fig. 1.29). Neutrophil accu-
mulations within herniated crypts in the GALT are

more pronounced in marmosets (Fig. 1.30). No
etiological agent has been identified, and the cause
remains unknown, although diet and nutrition are
thought to play a major role in this and in other
marmoset pathological syndromes.
FIGURE 1.29  Chronic colitis in the large intestine of a
marmoset. ×200.
FIGURE 1.30  Colitis with glandular herniation in the colon
of a marmoset. ×100.
There is a large difference between the natural
diet of free-living marmosets, which is mainly
composed of insects, fruit and sap, to that offered
to animals in captivity (commercial diet supple-
mented by fruits), and this difference has been
postulated as the main contributing factor to the
development of enteritis in this species (Kaspareit
et al 2006, Layne & Power 2003). Marmosets, like
most callithrids, also have a very high requirement
for protein, which is nearly twice that of rhesus
macaques (Bennett et al 1998), and a higher
requirement than macaques for vitamin E, which
makes them prone to hypoproteinaemia, hypoalbu-
minaemia, Heinz body anemia and, ultimately, to
stress. This, in turn, makes marmosets even more
selective in their eating habits, leading to patho-
logical manifestations of wasting marmoset syn-
drome, such as colitis and glomerulonephritis.
Parasitic granuloma(ta), with sections of nema-
tode parasites, are observed more commonly along
the gastrointestinal tract than elsewhere in the
body in macaques used in toxicity studies.
Oesophagostomum species are by far the most
commonly encountered nematode parasite in
laboratory-reared, non-human primates. Oesopha­
gostomum infections are usually asymptomatic
even though diarrhea and anemia may develop in
heavy infestations. Adult worms live freely in the
large intestine and do not cause lesions, but larvae
can cause characteristic submucosal or tunica
muscularis nodules consisting of a central abscess
with necrosis and mixed inflammatory infiltrates
that may include multinucleate giant cells encir-
cled by fibrosis. The nodules are found mainly in
the large intestine (Fig. 1.31), but may also be
found in other ectopic sites such as the stomach
wall, the peritoneum, kidney, liver and the sub-
cutis (Fig. 1.32).
FIGURE 1.31  Parasitic granuloma caused by Oesophagos-
tomum sp. in the large intestine in a cynomolgus monkey.
×50.
FIGURE 1.32  Parasitic granuloma caused by Oesophagos-
tomum sp. in the subcutis in a cynomolgus monkey.
×200.
Brunner’s glands are compound tubular mucous
glands located in the submucosa of the proximal
duodenum and are concentrated in the region
between the pylorus and the sphincter of Oddi.
They are anatomically, but not functionally, a con-
tinuation of the pyloric glands, the main function
of which is to produce an alkaline secretion (bicar-
bonate) that neutralizes acid from the stomach.
Some chemical compounds are known to produce
changes such as vacuolation, dilatation or cystic
changes of Brunner’s glands in other species
(Betton 1998) and in non-human primates, and
this lesion should be differentiated from sponta-
neous Brunner’s gland dilatation, hyperplasia or
cystic Brunner’s glands (Fig. 1.33). Cystic Brun-
ner’s glands are a rather common findings in mar-
mosets (Kaspareit et al 2006). The dilatation or
hyperplasia may result in slight distortion of the
overlying villi or the intestinal crypts into which
the secretion exits. This is a finding of unknown
pathological significance.
Non-human primates 7
FIGURE 1.33  Dilated Brunner’s glands in the duodenum
of a marmoset. ×100.
An accessory spleen is described as a congenital
duplication of splenic tissue in an ectopic location.
Accessory spleens can be located in various sites
within the abdominal cavity, but the tail of the
pancreas, or the adipose tissue surrounding this
area, is one of the most common locations for
accessory spleen in many animal species. This
occurs during organogenesis, because the spleen
originates from the mesenchymal primordium
adjacent to that of the pancreas in the embryonic
dorsal mesogastrium (Arey 1954). Interestingly, it
has been demonstrated that, in man, the incidence
of accessory spleen is increased in certain hema-
tological conditions (Olsen & Beaudoin 1969).
In non-human primates, accessory spleen within
the pancreas has been reported in a capuchin
monkey (Lau 1973) and in laboratory non-human
primates used in toxicity studies (Chamanza et al
2010). The finding also occurs commonly in other
animal species such as rabbits (Fox et al 1976,
Weisbroth et al 1976), chickens (Glick & Sato
1964), mice (Gad et al 2007) and Chinese ham-
sters (Yoon et al 2000), and is almost always
found within the tail of the pancreas. The true
incidence of accessory spleen within the pancreas
in laboratory non-human primates may therefore
be influenced by the sampling and trimming pro-
cedures employed by the laboratory (whether the
tail of the pancreas is consistently sampled),
because this finding is not easily recognized at
necropsy (Chamanza et al 2010).
Histologically, an intra-pancreatic accessory
spleen is usually well-demarcated from pancreatic
tissue and may be encapsulated by a thin layer of
collagenous connective tissue and smooth muscle
fibres with or without evidence of trabeculae
and reticular fibre support. The parenchyma is
composed of mature splenic tissue with recogniz-
able red and white pulp areas resembling those of
the spleen (Fig. 1.34). The reticular network is
usually poorly developed, and trabeculae may be
scarce or absent, but the presence of lymphoid
nodules or recognizable periarteriolar lymphoid
sheaths, deposits of hemosiderin and hematopoi-
etic tissue, including megakaryocytes, is helpful in
the diagnosis.
8 BACKGROUND LESIONS IN LABORATORY ANIMALS

Hepatic lipid vacuolation or diffuse lipidosis

FIGURE 1.34  Accessory spleen within the pancreas in a
cynomolgus monkey. ×100.
Liver and biliary system
Focal, random necrosis of the liver is a common,
spontaneous finding in marmosets (Chalmers
1983, Tucker 1984), but it occurs very rarely in
macaques. Small to large patches of coagulative
necrosis with hemorrhage and a minimal inflam-
matory response are more often observed in the
subcapsular areas of livers in conjunction with
underlying pathological entities such as lipid vacu-
olation and severe glycogen accumulation (Fig.
1.35). Most of these necrotic areas are not associ-
ated with any etiological agents, but organisms
known to cause focal necrosis in laboratory pri-
mates include Listeria, Yersinia species (causal
agent of pseudotuberculosis) (McClure et al
1978) and the causal agent of Tyzzer’s disease
(Tucker 1984). Marmoset livers with focal, non-
zonal necrosis may also have several foci of micro-
granulomata which may be confused with foci of
extramedullary hematopoiesis (Fig. 1.36).
of minimal to severe grades (Fig. 1.37) is often
observed in marmosets, but is seen very rarely in
young laboratory macaques, which usually present
with milder forms of the condition. Severe lipid
vacuolation in marmosets should be distinguished
from marked glycogen accumulation, which is a
common finding in this species, and the two find-
ings may be encountered in the same organ (Fig.
1.37). Grossly, the liver is usually pale yellow
with rounded edges or the liver may be enlarged.
The causes of lipid vacuolation in marmosets are
varied but may include anemia, hypoproteinemia,
hypoalbuminemia, renal disease, intestinal inflam-
mation or colitis and starvation (Chalmers et al
1983, Lowenstine 2003, Tucker 1984), most of
which are pathological conditions associated with FIGURE 1.38  Tension lipidosis or lipid vacuolation in the
wasting marmoset syndrome. Lipid vacuolation
can also occur in overweight macaques of a mean
liver in a cynomolgus monkey. ×50. Note the ligament
attachments.
age of nine years as part of the fatal fatty liver
syndrome, also known as the fatal fasting syn- Hepatic pigment is generally not a common,
drome (Bennett 1998, Lowenstine 2003), but is spontaneous finding in macaques, but it occurs
rarely observed in animals on a study, because with variable frequency in marmosets, depending
study animals tend to be much younger. on the diet and other pathological factors (Miller
et al 1997). Hepatic hemosiderosis, the deposi-
tion of the iron-containing pigment hemosiderin
in Kupffer cells, is the most common pigment
observed in the marmoset liver (Fig. 1.39) and is
thought to be associated with several factors such
as excessive amounts of iron in the diet, endog-
enous elevations of glucocorticoids due to stress,
as well as the presence of chronic inflammatory
processes. In the case of chronic inflammatory
diseases, such as those associated with WMS,
hemosiderosis is thought to occur as a result of
the sequestration of iron and poor iron transfer
from siderophages to erythrocytes in the anemia
of chronic disease (Lowenstine 2003).
FIGURE 1.37  Lipid vacuolation, glycogen and extramed-
ullary hemopoiesis in the liver of a marmoset. ×100.

Tension lipidosis is a form of focal, lipid vacuola-

FIGURE 1.35  Necrosis and glycogen accumulation in the
liver of a marmoset. ×100.
FIGURE 1.36  Necrosis and fibrosis in the liver of a
marmoset. ×100.
tion that occurs in subscapsular hepatocytes lying
adjacent to the insertion areas of ligament attach-
ments (Fig. 1.38). It is a commonly observed
lesion present at necropsy, and a common histo-
logical finding of the liver in non-human primates
and other laboratory animal species. The lesion is
seen at necropsy as a discrete, pale yellow area
near the hilus or other areas of the liver margin.
The focal accumulation of fat in the cytoplasm of
subcapsular hepatocytes lying adjacent to liga-
ment insertion points is thought to be due to
hypoxia. It is proposed that the contraction of
these ligaments exerts tension on the liver capsule,
leading to the impediment of the blood supply to
the adjacent hepatocytes, resulting in hypoxia and
lipid vacuolation. Besides lipid accumulation, cap-
sular or subcapsular fibrosis may also be observed.
The lesion is of no known functional significance
but it has to be differentiated from treatment-
related lipid vacuolation.
FIGURE 1.39  Hemosiderosis in the liver of a marmoset.
×200.
One very common feature of the marmoset
liver which sets it apart from that of macaques is
the high amount of glycogen which accumulates
in hepatocytes (Chalmers et al 1983, Foster 2005,
Kaspareit et al 2006, Tucker 1984). Glycogen
accumulation in marmosets is known to be diet-
and age-dependent (Foster 2005), but it can also
vary considerably between studies and among
animals in a study. It is seen histologically as
diffuse or pan-lobular vacuolation that lends a lacy
appearance or rarefaction to the hepatocyte cyto-
plasm (Figs 1.35 & 1.37).
Focal or localized subcapsular fibrosis, inflam-
mation, necrosis or hemorrhage associated with
minimal to slight bile-duct proliferation in the
adjacent periportal area, can occur commonly in a

group of young adult cynomolgus macaques or
marmosets without any associated clinical or
clinico-pathological abnormalities (Chamanza
et al 2010). The lesion is characterized by the
minimal to moderate deposition of reticular and
collagen fibres in periportal areas adjacent to the
capsule, or just beneath the capsule, and affects
two to more than five hepatic lobules. Since gross
findings associated with these subcaspular findings
are commonly observed on the parietal surface of
the liver, which lies against the diaphragm and
costal wall, they are considered to be associated
with mechanical handling of animals during treat-
ment (Shimoi et al 1998).
Focal mononuclear inflammatory cell infiltra-
tion in the liver is among the most common find-
ings observed in laboratory non-human primates
(Chamanza et al 2010, Drevon-Gaillot et al 2006,
Foster et al 2005). The infiltrates are often located
periportally, where they are usually not associated
with injury to surrounding tissues. Inflammatory
cell infiltrates located within the liver parenchyma
may or may not be associated with single-cell
necrosis (Fig. 1.40). Lesions with associated hepa-
tocyte damage often contain other inflammatory
cells such as neutrophils and macrophages. Spon-
taneous inflammatory cell foci in the liver, of
minimal or moderate grades, should be differenti-
ated from treatment-related findings caused by
pro-inflammatory test compounds. Chronic peri-
portal inflammation or bile-duct hyperplasia may
be associated with intestinal tract inflammation
(Chalmers et al 1983).
FIGURE 1.40  Inflammatory cell infiltrate and focal necro-
sis in the liver in a cynomolgus monkey. ×200.
Ectopic adrenal gland attached to the liver, and
lying within the liver capsule (Fig. 1.41), has been
observed sporadically in macaques (Chamanza
2010). The lesion occurs less commonly than
adreno-hepatic fusion/adhesion, which involves
ectopic hepatic tissue lying within or attached to
the right adrenal gland. Ectopic adrenal tissue
within the liver consists of one or more of the
three cortical zones lying just below the liver
capsule with a fusion of the adrenal and hepatic
capsule and co-mingling of hepatic and adrenal
tissue without intervening fibrous tissue. Ectopic
adrenal cortical tissue is also observed within the
kidney (Fig. 1.42a) and epididymis (Fig. 1.42b) in
non-human primates.
FIGURE 1.41  Liver with ectopic adrenal gland in a
cynomolgus monkey. ×100.
FIGURE 1.42a  Ectopic adrenal tissue in the kidney in a
cynomolgus monkey. ×100.
FIGURE 1.42b  Ectopic adrenal tissue in the epididymis in
a cynomolgus monkey. ×100.
Urinary system
Interstitial nephritis, characterized by minimal to
slight accumulation of lymphoplasmacytic cells in
the interstitium of mainly the outer cortex of the
kidney, is one of the most common incidental find-
ings in macaques (Chamanza at al 2010, Drevon-
Gaillot et al 2006). Interstitial nephritis is also
listed as the most frequently observed lesion in a
survey of lesions seen in non-human primates in
zoos (Hubbard et al 1987). Findings range from
minimal, focal, lymphoplasmacytic infiltration,
without associated damage of renal tubular epithe-
lium, to moderate lesions composed of large areas
of inflammatory-cell infiltration, renal tubular
necrosis and minimal peritubular fibrosis (Fig.
1.43).
Non-human primates 9
FIGURE 1.43  Interstitial lymphoplasmacytic infiltration of
the cortex of a kidney in a cynomolgus monkey. ×100.
Tubular or cuboidal metaplasia of the parietal
layer of the Bowman’s capsule of the renal glomer-
uli occurs as an incidental finding of no known
pathological significance in both male and female
cynomolgus monkeys (Kaspareit et al 2004).
Cuboidal metaplasia occurs in other species where
it has a different significance.
In cynomolgus macaques, where it has been
observed, cuboidal metaplasia is characterized by
replacement of the normal flattened epithelial
lining of the parietal layer of the Bowman’s
capsule by a cuboidal epithelium that structurally
resembles the epithelium of the proximal convo-
luted tubules (Fig. 1.44). The degree of metapla-
sia varies from affecting only a part of the parietal
layer to affecting the full circumference, and the
number of glomeruli affected per kidney also
varies. Usually a few glomeruli are affected per
kidney, and in some affected glomeruli the cuboi-
dal parietal layer epithelium is continuous with
the epithelium of the proximal tubules. No sex
difference has been observed with this phenom-
enon in cynomolgus macaques (Kaspareit et al
2004) and it does not appear to be age-related.
FIGURE 1.44  Cuboidal metaplasia of the Bowman’s cap-
sular epithelium of a kidney in a cynomolgus monkey.
×200.
Various forms of spontaneous glomeruloneph-
ropathy have been described in non-human pri-
mates. Lesions range from mesangioproliferative
glomerulonephritis, which is more commonly
seen in marmosets, to those resembling membran-
oproliferative glomerulonephritis and sclerotic
lesions similar to collagenofibrotic glomeruloneph-
ropathy (CFGN), occasionally seen in cynomolgus
macaques (Adachi 2005, Chamanza 2010). Most
spontaneous glomerular lesions in laboratory
macaques are focal lesions that involve part of
(segmental) or the whole (global) glomerulus and
10 BACKGROUND LESIONS IN LABORATORY ANIMALS
do not occur as part of a systemic disease state.
Idiopathic glomerulosclerosis, characterized by
segmental or global deposition of a pale Congo-
red-negative, but PAS-positive eosinophilic sub-
stance within an expanded mesangium, is the most
common form observed in macaques (Chamanza
et al 2010). There is minimal to marked enlarge-
ment of the glomeruli and a decrease in the
number of nuclei in the mesangium, with or
without narrowing of the capillary lumina (Fig.
1.45). Minimal thickening of the Bowman’s
capsule and periglomerular and peritubular fibrosis
may be present. Other glomeruli within the same
kidney may also show mesangial proliferation with
glomerular enlargement but less collagen deposi-
tion and periglomerular fibrosis. The findings
differ from the CFGN which has been reported in
cynomolgus macaques (Adachi 2005) in that there
is focal random involvement of glomeruli with
little or no tubular and interstitial involvement,
unlike CFGN, which has a diffuse distribution of
glomerular lesions, no mesangial proliferation and
is associated with clinical disease. The cause of
most glomerular lesions in untreated, laboratory
non-human primates is unknown, but some disease
states such as diabetes, malaria, dehydration asso-
ciated with colitis (and diarrhea) and infection
with the simian immunodeficiency virus (SIV) are
known to result in glomerular lesions.
FIGURE 1.45  Segmental glomerulosclerosis of the kidney
of a cynomolgus monkey. ×200.
Marmosets, which suffer more cases of natu-
rally occurring glomerulonephropathy than do
macaques, have more diffuse and severe lesions
which have clinical implications and which may be
an important cause of death. The most common
form of glomerulonephropathy in marmosets is
diffuse, mesangioproliferative glomerulonephritis
which occurs as part of the wasting marmoset
syndrome and has been demonstrated to be linked
to deposition of immune complexes directed
against parasitic, bacterial (dental disease) and
dietary antigens (Lowenstine, 2003). This form
of immune-mediated glomerulonephropathy is
thought to be either IgM- or IgA-mediated or both
(Borda et al 2004, Brack et al 1999) in marmosets
and other New World, non-human primates. Food
allergy, colitis and production of IgA-gliadin anti-
bodies are thought to be linked to glomerulone-
phropathy in WMS (Brack et al 1999). Diffuse
mesangioproliferative glomerulonephritis in mar-
mosets (Fig. 1.46) is characterized by enlargement
of glomerular corpuscles due to proliferation of
the mesangium (cellular and matrix proliferation),
slight thickening of the Bowman’s capsule accom-
panied by inflammation and fibrosis of the
interstitium as well as tubular damage with accu- and urinary bladder is observed as a background
mulation of tubular hyaline casts. change in the cynomolgus monkey (Fig. 1.48b).
FIGURE 1.46  Diffuse glomerulonephritis in the kidney of
a marmoset. ×200. FIGURE 1.48a  Eosinophilic cytoplasmic inclusions in the
urothelium in a cynomolgus monkey. ×400.
Subcapsular ectopic adrenal tissue is sometimes
observed in both rhesus and cynomolgus macaque
kidneys. Microscopically, the lesions appear as
poor to well-circumscribed but non-encapsulated
nodules composed of cells resembling the three
zones of the adrenal cortex in varying proportions
and arranged in any order (Fig. 1.42a). Medullary
cells are usually absent and, in some cases, a zone
of the adrenal cortex, usually zona glomerulosa or
reticularis, may also be absent. Ectopic adrenal
tissue lying below the kidney capsule has been
described in man, mice and macaques (Chamanza
et al 2010, Prentice & Jorgeson 1979). This is
considered to be an incidental finding of no known
clinical significance, but should be differentiated
from proliferative lesions of the renal tubular epi-
FIGURE 1.48b  Vacuolation of the transitional epithelium
thelium. Ectopic adrenal cortical tissue can also be
or urothelium of the urinary bladder of the cynomolgus
found outside the kidney capsule within the peri-
monkey. ×200.
renal fat (Kaspareit 2009).
Multinucleate cells of the renal tubular epithe- Mineralization of embryonic, non-patent, vas-
lium of the collecting ducts are a frequent occur- cular remnants, thought to be remnants of umbili-
rence in the renal papilla of macaques (Fig. 1.47). cal arteries, in the adventitia of the bladder is a
They are considered an incidental finding of no common finding in both non-human primates and
known etiology and of little or no pathological dogs used in preclinical studies (Fig. 1.49). The
significance (Lowenstine 2003). non-patent, vascular structures are commonly
present with or without mineralization in associa-
tion with fibrosis, hemosiderosis and thrombosis.
Mineralization of the functional blood vessels may
also be observed.
FIGURE 1.47  Multinucleate cells in the kidney tubules of
a cynomolgus macaque. ×200.
Urinary bladder FIGURE 1.49  Mineralization of embryonic remnants in
Intracytoplasmic, eosinophilic, (pseudo)inclusion the urinary bladder in a cynomolgus monkey. ×200.
bodies are often present in the transitional epithe-
lium of the renal pelvis and the urinary bladder
of macaques (Fig. 1.48a). These are known to
Endocrine system
consist of cytokeratin (tonofilaments) and are of Non-human primates have a fetal adrenal cortex
no pathological significance (Lowenstine 2003). during fetal life which is gradually replaced post-
Vacuolation of the urothelium of the renal pelvis natally by an adult cortex with three zones.

Involution of the fetal cortex is accompanied by
hemorrhage and necrosis which is thought to be
partly responsible for the mineralized foci often
seen at the corticomedullary junction (Kast et al
1994, Majeed & Gopinath 1980). Calcified foci
or concentrically arranged concretions of calcified
material, are often found at the junction of the
medulla and cortex or occasionally within the zona
reticularis. They vary in size from small foci of less
than 0.1 mm in diameter to larger foci occupying
a third or more of the medulla. A thin layer of
fibrous connective tissue is usually present around
the edges of the foci (Fig. 1.50). The lesions are
thought to develop from either stress-related,
focal degeneration and necrosis of medullary cells
followed by a dystrophic mineralization or from
apoptosis of fetal adrenal tissue located in the
corticomedullary region. Residual cells or nuclei
can be seen in the centre of mineralization in early
lesions, supporting the dystrophic calcification
theory. Rhesus macaques are far more greatly
affected than cynomolgus monkeys but no sex
difference has been reported.
FIGURE 1.50  Mineralization of the adrenal cortex of a
rhesus macaque. ×100.
Adreno-hepatic fusion (AHF) and adreno-
hepatic adhesions (AHA) on the right side of the
abdominal cavity have been observed with some
frequency in young adult cynomolgus macaques
(Chamanza et al 2010, Mousa & van Esch 2004).
Adreno-hepatic fusion is defined as the union of
liver tissue with that of the right adrenal gland
with close intermingling of their respective paren-
chymal cells without an intervening connective
tissue capsule between them (Honma 1991). This
is in contrast to adreno-hepatic adhesions in which
a capsule between the two parenchymal tissue
types exists and there is no close intermingling of
the two different types of tissues. Both condi-
tions, which are considered to be congenital in
nature, in non-human primates (Chamanza et al
2010, Mousa & van Esch 2004), can be detected
at necropsy. Due to the difficulties involved in
sampling and separating the attached adrenal
gland from the larger liver lobe, histological inci-
dences of these conditions are most likely under-
estimated, and cases of pieces of adrenal tissues
left attached to the right liver lobe are known to
occur. Since the affected adrenal gland remains in
its expected anatomical position, and does not lie
within the liver capsule, these conditions are con-
sidered to be different from cases of ectopic
adrenal gland in the liver (see Fig. 1.41).
Histologically, typical liver parenchymal cells
are observed lying adjacent to cortical or medul-
lary adrenal cells with close intermingling of the
two tissue types without any intervening fibrous
tissue between them in cases of AHF (Fig. 1.51).
Reticulin stains can be employed to demonstrate
the merging of the reticular network of the liver
trabeculae with that of adrenal medullary cell
nests.
FIGURE 1.51  Adreno-hepatic fusion in the liver in a
cynomolgus monkey. ×100.
Histological evidence of ectopic liver tissue
completely surrounded by adrenal tissues or
embedded between cortical and medullary tissue
has also been observed (Chamanza et al 2010).
The clinical significance of these conditions
has been described in man (Woo et al 2007) in
which an adrenal cortical adenoma developed
in AHF tissue and mimicked malignant hepatic
tumor at computerized tomographic scanning and
angiography-assisted computerized tomographic
scanning.
In marmosets, it is common to observe ectopic
liver tissue consisting of a cluster of bile ducts,
with or without islands of hepatocytes, in extra-
hepatic areas such as at the hilus of the liver (Fig.
1.52) and in connection with ligaments derived
from embryonic remnants such as the round
ligament.
FIGURE 1.52  Ectopic liver and bile duct structures in a
marmoset. ×100.
Minimal to slight grades of age-related lipofus-
cin pigment accumulation in the adrenal gland
cortex is infrequently detected in young adult
cynomolgus macaques used in toxicity studies
(Kaspareit 2009). The initial stages of the lesion,
seen in the three- to four-year old animals used in
preclinical studies, is characterized by yellow to
light-brown, granular pigment in the zona reticu-
laris and fascicularis, and these regions turn dark
brown as the animals get older. The pigment has
to be differentiated from hemosiderin which may
be present in macrophages in the same region
Non-human primates 11
(Kaspareit 2009). A common finding in the
adrenals of non-human primates is vacuolation of
cortical cells, which can be either focal, and
restricted to one cortical zone, or diffuse, and
associated with cortical hypertrophy. Moderate
grades of vacuolation can be seen in marmosets,
and the vacuoles stain positive for lipid (Kaspareit
2009). Extramedullary hemopoiesis is commonly
observed within the adrenal gland of marmosets
(Fig. 1.53) (Okazaki et al 1996).
FIGURE 1.53  Extramedullary hemopoiesis in the adrenal
gland of a marmoset. ×100.
C-cell hyperplasia is a very common finding in
the thyroid glands of marmosets (Kaspareit et al
2006, Tucker 1984). It may occur as one or more
nodules of concentrated C-cell areas in the middle
of the organ (Fig. 1.54) or an increased number
of C-cells may be found diffusely distributed
throughout the organ, causing follicles to appear
smaller and fewer in number and giving the organ
a more solid appearance. The significance of this
finding is not fully known because no C-cell ade-
nomas have been reported in marmosets to date
and the condition does not appear to be sex-
dependent as in the rat (Kaspareit et al 2006).
FIGURE 1.54  C-cell hyperplasia in the thyroid of a mar-
moset. ×100.
Ectopic thymus tissue in the thyroid gland or
the parathyroid gland is a very common finding of
no known pathological or clinical significance in
non-human primates used in toxicity studies
(Chamanza et al 2010, Tucker 1984). In man, due
to the link between myasthenia gravis and the
thymus, ectopic thymic tissue has a greater clini-
cal significance. Ectopic thymic tissue is usually
located at the periphery of the thyroid or parathy-
roid gland (Fig. 1.55) as a single, small nodule that
may contain both the medulla (with epithelial
components) and the cortex, or may just be made
up of the cortical tissue. Embryologically, the
12 BACKGROUND LESIONS IN LABORATORY ANIMALS
thymus, parathyroid and thyroid gland have a
common pharyngeal pouch origin, hence the high
prevalence of ectopic thymus in the parathyroid
and thyroid or vice versa.
FIGURE 1.55  Ectopic thymus in the parathyroid gland of
a marmoset. ×100.
Eosinophilic intranuclear inclusions are visible
in the pars adenohypophysis of the pituitary.
These inclusions are thought to be cytoplasmic
invaginations similar to those observed in the
mouse hepatocytes. This lesion is reported to
occur in approximately 50% of all cynomolgus
monkeys and should be distinguished from viral
inclusions by using a phloxine tartrazine special
stain.
Muscles, bones and joints
Aggregates of large, foamy macrophages with pale
basophilic staining cytoplasm (Fig. 1.56), repre-
senting vaccination or alum granuloma (sites of
previous intramuscular vaccinations), are usually
observed within the thigh muscle that is routinely
collected for histology in toxicity studies. They are
an incidental finding of no clinical or pathological
significance.
FIGURE 1.56  Vaccine granuloma in the skeletal muscle
in a cynomolgus monkey. ×100.
Cysts of Sarcocystis parasites are occasionally
observed in skeletal muscle, heart or smooth
muscle of both wild-caught and captive-bred
rhesus macaques. While they occur commonly in
wild-caught animals, they are relatively uncom-
mon in laboratory-raised macaques. They are
generally regarded as incidental and of no patho-
logical significance, even though some reports of
generalized and fulminating disease associated
with Sarcocystis species have been reported in
rhesus macaques (Gozalo et al 2007, Lane et al
1998). Cysts are commonly found within the
thigh, tongue and esophageal muscles (Fig. 1.57),
and extremely rarely within the heart. They
consist of cross sections of tissue cysts containing
crescent-shaped protozoal zoites that are not asso-
ciated with an inflammatory infiltrate.
FIGURE 1.57  Sarcocystis cysts present in the skeletal
muscle of the tongue of a cynomolgus monkey. ×100.
Skeletal abnormalities occur not uncommonly
in young macaques and marmosets on studies;
they include bone fractures, developmental carti-
lage and bone abnormalities, hyperostosis and
periosteal reaction, growth-plate fractures associ-
ated with metaphyseal dysplasia (Chamanza et al
2010) in cynomolgus macaques, and spontaneous
osteomyelitis (Fig. 1.58) and nutritional osteodys-
trophy of young marmosets (Chalmers et al
1983). Fibrous osteodystrophy (FOD) of marmo-
sets has generally been attributed to the inability
of vitamin D2 to promote the intestinal absorption
of calcium in New World monkeys, which can be
ameliorated by substituting vitamin D2 with
vitamin D3 or exposure to sunlight or artificial
light of appropriate wavelength (National Research
Council 2003). Rickets and osteomalacia (in older
animals) are syndromes associated with feeding
diets containing only vitamin D2 with insufficient
exposure to ultraviolet light. However, FOD,
osteomalacia and rickets in marmosets have
almost disappeared in modern facilities because
vitamin D3 is now used to substitute vitamin D2
in commercial diets.
FIGURE 1.58  Osteomyelitis in the tibia of a marmoset.
×100.
FOD, which is a common cause of fractures in
marmosets, is characterized by thin bone cortices
and trabeculae surrounded by fibrous stroma.
Thinning of bone is brought about by marked
resorption of spongy and cortical bone. Numerous
osteoclasts are therefore observed within How-
ship’s lacunae and there is an invasion of Haver-
sian spaces and the marrow canal by fibrous tissue
(Fig. 1.59). The lesions are a result of direct action
of parathyroid hormone (PTH) on bone, and
excess PTH production associated with this syn-
drome may also contribute to soft-tissue minerali-
zation seen in marmosets. Oversupply of vitamin
D is also known to lead to soft-tissue minerali­
zation and nephrocalcinosis (National Research
Council 2003).
FIGURE 1.59  Fibrous osteodystrophy in a marmoset.
×50.
Spontaneous growth-plate fractures or micro-
fractures in animals with physeal retention or
metaphyseal dysplasia have been observed in a
number of young cynomolgus macaques and the
causes remain unclear (Chamanza et al 2010).
They are characterized by foci of persistent hyper-
trophic chondrocytes lying above a fracture line,
an accumulation of primary bone spongiosa and a
large number of osteoclasts (Fig. 1.60). Distin-
guishing these lesions from rickets can prove to be
a major diagnostic challenge.
FIGURE 1.60  Growth plate dysplasia with fracture in a
cynomolgus monkey. ×25.
Brain and nervous system
Mineralized bodies or vascular mineralization
occur commonly as a spontaneous finding in the
brain of apparently normal non-human primates
(Wadsworth et al 1995, Yanai et al 1994). This
finding has been reported in most non-human
primate species used in toxicity studies and does
not appear to be age-related. The mineralized
bodies are commonly found in association with
small blood vessels in the globus pallidus, putamen
or, occasionally, the caudate nucleus, and are
therefore usually observed on coronary sections of
the brain that pass through the optic chiasma and
mammillary bodies (Wadsworth et al 1995). They
appear as irregular and often elongated structures
composed of smaller globoid bodies formed

by concentric lamellar parts that stain strongly
basophilic or a deep purple colour with hematoxy-
lin and eosin (Fig. 1.61). The margins of some of
the larger mineralized structures often show eosi-
nophilic staining, while smaller eosinophilic struc-
tures may be observed in association with small
arteries. The nature and the exact origin of these
structures is not known; they are considered to be
associated with iron metabolism by perivascular
and perineuronal oligodendrocytes because they
are found in regions of high iron metabolism and
stain positive with Perl’s stain (Wadsworth et al
1995).
FIGURE 1.61  Brain mineralization in a cynomolgus
monkey. ×400.
Various types of pigmentation are observed in
the brain of the macaque. While age-related and
non age-related, ceroid-lipofuscin pigments are
occasionally encountered, perivascular melanin
pigmentation is the most commonly encountered
form of pigmentation in the brain of cynomolgus
macaques used on toxicity studies (Chamanza
et al 2010). The finding can be observed as black
discoloration at post mortem, and is observed as
Fontana–Masson-positive, perivascular pigment
on histopathological examination (Fig. 1.62).
FIGURE 1.62  Perivascular melanin in the brain in a
cynomolgus monkey. Fontana-Masson stain. ×100.
Minimal focal gliosis and glial scarring are
unusual, but not uncommon findings in cynomol-
gus monkey brains (Chamanza et al 2010). These
lesions are often associated with perivascular
cuffs, neuronal necrosis and neuronophagia (Fig.
1.63a) and macrophage accumulation with lipo-
fuscin pigment. The lesions can be observed in any
part of the brain, are usually minimal in severity,
and are often not associated with any clinical or
necropsy abnormalities. No etiological agent has
been identified. Perivascular cuffing occurs far
more commonly than gliosis and may or may not
Non-human primates 13
be associated with this lesion. Perivascular cuffs
are characterized by the accumulation of a uniform
layer of lymphocytes in the Virchow–Robin space
around small blood vessels (Fig. 1.63b), and are
usually multifocal in distribution. Besides blood
vessels in the brain parenchyma, choroidal, menin-
geal and spinal-cord blood vessels are also affected
to a similar or greater extent. Vater Pacini corpus-
cles are large mechanoreceptors that sense pres-
sure and stretch and are observed occasionally
within the skin or the pancreas. The structure has
a unique ‘onion-skin’ appearance (Fig. 1.63c).
FIGURE 1.64a  Lymphocytic infiltration into the choroid of
the eye in a cynomolgus monkey. ×200.
FIGURE 1.63a  Glial scar in the brain in a cynomolgus
monkey. ×200.
FIGURE 1.64b  Optic nerve neuritis in a cynomolgus
monkey. ×100.
References
Adachi, K., Mori, T., Ito, T., et al., 2005.
Collagenofibrotic glomerulonephropathy in
a cynomolgus macaque (Macaca fascicularis).
Vet. Pathol. 42, 669–674.
Adler, R.R., Moore, P.F., Schmucker, D.L., et al., 1993.
Chronic colitis, juvenile macaca mulatta. In: Jones,
FIGURE 1.63b  Gliosis and perivascular cuffing in the T.C., Mohr, U., Hunt, R.D. (Eds.), Monographs on
brain in a cynomolgus monkey. ×100. the pathology of laboratory animals, nonhuman
primates, vol. 2. Springer-Verlag, Berlin, pp. 20–32.
Albassam, M.A., Lillie, L.E., Smith, G.S., 1993.
Asymptomatic polyarteritis in a cynomolgus monkey.
Lab. Anim. Sc. 43, 628–629.
Arey, L.B., 1954. Developmental anatomy; a textbook
and laboratory manual of embryology, seventh ed.
Saunders, Philadelphia, pp. 342–374.
Bennett, B.T., Abee, C.R., Henrickson, R., 1998.
Nonhuman primates in biomedical research.
Diseases. American College of Laboratory Animal
Medicine Series. Academic Press, San Diego.
Betton, G.R., 1998. The digestive system 1: the
gastrointestinal tract and exocrine pancreas. In:
Turton, J., Hooson, J. (Eds.), Target organ pathology.
Taylor & Francis, London, p. 51.
Borda, J.T., Pauley, D.R., Mackey, J.J., et al., 2004.
Immunoglobulin-A nephropathy with crescentic
FIGURE 1.63c  Vater Pacinian corpuscle in the mammary glomerulonephritis in a pigtailed macaque (Macaca
nemestrina). Vet. Pathol. 41, 44–49.
gland of a cynomolgus monkey. ×100.
Brack, M., Schroeder, C., Fooke, M., et al., 1999.
A cluster of uniform small lymphocytes are IgM/IgA nephropathy in callitrichids: antigen
studies. Nephron. 82, 221–231.
occasionally seen within the choroid or ciliary
Bundza, A., Dukes, T.W., 1978. Some heterotropic
body of the eyes of cynomolgus macaques (Sinha
tissue remnants in domestic animals. Can. Vet. J.
et al 2006). The lesion is of no consequence and 19, 322–324.
is not associated with an inflammatory reaction
Chalmers, D.T., Murgatroyd, L.B., Wardsworth, P.F.,
(Fig. 1.64a). Optic neuritis is also observed in the 1983. A survey of the pathology of marmosets
optic nerves of cynomolgus monkeys with no (Callithix jacchus) derived from a marmoset
other evidence of disease (Fig. 1.64b). breeding unit. Lab. Anim. 17, 270–279.
14 BACKGROUND LESIONS IN LABORATORY ANIMALS
Chamanza, R., Parry, N.M., Rogerson, P., et al., 2006.
Spontaneous lesions of the cardiovascular system in
purpose-bred laboratory nonhuman primates.
Toxicol. Pathol. 34, 357–363.
Chamanza, R., Marxfeld, H., Blanco, A., et al., 2010.
Incidences and range of spontaneous lesions in
control cynomolgus monkeys (Macaca fascicularis)
used in toxicity studies. Toxicol. Pathol. 38,
642–657.
David, J.M., Dick, E.J. Jr, Hubbard, G.B., 2009.
Spontaneous pathology of the common marmoset
(Callithrix jacchus) and tamarins (Saguinus
oedipus, Saguinus mystax). J. Med. Primatol. 38(5),
347–359.
De Lacroix, W.F., Hübner, G., 1974. Ciliated epithelial
inclusion cyst of the heart. Beitr. Pathol. 151,
103–110.
Drevon-Gaillot, E., Perron-Lapage, M., Clement, C.,
et al., 2006. A review of background findings in
cynomolgus monkeys (Macaca fascicularis) from
three different geographical origins. Exp. Toxicol.
Pathol. 58, 77–88.
Dubois, A., Tarnawski, A., Newell, D.G., et al., 1991.
Gastric injury and invasion of parietal cells by spiral
bacteria in rhesus monkeys. Are gastritis and
hyperchlorhydria infectious diseases?
Gastroenterology 100, 884–891.
Dubois, A., Fiala, N., Heman-Ackah, L.M., et al.,
1994. Natural gastric infection with Helicobacter
pylori in monkeys: a model for spiral bacteria
infection in humans. Gastroenterology 106,
1405–1417.
Elwell, M.R., Mahler, J.F., 1999. Heart, blood vessels
and lymphatic vessels. In: Maronpot, R.R., Boorman,
G.A., Gaul, B.W. (Eds.), Pathology of the mouse.
Cache River Press, Vienna, pp. 361–380.
Engedal, H., Jensen, H., Saetersdal, T.S., 1977.
Ultrastructure of abnormal membrane inclusions in
nuclei of human myocardial cells. Br. Heart J. 39,
145–151.
Fikes, J.D., O’Sullivan, M.O., Bain, M.T., et al., 1996.
Gastric infarction in cynomolgus monkeys (Macaca
fascicularis). Vet. Pathol. 33, 171–175.
Foster, J.R., 2005. Spontaneous and drug-induced
hepatic pathology of the laboratory beagle dog, the
cynomolgus macaque and marmoset. Toxicol. Pathol.
33, 63–74.
Fox, R.R., Weisbroth, S.S., Crary, D.D., et al., 1976.
Accessory spleens in domestic rabbits I. Frequency
description and genetic factors. Teratology 13,
243–252.
Gad, S.C., Frith, C., Goodman, D.G., et al., 2007.
The mouse. In: Gad, S.C. (Ed.), Animal models in
toxicology, second ed. Taylor & Francis, Boca Raton,
pp. 94.
Glick, B., Sato, K., 1964. Accessory spleens in the
chicken. Poultry Sci. 43, 1610–1612.
Gozalo, A.S., Montali, R.J., St Claire, M., et al., 2007.
Chronic polymyositis associated with disseminated
sarcocystosis in a captive-born rhesus macaque. Vet.
Pathol. 44, 695–699.
Greaves, P., 2000. Patterns of cardiovascular pathology
induced by diverse cardiovascular drugs. Toxicol.
Lett. 112, 547–552.
Guzman, R.E., Kerlin, R.L., Zimmerman, T.E., 1999.
Histologic lesions in cynomolgus monkeys (Macaca
fascicularis) naturally infected with simian retrovirus
type D: comparison of seropositive, virus-positive,
and uninfected animals. Toxicol. Pathol. 27,
672–677.
Honma, K., 1991. Adreno-hepatic fusion: an autopsy
study. Zentralbl. Pathol. 137, 117–122.
Hubbard, G.B., Schmidt, R.E., Fletcher, K.C., 1987.
Systematic survey of lesions from animals in a
zoological collection. J. Zoo Anim. Med. 18,
14–46.
Ito, T., Chatani, F., Sasaki, S., et al., 1992.
Spontaneous lesions in cynomolgus monkeys
used in toxicity studies. Jikken Dobutsu. 41,
455–469.
Jasty, V., Jamison, J.R., Hartnagel, R.E., 1984. Three
types of cytoplasmic granules in cardiac muscle cells
of cynomolgus monkeys (Macaca fascicularis). Vet.
Pathol. 21, 505–508.
Kaspareit, J., 2009. Adrenal gland background
pathology of primates in toxicological studies.
In: Harvey, P.W., Everett, D.J., Springall, C.J.
(Eds.), Adrenal toxicology. Target Organ Toxicology
Series, vol. 26. Informa Healthcare, New York,
pp. 139–160.
Kaspareit, J., Friderichs-Gromoll, S., Buse, E., et al.,
2003. Spontaneous squamous cysts and squamous
epithelial plaques in the heart of cynomolgus
monkeys (Macaca fascicularis). Exp. Toxicol. Pathol.
54, 301–303.
Kaspareit, J., Friderichs-Gromoll, S., Buse, E., et al.,
2004. Spontaneous tubular (cuboidal) metaplasia of
the parietal layer of Bowman’s capsule in
cynomolgus monkeys (Macaca fascicularis). J. Exp.
Anim. Sci. 43, 13–17.
Kaspareit, J., Friderichs-Gromoll, S., Buse, E., et al.,
2006. Background pathology of the common
marmoset (Callithrix jacchus) in toxicological
studies. J. Exp. Anim. Sci. 57, 405–410.
Kast, A., 1994. Pulmonary hair embolism in monkeys.
Exp. Toxicol. Pathol. 46, 183–188.
Kast, A., Peil, H., Weisse, I., 1994. Calcified foci at the
junction between adrenal cortex and medulla of
rhesus monkeys. Lab. Anim. 28, 80–89.
Keenan, C.M., Vidal, J.D., 2006. Standard
morphologic evaluation of the heart in the
laboratory dog and monkey. Toxicol. Pathol. 34,
67–76.
Khullar, M., Datta, B.N., Wahi, P.L., et al., 1989.
Catecholamine-induced experimental
cardiomyopathy; a histopathological, histochemical
and ultrastructural study. Indian Heart J. 41,
307–313.
Lane, J.H., Mansfield, K.G., Jackson, L.R., et al., 1998.
Acute fulminant sarcocystosis in a captive-born
Rhesus macaque. Vet. Pathol. 35, 499–505.
Lau, D.T.L., 1973. Ectopic splenic nodules in the
pancreas of a capuchin monkey (Cebus albifrons).
J. Med. Primatol. 2, 67–70.
Layne, D.G., Power, R.A., 2003. Husbandry, handling
and nutrition for marmosets. Comp. Med. 53,
351–359.
Lilbert, J., Burnett, R., 2003. Main vascular changes
seen in the saline controls of continuous infusions
studies in the cynomolgus monkey over an 8-year
period. Toxicol. Pathol. 31, 273–280.
Lindsay, S., Chaikoff, I.L., 1966. Naturally occurring
arteriosclerosis in nonhuman primates. J. Atherosc.
Res. 6, 36–61.
Losco, P.E., Kaminska-McNamara, G.Z., Johnson, J.,
et al., 2002. Common incidental histopathological
findings in cynomolgus monkeys. Poster presentation
to Society for Toxicological Pathology.
Lowenstine, L.J., 1993. Type D retrovirus infection,
macaques. In: Jones, T.C., Mohr, U., Hunt, R.D.
(Eds.), ILSI Monographs on the pathology of
laboratory animals, nonhuman primates, vol. 1.
Springer-Verlag, Berlin, pp. 20–32.
Lowenstine, L.J., 2003. A primer of primate
pathology: lesions and nonlesions. Toxicol. Pathol.
31, 91–102.
Majeed, S.K., Gopinath, C., 1980. Calcification in the
adrenals and ovaries of monkeys. Lab. Anim. 4,
363–365.
McClure, H.M., Chapman, W.L., Hooper, B.E., et al.,
1978. The digestive system. In: Benirschke, K.,
Garner, F.M., Jones, T.C. (Eds.), Pathology of
laboratory animals, vol. 1. Springer-Verlag, New York,
pp. 1–56.
McKeag, S., Mcinnes, E.F., 2012. The incidence of
lymphoplasmacytic gastritis in the fundus and
antrum of cynomolgus monkey (Macaca fascicularis)
stomachs. Journal of Toxicologic Pathology (In
press).
Miller, G.F., Barnard, D.E., Woodward, R.A., et al.,
1997. Hepatic hemosiderosis in common marmosets,
Callithrix jacchus: effect of diet on incidence and
severity. Lab. Anim. Sci. 47, 138–142.
Mousa, S., van Esch, E., 2004. Two cases of
adreno-hepatic fusion in cynomolgus monkeys
(Macaca fascicularis). Toxicol. Pathol. 32,
511–513.
National Research Council, 2003. Nutritional
requirements of nonhuman primates, second ed.
National Academies Press, Washington DC,
pp. 116–121.
Okazaki, Y., Kurata, Y., Makinodan, F., et al., 1996.
Spontaneous lesions detected in the common
cotton-eared marmosets (Callithrix jacchus). J. Vet.
Med. Sci. 58 (3), 181–190.
Olsen, W.R., Beaudoin, D.E., 1969. Increased
incidence of accessory spleens in hematological
disease. Arch. Surg. 198, 762–763.
Porter, B.F., Frost, P., Hubbard, G.B., 2003.
Polyarteritis nodosa in a cynomolgus macaque
(Macaca fascicularis). Vet. Pathol. 40, 570–573.
Prentice, D.E., Jorgensen, W., 1979. Ectopic adrenal
tissue in the kidney of rhesus monkeys (Macaca
mulatta). Lab. Anim. 13, 221–223.
Qureshi, S.R., 1979. Chronic interstitial myocarditis in
primates. Vet. Pathol. 16, 486–487.
Reindel, J.F., Fitzgerald, A.L., Breider, M.A., et al.,
1999. An epizootic of lymphoplasmacytic gastritis
attributed to Helicobater pylori infection in
cynomolgus monkeys (Macaca fascicularis). Vet.
Pathol. 36, 1–13.
Scott, G.B.D., 1982. Mucosal microhernias in the
nonhuman primate colon: their role in the
pathogenesis of colonic disease. J. Vet. Path. (supp.)
7, 130–140.
Scott, G.B.D., 1999. Comparative primate pathology.
Blackwell Science, Oxford.
Shimoi, A., Kakinuma, C., Kukayama, C., et al., 1998.
Comparison of spontaneous minor lesions in
wild-caught and laboratory-bred monkeys. J. Toxicol.
Pathol. 11, 85–94.
Sinha, D.P., Cartwright, M.E., Johnson, R.C., 2006.
Incidental mononuclear cell infiltrate in the uvea
of cynomolgus monkeys. Toxicol. Pathol. 34,
148–151.
Thomas, R., van Wesep, R., 1990. Intracardiac
epithelial cyst in association with an atrioventricular
canal defect. Am. J. Cardiovasc. Pathol. 3,
325–328.
Tucker, M.J., 1984. A survey of the pathology of
marmosets (Callithrix jacchus) under experiment.
Lab. Anim. 18, 351–358.
Vidal, J.D., Mirabile, R.C., Thomas, H.C., 2008.
Evaluation of the cynomolgus monkey stomach:
recommendations for standard sampling procedures
in nonclinical safety studies. Toxicol. Pathol. 36,
250–255.
Vidal, J.D., Drobatz, L.S., Holliday, D.F., et al., 2010.
Spontaneous findings in the heart of Mauritian-origin
macaques (Macaca fascicularis). Toxicol. Pathol.
238, 297–302.
Vogel, P., Fritz, D., 2003. Cardiomyopathy associated
with angiomatous pheochromocytoma in a rhesus
macaque (Macaca mulatta). Vet. Pathol. 40,
468–473.
Wadsworth, P.F., Jones, H.B., Cavanagh, J.B., 1995.
The topography, structure and incidence of
mineralized bodies in the basal ganglia of the brain

of cynomolgus monkeys (Macaca fascicularis). Lab.
Anim. 29, 276–281.
Weisbroth, S.H., Fox, R.R., Scher, S., et al., 1976.
Accessory spleens in domestic rabbits (Oryctolagus
suniculus). II. Increased frequency in hematological
diseases and experimental induction with
phenylhydrazine. Teratology 13, 253–262.
Woo, H.S., Lee, K.H., Park, S.Y., et al., 2007. Adrenal
cortical adenoma in adrenohepatic fusion tissue: a
mimic of malignant hepatic tumor at CT. Am. J.
Roentgenol. 188, 246–248.
Yanai, T., Masegi, T., Ueda, K., et al., 1994. Vascular
mineralisation in the monkey brain. Vet. Pathol. 34,
546–552.
Non-human primates 15
Yoon, Y.S., Shin, J.W., Park, C.B., et al., 2000.
Morphological structure of accessory spleen in
Chinese hamsters. J. Vet. Sci. 1, 73–75.
Zabka, T.S., Irwin, M., Albassam, M.A., 2009.
Spontaneous cardiomyopathy in cynomolgus
monkeys (Macaca fascicularis). Toxicol. Pathol. 37,
814–818.
 Wistar and Sprague–Dawley rats 17
CHAPTER 2 
Elizabeth F McInnes

Wistar and Sprague–Dawley rats

the cartilaginous foci may mineralize (MacKenzie

Introduction & Alison 1990). Adipose tissue infiltration in the
Almost all laboratory and domestic pet rats belong heart is characterized by the presence of sub­
to a single species, the Norway rat (Rattus nor- epicardial adipose cells and is often observed in
vegicus). The rat is the most commonly used older, obese animals (Ruben 2000). Lipofuscinosis
rodent species for toxicology studies. A number results in golden brown pigment granules at the
of rat species are used in research, contract poles of the nuclei of myocytes (Ruben 2000).
research organizations and in the pharmaceutical
industry including the Wistar, Sprague Dawley
and Fischer 344 rat. Different strains of rat differ
in their sensitivity to various compounds and in
their incidence of spontaneous tumors. Differ-
ences in age and sex of rats should also be taken
into consideration when designing toxicological
and experimental studies (Johnson & Gad 2007).
Background lesions in rats are made up of degen- FIGURE 2.1  Cardiomyopathy in left ventricle. ×100.
erative, inflammatory, proliferative and congenital
changes. Although aging rats produce many inter-
esting lesions, there are also a number of lesions
that occur spontaneously in young animals.
The pathology of the aged rat becomes impor-
tant in the latter stages of carcinogenicity studies
(Chandra & Frith 1992). The nonneoplastic
lesions and proliferations noted in aging rats are FIGURE 2.4  Cartilage at base of aorta in a rat. ×100.
problematic as they can be confused with preneo-
plastic and neoplastic disease (Johnson & Gad Endocardial myxomatous change occurs in
2007). This chapter will give an overview of back- older rats and is recognized by focal or diffuse
ground lesions encountered in both young and thickening in the subendocardium due to the pres-
older Wistar and Sprague-Dawley rats. ence of myxomatous tissue (Ruben 2000). Hyper-
trophy and hyperplasia of the endocardium may
be seen in conjunction with the myxomatous val-
Cardiovascular system vular changes. In addition, the lesion is character-
FIGURE 2.2  Myocardial lymphoid cells. ×10. ized by focal areas of hemorrhage, hemosiderin
Cardiomyopathy (Fig. 2.1) in aging rats (spontane-
ous cardiomyopathy) is a common lesion of older pigmentation, hyalinization, thrombi formation,
Mesenchymal proliferation in the subendocar- inflammatory and mast cells and polypoid projec-
rats. It is found most frequently in the subintimal
dium (Fig. 2.3) is also known as endocardial tions (Ruben, 2000). Heart valve cysts may occa-
myocardium of the left ventricular wall and is
hyperplasia, endocardial disease, endomyocardial sionally be observed in the heart valves of rats
more common in male rats (MacKenzie & Alison
disease or subendocardial fibrosis. Although endo- (Fig. 2.5).
1990). The etiology of the disease is unknown,
cardial schwannomas can be found in the same
but severity and age of onset are influenced by
location, it is not known whether the mesenchy-
diet, environment and stress (MacKenzie & Alison
mal proliferation in subendocardium always
1990). The lesion is characterized initially by
progresses to neoplasia (Johnson & Gad 2007).
necrotic, eosinophilic cardiomyocytes surrounded
The cells are distinct from the bordering myocar-
by fibroblasts and inflammatory cells (Fig. 2.2),
dial cells, but may extend into muscle bundles.
predominantly lymphocytes. Later, fibrosis is
prominent in aging rats and is generally present
in the left ventricle, intraventricular septum and
papillary muscle. Cartilagenous and osseous meta-
plasia of the chorda tendinae may occur at the end
stage of the disease (Ruben 2000). Atrial (left)
and ventricular thrombosis are occasionally associ-
ated with cardiomyopathy in aging rats.

FIGURE 2.5  Heart valve cyst in the rat. ×100.

Vascular lesions in the blood vessels of Sprague-

FIGURE 2.3  Endocardial mesenchymal proliferation in
the rat heart. ×100.
Cartilaginous foci at the base of the aorta are
common in both young and old Sprague–Dawley
rats (Fig. 2.4) (Johnson & Gad 2007). In older rats
Dawley and Wistar rats include medial degenera-
tion and medial hypertrophy. Medial degeneration
is common in the coronary arteries of rats and
the affected vessels display enlarged, irregularly
arranged and sparse nuclei present in a PAS-
positive matrix (Ruben 2000). Medial hyper­
trophy of the blood vessels is rare and tends to
occur in pulmonary arteries. This lesion consists
18 BACKGROUND LESIONS IN LABORATORY ANIMALS
of thickening of the tunica media due to hyper-
trophy of the smooth muscle cells (Ruben 2000)
(Fig. 2.6).
FIGURE 2.6  Medial hypertrophy in heart artery. ×100.
Arteritis (polyarteritis, periarteritis, panarteritis
nodosa) (Fig. 2.29) is found occasionally in
Wistar rats, but is more common in Fischer rats
(MacKenzie & Alison 1990). The occurrence of
the lesion is higher in males and increases with
age. Pancreatic, mesenteric and spermatic arteries
are more commonly affected. The lesion consists
of fibrinoid degeneration of the media with
inflammatory cell infiltration progressing to fibro-
sis, vascular dilatation, occlusion and aneurysm
(Mackenzie & Alison 1990). The pathogenesis is
unknown but may be related to immune complex
disease. The vaso vasorum of the mesenteric arter-
ies of the rat can occasionally be confused with a
treatment-related finding (Fig. 2.7a). Age-related
vascular medial cartilage and mineralization may
be encountered in the blood vessels of aging rats
(Fig. 2.7b).
FIGURE 2.7a  Vaso vasorum in adventitia of a mesenteric
artery blood vessel in the rat. ×100.
FIGURE 2.7b  Age-related vascular medial cartilage and
mineralization in the tongue of a rat. ×100.
Arterial thrombosis (not caused by arteritis)
occurs occasionally in the lung, kidney, liver and
adrenal (Ruben 2000). These thrombi may be
emboli originating from atrial thromboses in older
rats. Atherosclerosis is a rare background lesion
consisting of focal areas of foam cells within the
tunica intima or tunica media (Ruben 2000).
Intimal and/or medial sclerosis with mineraliza-
tion of the elastic fibres occurs in the aorta (Ruben
2000). Vascular mineralization in arteries and
veins in various organs such as the lung, is occa-
sionally observed as a background lesion in rats.
Hemolymphoreticular system
Pigmented macrophages in the sinuses (Fig. 2.8)
(particularly the medullary or subscapular sinuses)
of the lymph nodes are observed commonly in
rats. Generally this pigment is ceroid or lipofuscin
material (Stefanski et al 1990). Special stains
(such as Schmorl’s for lipofuscin) should be used
to distinguish ceroid pigment from iron (Perl’s-
positive) or melanin (Masson Fontana-positive).
FIGURE 2.8  Macrophages with brown pigment in the
mesenteric lymph node. ×100.
Cystic degeneration (Fig. 2.9) of various differ-
ent lymph nodes (also known as sinus dilatation,
lymphangiectasia, cystic ectasia, lymphatic sinus
ectasia, lymphangiectasis, lymphatic cysts) is
observed as a background lesion in rat lymph
nodes. Generally the subcapsular and medullary
sinuses are dilated. The sinuses are dilated with
lymph and occasional lymphocytes or macro-
phages and erythrocytes or proteinaceous fluid
can be observed within the lumen (Stefanski et al
1990). Cystic dilatation of the lymph nodes may
be so prominent that it is noted at necropsy. The
mesenteric, mediastinal and paralumbar lymph
nodes are often affected (Frith et al 2000b). Vas-
cular ectasia (also known as angiectasia, vascular
sinus dilatation, peliosis, telangiectasis) is the dila-
tation of vascular channels within the medullary
sinuses (Frith et al 2000b). Thrombosis is occa-
sionally observed in the blood vessels in the lymph
node (Frith et al 2000b).
FIGURE 2.9  Rat lymph node with sinus dilatation. ×200.
Lymph node atrophy (also known as senescent
atrophy) is a common lesion of aging rats. The
atrophy is generally characterized by a reduction
in germinal centres and follicles and depletion of
lymphocytes in the paracortex may also be seen
(Stefanski et al 1990). Senescent atrophy is often
nearly ubiquitous in carcinogenicity studies. Mod-
erate amounts of intracellular pigment (generally
lipofuscin) are often noted in the medullary area.
Mast cells in lymph nodes (Fig. 2.10), particu-
larly the mandibular lymph node, are observed
frequently in the lymph nodes of both young and
old rats. Plasmacytosis of mandibular lymph node
(Fig. 2.11) is found predominantly in the medul-
lary cords (Stefanski et al 1990). Plasmacytosis of
lymph nodes may be so severe as to be noted at
necropsy as enlargement. The etiology and signifi-
cance of the lesion are unknown, but the lesion is
particularly common in mandibular lymph nodes,
particularly in conjunction with oral cavity lesions.
Plasmacytosis may be prominent in animals with
T cell atrophy (Frith et al 2000b). In addition,
accumulations of Russell bodies may also be
observed in a lymph node with plasmacytosis (Fig.
2.12).
FIGURE 2.10  Mast cells in the sinuses of a mandibular
lymph node. ×200.

FIGURE 2.11  Plasmacytosis in the mandibular lymph
node. ×200.
FIGURE 2.12  Russell bodies in a lymph node. ×200.
Mineralization of Peyer’s patches or other
lymph nodes is occasionally observed in the ileum
and is likely to be secondary to tissue damage
(Frith et al 2000b) (Fig. 2.13). Small capsular
cysts may occur on the surface of the rat spleen
(Fig. 2.14). The cysts are lined by endothelial cells
and con­tain a pink, proteinaceous fluid (Stefanski
et al 1990).
FIGURE 2.13  Mineralization of Peyer’s patches in the
small intestine of the rat. ×200.
Wistar and Sprague–Dawley rats 19
FIGURE 2.14  Capsular cyst in rat spleen. ×100.
FIGURE 2.16  Perl’s-positive hemosiderin pigment pre­
sent in the red pulp of a rat spleen. ×200.
Sinus erythrocytosis is characterized by the
presence of erythrocytes within the lymph node
sinuses (Stefanski et al 1990). This lesion is
common and generally occurs due to the presence
of hemorrhage in the organs draining into the
lymph nodes. Erythrophagocytosis by macro-
phages may also be visible within the sinuses con-
taining red blood cells.
Increased extramedullary hematopoiesis is a
common lesion in the rat spleen (Fig. 2.15). The
rat spleen normally has a minimal level of extra­
medullary hematopoiesis; however, it is very dif-
ficult to distinguish between normal and excessive
levels of extramedullary hematopoiesis. Conges-
tion is a common background lesion of the spleen
and may be linked to the method of euthanasia
used to kill the animals (Frith et al 2000b). FIGURE 2.17  Tatto pigment in a rat popliteal lymph node.
×200.
Accessory spleen is an uncommon background
finding in the rat abdomen. Generally it presents
with nodules of splenic tissue present within the
mesenteric adipose tissue. The cause is either con-
genital or traumatic injury (Stefanski et al 1990).
Mineralization may occasionally be seen in the
walls of the blood vessels of the spleen as well as
in the capsule of aging rats (Frith et al 2000b).
Parenchymal fibrosis of the red pulp of the spleen
can occasionally be observed in aging rats.
Ectopic thymus is a common finding adjacent
to the thyroid. Ectopic thymus is often associated
with the thyroid glands due to the fact that par-
athyroid glands arise from the same pharyngeal
FIGURE 2.15  Minimal extramedullary hemopoiesis in a pouches as the thymus (Stefanski et al 1990).
rat spleen. ×200. Thymic cysts (Fig. 2.18) or epithelial remnants
are observed as a background change in the rat
Increased hemosiderin (Fig. 2.16) is observed thymus. These may be made up of epithelial
as a background lesion in rats. The rat spleen, tubules or nests in the medullary region of the
particularly in female animals, normally has thymus in older rats. Occasionally these remnants
minimal to slight amounts of hemosiderin. In can give rise to tumors. Some of the cysts are
common with extramedullary hematopoiesis, it remnants of the thymopharyngeal duct, others are
is often difficult to distinguish between normal dilatations of thymic tubular structures (Stefanski
and excessive amounts of hemosiderin. Lipofuscin et al 1990). Thymic cysts are often lined by squa-
pigment may also be present in the red pulp of mous cells or ciliated epithelium.
older rats (Frith et al 2000b). In addition, tattoo
pigment which has drained to the peripheral
lymph nodes may occasionally be observed in rat
lymph nodes (Fig. 2.17).
20 BACKGROUND LESIONS IN LABORATORY ANIMALS

FIGURE 2.18  Thymic cysts lined by squamous epithe-
lium. ×200.
Ectopic parathyroid tissue can occasionally be
encountered adjacent to the thymus (Frith et al
2000b). Multifocal hemorrhage (and occasionally
eosinophilic crystals) is very common in the thymus
of rats, particularly as an agonal lesion due to
carbon dioxide euthanasia (Frith et al 2000b) (Fig.
2.19). Thymic atrophy (Fig. 2.20) (also known as
involution) is a common background finding in
aged rats. Although the thymus continues to
increase in size initially, it begins to regress and at
the end of a carcinogenicity study of two years
duration the thymus will be severely depleted
(Stefanski et al 1990). As the atrophy progresses,
the epithelial components of the thymus become
more prominent and often form tubular structures
(Stefanski et al 1990).
Thymic cortical apoptosis is a common back-
ground finding (Fig. 2.21). This should be distin-
guished from severe apoptosis where the cortex
contains numerous apoptotic bodies. Severe apop-
tosis is often observed when animals are placed
under severe stress or treated with immunosup-
pressive drugs (Stefanski et al 1990).
FIGURE 2.21  Minimal apoptosis of the rat thymic cortex.
×100.
Bone marrow atrophy is observed in aging rats
and the lesion is characterized by a decrease in
hemopoietic cells, an increase in adipocytes and
an increase reticular stroma (Frith et al 2000b).
Hemosiderin pigment is occasionally observed in
the bone marrow and may be associated with pre-
vious hemorrhage (Frith et al 2000b).
Germinal centres and lymphoid follicles can
occasionally be present in the bone marrow of
normal rats (Frith et al 2000b).
Foci of mineralized concretions or corpora amy-
lacea (Fig. 2.23) are observed commonly in nasal
epithelia lining the nasal turbinates (Monticello
et al 1990). Osteopetrosis of the nasal turbinates
may present as a spontaneous lesion (Renne et al
2003). Globule leukocytes are present in the res-
piratory epithelium of the larynx and trachea in
rats (Renne et al 2003, 2009). Mineralized hair,
food particles or debris are often present in the
ventral pouch of the rat larynx, particularly in
older rats and squamous metaplasia may be
recorded in the respiratory epithelium overlaying
the ventral gland as a spontaneous lesion in male
Wistar rats on inhalation studies (Renne et al
2003). Fischer 344 rats, especially the female sex,
exposed to chronic daily irritation by gavage, are
predisposed to high mortality rates on chronic
studies due to the high incidence of cartilage
degeneration, which presumably leads to a dys-
function of the larynx (Germann et al 1998).

Respiratory system FIGURE 2.23  Mineralization (corpora amylacea) of the

FIGURE 2.19  Thymus hemorrhage and eosinophlilic
crystals. ×200.
olfactory epithelium of the rat nasal turbinate. ×200.
Eosinophilic inclusions (also known as globules
and droplets) are observed commonly in aging
rats in the nasal epithelium (Renne et al 2003). A common background lesion in the rat lung is
The inclusions occur in the olfactory, respiratory mineralization of pulmonary arteries (Fig. 2.24).
epithelium and mucous glands. The inclusions are The lesion consists of focal or multifocal minerali-
thought to be proteinaceous and the lesion may zation situated beneath the intima or within the
also be induced by irritant compounds (Renne muscular walls of the arteries. The lesion is not
et al 2003) (Fig. 2.22). Eosinophilic inclusions associated with metastatic calcification.
may occur in association with the loss of sensory
cells (Renne et al 2009). These inclusions are
negative for periodic acid–Schiff (PAS), Alcian
Blue, Von Kossa, mucicarmine, phosphotungstic
acid hematoxylin (PTAH), Masson’s trichrome,
Congo Red, and toluidine blue stains (Monticello
et al 1990). Ultrastructurally, the inclusions
appear to be amorphorous, flocculent material in
membrane-bound vacuoles (Renne et al 2009).

FIGURE 2.24  Mural mineralization of an artery in the rat

lung. ×100.

FIGURE 2.20  Thymic atrophy or involution in the rat FIGURE 2.22  Eosinophilic inclusions in the olfactory epi- Small areas of alveolar hemorrhage (Fig. 2.25)
thymus. ×100. thelium of the rat nasal turbinate. ×200. are often present in the rat lung and represent an
agonal event, particularly in rats killed with carbon
dioxide or those that die spontaneously (Renne
 Wistar and Sprague–Dawley rats 21

et al 2003). Occasionally, small foci of neuroen-

docrine cells in the lung (Fig. 2.26) may be visible
in aging rats. Pulmonary neuroendocrine cells are
found as clusters called neuroepithelial bodies or
as single cells scattered in the respiratory epithe-
lium (Haworth et al 2007). Pulmonary neuroen-
docrine cells are defined as foci of neuroendocrine
cells with greater than 40 nuclei (Haworth et al
2007). The cause of the hyperplasia is not known
and the lesion is not thought to progress to neo-
plasia (Haworth et al 2007). Morphologically the
lesion resembles chemoreceptors such as carotid
bodies (Haworth et al 2007).

FIGURE 2.27  Alveolar histiocyte or macrophage aggre- FIGURE 2.30  Pleural lung tag on surface of rat lung.
gates in the rat lung. ×100. ×100.

FIGURE 2.25  Alveolar hemorrhage with erythrophagocy-

tosis in the rat lung. ×200.

FIGURE 2.28  Cholesterol clefts in the rat lung. ×400. FIGURE 2.31  Mineralization of tracheal cartilage in the
rat. ×100.

FIGURE 2.26  Proliferation of neuroendocrine cells in

bronchiole of rat lung. ×100.

Multifocal aggregates of alveolar macrophages
(Fig. 2.27) (also known as alveolar histiocytosis)
within alveoli and terminal airways are observed
in both young and old rats. The alveolar macro-
phages often contain abundant, foamy cytoplasm
(Boorman & Eustis 1990) and the lesion may be
visible at necropsy as white areas on the lung
surface (Johnson & Gad 2007). The aggregates are
often subpleural or located in the more peripheral
regions of the lung (Boorman & Eustis 1990).
Cholesterol clefts (Fig. 2.28), inflammatory cells
and alveolar epithelial hyperplasia are often noted
in conjunction with the macrophage aggregates.
The aggregates may represent resolved inflamma-
tory foci and may represent a focal deficit in the
pulmonary clearance mechanisms. Some macro-
phages contain brown pigment which may be
hemosiderin (Renne et al 2003) or lipofuscin (Fig.
2.29) or carbon from the atmosphere.
FIGURE 2.29  Brown pigmented alveolar macrophages in
the rat lung. ×200. FIGURE 2.32  Eosinophilic inclusion wthin a Clara cell in
a bronchiole of rat lung. ×100.
Occasionally pleural tags (Fig. 2.30) are
observed in rat lungs. These consist of pleural
extensions which often contain mononuclear cells
such as lymphocytes. Mineralization of tracheal
cartilage (Fig. 2.31) is a common aging change
in rats and is observed in most two-year-old rats
on carcinogenicity studies. Large eosinophilic
cytoplasmic inclusions are occasionally seen in
untreated rat (Kambara et al 2009) Clara cells
(Fig. 2.32) and are generally found in the bronchi-
olar epithelium. The presence of this deeply eosi-
nophilic solitary cell in the lung is often confusing.
Perivascular eosinophilic infiltration (Fig. 2.33) is
observed commonly in the lungs of young and old
rats. The etiology of this finding is unknown.
FIGURE 2.33  Eosinophilic perivascular infiltration in the
rat lung. ×100.
22 BACKGROUND LESIONS IN LABORATORY ANIMALS

Hair shaft emboli or skin fragment emboli are
seen in the lungs of animals on intravenous injec-
tion studies. These foreign bodies generally induce
a granulomatous reaction consisting of multinucle-
ated giant cells and other inflammatory cells
(Renne et al 2003) (Fig. 2.34). Eosinophilic crys-
tals are occasionally observed free within the
alveoli (Renne et al 2003) of rat lungs. The crys-
tals may be associated with areas of alveolar hem-
orrhage (Fig. 2.35) or dilated submucosal glands
in the larynx. Hyperinflation (Renne et al 2003)
or pulmonary acinar ectasia consists of areas of
alveolar dilatation and may be observed in the
subpleural region in aged rats (Renne et al 2003)
(Fig. 2.36a). Isolated cystic spaces in lung paren-
chyma are rarely seen in rats and are of uncertain
origin. These cysts are lined only by fibrous tissue,
with no evidence of inflammation (Renne et al
2009). Apparent dilatation of alveoli can also
occur due to excessive instillation of fixative into
the lung at necropsy (Renne et al 2003).
Dilatation of submucosal glands of the larynx or
trachea may also occur as a background lesion in
rats (Renne et al 2009) (Fig. 2.36b). Foci of
osseous metaplasia are commonly observed in the
lungs of both young and old rats (Renne et al
2009) (Fig. 2.37). In addition, ossification of the
cartilage in the larynx is also occasionally noted in
aging rats (Fig. 2.38). Furthermore, mineralization
of mucus is rarely noted within bronchioles of the
lungs and within the lumena of the nasal tur-
binates (Figs 2.39, 2.40). Foci of minimal, gener-
ally mononuclear cell inflammation may be noted
below the pleura in the rat lung (Fig. 2.41).
FIGURE 2.39  Mineralization of mucus within a bronchiole
in the rat lung. ×100.

FIGURE 2.36b  Dilated submucosal glands and eosi-

naphilic crystals in the larynx of the rat. ×200.

FIGURE 2.40  Mineralization of mucus within the lumen

of the nasal turbinates of the rat. ×100.

FIGURE 2.34  A hair embolus within an artery in the rat

lung, surrounded by inflammatory cells. ×100.

FIGURE 2.37  Osseus metaplasia in the rat lung. ×400.

FIGURE 2.41  Minimal, multifocal, subpleural inflamma-

FIGURE 2.35  Eosinophilic crystals in the rat lung. ×200.
FIGURE 2.38  Ossification of laryngeal cartilage in the rat.
×100.
tion in the rat lung. ×100.
Researchers have noticed inflammatory lesions
in the lungs of rats used in research in recent
years. The lesions consist of perivascular lym-
phocytes, inflammatory cells and alveolar macro-
phages in the lung alveoli and the presence of
epithelial hyperplasia. These lesions may mask or
confuse research findings, particularly in inhala-
tion studies. Initially a virus was suggested as the
cause (rat respiratory virus) (Albers et al 2009),
but recently researchers have demonstrated a cor-
relation between the presence of inflammatory
lung lesions and Pneumocystis carinii DNA
detected by PCR (Livingston et al 2011).

FIGURE 2.36a  Pulmonary acinar ectasia in the rat lung.

×200.

Gastrointestinal system
Dental dysplasia (Fig. 2.42) is a congenital lesion
generally involving the incisor teeth and it may be
related to malocclusion or food impaction (Fig.
2.43) (Monticello et al 1990). Dental dysplasia is
the abnormal development of odontogenic tissues
(Bertram et al 1996). Dental dysplasia is often
observed in conjunction with inflammation and
trauma. The lesion consists of a large mass of
dentine-like material with fragments of tooth and
bone (Bertram et al 1996). Cleft palate is a con-
genital lesion encountered in the midline of the
hard palate of rats (Renne et al 2009). A pulp
stone is a focal area of dentine in the pulp cavity
which is often mineralized. Pulp stones often form
around cellular debris (Bertram et al 1996).
FIGURE 2.42  Dental dysplasia visible in the rat oral
cavity. ×100.
FIGURE 2.43  Food accummulation around the molar in
the rat oral cavity. ×400.
Sebaceous gland ectopia in the oral cavity is
observed at the base of the molar teeth (male rats)
and between the upper incisors (Bertram et al
1996). This finding, referred to as Fordyce’s
granules, which displays morphologically normal
sebaceous glands, is described in several species
including humans and rats. Histologically,
Fordyce’s granules are identical to their cutaneous
counterpart but are not associated with hair
follicles.
In rats, there are, at present, only four reports
of intraoral ‘ectopic’ sebaceous glands in the lit-
erature (Bernick & Bavetta 1962, Frandsen 1962,
Rulli & Martinelli 1971, Yoshitomi et al 1990). In
Holtzman and Long–Evans rats, as well as in
Wistar rats, Fordyce’s granules are found in the
molar gingiva. The most common site in the rat
molar gingiva is probably adjacent to the first
Wistar and Sprague–Dawley rats 23
molar. Ultrastructurally, Fordyce’s granules consist
primarily of typical lipid-filled sebaceous cells. In
addition to the incisor and molar area, the buccal
mucosa as well as the hard palate and tongue
should also be considered as possible sites of
occurrence of Fordyce’s granules in rats.
Squamous cysts (Fig. 2.44), often containing
keratin, are common in the stomach, particularly
in the region of the limiting ridge and antral
mucosa. In addition, squamous cysts may also be
encountered in the gingiva or associated with the
teeth in the oral cavity (Bertram et al 1996) as
well as in the esophagus (Bertram et al 1996).
Glandular cysts (Fig. 2.45) lined by cuboidal
or columnar epithelium may be observed in the
glandular stomach (Bertram et al 1996). Meg- FIGURE 2.46  Cystic dilatation of gastric glands in the rat
aesophagus is a congenital condition observed in glandular stomach. ×200.
young rats (Bertram et al 1996).
FIGURE 2.47  Focal ulceration of the non-glandular
stomach in the rat. ×100.
FIGURE 2.44  Squamous cyst filled with keratin near lim-
iting ridge in the rat stomach. ×20. There is often a minimal eosinophilic infiltra-
tion in the submucosa of the stomach (Fig. 2.48).
The cause of this finding is unknown. Ectopic
hepatocytes and pancreatic exocrine cells may be
encountered in the lamina propria of the stomach
(Bertram et al 1996). Mesenteric adipose tissue
necrosis (Fig. 2.49) is a common lesion in aging
rats and consists of nodular masses of necrotic
mesenteric adipose tissue generally surrounded by
inflammatory cells, particularly in the intersti-
tium. The lesion is often noted at necropsy in the
peritoneal adipose tissue or adjacent to the testes.
The cause of the lesion is thought to be a foreign-
body response against released lipid (Brown &
Hardisty 1990) which results from torsion or
ischemic pressure (Brown & Hardisty 1990). The
ischemic adipose tissue may later undergo miner-
FIGURE 2.45  Cyst lined by ciliated epithelium in the non- alization (Fig. 2.50).
glandular stomach of the rat. ×200.
Iatrogenic gavage injury is not a background
change as such, but is commonly encountered in
rats. Gavage injuries include esophagitis, pleuritis,
periesophageal abscessation and fibrosis (Bertram
et al 1996). Cystic dilatation of gastric glands
(Fig. 2.46) is a common finding in the rat stomach.
The lesion is not accompanied by inflammation
or any other deleterious lesions. Gastric erosion
and ulceration of the glandular or non-glandular
stomach (Fig. 2.47) is commonly observed in aged
rats. It is more common in gavage studies, pre-
sumably due to trauma during dosing. The edges
of the ulcers in the non-glandular stomach often
display epithelial hyperplasia. It is always impor- FIGURE 2.48  Submucosal eosinophilis present in the
tant to distinguish ulceration from autolysis of the submucosa of the rat glandular stomach. ×200.
superficial mucosa.
24 BACKGROUND LESIONS IN LABORATORY ANIMALS
FIGURE 2.49  Mesenteric adipose tissue necrosis in the
rat. ×100.
FIGURE 2.50  Strangulated or ischemic adipose tissue
displays mineralization. ×100.
Lobular atrophy of various salivary glands (Fig.
2.51) with fibroplasia, inflammatory cells and
occasional adipocytes in the surrounding intersti-
tium and tinctorial alteration are common back-
ground changes. Ectopic foci of parotid acini can
occur in the sublingual gland (Bertram et al 1996)
(Fig. 2.52a). In addition, areas of hypertrophied
basophilic cells may be encountered in the parotid
salivary gland of the rat (Fig. 2.52b).
FIGURE 2.51  Acinar atrophy of the salivary gland with
concommitant inflammation. ×200.
FIGURE 2.52a  Serous focus within mucous salivary FIGURE 2.53  Plasmacytic infiltrates into the salivary
gland in the rat. ×200. gland of the rat. ×200.
FIGURE 2.52b  Basophilic hypertrophic focus in the rat FIGURE 2.54  Ectopic small intestinal tissue present in
salivary gland. ×400. the rat stomach. ×200.
Oncocytic cells (also known as eosinophilic
cells, oxyphil cells) contain large amounts of gran-
ular eosinophilic cytoplasm due to mitochondrial
accumulation. These cells occur in the epithelium
of the intercalated ducts and in the secretory
ducts of older rats (Neuenschwander & Elwell
1990). Foci of mineralization are occasionally
observed in the duct lumena and within the acini
of the rat salivary gland (Bertram et al 1996).
Adipose tissue infiltration or lipomatosis is char-
acterized by the presence of adipocytes between
normal acini (Bertram et al 1996) or within areas
of atrophy and inflammation (Neuenschwander &
Elwell 1990).
Plasmacytic infiltration into the parotid salivary
gland is common (Fig. 2.53). Foci of ectopic small FIGURE 2.55  Diverticulum of the duodenum in the rat.
and large intestine (Fig. 2.54), as well as focal epi- ×40.
dermal cysts may be observed in the glandular area
of the rat stomach. In addition, diverticula may be Focal areas of moderate to severe hemorrhage,
encountered in the small intestine (Fig. 2.55). fibrosis, myodegeneration and inflammatory cell
infiltration are noted routinely in the skeletal
muscle of tongues from rats where lingual blood
collection has been performed (Fig. 2.56). Ectopic
hepatocyte islands may be noted in the pancreas,
particularly situated around the islets. These foci
consist of normal hepatocytes with Kupffer cells
as well as bile ducts (Detilleux et al 1995). Pan-
creatic hepatocytes should not be confused with
the large eosinophilic acinar cells which are some-
times encountered around the islets (Detilleux
et al 1995).

FIGURE 2.56  Focal myopathy, fibrosis and inflammatory-
cell infiltration is noted in the tongue when lingual blood
collection is performed. ×200.
Acinar atrophy of the pancreas (Fig. 2.57) is
observed in young and old rats. The lesion consists
of atrophy of the acinar elements with persistence
of the duct, a mild inflammatory response and
fibrosis. The islets are not affected (Kendrey &
Roe 1969). The cause of this lesion is not known
(Detilleux et al 1995). Hemosiderin pigment
deposition and mineralization can also be features
in these areas, particularly in older rats (Detilleux
et al 1995).
FIGURE 2.57  Acinar atrophy of the exocrine pancreas
with associated inflammation. ×200.
Pancreatic vacuoles (Fig. 2.58) are noted in the
pancreas of young and old rats. This finding con-
sists of clear vacuoles in acinar cells and is a
common incidental finding of no toxicological sig-
nificance (Detilleux et al 1995). The lesion may
be an early manifestation of autolysis. Occasional
apoptotic pancreatic acinar cells may be noted in
the rat pancreas and this change is not considered
to be significant (Detilleux et al 1995)
FIGURE 2.58  Vacuolation of the acinar cells in the exo-
crine pancreas. ×200.
Pancreatic acinar hyperplasia (Fig. 2.59) is a
focal or multifocal lesion that is observed as an
occasional background finding in rats. The area
consists primarily of tinctorial alteration. In addi-
tion, foci of cellular alteration (basophilic cyto-
plasmic change, basophilic foci, focal dysplasia)
are a common finding in the pancreas of rats. The
lesion is characterized by small, non-compressive
foci of acinar cells with moderate to severely
basophilic cytoplasm and decreased zymogen
granules (Fig. 2.60a) (Detilleux et al 1995).
Ectopic rests of acinar pancreatic tissue may be
encountered in the duodenum, jejunum, stomach,
liver, spleen or mesentery (Detilleux et al 1995)
(Fig. 2.60b).
FIGURE 2.59  Basophilic tinctorial change and hyperpla-
sia in the exocrine pancreas. ×200.
FIGURE 2.60a  Focus of zymogen degranulation in the rat
pancreas. ×100.
Wistar and Sprague–Dawley rats 25
FIGURE 2.60b  Ectopic pancreas present in the submu-
cosa and tunica muscularis of the small intestine of a rat.
×100.
Foci of perivascular or periductular lymphocytes
and plasma cells may be observed as a normal
background finding in the pancreas of young and
older rats (Detilleux et al 1995). Acinar and duct
dilatation and cyst formation are spontaneous
lesions encountered in the rat pancreas, often in
association with pancreatic acinar atrophy and
inflammation (Detilleux et al 1995). The dilated
acini and ducts are lined by flattened cuboidal
epithelium and many contain necrotic debris
(Kendrey & Roe 1969).
Pancreatic adipocyte infiltration (fatty infiltra-
tion, lipomatosis) may be observed in the older rat
pancreas (Detilleux et al 1995). This lesion is
associated with obesity, atrophy and inflamma-
tion. Lymphoid hyperplasia of the gut-associated
lymphoid tissue (GALT) (Fig. 2.61) in the rat
large intestine is a common background finding in
young and older rats.
FIGURE 2.61  Lymphoid hyperplasia of the rat gut-
associated lymphoid tissue in the large intestine. ×40.
Liver and biliary system
Tension lipidosis in the median cleft area of liver
(Fig. 2.62) is often recognized at necropsy as a
circumscribed, pale yellow area. It usually occurs
at the periphery of the lobe near the attachments
to the adjacent lobes or tissues. The lesion is char-
acterized by the presence of focal, minimal to
slight, generalized hepatocyte vacuolation. This
lesion is thought to be caused by local hypoxia,
i.e. either the adhesions or attachments between
the liver lobes causing constriction and thereby
causing interference with the local perfusion and
consequent lipidosis or vacuolation of the underly-
ing hepatocytes (Stalker & Hayes 2007).
26 BACKGROUND LESIONS IN LABORATORY ANIMALS
FIGURE 2.62  Tension lipidosis in the median cleft area
of the rat liver. ×400.
Bile duct hyperplasia of the liver (Fig. 2.63) is
observed in Sprague–Dawley rats and is a common
aging lesion. The lesion is made up of hyperplasia
of the bile ducts accompanied by a minimal
inflammatory-cell infiltrate as well as fibrosis in
the portal area. Occasionally the bile duct epithe-
lium may be atrophic or degenerate (Eustis et al
1990). The lesion does not appear to progress to
neoplasia (Eustis et al 1990).
FIGURE 2.63  Bile ductule hyperplasia in the liver of the
rat. ×200.
Periportal lipid vacuolation in liver (Fig. 2.64)
is a spontaneous change seen in older rats. particu-
larly Sprague–Dawley rats. Pathologists need to be
careful to distinguish this from a treatment-
related change. Hepatic angiectasis (telangiecta-
sia, peliosis hepatis) (Fig. 2.65) is a common lesion
in the rat liver characterized by focal or multifocal
areas of sinusoidal dilatation. The sinusoids in
these areas are filled with erythrocytes and are
lined by a single layer of endothelial cells. Occa-
sional, subcapsular areas of sinusoidal dilatation
result in depression of the liver surface. The lesion
is occasionally noted as a dark red area at necropsy.
The lesion is more common in male rats (Eustis
et al 1990).
FIGURE 2.64  Periportal lipid vacuolation in the rat liver.
×100.
FIGURE 2.65  Hepatic angiectasis with dilatation of the
sinusoids in the rat liver. ×200.
Cystic degeneration of the liver (spongiosis
hepatis) is a multilocular, cystic lesion wherein the
cyst-like structures contain an eosinophilic mate-
rial (Fig. 2.66). The cyst-like structures are not
lined with endothelial cells (in contrast to hepatic
angiectasis). This background lesion is observed at
a low incidence in aging rats (Eustis et al 1990).
Areas of cystic degeneration may be found inde-
pendently or within areas of altered hepatocyte
foci.
FIGURE 2.66  Cystic degeneration of the rat liver. ×100.
Intracytoplasmic, eosinophilic, inclusion bodies
(Fig. 2.67) may develop spontaneously in rat hepa-
tocytes (Detilleux et al 1995). Hepatocyte vacu-
olation is a common background change in rats
which are not exsanguinated at necropsy and
where there is a delay between death and removal
of the liver. Histologically, the affected hepato-
cytes contain a single cytoplasmic vacuole filled
with eosinophilic material, although occasionally
the vacuoles are clear. The hypothesis is that the
vacuoles contain blood plasma. This change is asso-
ciated with an increase in liver weight (Detilleux
et al 1995). Intranuclear inclusions resulting from
the invagination of cytoplasm into the nucleus are
occasionally observed in rat hepatocytes and are
similar to those seen in the mouse liver. Ectopic
hepatocytes are rarely encountered in the submu-
cosa of the stomach and gallbladder (Fig. 2.68).
Islands of pancreatic tissue localized within the
hepatic parenchyma are a rare spontaneous finding
in rats (Thoolen et al 2010). In addition, intravas-
cular hepatocytes which protrude into hepatic
veins or occur within the contour of the vessel are
reported (Thoolen et al 2010).
FIGURE 2.67  Eosinophilic, intracytoplasmic inclusions in
the rat liver. ×200.
FIGURE 2.68  Hepatocytes in the submucosa of the non-
glandular rat stomach. ×200.
Hemopoiesis in the liver is normal in the fetal
rat liver, but occurs only under pathological con­
ditions in the adult rat (Detilleux et al 1995).
Hematopoiesis of the erythroid and myeloid pre-
cursors is generally always accompanied by similar
lesions in the spleen and other sites (Fig. 2.69).
Parenchymal and portal lymphoid or inflammatory
cell aggregates in the liver are observed commonly
in the majority of rat livers. The cause of this lesion
is likely to be due to bacterial showering from the
intestine. The lesion may be acute, subacute or
granulomatous. Individual cell necrosis of occa-
sional hepatocytes, accompanied by inflammatory-
cell infiltration, is often observed in rat livers
(Detilleux et al 1995) and focal areas of necrosis,
thought to be related to infective processes in
the gastrointestinal tract, are also common in rat
livers (Detilleux et al 1995) (Fig. 2.70). Cholan-
giofibrosis (Fig. 2.71) is a variant of bile-duct

hyperplasia in which the bile duct epithelium
assumes the characteristics of intestinal epithe-
lium and produces large cystic areas filled with
mucus (Gopinath 1987). This is a controversial
lesion in the rat liver and is generally induced by
xenobiotics (Thoolen et al 2010). Biliary cysts are
single or multilocular cysts lined by flattened
cuboidal epithelium and are common in older rats
(Detilleux et al 1995) (Fig. 2.72).
FIGURE 2.69  Minimal foci of hemopoiesis in the rat liver.
×100.
FIGURE 2.70  Hepatocyte necrosis and fibrosis in the rat
liver. ×200.
FIGURE 2.71  Cholangiofibrosis in the rat liver. ×100.
Wistar and Sprague–Dawley rats 27
FIGURE 2.74  Multinucleate hepatocytes in the rat liver.
FIGURE 2.72  Focal biliary cyst in the rat liver. ×100. ×200.
Hepatodiaphragmatic nodules (Fig. 2.73) caus­
ing nodular protrusions on the anterior surface of
the liver are associated with small diaphragmatic
hernia. These are common in rats of all ages,
including fetuses (Detilleux et al 1995) and the
hepatocyte tissue is normal both macroscopically
and microscopically, in the areas that have pro-
truded through the hernia. These nodules can be
confused with hepatic adenomas.
FIGURE 2.75  Rat liver lobe torsion with coagulative
necrosis. ×100.
The spontaneous accumulation of lipofuscin
pigment (Fig. 2.76) is noted in the hepatocytes
of older rats (Detilleux et al 1995), and bile
pigments may be present in the bile ducts of
aged rats (Detilleux et al 1995). In addition, por-
phyrin is a dense, dark brown pigment which may
rarely be noted within bile ductules and bile canal-
FIGURE 2.73  Herniated liver lobe with fibrosis and iculi (Thoolen et al 2010). Prominent ploidy is
inflammation in the pedicle. ×100. occasionally observed in the rat liver (Fig. 2.77).
Foci of cellular alteration are common in rat
Mitotic figures and multinucleated hepatocytes studies greater than twelve months (Thoolen et al
are observed as a background finding in small 2010). Hypertrophy and hyperplasia of Kupffer
numbers of rat livers. Multinucleate hepatocytes cells is a rare spontaneous finding in the rat liver
(Fig. 2.74) occur spontaneously in older rats and is characterized by the proliferation of the
(Burek 1978). If these lesions are noted in spindled cells lining the sinusoids (Thoolen et al
increased numbers, then the phenomenon may be 2010).
related to treatment. Apoptosis of hepatocytes is
observed at very low levels in rat livers (Detilleux
et al 1995). Torsion of the liver (Fig. 2.75) lobes
is observed occasionally as a background, sponta-
neous lesion. Although torsion may occur in any
lobe, the papillary process of the caudate lobe is
generally the most commonly affected. The lesion
is characterized by focal areas of coagulative
necrosis with a variable inflammatory cell infil-
trate. Later, the lesion may progress to fibrosis as
well as areas of nodular regeneration. Mineraliza-
tion is rarely observed in the liver of rats, but
dystrophic mineralization may be noted in con-
junction with hepatic necrosis (Thoolen et al
2010).
FIGURE 2.76  Hepatocyte lipofuscin pigment present in
rat liver. ×400.
28 BACKGROUND LESIONS IN LABORATORY ANIMALS

hydronephrosis or pelvic dilatation is seen com-
monly in rat kidneys. It is often unilateral and the
right kidney is more commonly affected than the
left (Hard et al 1999) (Fig. 2.80).
commonly in aging rats is likely to be lipofuscin
(Schmorl’s-positive) (Fig. 2.82). Cystic tubules
can manifest in different forms and are generally
found in the cortex. They can be single or multiple
and are generally lined with a single layer of flat-
tened epithelium (Hard et al 1999). Cysts may
be empty or may contain proteinaceous material
(Fig. 2.83).

FIGURE 2.77  Prominent ploidy in the rat liver. ×400.

Urinary system
Chronic progressive nephropathy is a common
spontaneous disease of aging rats including Fischer
344 rats (Dixon et al 1995). Chronic progressive
nephropathy is more common in male rats and
is directly influenced by a high protein diet.
The lesion is characterized by basophilic cortical
tubules, hyaline casts, glomerulosclerosis and
atrophy, interstitial inflammatory cell infiltration
and interstitial fibrosis (Montgomery & Seely
1990) (Figs 2.78, 2.79).
FIGURE 2.78  Chronic progressive nephropathy with
dilated tubules, hyaline casts, basophilic tubules and
interstitial mononuclear cell in the rat kidney. ×200.
FIGURE 2.80  Hydronephrosis or pelvic dilatation in the
rat kidney. ×100.
The single layer of the parietal epithelium lining
FIGURE 2.82  Lipofuscin pigment in the cortical tubular
the Bowman’s capsule can transform from normal
epithelium in the rat kidney. ×400.
squamous epithelium to cuboidal cells similar to
those of the cortical tubules (Hard et al 1999).
This change is observed in older male rats and is
referred to as Bowman’s capsule metaplasia. In
rats, Bowman’s capsule metaplasia is associated
with age (Haley & Bulger 1983), hypertension and
unilateral nephrectomy (Andrews 1981).
Osseous metaplasia, unrelated to renal miner-
alization, is occasionally seen in the interstitium of
the renal cortex (Hard et al 1999). Hyaline drop-
lets (Fig. 2.81) are eosinophilic, intracytoplasmic
inclusions that generally occur in the cortical
tubules. They are thought to represent liposomes
containing protein (Hard et al 1999). Hyaline
droplets occur normally in the mature, male rat
where they represent reabsoprtion of alpha2u-
globulin. Hyaline droplet nephropathy (i.e. accu-
mulation of large numbers of hyaline droplets) can FIGURE 2.83  Polycystic rat kidney with multiple cysts in
be induced by the presence of histiocytic sarcoma the cortex. ×100.
as well as xenobiotics. Mallory Heidenhain stain
is used to visualize hyaline droplets (Hard et al Mineralization (Fig. 2.84) is a spontaneous
1999). background finding present in young and old rats
(Hard et al 1999). The lesion may occur in the
cortex, corticomedullary junction, medulla and
papilla. Female rats tend to be more affected than
male rats and the lesion is thought to be influ-
enced by diet (Hard et al 1999). Renal calculi
may occur in older rats (Hard et al 1999). Miner-
alization of the basement membrane of the
glomerulus may occur during chronic progressive
nephropathy in the rat (Fig. 2.85). Basophilic cor-
tical tubules may be a spontaneous background
finding, although an increase in basophilic tubules
is likely to be related to regeneration after an
insult to the cortical tubules.

FIGURE 2.81  Hyaline droplets in the rat kidney associ-

FIGURE 2.79  Glomerulosclerosis and hyaline casts in
chronic progressive nephrotophay in the rat kidney. ×200.
Occasionally a focal area of adrenal cortical
cells may be observed outside or immediately
below the kidney capsule (Hard et al 1999). This
is referred to as an adrenal rest. Congenital
ated with histiocytic sarcoma. ×400.
Pigmentation characterized by the intracyto-
plasmic presence of yellow to brown granular
material is observed in proximal convoluted
tubules (Hard et al 1999). Although the pigment
may be iron (Perl’s-positive) or bile (Fouchet’s-
positive), the brown pigmentation observed
 Wistar and Sprague–Dawley rats 29

FIGURE 2.84  Corticomedullary mineralization in the rat FIGURE 2.86  Colloidal plug in the urinary bladder of the FIGURE 2.89  Lipomatous transformation of the rat renal
kidney. ×200. rat. ×200. cortex. ×200.

FIGURE 2.85  Mineralization of the basement membranes FIGURE 2.90  Renal tubular hypertrophy in the rat kidney
of the glomeruli and tubules in the rat kidney due to FIGURE 2.87  Hyperplasia of the rat transitional epithe- cortex. ×400.
chronic progressive nephropathy. ×400. lium in the rat kidney pelvis. ×200.

Artificial separation between the urothelium

and submucosa in the urinary bladder occurs after
inflation with formalin. The fixative may be inad-
vertently injected into the wall of the bladder
during inflation at necropsy (Hard et al 1999).
This lesion may be mistaken for submucosal
edema (Hard et al 1999). In addition, vacuolation
of the transitional epithelium of the urinary
bladder is a background change usually attributed
to autolysis (Hard et al 1999). Calculi may occur
spontaneously in the rat urinary bladder (Hard
et al 1999).
Proteinaceous or colloidal plugs in the bladder
or urethra are observed commonly in male rats
(Fig. 2.86). They are made up of deeply eosi- FIGURE 2.91  Hyperplasia of rat pelvic urothelium with
nophilic material, occasionally containing sperma- FIGURE 2.88  Lymphocytic infiltration into urinary bladder vascular ectasia. ×200.
tozoa or cellular debris. This lesion is thought to in the rat. ×200.
be caused by ejaculation during euthanasia and is
considered to be an insignificant background Lipomatous transformation of the rat renal
lesion (Hard et al 1999). Perfusion of the rat cortex is observed in older rats and may be a
kidney with glutaraldehyde can produce vacuola- precursor to lipoma and liposarcoma (Fig. 2.89).
tion of the proximal tubular cells which may not Occasional enlarged, hypertrophic renal tubules
be distinguished from genuine pathological vacu- are observed within the rat renal cortex (Fig.
olation (Hard et al 1999). Hyperplasia of the tran- 2.90). Hyperplasia of the rat transitional epithe-
sitional epithelium of the rat urinary bladder and lium lining the kidney pelvis, often accompanied
renal pelvis is observed commonly (Fig. 2.87). by vascular ectasia, is observed in the older rat
Lymphocyte infiltration into the submucosa of kidney (Fig. 2.91). Mineralization of the transi-
the rat urinary bladder is noted in older rats tional epithelium of the rat kidney pelvis may also
(Fig. 2.88). be observed in older rats (Fig. 2.92). A unique
form of autolysis is occasionally observed in the
kidney of the Wistar rat and may cause confusion
in carcinogenicity studies if it is mistaken for a
treatment-related finding (Fig. 2.93). FIGURE 2.92  Mineralization of the transitional epithelium
of the pelvis of the rat kidney. ×200.
30 BACKGROUND LESIONS IN LABORATORY ANIMALS

single type of cell may occur in the rat pituitary

(Fig. 2.97). The focus is slightly delineated from
the normal cell population with little or no evi-
dence of compression. The vascular channels
within the region are normal in comparison to the
pattern seen in adenomas, in which they are often
ectatic (Fig. 2.98). Portions of the margins of the
focus may blend with the surrounding paren-
chyma. The uniform population of cells forming
these foci vary little in size and character from
normal but tend to be pale pink in colour (Fig.
2.99). Individual eosinophilic or basophilic cells
that appear normal may be present in scattered
regions of the focus (Frith et al 2000a).

FIGURE 2.93  Autolysis artifact observed in Han Wistar rat FIGURE 2.99  Pituitary gland adenohypophysis prolifera-
kidney. ×100. tion with clear cells. ×400.

Cortical vacuolation (Fig. 2.100) of the adrenal

Endocrine glands is very common and may be focal or multifocal
Pituitary cysts (Fig. 2.94) are very common and (Frith et al 2000a). The cells of the zona fascicu-
are occasionally lined by cuboidal, ciliated epithe- laris have clear cytoplasmic vacuoles. Cystic
lium. Pituitary cysts containing eosinophilic mate- degeneration (Fig. 2.101) of the zona fascicularis
rial are likely to represent cystic remnants of is also commonly encountered in aging, female
Rathke’s pouch. Here, the pars intermedia and the rats (Frith et al 2000a). The lesion consists of
pars nervosa of the pituitary may contain focal, large, clear or blood-filled spaces. Often the sur-
unremarkable, glandular structures which do not rounding cortical cells display vacuolation or the
display neoplastic transformation (Fig. 2.95). The cystic lesion may be part of a hyperplastic (Fig.
epithelium lining the glandular structures as well 2.102) or neoplastic area. The lesion is most
as the cyst is largely cuboidal and the cyst lining common in female rats and may be noted as a red
FIGURE 2.96  Angiectasis in the rat pituitary. ×40. mass at necropsy. Foci of cortical hyperplasia,
is ciliated (Frith et al 2000a).
often vacuolated, may be observed in the rat
adrenal (Fig. 2.106) (Frith et al 2000a).

FIGURE 2.94  Pituitary cyst in the pars intermedia in the FIGURE 2.97  Focal hyperplasia of the pars distalis of the
rat. ×200. rat pituitary. ×100.
FIGURE 2.100  Cortical vacuolation in the rat adrenal
gland. ×200.

FIGURE 2.95  Rathke’s pouch remnants in the rat pitui- FIGURE 2.98  Hyperplasia of pars distalis in rat pituitary
tary gland. ×100. with an unremarkable vascular pattern. ×200.
FIGURE 2.101  Cortical cystic degeneration in the rat
Hemangiectasis, i.e. the dilatation of blood- adrenal gland. ×200.
filled spaces in the adenohypophysis (Fig. 2.96) of
the pituitary, is common in older rats (Frith et al
2000a). One or more focal proliferations of a
 Wistar and Sprague–Dawley rats 31

FIGURE 2.102  Cortical hyperplasia and vacuolation in the FIGURE 2.104  Extramedullary hemopoiesis in the adrenal FIGURE 2.107  Cortical ossification in the rat adrenal
rat adrenal gland. ×400. cortex of the rat. ×200. gland. ×200.

Angiectasis (hemangiectasis, capillary dilata-

tion, peliosis) (Fig. 2.103) is also very common in
the adrenal zona fascicularis as well as occasionally
in the medulla. The lesion consists of dilatation of
the vascular spaces lined by endothelium and may
also be linked to the areas of cystic degeneration
and cortical vacuolation. In addition, foci of corti-
cal hyperplasia (foci of cellular alteration) may
also be present within these degenerative lesions.
The cells are variably sized and may be vacuolated.
Single or multiple well-demarcated lesions are
most commonly found in the zona fasciculata and
zona reticularis. The foci maintain architectural
relationships and produce minor or no compres-
sion of the adjacent parenchyma. The cells within
the foci may resemble those of surrounding FIGURE 2.105  Extracapsular cortical tissue in the rat FIGURE 2.108  Ceroid or lipofuscin pigment present in the
cortex, be tinctorially distinct and smaller in size, adrenal gland. ×100. cortex of the rat adrenal gland. ×200.
or display vacuolated cytoplasm. Highly vacu-
olated cells may be more numerous in the inner Aggregates of medullary cells with minimal
region of the foci of hyperplasia. Mitotic activity altered cellular arrangement and cytological fea-
is typically low and cellular atypia is not present tures may also be encountered in the rat adrenal
(Frith et al 2000a). gland (Fig. 2.109). The affected cells may be
slightly enlarged, with a round, vesicular nucleus,
or smaller than normal cells with a hyperchro-
matic nucleus. The cytoplasm may show increased
basophilia (Frith et al 2000a). Occasionally foci of
hypertrophy may be noted in the zona glomeru-
losa of the rat adrenal gland (Fig. 2.110).

FIGURE 2.106  Cortical hyperplasia in the rat adrenal

gland. ×200.

Adrenal mineralization may occur in the adrenal

cortex following inflammation or hemorrhage
FIGURE 2.103  Angiectasis in the rat adrenal cortex.
(Frith et al 2000a). In addition, adrenal ossifica-
×200.
tion is occasionally observed in the cortex of older
rats (Fig. 2.107). Lipofuscin pigmentation (Fig.
Foci of extramedullary hemopoiesis are occa- 2.108) or ceroid pigmentation is characterized by
sionally observed in the rat adrenal (Fig. 2.104), the presence of yellow brown pigment in the zona
but these are generally linked to animals respond- reticularis and this background change is observed
ing to a regenerative anaemia. In these cases, in older rats (Frith et al 2000a). FIGURE 2.109  Medullary hyperplasia of the rat adrenal
extramedullary hemopoiesis is likely to be encoun- gland. ×200.
tered at multiple sites such as the liver and the
spleen. Ectopic adrenal tissue (Fig. 2.105) is a
common finding in the rat adrenal. The tissue is
generally present immediately outside the adrenal
capsule. Foci of cortical tissue may occasionally be
found in the medulla, and medullary tissue may
extend into the cortex.
32 BACKGROUND LESIONS IN LABORATORY ANIMALS
FIGURE 2.110  Focal hypertrophy of zona glomerulosa of
the rat adrenal gland. ×200.
A common background lesion in the rat thyroid
is the presence of cysts. These are generally lined
by squamous epithelium and filled with keratin
and cellular debris. Follicular cysts and dilated
follicles, filled with colloid, may also be observed.
Ultimobranchial cysts are the remnants of embry-
onic ultimobranchial bodies that originate from
the third pharyngeal pouch (Frith et al 2000a).
These cysts are lined by flattened squamous epi-
thelium and often contain keratin (Frith et al
2000a). Persistent thyroglossal ducts originate
from a ventral-down growth of endoderm and are
lined with cuboidal or ciliated columnar epithe-
lium (Frith et al 2000a).
Basophilic concretions in the colloid are also
observed commonly in aging rats. This change
appears to represent mineralization and occurs in
aging rats. C-cell hyperplasia (Fig. 2.111) is occa-
sionally observed in the rat thyroid, but is not as
common as the C-cell hyperplasia noted in the
dog thyroid. Areas of ectopic thymus are common
adjacent to the thyroid in the rat (Johnson & Gad
2007).
FIGURE 2.111  Focal C-cell hyperplasia in thyroid of the
rat. ×200.
Brown pigment may be observed within the
follicular epithelial cells in aging rats (Frith et al
2000a). In addition, foci of lymphocytes and
plasma cells may be observed in the interstitium
of the thyroid of older rats (Frith et al 2000a).
Intracytoplasmic colloid (Fig. 2.112) is occasion-
ally observed in the rat thyroid. This lesion may be
related to intracytoplasmic microfollicular vacu-
oles described by Neve & Rondeaux (1991). Juve-
nile thyroid follicular epithelium may appear
hypertrophied, but this is a normal change in young
rats (Rao-Rupanagudi et al 1992) (Fig. 2.113).
FIGURE 2.112  Intracytoplasmic colloid in the rat thyroid.
×200.
FIGURE 2.113  Follicular epithelial hypertrophy in the
juvenile rat thyroid. ×200.
Thickened trabeculae, made up of fibrous con-
nective tissue, are observed commonly in the par-
athyroid of aging rats. Cysts may also be present
in the parathyroid. The cysts are thought to derive
from the third and fourth pharyngeal pouches
(Frith et al 2000a). Multinucleate giant cells are
occasionally observed at the periphery of the par-
athyroid. The significance of these cells is not
known (Frith et al 2000a). Intracytoplasmic inclu-
sions are rarely observed in the parathyroid of the
rat, but are though to be a background change.
Focal areas of hypertrophy may be observed in the
rat parathyroid gland (Fig. 2.114).
FIGURE 2.114  Focal hypertrophy in the rat parathyroid
gland. ×400.
Aging Wistar rats develop fibrosis, inflammation
and pigmentation of the islets of Langerhans in the
pancreas. This may produce a lobular appearance
in the islets of older rats (Frith et al 2000a).
Hemangiectasis, i.e. blood-filled spaces, may
occur in the islets of aging rats (Frith et al 2000a).
Pancreatic hepatocytes are occasionally observed
in the periphery around an islet in the pancreas
and are thought to derive from pancreatic acinar
cells (Frith et al 2000a). Peri-insular halos (juxta-
insular cells) consist of large, acinar cells adjacent
to the islets with enlarged nuclei and granular,
hypereosinophilic cytoplasm due to an increase in
zymogen granules (Kramer & Tan 1968). The
cause is thought to be due to the greater concen-
trations of insulin in the cells immediately adja-
cent to the islets (Kramer & Tan 1968) (Fig.
2.115). Variation in the size of islets is commonly
encountered, but it is not as extreme as it is in
mice (Fig. 2.116a).
FIGURE 2.115  Increased zymogen granules around the
islets in rat pancreas. ×100.
FIGURE 2.116a  Islet cell hyperplasia in the rat pancreas.
×200.
Skin and appendages
Staphylococcal infections causing ulcerative tarsal
dermatitis (also known as tarsal granulomas) are
common in male rats; however, it is likely that
initial trauma caused by wire cages, persistent irri-
tation and fighting are important contributing
factors. The presence of the lesion on the under-
side of the tarsus suggests that wire cages may
cause the initial trauma. The disease is common in
heavy, aged, male rats and may be related to an
underlying osteoarthritis (Percy & Barthold 2007)
(Fig. 2.116b). Epidermal, keratin or follicular
cysts, especially on the tail, are common in the rat
skin. The lesion consists of a cyst filled with keratin
and lined by squamous epithelium (Fig. 2.117).

FIGURE 2.116b  Tarsal ulceration and dermatitis in the
rat skin. ×200.
FIGURE 2.117  Epidermal or follicular cyst filled with
keratin in the rat skin. ×40.
Muscle, bones and joints
Atrophy of skeletal muscles occurs spontaneously
in aging rats (Fig. 2.118) (Greaves et al 1996) and
the fibres assume a compressed angular profile
(Greaves et al 1996), particularly in the hind
limbs. In addition, the atrophic muscle fibres are
accompanied by variation in muscle fibre size,
fibrous tissue and the inclusion of lipofuscin and
lipid droplets within the fibres (Greaves et al
1996). Spontaneous synovitis may be detected in
the stifle joints of control Wistar rats in four-week
and two-year toxicity studies (Sasaki et al 1998).
The severity is not increased with aging. The
pathogenesis is thought to be joint instability
which induces mechanical damage of the syn-
ovium (Sasaki et al 1998).
FIGURE 2.118  Atrophy of myofibres in rat skeletal
muscle. ×400.
Wistar and Sprague–Dawley rats 33
Spontaneous age-related atrophy of bone is
reported in rats (Leininger & Riley 1990). Bone
cysts (Fig. 2.119) are rare in rats (Long et al
1996). Chondromucinous degeneration (Fig.
2.120) is observed frequently in the sternum and
growth plate and articular cartilage of long bones
(Leininger & Riley 1990). The lesion increases as
the rat becomes older. The etiology of the condi-
tion is not known and the lesion does not appear
to progress.
FIGURE 2.121  Joint mouse in the knee joint of the rat.
×200.
FIGURE 2.119  Subarticular bone cyst in the rat femur.
×200.
FIGURE 2.122  Keratin cyst with associated inflammation
within the skeletal muscle of the rat. ×200.
Brain and nervous system
Axonal degeneration is commonly observed in the
cervical, thoracic and lumbar spinal cords of older
rats (McMartin et al 1997). Sciatic nerve demy-
elination is common in aging rats. Myelopathy in
the thoracic and lumbar spinal cord, characterized
FIGURE 2.120  Chondromucinous degeneration of the
by distended axon sheaths and myelinophages,
bone in the rat sternum. ×100.
may be linked to the demyelination in the sciatic
nerve (Fig. 2.123). Lipofuscin pigmentation of
Degenerative joint disease is observed in the neurones is common in the aging rat brain (Knox
femur-tibial joint (knee) of aging rats. The lesion et al 1980). Lipofuscin can be identified with the
is characterized by irregularity and thickening of use of Schmorl’s or carbol fuchsin stains. Loss of
the articular cartilage and the formation of ventral horn motor cells has been reported in aging
chondrocyte clusters (Long et al 1996). Later, rats (McMartin et al 1997).
cartilage necrosis, cyst formation, synovial hyper-
plasia and thickening of the joint capsule are
observed (Long et al 1996). Joint mice may occa-
sionally be observed within the affected joint (Fig.
2.121). Osteochondrosis occurs spontaneously in
Sprague–Dawley rats. Occasional keratin or epi-
dermal cysts may be encountered in the skeletal
muscle of the rat (Fig. 2.122).
FIGURE 2.123  Sciatic nerve demyelination in the rat.
×200.
34 BACKGROUND LESIONS IN LABORATORY ANIMALS
Epidermoid cysts (Fig. 2.124) originate from
embryological remnants of ectoderm and are
made up of keratin-filled cysts lined by squamous
epithelium (McMartin et al 1997). Epidermoid
cysts are found occasionally on the surface of the
spinal cord or the brain. Brain mineralization (Fig.
2.125) is occasionally encountered in the brains of
older animals, especially in the thalamus (Burek
1978). Spontaneous globoid mineralization has
been noted in the cerebellum of rats (Yanai et al,
1993). Age-related mineralization is generally
not observed in conjunction with glial reactions
(McMartin et al 1997).
FIGURE 2.124  Epidermal or keratin cyst in the spinal
cord of the rat. ×200.
FIGURE 2.125  Brain mineralization in areas of malacia in
the rat brain. ×200.
Gliosis is a response to nervous system injury,
but may be observed as a background lesion in rats
(McMartin et al 1997). The term refers to the
increase in the number of astrocytes and/or
microglia and their processes (McMartin et al
1997). Hemorrhage into the Virchow–Robin
space around a blood vessel may be observed in
the rat. Artifactual hemorrhage into the brain,
spinal cord and cerebral spinal spaces is common
when the nervous system is not handled carefully
(McMartin et al 1997). Hydrocephalus, i.e. the
expansion of the ventricles with cerebrospinal
fluid, may be encountered in younger animals as
an incidental finding or it may occur due to sec-
ondary pressure (e.g. tumors).
Vacuolation is occasionally observed in the large
neurones of aged rats as a background change
(McMartin et al 1997). Artifactual vacuolation in
the brain is common and should be distinguished
from vacuolation associated with pathology. The
lesion may occur unilaterally or bilaterally. Gener-
ally, vacuolation associated with genuine pathology
is accompanied by inflammatory cells and reduced
staining of the myelin (McMartin et al 1997). Cer-
ebellar hypoplasia is a congenital lesion which is
rarely observed in the rat brain (Fig. 2.126). Lym-
phocyte aggregates may be noted in the pineal
gland of young rats. The nucleus circularis is a focus
of neurones in the neonatal rat brain which may be
confused with a treatment-related finding (Fig.
2.127). The nucleus circularis, in the anterior
hypothalamus, is a group of magnocellular ele-
ments arranged in a ring around a capillary bed. The
entire nucleus is surrounded or encapsulated by
myelinated fibers (Hatton 1976). The neonatal rat
brain also contains focal areas of neuro­blasts, which
may be confused with gliosis (Fig. 2.128). In addi-
tion, the neonatal rat brain may also display foci of FIGURE 2.129  Extramedullary hemopoiesis in the choroid
extramedullary hemopoiesis in the choroid plexus plexus of the neonatal rat. ×200.
(Fig. 2.129), which is a normal feature at day 12.
Furthermore, the neonatal brain also displays the
presence of cysts (Fig. 2.130) and large, bizarre
mitotic figures (Fig. 2.131). Ectopic granule cells
are also visible in the neonatal rat brain (Fig. 2.132).
FIGURE 2.130  Cyst present in neonatal rat brain. ×100.
FIGURE 2.126  Cerebellar hypoplasia in the rat cerebel-
lum. ×100.
FIGURE 2.131  Bizarre mitotic figures present in the neo-
natal rat brain at day 12. ×100.
FIGURE 2.127  Nucleus circularis in the cerebrum of a
neonatal rat. ×200.
FIGURE 2.132  Ectopic granule cells in the neonatal rat
brain. ×100.
FIGURE 2.128  Foci of neuroblasts in the neonatal rat
brain. ×100.

Eye and ear
Porphorin accumulations in the Harderian gland
are considered to be a normal background in
nearly every rat. Inflammation of the Harderian
gland is common, and squamous hyperplasia is
also observed occasionally within the rat Harde-
rian gland (Fig. 2.133). Cellular pleomorphism is
a normal background change in the lacrimal gland.
In addition, one can often see areas of Harderiani-
zation in the lacrimal gland (Fig. 2.134) and this
is an example of sexual dimorphism of the
extraorbital lacrimal gland in the rat (Ferrara et al
2004). Acinar atrophy and inflammation may be
noted in the lacrimal gland (Fig. 2.135). Acinar
hypertrophy may also be present in the lacrimal
gland (Fig. 2.136).
FIGURE 2.133  Squamous hyperplasia of the Harderian
gland in the rat. ×400.
FIGURE 2.134  Harderianization of the lacrimal gland of
the rat. ×200.
FIGURE 2.135  Acinar atrophy and inflammation in lac-
rimal gland of the rat. ×200.
Wistar and Sprague–Dawley rats 35
FIGURE 2.136  Acinar hypertrophy in the lacrimal gland FIGURE 2.138  Optic nerve atrophy and fibre degenera-
in the rat. ×200. tion in the rat. ×100.
In the rat eye, retinal rosettes are often noted
as a background change (Fig. 2.137). Optic nerve
atrophy and degeneration with areas of vacuola-
tion is observed as an aging change in the rat
(Fig. 2.138). Corneal opacity is a distinct entity in
aging rats and occurs predominantly in male rats
(Taradach & Greaves 1984). Increased fibrosis of
the ciliary body and iris is reported in aging rats
(Taradach & Greaves 1984). Synechia between
the iris and cornea, and iris and lens may be a
congenital or inflammatory lesion in young rats
and coloboma is a congenital condition occasion-
ally reported in rats (Taradach & Greaves 1984).
Lenticular degeneration (Fig. 2.139) and cataracts
are common in older rats, as is retinal atrophy
(Fig. 2.140) which may be a background lesion FIGURE 2.139  Lenticular degeneration of the lens of the
rat eye. ×200.
or treatment-related or caused by excessive light
and other causes. Interestingly, autolysis may also
cause a lesion in the lens of the rat which appears
to be similar to lenticular degeneration (Fig.
2.141).
FIGURE 2.140  Rat eye retinal atrophy loss of outer
nuclear layer. ×200.
FIGURE 2.137  Retinal rosettes in the eye of the rat.
×100.
Lens
Artefact
Cornea
Iris
Lens capsule
FIGURE 2.141  Lenticular autolysis which is similar to
lenticular degeneration in the rat eye. ×100.
36 BACKGROUND LESIONS IN LABORATORY ANIMALS
Auricular chondritis (Fig. 2.142) is a lesion
noted in the ears of older rats which consists of
cartilage degeneration, fibrosis and inflammatory
cell infiltration.
FIGURE 2.142  Auricular chondritis in the rat ear with
cartilage degeneration and mineralization. ×100.
References
Albers, T.M., Simon, M.A., Clifford, C.B., 2009.
Histopathology of naturally transmitted “rat
respiratory virus”: progression of lesions and proposed
diagnostic criteria. Vet. Pathol. 46, 992–999.
Andrews, P.M., 1981. The presence of proximal tubule
like cells in the kidney parietal epithelium in response
to unilateral nephrectomy. Anat. Rec. 200, 61–65.
Bernick, S., Bavetta, L.A., 1962. The development of
gingival sebaceous-like glands and cysts in rats of the
Holtzman strain. Oral Surg. Oral Med. Oral Pathol.
15, 351–354.
Bertram, T.A., Markovits, J.E., Juliana, M.M., 1996.
Non-proliferative lesions of the alimentary canal in
rats. In: GI-1 guides for toxicologic pathology. STP/
ARP/AFIP, Washington DC.
Boorman, G.A., Eustis, S.L., 1990. Lung. In: Boorman,
G.A., Eustis, S.L., Elwell, M.R., et al. (Eds.),
Pathology of the Fischer rat. Academic Press, San
Diego, pp. 339–367.
Brown, H.R., Hardisty, J.F., 1990. Oral cavity,
esophagus, and stomach. In: Boorman, G.A., Eustis,
S.L., Elwell, M.R., et al. (Eds.), Pathology of the
Fischer rat. Academic Press, San Diego, pp. 9–29.
Burek, J.D., 1978. Pathology of aging rats. CRC Press,
West Palm Beach, FL.
Chandra, M., Frith, C.F., 1992. Spontaneous
neoplasms in aged control Fischer 344 rats. Cancer
Lett. 62, 49–56.
Detilleux, P.G., Gruebbel, M.M., Botts, S., et al.,
1995. Non-proliferative changes of the liver,
exocrine pancreas and salivary glands of the rat. In:
GI-2 guides for toxicologic pathology. STP/ARP/
AFIP, Washington DC.
Dixon, D., Heider, K., Elwell, M.R., 1995. Incidence
of nonneoplastic lesions in historical control male
and female Fischer-344 rats from 90-day toxicity
studies. Toxicol. Pathol. 23, 338–348.
Eustis, S.L., Boorman, G.A., Harada, T., et al., 1990.
Liver. In: Boorman, G.A., Eustis, S.L., Elwell, M.R.,
et al. (Eds.), Pathology of the Fischer rat. Academic
Press, San Diego, pp. 71–92.
Ferrara, D., Monteforte, R., Baccari, G.C., et al., 2004.
Androgen and estrogen receptors expression in the
rat exorbital lacrimal gland in relation to
“harderianization”. J. Exp. Zool. 301A, 297–306.
Frandsen, A.M., 1962. Sebaceous glands in the gingiva
of the rat. Oral Biol. 7, 247–248.
Frith, C.H., Bott, S., Jokinen, M.P., et al., 2000a.
Non-proliferative lesions of the endocrine system in
rats. In: E-I guides for toxicologic pathology. STP/
ARP/AFIP, Washington DC.
Frith, C.H., Ward, J.M., Chandra, M., et al., 2000b.
Non-proliferative lesions of the hematopoietic
system in rats. In: HL-1 guides for toxicologic
pathology. STP/ARP/AFIP, Washington DC.
Germann, P.G., Ockert, D., Heinrichs, M., 1998.
Pathology of the oropharyngeal cavity in six strains
of rats: predisposition of Fischer 344 rats for
inflammatory and degenerative changes. Toxicol.
Pathol. 26, 283–289.
Gopinath, C., Prentice, D.E., Lewis, D.G., 1987. The
liver. In: Gresham, G.A. (Ed.), Atlas of
experimental toxicological pathology, vol. 13. MTP
Press, Lancaster, pp. 43–60.
Greaves, P., Seely, J.C., 1996. Non-proliferative lesions
of soft tissues and skeletal muscle in rats. In: MST-1
guides for toxicologic pathology. STP/ARP/AFIP,
Washington DC.
Haley, D.P., Bulger, R.E., 1983. The aging male rat:
structure and function of the kidney. Am. J. Anat.
67, 1–13.
Hard, G.C., Alden, C.L., Bruner, R.H., 1999.
Non-proliferative lesions of the kidney and lower
urinary tract in rats. In: URG-1 guides for
toxicologic pathology. STP/ARP/AFIP, Washington
DC.
Hatton, G.I., 1976. Nucleus circularis: is it an
osmoreceptor in the brain? Brain research bulletin 1,
123–131.
Haworth, R., Woodfine, J., McCawley, S., et al., 2007.
Pulmonary neuroendocrine cell hyperplasia:
identification, diagnostic criteria and incidence in
untreated ageing rats of different strains. Toxicol.
Pathol. 35, 735–740.
Johnson, M.D., Gad, S.C., 2007. The rat. In: Gad S.C.
(Ed.), Animal models in toxicology, second ed.
Taylor & Francis, Boca Raton, pp. 147–217.
Kambara, T., McKevitt, T.P., Francis, I., et al., 2009.
Eosinophilic inclusions in rat Clara cells and the
effect of an inhaled corticosteroid. Toxicol. Pathol.
37, 315–323.
Kendrey, G., Roe, F.J., 1969. Melanotic lesions of the
eye in August hooded rats induced by urethan or
N-hydroxyurethan given during the neonatal period:
a histopathological study. J. Natl. Cancer Inst. 43,
749–762.
Knox, C.A., Yates, R.D., Chen, I., et al., 1980. Effects
of aging on the structural and permeability
characteristics of cerebrovasculature in normotensive
and hypertensive strains of rats. Acta. Neuropathol.
51, 1–13.
Kramer, M.F., Tan, H.T., 1968. The peri-insular acini of
the pancreas of the rat. Z. Zellforsch Mikrosk Anat.
86, 163–170.
Leininger, J.R., Riley, M.G., 1990. Bones, joints and
synovia. In: Boorman, G.A., Eustis, S.L., Elwell,
M.R., et al. (Eds.), Pathology of the Fischer rat.
Academic Press, San Diego, pp. 209–225.
Livingston, R.S., Besch-Williford, C.L., Myles, M.H.,
et al., 2011. Pneumocystis carinii infection causes
lung lesions historically attributed to rat respiratory
virus. Comparative Medicine 61, 45–52.
Long, P.H., Leinninger, J.R., Ernst, H., 1996.
Non-proliferative lesions of bone, cartilage, tooth
and synovium of rats. In: MST-2 guides for
toxicologic pathology. STP/ARP/AFIP,
Washington DC.
MacKenzie, W.F., Alison, R.H., 1990. Heart. In:
Boorman, G.A., Eustis, S.L., Elwell, M.R., et al.
(Eds.), Pathology of the Fischer rat. Academic Press,
San Diego, pp. 461–471.
McMartin, D.N., O’Donoghue, J.L., Morissey, R.,
1997. Non-proliferative lesions of the nervous
system in rats. In: NS-1 guides for toxicologc
pathology. STP/ARP/AFIP, Washington DC.
Montgomery, C.A. & Seely, J.C., 1990. Kidney. In:
Boorman, G.A., Eustis, S.L., Elwell, M.R., et al.
(Eds.), Pathology of the Fischer rat. Academic Press,
San Diego, pp. 127–152.
Monticello, T.M., Morgan, K.T., Uriah, I., 1990. Non
neoplastic lesions nasal lesions in rats and mice.
Environ. Health Perspect 85, 249–274.
Neuenschwander, S.B., Elwell, M.R., 1990. Salivary
glands. In: Boorman, G.A., Eustis, S.L., Elwell,
M.R., et al. (Eds.), Pathology of the Fischer rat.
Academic Press, San Diego, pp. 31–41.
Nève, P., Rondeaux, P., 1991. Age-related accumulation
of lysosomes and other cytological features in active
thyroid follicles of the CBA mouse. Cell Tissue Res.
65, 275–285.
Percy, D.H., Barthold, S.W., 2007. Pathology of
laboratory rodents and rabbits, third ed. Iowa State
University Press, IA, p. 133.
Rao-Rupanagudi, S., Heywood, R., Gopinath, C., 1992.
Age-related changes in thyroid structure and
function in Sprague-Dawley rats. Vet. Pathol. 29,
278–287.
Renne, R.A., Dungworth, D.L., Keenan, C.M., 2003.
Non-proliferative lesions of the respiratory tract in
rats. In R-1 guides for toxicologic pathology. STP/
ARP/AFIP, Washington, DC.
Renne, R., Brix, A., Harkema, J., et al., 2009.
Proliferative and non proliferative lesions of the rat
and mouse respiratory tract. Toxicol. Pathol. 37,
5S-73S.
Ruben, Z., Arceo, R.J., Bishop, S.P., et al., 2000.
Non-proliferative lesions of the heart and
vasculature in rats. In: CV-1 guides for toxicologic
pathology. STP/ARP/AFIP, Washington DC.
Rulli, M.A., Martinelli, C., 1971. Free sebaceous glands
in the gingival of some rats. Archs Oral Biol. 16,
831–832.
Sasaki, S., Nagai, H., Mori, I., et al., 1998.
Spontaneous synovitis in Wistar rats. Toxicol. Pathol.
26, 687–690.
Stalker, M.J., Hayes, M.A., 2007. Liver and biliary
system. In: Maxie MG (Ed.), Jubb, Kennedy &
Palmer’s pathology of domestic animals, vol. 2. fifth
ed. Saunders, Edinburgh, p. 312.
Stefanski, S.A., Elwell, M.R., Stromberg, P.C., 1990.
Spleen, lymph nodes and thymus. In: Boorman,
G.A., Eustis, S.L., Elwell, M.R., et al. (Eds.),
Pathology of the Fischer rat. Academic Press, San
Diego, pp. 369–393.
Taradach, C., Greaves, P., 1984. Spontaneous eye
lesions in laboratory animals: incidence in relation to
age. Crit. Rev. Toxicol. 12, 121–147.
Thoolen, B., Maronpot, R.R., Harada, T., et al., 2010.
Proliferative and nonproliferative lesions of the rat
and mouse hepatobiliary system. Toxicol. Pathol. 38,
5S–81S.
Yanai, T., Masegi, T., Ueda, K., et al., 1993.
Spontaneous globoid mineralization in the
cerebellum of rats. J. Comp. Pathol. 109,
447–451.
Yoshitomi, K., Brown, H.R., Eustis, S., 1990. Fordyce’s
granules of the incisor and molar gingiva in F344
rats. Vet. Pathol. 27, 432–438.
 Beagle dog 37
CHAPTER 3 
Cheryl Scudamore

Beagle dog

Introduction
The majority of dogs used in toxicity studies are
young beagles (generally less than one year old at
the start of study), and therefore a limited range
of background findings is recognized compared to
the ageing population of dogs generally seen in
veterinary pathology diagnostic facilities. The most
common observations are minimal inflammatory-
cell foci, usually predominantly mononuclear cells,
of unknown etiology. Inflammatory-cell foci are
seen in many organs but are particularly common
in the liver (Foster 2005), lung, salivary glands and
tongue. Other common findings include granular
brown pigment in the spleen (usually hemosiderin)
and liver (usually lipofuscin) and agonal congestion FIGURE 3.3  Focal mineralization of media of aorta. ×100.
FIGURE 3.1  Haematocyst at the base of heart valve,
of the vasculature, particularly in the gastro­ characterized by blood-filled channels surrounded by
intestinal tract (Haggerty et al 2007). Other back- Intimal thickening as the result of musculoelas-
connective tissue. ×400. Image courtesy of A de Molina,
ground and incidental findings are seen with tic proliferation of intramural vessels in the myo-
HLS.
varying incidence depending on the strain, supplier cardium (Grant Maxie & Robinson 2007) and
and test facility, and are illustrated below. pulmonary vessels (Fig. 3.4) can be found in nor-
motensive laboratory dogs. The change needs to
be distinguished from oblique planes of section
Cardiovascular system through blood vessel walls, particularly where
Young dogs of all breeds – including beagles – there is the possibility that there are drug-induced
may show spontaneous lesions including focal changes in vascular tone or blood pressure. There
telangiectasia (hematocysts) of the heart valve is little literature on the spontaneous incidence of
(Fig. 3.1) and endocardiosis (Takeda et al 1991). these changes (Detweiler et al 1961).
Hematocysts are seen macroscopically as dark red
or black cysts on valve cusps. The incidence of
endocardiosis increases with age, with a frequency
of 11–25% reported in lifetime studies of beagles
(Van Fleet 2001). Arteritis is seen sporadically,
reportedly in 5–10% of young beagles (Grant
Maxie & Robinson 2007), and occurs more com-
monly in males, affecting either single or multiple
vessels, most commonly in the epididymides,
thymus, and the coronary arteries of the heart
(Fig. 3.2) (Son 2004). In the majority of cases in FIGURE 3.2  Arteritis and periarteritis of extramural coro-
young dogs the lesions are idiopathic (Hartman nary artery. ×200. Image courtesy of E McInnes.
1987) and occur in the absence of any clinical
signs, but they can also occur as part of the ‘beagle Mineralization of the root of the aorta (Glaister
pain syndrome’. Distinguishing spontaneous 1986, Kelly et al 1982) (Fig. 3.3) and other vessels
lesions from those induced by drugs can be diffi- is occasionally seen in young dogs, although the
cult, and pathologists may need to use a weight- incidence increases with age (Schwarz et al 2002).
FIGURE 3.4  Focal villous intimal proliferation in an intra-
of-evidence approach based on the incidence, In beagles, this lesion can occur in animals with
mural myocardial artery. Proliferation may be concentric
distribution of lesions, morphological appearance, normal serum calcium and phosphorus levels, in
or focal forming arterial ‘cushions’. ×100.
and associated pathology (Clemo et al 2003) to the absence of any other pathology, and may only
reach a diagnosis. be visible microscopically.
Villous mesothelial proliferations are occasion-
ally seen attached to the epicardium and pleura
(Mesfin 1990) (Fig. 3.5), possibly resulting from
local friction or other stimulation. These pro­
jections consist of an extracellular matrix core
surrounding capillaries covered by a cuboidal mes-
othelial lining.
38 BACKGROUND LESIONS IN LABORATORY ANIMALS

FIGURE 3.5  Villous mesothelial projections from epicar-
dium of atrium characterized by a connective tissue core
covered by mesothelium. Atrial wall also shows extensive
adipocyte infiltration. ×100.
FIGURE 3.7  Prominent Hassall’s corpuscles composed of
concentric whorls of keratin are a normal feature of
canine thymus. ×100.
Respiratory system
Normal histological variations occur throughout
the respiratory tract and include foci of squamous
epithelium (Fig. 3.10) in the trachea. Pulmonary
mineralization can occur in two forms: either asso-
ciated with the alveolar interstitium, when it
tends to be diffuse or multifocal, or as focal osteo-
phyte formation (Fig. 3.11). Diffuse pulmonary
mineralization is reported in dogs with uraemia
and Cushing’s syndrome (Berry et al 2005)
but can also be an idiopathic finding, and subepi-
thelial mineralization of peribronchiolar smooth
muscle (Fig. 3.12) can be seen as a spontaneous
finding in control beagles (D J Lewis, personal
communication).

Hemolymphoreticular system
Few lesions are recorded in the lymphoid tissues
of young beagles. Thymic cysts or epithelial rem-
nants from the branchial pouches (Fig. 3.6) are
commonly seen in the medulla (Newman 1971),
as in other species, and may increase in promi-
nence with involution. Prominent Hassall’s cor-
puscles may also be seen in the thymic medulla
(Snyder et al 1993) and are composed of concen-
tric layers of keratin (Fig. 3.7). In the spleen of
young dogs haemosiderotic foci or plaques may
not be obvious macroscopically, but may be seen
microscopically, usually associated with connec-
tive tissue trabeculae or in connective tissue
around the hilar vessels (Fig. 3.8). In older animals
these are recognized macroscopically as yellow or
grey encrustations or nodules along the capsular
borders, sometimes referred to as Gandy–Gamna
bodies.
FIGURE 3.10  Squamous epithelium is a normal feature
of the epithelial lining of the dorsal trachea where the
cartilage is discontinuous. ×100.
FIGURE 3.8  Hemosiderotic plaque in the spleen. Mineral
and brown pigment deposits on connective tissue trabec-
ulae. ×100.
Accessory spleens may be present in the
omentum or attached to the surface of the spleen.
They appear as brown nodules, usually less than
2mm in diameter (Kelly et al 1982, Patnaik
1985), and they have their own independent
blood supply. Hyalinized material may occasion-
ally be seen in the follicles of lymph nodes and
spleen (Fig. 3.9). In some cases this may be
amyloid deposition but may alternatively be
accumulations of immunoglobulin.

FIGURE 3.11  Focal osseous metaplasia or osteophyte

formation in lung parenchyma. Image courtesy of DJ
Lewis. ×200.

FIGURE 3.6  Prominent branchial cyst in involuting

thymus. ×100.

FIGURE 3.9  Hyaline deposits in the germinal centre of a

lymphoid follicle. ×100. FIGURE 3.12  Mineralization of peribronchiolar smooth
muscle. Image courtesy of DJ Lewis. ×200.
 Beagle dog 39

Fibrosing alveolitis, also known as segmental

fibrosis, may be seen macroscopically as pale,
white, focal thickenings of the pleural surface,
often at the edges of the lung lobes. Microscopi-
cally (Figs 3.13, 3.14) these lesions are wedge-
shaped areas of alveolar fibrosis extending into
the lung parenchyma from the pleura, with associ-
ated epithelial hyperplasia and occasional small
numbers of chronic inflammatory cells (Glaister
1986, Hahn & Dagle 2001). These lesions are well
recognized in beagle tissues but do not seem to be
reported in the veterinary literature. There is
some confusion about the cause of these lesions. FIGURE 3.17  Dilated subpleural lymphatics. ×100.
Almost invariably no etiological agent is seen in
the sections, although they may represent damage
caused by previously migrating parasites such as FIGURE 3.14  Fibrosing alveolitis at higher magnification
Filaroides and Toxocara species. Filaroides hirthi characterized by fibrosis of the alveolar septae with a Gastrointestinal system
lung infestations have been reported in beagle hyperplastic epithelial lining with or without an associated
inflammatory reaction. ×100. Subepithelial inflammatory deposits are common
with demonstrable parasites (Bahnemann & Bauer
in the dog tongue, and occasional granulomas and
1994, Hottendorf & Hirth 1974), but the current
focal ulcers may be associated with particles of
incidence is unknown in laboratory colonies. In
foreign material, for example sawdust bedding
active Filaroides infections macroscopic tan, grey
(Fig. 3.18). These infiltrates need to be distin-
or green nodules may be seen on the pleural
guished from the cellular arteriovenous anastomo-
surface associated microscopically with granulo-
ses (Fig. 3.19) which lie below the epithelium
mas with eosinophils with or without cross sec-
(Pritchard & Daniel 1953). Adipocyte infiltration
tions of nematodes (Fig. 3.15). These granulomas
of the salivary glands may be seen incidentally
should be distinguished from those which occur
without clinical correlation or inflammation (Fig.
secondarily to the inhalation of foreign material or
3.20), although it is reported to cause palpable
due to emboli of keratin from intravenous injec-
glandular enlargement in older animals (Bindseil
tion sites (Fig. 3.16) (Hottendorf & Hirth 1974).
& Madsen 1997, Brown et al 1997).
Dilated pleural lymphatics are occasionally seen
and may be artefactual, related to changes in pul-
monary pressure due to clamping or infusion with
fixative (Fig. 3.17) (Colby et al 2007).

FIGURE 3.15  Cross sections of Filaroides larvae with

associated eosinophilic and granulomatous reaction.
Image courtesy of A Bradley. ×200.

FIGURE 3.18  Subepithelial inflammation of tongue.

×100.

FIGURE 3.13  Fibrosing alveolitis is often seen as a pale

depression in the pleural surface which corresponds with
a wedge-shaped lesion extending from the pleural
surface, often towards a pulmonary vessel. ×200.

FIGURE 3.16  Focal granuloma in the lung. Granuloma-

tous reaction surrounds refractile foreign material, pos-
sibly sawdust bedding. ×200.

FIGURE 3.19  Subepithelial arteriovenous anastamosis of

the tongue. ×100.
40 BACKGROUND LESIONS IN LABORATORY ANIMALS
FIGURE 3.20  Adipocytes separating glandular acini in the
salivary gland. ×200.
Mineralization of the gastric mucosa (Fig. 3.21)
is observed with varying incidence (5–10%) in
beagles with no underlying pathological lesions
(Glaister 1986, Majeed 1985). In some dogs occa-
sional small microscopic foci of mineralization
may be observed, but in other animals the depos-
its may be considerably more extensive leading to
a gritty feel when the mucosa is handled. It was
previously suggested (Majeed 1985) that this may
relate to dietary imbalances of calcium and phos-
phorus, but mineralization is now found in dogs
with normal serum calcium or phosphate levels
fed on balanced modern diets, and so the underly-
ing cause remains unknown.
FIGURE 3.21  Focal mineralization in gastric mucosa.
Note absence of inflammatory response. Image courtesy
of C Lopez. ×100.
A number of spiral-shaped bacteria (gastros-
pirilla) have been identified in the gastric mucosa
of dogs; they include Helicobacter felis, H. biz-
zozeronii, H cynogastricus and Candidatus Heli-
cobacter heilmanniii (Haesebrouck et al 2009).
Spiral organisms may be found in up to 86% of
clinically normal dogs and are frequently seen in
the surface mucus, in the lumina of gastric glands
(Fig. 3.22) and in parietal cells (Recordati et al
2009) of beagles with no associated pathology
(Prachasilpchai et al 2007). However spiral organ-
isms, in particular H. felis (Lee et al 1992), may
also be associated with a lymphocytic mucosal
infiltrate and/or organized lymphoid follicles
which may be seen as macroscopic pale white
nodules.
FIGURE 3.22  Spiral organisms in crypt of gastric gland. FIGURE 3.24  Brick inclusion in a hepatocyte nucleus.
×200. ×200.
Occasional developmental anomalies may also
occur in the gastrointestinal tract; for example
Meckel’s diverticulum, a remnant of the vitelline
duct, may be seen attached to the distal small
intestine or separated from the intestine associ-
ated with the serosa or mesentery (Brown et al
2007). Gastric heteropia may be seen within
diverticular or as isolated foci within the ileal
mucosa. These foci of histologically normal gastric
mucosa may be surrounded by mucous-cell hyper-
plasia considered to be an adaptive response to
local acid secretion (Fig. 3.23) (Iwata et al 1990,
Rest 1987).
FIGURE 3.25  Brick inclusion in nucleus of renal tubular
epithelial cell. ×200.
Lipid vacuolation of the gall bladder epithelium
(Boyd 1923) (Fig. 3.26) is a common incidental
finding, although it can also be increased following
treatment with a variety of compounds. Cystic
mucoid hyperplasia of the gall bladder is reported
to increase with age and to be related to progesta-
gen treatment, but it can also been seen in
untreated young beagles (Kelly et al 1982,
Murakoshil et al 2000) (Fig. 3.27).
FIGURE 3.23  Gastric heteropia in the intestine with adja-
cent mucus cell hyperplasia. ×100.
Liver and biliary system
Compared to other organs in the beagle, the liver
has the highest incidence of microscopic patho-
logical lesions (Foster 2005). In common with
other species, the majority of these lesions are
minimal, focal, inflammatory lesions which may
involve the parenchyma, perivascular or periportal
compartments. Acidophilic or hyaline rectangular
intranuclear periodic acid–Schiff(PAS)-positive
‘brick inclusions’ are occasionally observed in
hepatocytes (Fig. 3.24) (Harlemann et al 1987,
Maita et al 1977) and renal tubular epithelium FIGURE 3.26  Apical vacuolation of gall bladder
(Fig. 3.25) (Stalker & Hayes 2007). They may be epithelium. ×100.
large enough to displace the nucleus but are
believed to be of no significance, although their
composition and pathogenesis are unknown.
 Beagle dog 41

In the urinary bladder, cystitis (Fig. 3.33) with

or without pyelitis (Hottendorf & Hirth 1974) can
be seen incidentally or following catheterization
(Thomas 1979). Other incidental lesions in the
wall of the urinary bladder include lymphoid fol-
licles in the submucosa (Fig. 3.34) and, the throm-
bosed or mineralized remnants of the umbilical
arteries in the muscular wall of the urinary bladder
(Fig. 3.35) (Grant Maxie & Newman 2007b).

FIGURE 3.27  Mucoid hyperplasia of the gall bladder FIGURE 3.29  Focal tubular basophilia with minimal
epithelium. ×100. inflammation of beagle kidney. ×100.

Urinary system
Papillary mineralization in the kidney (Fig. 3.28)
is commonly observed in the absence of imbal-
ances in calcium and phosphorus. Individual
basophilic tubules (focal nephropathy) (Fig. 3.29)
(Morishima et al 1990) and focal interstitial fibro- FIGURE 3.33  Cystitis. Thickening of the transitional epi-
sis and inflammation are seen at a low incidence thelium with inflammatory cells within the epithelium and
and severity in young beagles with increasing in the submucosa. ×100.
numbers of cysts and glomerulosclerosis observed
with increasing age (Pomeroy & Roberston 2004).
Cortical tubular vacuolation may be visible as a
background finding in beagles and is seen more
commonly in females (Morishima et al 1990)
(Fig. 3.30); it is thought to be largely due to the FIGURE 3.30  Vacuolation of cortical tubular epithelium in
accumulation of lipid. Lipid may also accumulate the inner cortex of the kidney. ×100.
in fine vacuoles in the mesangial cells of the
glomeruli giving rise to ‘foam cells’ (Fig. 3.31)
(Grant Maxie & Newman 2007a). Small, densely
cellular glomeuli and tubules may be seen, par-
ticularly close to the capsular surface of the
kidney, and are thought to represent underdevel-
oped fetal or juvenile remnants in the subcapsular
nephrogenic zone (Fig. 3.32) (Eisenbrandt &
Phemister 1978).

FIGURE 3.34  Lymphoid follicles in urinary bladder wall.

×100.

FIGURE 3.31  Lipid accumulation in mesangial cells of the

glomerulus. ×200.

FIGURE 3.28  Mineralization of renal papilla of beagle

kidney. ×100.

FIGURE 3.35  Thrombosed urachal artery in muscular

wall of the urinary bladder. ×100.

FIGURE 3.32  Juvenile glomerulus centre with normal
glomerulus to left. ×100.
Endocrine glands
Ectopic thyroid tissue and cysts are the most
common incidental findings in the endocrine
system (Glaister 1986), usually originating from
remnants of embryonic ducts. Cysts originating
42 BACKGROUND LESIONS IN LABORATORY ANIMALS

from the craniopharyngeal duct are frequently

found in the pituitary between the pars distalis
and pars nervosa, and they can be quite extensive
(Fig. 3.36). The cysts are lined by a cuboidal to
columnar epithelium which may be ciliated.
Similar cysts arise in the parathyroid (Fig. 3.37)
and thyroid. In the thyroid, cysts originate from
the ultimobranchial duct and are lined by a kerati-
nized squamous epithelium (Capen 2007a).

FIGURE 3.38a  Extensive C-cell infiltrates in thyroid gland FIGURE 3.40  Vacuolation of the zona reticularis of the
with occasional trapped follicles. ×100. adrenal gland. ×200.

Focal areas of pancreatic atrophy are occasion-

ally seen in beagles. In these irregular foci acinar
cells may be small or completely replaced by
fibrous tissue and inflammatory cells (Fig. 3.41).
These lesions are thought to be secondary to local
inflammation or obstruction of the duct system
(Charles 2007).
FIGURE 3.36  Multiple cysts originating between pars
distalis and pars nervosa. ×100.

FIGURE 3.38b  Lymphocytic thyroiditis in the dog thyroid.

×100.

The presence of focal adrenal cortical vacuola-

tion is highly variable between animals, but tends
to be more extensive and occur at a greater fre-
quency in female beagles (Morishima at el 1990).
Vacuolation may be seen in all layers of the cortex
FIGURE 3.41  Pancreatic atrophy. Loss of acinar cells
(Figs 3.39, 3.40).
and replacement by fibrous tissue and inflammatory cell
infiltrate. ×200.

FIGURE 3.37  Parathyroid-associated cyst. ×100. Nesidioblastosis is an uncommon finding which

consists of a mixture of Islet cell (predominantly
beta cells) and ductular proliferation (Son et al
Focal to extensive infiltration of the thyroid by 2010). In beagles this finding has been reported
nests of C cells, which represent ultimobranchial in the absence of any biochemical or pathological
remnants, is a normal finding in dogs and is some- abnormalities, although in other breeds it has been
times called C-cell complex. The term C-cell associated with islet-cell neoplasia and diabetes
hyperplasia should only be used when there is a mellitus (Katsuta et al 1992) (Fig. 3.42).
clear increase in numbers compared to those in
age-matched control animals. The cells are pale-
staining and polygonal with haematoxylin and
eosin (H&E) staining and they express thyroglob-
ulin, calcitonin, calcitonin gene-related peptide
and somatostatin (Fig. 3.38a) (Capen 2007b).
Lymphocytic thyroiditis is present in a small per- FIGURE 3.39  Vacuolation of the zona glomerulosa of the
centage of beagle dogs used in toxicological studies adrenal. ×200.
(Fig. 3.38b) and may be a heritable condition
(Benjamin et al 1996) possibly associated with the
subsequent development of thyroid neoplasia.

FIGURE 3.42  Nesidioblastosis of the pancreas. ×400.


Skin and appendages
Few incidental skin lesions are reported in beagles,
with folliculitis being the most common. There is
often no apparent etiological agent, but occasion-
ally Demodex mites may be present (Fig. 3.43)
consistent with the self-limiting localized presen-
tation of demodicosis in young dogs. Macroscopi-
cally there may be no obvious lesions or there may
be a local area of scaling and alopecia.
FIGURE 3.43  Folliculitis associated with demodex mite
infestation. ×200.
Ear margin dermatosis is seen in laboratory
beagle dogs (and other hound breeds) as scaling
and alopecia of the ear margins. Microscopically
it is characterized by a localized marked orthok-
eratotic, epidermal and follicular hyperkeratosis
(Fig. 3.44) which may be complicated by second-
ary bacterial infection (Ginn et al 2007). It is
considered to be an idiopathic primary sebor-
rhoeic condition.
FIGURE 3.44  Epidermal and follicular hyperkeratosis
with minimal dermal inflammatory infiltrate. Ear margin
dermatitis. ×200.
Muscles, bones and joints
Focal chondromucinous degeneration may occa-
sionally be seen in the growth plates of young
beagles, particularly in the sternum. This change
includes focal degradation of the matrix with loss
of affinity for toluidine blue stain and degenera-
tion of chondrocytes, eventually leading to an
amorphous area of chondromalacia (Fig. 3.45)
(Yamasaki 1994). The cause of this lesion is
unknown, and it does not appear to result in any
abnormality in the mature skeleton. Adipocytes
may be seen infiltrating normal skeletal muscle
(Van Fleet & Valentine 2007) and may be
Beagle dog 43
particularly prominent in the myocardium where
they may extend through half the thickness of the
ventricular wall (Fig. 3.5) (Pick & Eubank 1965).
Amylase-resistant, periodic acid-Schiff (PAS)–
positive inclusions may be present in the skeletal
muscle of beagle dogs. Affected myofibers con-
tained amorphous material that stains lightly
basophilic with hematoxylin and eosin and is
strongly PAS–positive with amylase resistance.
Transmission electron micrographic examination
of the inclusions reveals granular, non-membrane–
bound, electron-dense material, consistent with
polysaccharide (Wancket et al 2011).
FIGURE 3.46  Focal mineralization of spinal cord menin-
ges. ×200.
FIGURE 3.45  Chondromucinous degeneration of the
sternum. ×200.
Brain and nervous system FIGURE 3.47  Perivascular cuffing in the central nervous
system (CNS). Unilateral, focal blood vessels surrounded
Incidental findings in the central nervous system by cuff of mononuclear cells. Image courtesy of J Bowles.
are rare in beagles. Although uncommon in young ×100.
animals, the most commonly observed lesions are
collagenous thickening of the meninges, ossifying
Renaut bodies (Elcock et al 2001) may be seen
pachymeningitis (Grant Maxie & Youssef 2007)
in sciatic nerve sections in the endoneurium or
and mineralization of spinal nerves (Fig. 3.46).
perineurium and consist of well demarcated,
These lesions probably all represent early onset of
sparsely cellular structures with a filamentous col-
degenerative changes. Perivascular cuffing of
lagen matrix; they are not associated with evi-
vessels (Fig. 3.47) characterized by minimal accu-
dence of axonal degeneration or inflammation and
mulations of predominantly lymphocytic cells can
are considered incidental findings (Fig. 3.48). The
be seen at a low incidence in normal beagle dogs.
cell rest of Hortega is visible in the canine cere-
The lesions affect only a few vessels, are not bilat-
brum and may be confused with an area of gliosis
eral or symmetrical, and have no associated clini-
(Fig. 3.49).
cal signs (Ueda et al 2004). The origins and
significance of these lesions are unknown, and
although they have been suggested to be post-
vaccinal or post-viral, there is no clear evidence
to support this. Focal accumulations of inflamma-
tory cells may be seen in the choroid plexus; these
may occur in the absence of any other brain
pathology and appear to be of no significance.
Non-suppurative or granulomatous meningitis
may also occasionally be seen, often without asso-
ciated clinical signs in control dogs. The underly-
ing cause of this lesion is not known, but may be
infectious (Ueda et al 2004).
FIGURE 3.48  Renaut body in sciatic nerve. ×100.
44 BACKGROUND LESIONS IN LABORATORY ANIMALS
FIGURE 3.49  Cell rest of Hortega in canine cerebellum.
×200.
Acknowledgments
The author is grateful for the help of J Bowles,
GSK for preparation of many of the images not
otherwise acknowledged.
References
Bahnemann, R., Bauer, C., 1994. Lungworm infection
in a beagle colony: Filaroides hirthi, a common but
not well-known companion. Exp. Toxicol. Pathol. 46,
55–62.
Benjamin, S.A., Stephens, L.C., Hamilton, B.F., et al.,
1996. Associations between lymphocytic thyroiditis,
hypothyroidism and thyroid neoplasia in Beagles.
Vet. Pathol. 33, 486–494.
Berry, C.R., Ackerman, N., Monce, K., 2005.
Pulmonary mineralization in 4 dogs with Cushing’s
syndrome. Vet. Radiol. Ultrasound 35, 10–16.
Bindseil, E., Madsen, J.S., 1997. Lipomatosis causing
tumour-like swelling of a mandibular salivary gland
in a dog. Vet. Rec. 140, 583–584.
Boyd, W., 1923. Studies in gall bladder pathology.
Br. J. Surg. 10, 337–356.
Brown, P.J., Lucke, V.M., Sozmen, M., Whitbread, T.J.,
Wyatt, J.M., 1997. Lipomatous infiltration of the
canine salivary gland. J. Small Anim. Pract. 38,
234–236.
Brown, C.C., Baker, D.C., Barker, I.K., 2007.
Alimentary system. In: Grant Maxie, M. (Ed.),
Jubb, Kennedy and Palmer’s pathology of domestic
animals, vol. 2, fifth ed. Saunders, Philadelphia,
p. 85.
Capen, C.C., 2007a. Endocrine glands. In: Grant
Maxie, M. (Ed.), Jubb, Kennedy and Palmer’s
pathology of domestic animals, vol. 2, fifth ed.
Saunders, Philadelphia, p. 360.
Capen, C.C., 2007b. Endocrine glands. In: Grant
Maxie, M. (Ed.), Jubb, Kennedy and Palmer’s
pathology of domestic animals, vol. 2, fifth ed.
Saunders, Philadelphia, p. 357.
Charles, J.A., 2007. Pancreas. In: Grant Maxie, M.
(Ed.), Jubb, Kennedy and Palmer’s pathology of
domestic animals, vol. 2, fifth ed. Saunders,
Philadelphia, pp. 396–397.
Clemo, F.A., Evering, W.E., Snyder, P.W., Albassam,
M.A., 2003. Differentiating spontaneous from
drug-induced vascular injury in the dog. Toxicol.
Pathol. 31(suppl):25–31.
Colby, T.V., Leslie, K.O., Yousem, S.A., 2007. Lungs.
In: Sternberg, S.S. (Ed.), Histology for pathologists,
third ed. Lippincott, Williams & Wilkins,
Philadelphia, p. 489.
Detweiler, D.K., Patterson, D.F., Hubben, K., Botts,
R.P., 1961. The prevalence of spontaneously
occurring cardiovascular disease in dogs. Am. J.
Public Health Nations Health 51, 228–241.
Eisenbrandt, D.L., Phemister, R.D., 1978. Postnatal
development of the canine kidney. Am. J. Anat.
154, 179–193.
Elcock, L.E., Stuart, B.P., Hoss, H.E., et al., 2001.
Renaut bodies in the sciatic nerve of beagle dogs.
Exp. Toxicol. Pathol. 53, 19–24.
Foster, J.R., 2005. Spontaneous and drug-induced
hepatic pathology of the laboratory Beagle dog, the
Cynomolgus macaque and the marmoset. Toxicol.
Pathol. 33, 63–74.
Ginn, P.E., Mansell EKL, Rakich, P.M., 2007. Skin
and appendages. In: Grant Maxie, M. (Ed.), Jubb,
Kennedy and Palmer’s pathology of domestic
animals, vol. 1, fifth ed. Saunders, Philadelphia,
p. 598.
Glaister, J., 1986. Laboratory animal pathology. In:
Principles of toxicological pathology. Taylor &
Francis, London, pp. 146–152.
Grant Maxie, M., Newman, S.J., 2007a. Urinary
system. In: Grant Maxie, M. (Ed.), Jubb, Kennedy
and Palmer’s pathology of domestic animals, vol. 2,
fifth ed. Saunders, Philadelphia, pp. 465–466.
Grant Maxie, M., Newman, S.J., 2007b. Urinary
system. In: Grant Maxie, M. (Ed.), Jubb, Kennedy
and Palmer’s pathology of domestic animals, vol. 2,
fifth ed. Saunders, Philadelphia, p. 505.
Grant Maxie, M., Robinson, W.F., 2007. Cardiovascular
system. In: Grant Maxie, M. (Ed.), Jubb, Kennedy
and Palmer’s pathology of domestic animals, vol. 2,
fifth ed. Saunders, Philadelphia, p. 71.
Grant Maxie, M., Youssef, S., 2007. Nervous system.
In: Grant Maxie, M. (Ed.), Jubb, Kennedy and
Palmer’s pathology of domestic animals, vol. 1, fifth
ed. Saunders, Philadelphia, p. 345.
Haesebrouck, F., Pasmans, F., Flahou, B., et al., 2009.
Gastric helicobacters in domestic animals and
nonhuman primates and their significance for human
health. Clin. Microbiol. Rev. 22, 202–223.
Haggerty, G.C., Peckham, J.C., Thomassen, R.W., Gad,
S.C., 2007. The dog. In: Gad, S.C. (Ed.), Animal
models in toxicology, second ed. Taylor & Francis,
Boca Raton, pp. 588–645.
Hahn, F.F., Dagle, G.E., 2001. Non-neoplastic
pulmonary lesions. In: Mohr, U., Carlton, W.W.,
Dungworth, D.L., et al. (Eds.), Pathobiology of
the aging dog, vol. 1, ISU Press/ILSI, Ames, IA,
pp. 60–61.
Harlemann, J.H., Suter, J., Fischer, M., 1987.
Intracytoplasmic eosinophilic inclusion bodies the in
the liver of beagle dogs. Lab. Animal Sci. 37, 229.
Hartman, L.A., 1987. Idiopathic extramural coronary
arteritis in beagle and mongrel dogs. Vet. Pathol. 24,
537–544.
Hottendorf, G.H., Hirth, R.S., 1974. Lesions of
spontaneous subclinical disease in Beagle dogs. Vet.
Pathol. 11, 240–258.
Iwata, H., Arai, C., Koike, Y., et al., 1990. Heretotopic
gastric mucosa of the small intestine in laboratory
beagle dogs. Toxicol. Pathol. 18, 373–379.
Katsuta, O., Tsuchitani, M., Narama, I., 1992.
Abnormal proliferation of pancreatic endocrine cells
in beagle dogs. J. Toxicol. Pathol. 5, 67–76.
Kelly, D.F., Lucke, V.M., Gaskell, C.J., 1982.
Incidental necropsy findings. Notes on pathology for
small animal clinicians. John Wright, Bristol, ch 6,
pp. 77–96.
Lee, A., Krakowa, S., Fox, J.G., et al., 1992. The role
of Helicobacter felis in chronic canine gastritis. Vet.
Pathol. 29, 487–494.
Maita, K., Masuda, H., Suzuki, Y., 1977. Spontaneous
lesions detected in the Beagles used in toxicity
studies. Jikken Dobutsu 26, 161–167.
Majeed, S.K., 1985. Mineralisation in kidney and
stomach of Beagle dogs. Vet. Quarterly 2, 162–164.
Mesfin, G.M., 1990. Spontaneous epicardial fibrous
fronds on the atria of Beagle dogs. Vet. Pathol. 27,
458–461.
Morishima, H., Nonoyama, T., Sasaki, S., Miyajima,
H., 1990. Spontaneous lesions in beagle dogs used in
toxicity studies. Jikken Dobutsu 39, 239–248.
Murakoshil, M., Ikeda, R., Tagawa, M., Iwasaka, T.,
Nakayama, T., 2000. Histopathological study of
female Beagle dogs for four year treatment with
subcutaneous implantation of chlomdinone acetate
(CMA). Tokai J. Exp. Clin. Med. 25, 87–91.
Newman, A.J., 1971. Cysts of branchial arch origin in
the thymus of the Beagle. J. Small Anim. Pract. 12,
681–685.
Patnaik, A., 1985. Splenosis in a dog. J. Small Anim.
Pract. 26, 23–31.
Pick, J.R., Eubank, J.R., 1965. A clinicopathologic
study of heterogenous and homogeneous dog
populations in North Carolina. Lab. Anim. Care 15,
11–17.
Pomeroy, M.J., Robertson, J.L., 2004. The relationship
of age, sex, and glomerular location to the
development of spontaneous lesions in the canine
kidney: analysis of a life-span study. Toxicol. Pathol.
32, 237–242.
Prachasilpchai, W., Nuanualsuwan, S., Chatsuwan, T.,
et al., 2007. Diagnosis of Helicobacter spp. infection
in canine stomach. J. Vet. Sci. 8, 139–145.
Pritchard, M.M., Daniel, P.M., 1953. Arterio-venous
anastomoses in the tongue of the dog. J. Anat. 87,
66–74.
Recordati, C., Gualdi, V., Craven, M., et al., 2009.
Spatial distribution of Helicobacter spp. in the
gastrointestinal tract of dogs. Helicobacter 3,
180–191.
Rest, J.R., 1987. Gastrointestinal anomalies in the
dog – two cases reports. Vet. Rec. 121, 426–427.
Schwarz, T., Sullivan, M., Störk, C.K., et al., 2002.
Aortic and cardiac mineralization in the dog. Vet.
Radiol. Ultrasound 43, 419–427.
Snyder, P.W., Kazacos, E.A., Felsberg, P.J., 1993.
Histologic characterisation of the thymus in canine
X-linked severe combined immunodeficiency. Clin.
Immunol. Immunopathol. 67, 55–67.
Son, W-C., 2004. Idiopathic canine polyarteritis in
control beagle dogs from toxicity studies. J. Vet. Sci.
5, 147–150.
Son, W.C., Faki, K., Mowat, V., Gopinath, C., 2010.
Spontaneously occurring extra-islet endocrine cell
proliferation in the pancreas of young Beagle dogs.
Toxicol. Lett. 193, 179–182.
Stalker, M.J., Hayes, M.A., 2007. Liver and biliary
system. In: Grant Maxie, M. (Ed.), Jubb, Kennedy
and Palmer’s pathology of domestic animals, vol. 2,
fifth ed. Saunders, Philadelphia, p. 307.
Takeda, T., Makita, T., Nakamura, N., Kimizuka, G.,
1991. Morphologic aspects and morphogenesis of
blood cysts on canine cardiac valves. Vet. Pathol. 28,
16–21.
Thomas, J.E., 1979. Urinary tract infection induced by
intermittent urethral catheterisation in dogs. JAVMA
174, 705–707.
Ueda, A., Ueda, M., Nakazawa, M., et al., 2004.
Nonsuppurative meningoencepahlitis in a Beagle
dog. Tox. Pathol. 1, 51–53.
Van Fleet, J.F., 2001. Age related non-neoplastic lesions
of the heart. In Mohr, U., Carlton, W.W., Dungworth,
D.L. et al. (Eds.), Pathobiology of the aging dog, vol.
2, ISU Press/ILSI, Ames, IA, pp. 102–104.
Van Fleet, J.F., Valentine, B.A., 2007. Muscle and
tendon. In: Grant Maxie, M. (Ed.), Jubb, Kennedy
and Palmer’s pathology of domestic animals, vol. 1,
fifth ed. Saunders, Philadelphia, p. 202.
Wancket, L.M., Quist, E.M., Le Net, J.L., Guzman,
R.E., Muravnick, K.B., 2011. Spontaneous complex
polysaccharide inclusions in the skeletal muscle of
purpose-bred beagle dogs. Toxicol. Pathol. 39,
410–413.
Yamasaki, K., 1994. Lesions of sternal and growth plate
cartilage in Beagles. Lab. Anim. Sci. 44, 389–392.

Introduction
The mouse is one of the most widely used experi-
mental models in pre-clinical toxicity and carcino-
genicity testing, and biomedical research in
general, and as such there are numerous sources
of information on the biology, physiology and
spontaneous pathology of the many stocks and
strains of mouse used in biomedical research.
There are a limited number of mouse stocks and
strains routinely used in pre-clinical toxicity
testing, however. This is, in part, because of the
regulatory requirement for the background biology
and pathology of these models to be well under-
stood, and for there to be sufficient background
data available for all the different parameters
measured during toxicity and carcinogenicity
studies.
It is important for toxicological pathologists
working with mice to be aware of the background
changes commonly seen in laboratory mice, and
the stock and strain differences that occur in their
background pathology. A knowledge of the spon-
taneous findings which occur in young mice, and
those changes associated with aging, is essential
if a proper interpretation of possible treatment-
related changes in toxicity and carcinogenicity
studies is to be made.
The current chapter aims to present those non-
neoplastic findings which occur spontaneously in
young and old mice, and which the toxicological
pathologist will encounter during evaluation of
routine toxicity studies using the mouse. The
pathologist should be aware that long-term admin-
istration of test materials may be associated with
an exacerbation of age-related non-neoplastic
pathology which might otherwise be considered
to be background changes. Careful comparison of
treated animals with contemporaneous controls,
and interpretation based on evaluation of all
the results in the study (including survival and
body weights), are required to properly identify
the significance of changes seen in pre-clinical
studies.
A detailed presentation of the embryology,
anatomy and histology of all the different organ
systems discussed is beyond the scope of the
current chapter, and the reader is referred to rel-
evant texts for each of the different organs dis-
cussed. A general review of the anatomy and
biology of the laboratory mouse and the use of
mice in biomedical research has been presented in
various publications (Cook 1965, Green 1966,
Hedrich & Bullock 2004, Hummel et al 1966,
Komárek 2004, Krinke 2004, Percy & Barthold
2007, Staats 1966).
Many sources of information on the spontane-
ous, non-neoplastic pathology of laboratory mice
are available. The aging patterns of mice and strain
variations in background pathology have been
reported (Brayton 2007, Cotchin & Roe 1967,
Hedrich & Bullock 2004, Mohr et al 1996a, b,
Russell 1966, Russell & Meier 1966). The pathol-
ogy of aging mice based on the studies performed
at different laboratories has been presented for
the BALB/c and B6C3F1 mouse (Frith et al 1985,
Frith & Ward 1988), the CD-1 mouse (Faccini
et al 1990), and those strains of mouse used in the
generation of genetically engineered mice (Haines
et al 2001, Mahler et al 1996, Percy & Barthold
2007, Ward et al 2000). Descriptions of spontane-
ous, murine, non-neoplastic pathology are also
included in other publications reviewing the spon-
taneous and induced lesions in different organ
systems in rodents in pre-clinical toxicity and car-
cinogenicity studies (Gad 2007, Jones et al 1983,
Jones et al 1985a,b, Jones et al 1986, 1988, 1991,
Maronpot et al 1999, Mohr 2001).
The National Toxicology Program (NTP) rou-
tinely uses the B6C3F1 mouse, an F1 hybrid of
two inbred strains, the C57BL/6 and C3H strains
(Rao & Boorman 1999), in their toxicity and car-
cinogenicity studies. The outbred CD-1 mouse is
also routinely used throughout the world in pre-
clinical testing of pharmaceuticals, agrochemicals
and industrial chemicals (Gad 2007).
The NTP publishes on their websites the obser-
vations recorded in the numerous toxicity and
carcinogenicity studies performed by them every
year, and maintain a historical control database of
neoplastic findings classified by species, strain,
route of administration, vehicle and diet (http://
ntp.niehs.nih.gov/).
The NTP also summarize the information
they have gathered to date on their evaluation
of transgenic models for use in carcinogenicity
testing. The historical control tumor rates for
these models are also provided on their web pages.
Animal suppliers also routinely maintain histori-
cal control pathology data, and this information
can usually be retrieved from their web sites
(Charles River: http://www.criver.com/; Harlan:
http://www.harlan.com/) although this may be
limited to neoplastic findings for some strains of
animals.
There is a need to maintain an up-to-date source
of information on the ever increasing numbers of
inbred and genetically modified strains of mice
used in biomedical research and those differences
in biology associated with these new genotypes
(Anagnostopoulos et al 2001, Bolon 2007, Brayton
et al 2001, Linder 2003, 2006, Linder & Davisson
2004, Yoshiki & Moriwaki 2006). The Jackson
Laboratory maintains the Mouse Genome Infor-
matics web site, a database designed to act as an
international resource for the laboratory mouse,
integrating genetic, genomic and biological data
(http://www.informatics.jax.org/). This site offers
resources and links to multiple databases, many of
which provide valuable information for the toxi-
cological pathologist involved in the evaluation of
preclinical studies in the mouse. Other internet
sources of information on genetically engineered
rodents have been reviewed by Bolon (2006) and
Linder & Davisson (2004).
The data presented in this chapter are based
in the main on findings observed in the outbred
Swiss mouse, obtained from Charles River, United
Kingdom (Crl:CD1(ICR) (outbred)). Where
specific strain differences in the occurrence of
Mouse 45
CHAPTER 4 
Ian Taylor
Mouse
spontaneous pathology occur, these will be referred
to and reference will be made to the sources of
information available in the scientific literature.
Cardiovascular system
The embryology, anatomy, histology and physiol-
ogy of the heart and blood vessels are reviewed by
Elwell & Mahler (1999) and Michael et al (2004)
and an extensive review of myocardial diseases of
animals has been presented by Van Vleet & Ferrans
(1986).
Congenital lesions in the heart are rare (Hagi-
wara et al 1996). Cardiomyopathy is a diagnostic
term used to describe a spectrum of spontaneous,
age-related, degenerative changes, including
degeneration, necrosis and increased interstitial
fibrous tissue. The inflammatory component of
these changes varies (Berdanier 2004, Elwell &
Mahler 1999, Frith et al 2007). The reported inci-
dence of this finding is considered to vary partly
because of variation in reporting levels and termi-
nology between pathologists (Elwell & Mahler
1999). These changes can occur at an early age,
and are occasionally responsible for the early
death of mice on long-term studies (Son 2003a)
(Fig. 4.1).
FIGURE 4.1  Heart: cardiomyopathy. Two-year-old, CD-1,
male mouse. Degenerate cardiomyocytes are replaced by
fibrous tissue. ×100.
Atrial thrombosis is described as an uncommon
lesion by some authors (Frith & Ward 1988, Frith
et al 2007), but as common by others (Berdanier
2004, Faccini et al 1990). This probably reflects
the strain and stock differences in the occurrence
of this lesion. It does occur as a spontaneous
lesion, however, in CD-1 mice, more commonly
in the left atrium than in the right (Carlton &
Engelhardt 1991, Elwell & Mahler 1999, Frith &
Ward 1988, Frith et al 2007, Hagiwara et al 1996,
Meier & Hoag 1966, Percy & Barthold 2007,
Van Vleet & Ferrans 1986). The distended atrium
contains an organizing, mural thrombus which
may contain areas of cartilaginous metaplasia.
The character and features of the thrombus are
dependent on the age of the thrombus. There are
strain and sex differences in the occurrence of
atrial thrombi. Diet and parity also influence the
46 BACKGROUND LESIONS IN LABORATORY ANIMALS
occurrence. Atrial thrombosis was identified as
one of the major causes of death or morbidity in
a review of 11 long-term studies in CD-1 mice
(Maita et al 1988) (Fig. 4.2).
FIGURE 4.2  Atrial thrombus, 18-month-old, male, CD-1
mouse. The majority of the lumen of the left atrium is
filled with an organizing thrombus. An area of cartilagi-
nous metaplasia is apparent at the periphery of the throm-
bus and the atrial wall has a prominent inflammatory cell
infiltrate. ×20.
Dystrophic mineralization (dystrophic calcifica-
tion) is a strain-specific finding and occurs at a
high incidence and at an early age in susceptible
strains. There are sex differences in the occur-
rence of this finding within strains, and parity and
diet also influence its occurrence (Berdanier 2004,
Eaton et al 1978, Elwell & Mahler 1999, Frith &
Ward 1988, Frith et al 2007, Hagiwara et al 1996,
Maeda et al 1986, Meier & Hoag 1966, Percy &
Barthold 2007, Russell 1966, Van Vleet & Ferrans
1986, 1991a, Yamate et al 1987). The distribution
of the mineralization is also strain specific and
can be epicardial, myocardial or both. Myocardial
mineralization is an uncommon finding in CD-1
mice (Faccini et al 1990), but is common in
BALB/c mice (Fig. 4.3).
FIGURE 4.3  Dystrophic mineralization of the epicardium
of a 10-week-old, male, BALB/c mouse. ×100.
In old mice, cartilage cells can sometimes be
observed in the fibrous tissue at the base of the
heart valves (Hummel et al 1966). Endocardial
myxomatous change has been described as an age-
related lesion involving the generation of excess
myxomatous matrix within valvular interstitial
cells. This change is not associated with increased
morbidity or mortality. The low level of reporting
of this lesion is considered to be related to incon-
sistency of sampling in routine toxicity studies
(Donnelly 2008, Elangbam et al 2002).
Perivascular lymphoid infiltrates frequently
occur in older mice in salivary glands, kidneys and
other organs (Percy & Barthold 2007). Inflamma-
tory changes in the vasculature can occur as iso-
lated lesions, affecting the vessels of a single
organ, or can affect several systemic arteries. The
occurrence and severity of this change varies with
age, strain and sex, but the etiology is generally
considered to be unknown (Elwell & Mahler
1999, Faccini et al 1990, Frith & Ward 1988, Frith
et al 2007, Percy & Barthold 2007). There are
reports of correlations between elevated blood
pressure in mice and an increased incidence
of polyarteritis (Mullink & Haneveld 1979).
Histologically, the affected arteries typically
show hyaline degeneration of the walls of the
vessel with associated inflammatory cell infiltra-
tion affecting the intima, media and adventitia.
Aneurismal dilatation and hemorrhage can be seen
in more severe cases (Fig. 4.4).
FIGURE 4.5  Inflammatory changes involving the root
of the aorta (aortitis) in an 18-month-old, male, CD-1
mouse. ×40.
FIGURE 4.4  Arteritis/periarteritis of a renal artery in an
18-month-old, CD-1, male mouse. The affected artery
shows degeneration of the wall of the vessel with inflam-
matory cell infiltration affecting the adventitia and media.
×100.
Inflammatory changes involving the root of the
aorta (aortitis) have been described in BALB/c FIGURE 4.6  Focal angiectasis in the liver of an 18-month-
mice (Ramot et al 2009). These authors consid- old, male, CD-1 mouse, showing prominent dilatation of
ered that these changes have been under-reported the vascular channels. ×100.
because of variation in sampling of this area of the
heart in routine toxicity studies (Fig. 4.5). Sys-
temic polyarteritis has been reported as one of the Hemolymphoreticular system
major causes of death or morbidity in a review of
The embryology, anatomy, histology and physiol-
11 long-term studies in CD-1 mice (Maita et al
ogy of the different elements of the hematopoi-
1988). Angiectasis, the focal dilatation of vascular
etic system have been reviewed by a number of
channels, can occur in any organ, but usually
authors (Cesta 2006a, b, Pearse 2006a, Travlos
affects the liver, spleen and lymph nodes of mice.
2006a, van Rees et al 1996, Ward et al 1999,
The existing vascular spaces are dilated and prom-
Willard-Mack 2006). In addition, a recent mono-
inent, and lining endothelial cells are normal in
graph published by the Society of Toxicologic
appearance, number and size. The etiology of this
Pathology presented a number of papers covering
change is unknown (Elwell & Mahler 1999, Faccini
the normal structure, function, pathology and
et al 1990, Frith & Ward 1988, Frith et al 2007,
enhanced histopathology of the hematopoietic
Plendi et al 1996) (Fig. 4.6).
tissues (Maronpot 2006).
The organs of the hematopoietic system repre-
sent a complex and dynamic system, where
changes can occur in a single component of the
system or be manifested by changes in all of the
different organs. There is also a large variation in
the normal appearance of the different tissues,
dependent on the age and sex of the animals
examined, as well as differences in the individual
animal’s background of antigenic stimulation
(Frith et al 1985, Wijnands et al 1996).

A limited number of lymph nodes are routinely
examined in pre-clinical toxicity and carcino­
genicity studies, with additional nodes usually
being presented for examination because of abnor-
malities reported at necropsy. Accurate identifica-
tion and naming of all the lymph nodes of the
mouse is important to ensure that comparisons
between studies relating to particular anatomical
sites can be correctly made. An investigation into
the normal distribution of lymph nodes in the
mouse, with suggested standardized nomencla-
ture, has been published by Van den Broeck et al
(2006).
Sinus ectasia (lymphangiectasia, lymphatic
cysts, sinus dilatation) is typically associated with
lymphoid atrophy and can involve the medullary
and subcapsular sinuses. It is found commonly
in the mesenteric lymph nodes of aging mice.
Dilated sinuses are lined by endothelium and can
contain pale eosinophilic/amphophilic material.
Ectasia is probably related to obstruction of effer-
ent lymph vessels (Elmore 2006, Wijnands et al
1996) (Fig. 4.7).
FIGURE 4.7  Sinus ectasia of the mesenteric lymph node
of a two-year-old, male, CD-1 mouse. A large dilated
sinus containing eosinophilic material. ×20.
Angiectasis is most often seen in the mesenteric
lymph nodes. The significance of this change is
unknown (Elmore 2006, Frith et al 1985, Ward
et al 1999). Sinus erythrocytosis can result from
a lymph node draining an area of hemorrhage, but
this lesion can also be artifactual and related to
handling procedures at necropsy. Depending on
the duration of the initiating lesion, sinus erythro-
cytosis may be accompanied by hemosiderin-
laden macrophages, erythrophagocytosis and
inflammatory cells (Elmore 2006, Frith et al
1985, Wijnands et al 1996) (Fig. 4.8).
Mouse 47
FIGURE 4.8  Sinus erythrocytosis of the mesenteric lymph FIGURE 4.9  Plasmacytosis. The sinusoids of the
nodes of an 18-month-old, male, CD-1 mouse. The sinu- mesenteric lymph node of an 18-month-old, male, CD-1
soids are packed with erythrocytes and macrophages mouse are packed with plasma cells. ×200.
containing pigment, probably hemosiderin, are also
present. ×400. Atrophy or lipomatosis occurs in older mice.
Adipocytes can often be seen in the hilus region
Sinus histiocytosis is characterized by accumu- or medulla of the lymph nodes of old mice (Ward
lation of histiocytes in the subcapsular and medul- et al 1999, Wijnands et al 1996) (Fig. 4.10). The
lary sinuses. The histiocytes have a characteristic spleen is the principal site for extramedullary
eosinophilic cytoplasm and may contain pigment hemopoiesis, but this change can sometimes be
or other phagocytosed material (Faccini et al present in the lymph nodes of mice, in response
1990, Frith & Ward 1988, Frith et al 1985, 2001, to a physiological need such as hemorrhage or
2007). Pigment accumulation in sinusoidal mac- severe inflammation (Elmore 2006, Faccini et al
rophages is a common occurrence in aging mice. 1990, Frith et al 2001).
The pigments are most often hemosiderin or lipo-
fuscin. Hemosiderin is an iron-containing, golden-
brown, granular pigment, and is usually associated
with sinus erythrocytosis. Lipofuscin is a golden-
brown pigment derived from the breakdown
of lipids from cell membranes and organelles
(Berdanier 2004, Elmore 2006, Wijnands et al
1996) (Fig. 4.8).
Lymphocyte hyperplasia may be evident to a
certain degree in various lymph nodes, and can be
dependent on the location of the lymph node and
health status of the animal. These changes increase
in incidence with age and occur more often in
females than in males (Elmore 2006, Frith & Ward
1988, Frith et al 2001, Ward et al 1999, Wijnands
et al 1996). Hyperplasia can involve different
types of cell, most commonly plasma cells, but FIGURE 4.10  Lipomatosis. Adipocytes present in the
can include an increase in the number of mast medulla of the mesenteric lymph node of an 18-month-
cells. old, male, CD-1 mouse. ×20.
Plasma cell hyperplasia (plasmacytosis) is a
common finding in rodents, particularly in the Hyperplasia of dendritic reticular cells (inter-
submandibular lymph nodes, and is usually in digitating cells) is occasionally seen in young and
response to antigenic stimulation (Elmore 2006, old mice. These are seen in response to viral infec-
Faccini et al 1990, Frith & Ward 1988, Frith et al tions and other processes (Frith et al 1985, Tew
1985, 2001, 2007, Hummel et al 1966, Ward et al 1982); they can occur in a single lymph node
et al 1999, Wijnands et al 1996) (Fig. 4.9). Mas- or affect multiple sites, and can occur as focal
tocytosis is an increase in the number of mast cells lesions in the paracortex. More extensive lesions
within the sinuses of the lymph nodes. Normal involve the entire lymph node (Fig. 4.11).
background levels of mast cells in the lymph nodes
may vary by strain (Frith & Ward 1988, Frith et al
1985, 2001, 2007).
48 BACKGROUND LESIONS IN LABORATORY ANIMALS

FIGURE 4.11  Hyperplasia of dendritic reticular cells of
the cervical lymph nodes of an 18-month-old, female,
CD-1 mouse. Cells resembling histiocytes with eosi-
nophilic cytoplasm and elongated nuclei are present in
large numbers throughout the lymph node. ×40.
Accessory splenic tissue can be found occasion-
ally, either in the pancreas or abdominal adipose
tissue (Frith & Ward 1988, Frith et al 1985, 2007,
Krinke 2004, Hummel et al 1966, Ward et al
1999). Extramedullary hemopoiesis (EMH) is a
normal finding in the red pulp of the spleen in
mice, and is usually more prevalent in young
animals and females (Cesta 2006a, Faccini et al
1990, Frith & Ward 1988, Frith et al 1985,
2001, 2007, Hardy 1967, Krinke 2004, Percy &
Barthold 2007, Suttie 2006, van Rees et al 1996,
Ward et al 1999, Wijnands et al 1996). Extramed-
ullary hemopoiesis can consist of erythroid pre-
cursors, myeloid precursors, megakaryocytes or all
three. The predominant feature of EMH depends
on the initiating stimulus (the erythroid compo-
nent predominates in response to hemorrhage, the
myeloid component predominates in response to
inflammatory conditions) (Suttie 2006). Increased
EMH can often be seen in response to skin ulcera-
tion, abscesses, or the presence of large tumors
(Fig. 4.12).
accumulate in the spleens of older mice. It is more Embryonic thymic remnants can give rise to
common in females than in males. In pigmented ectopic thymic tissues in the thyroids or parathy-
mouse strains, pigmentation of the spleen can roids, and occasionally ectopic parathyroid tissue
occur as a result of accumulation of melanin is seen in the thymus (Faccini et al 1990, Frith &
(Faccini et al 1990, Frith & Ward 1988, Frith et al Ward 1988, Frith et al 1985, 2007, Pearse 2006a,
1985, 2007, Hardy 1967, Percy & Barthold 2007, Percy & Barthold 2007, Ward et al 1999). Lym-
Suttie 2006, van der Heijden et al 1995, Ward phoid hyperplasia of the thymus occurs in mice
et al 1999, Wijnands et al 1996). over six months of age, is more common in
Hyperplasia of the splenic white pulp can females, and may be seen in association with
increase in incidence with age. The lymphoid thymic involution (Faccini et al 1990, Frith et al
component of the spleen can increase in number 1985, 2001, Pearse 2006b, Ward et al 1999) (Fig.
and size, and enlarged follicles often coalesce 4.14). Lymphofollicular structures with germinal
(Faccini et al 1990, Frith & Ward 1988, Frith et al centres can be seen at the corticomedullary junc-
1985, 2001, 2007, Ward et al 1999, Wijnands tion in aged mice, in association with atrophic
et al 1996). cortical changes and hyperplastic medullary
Lymphoid hyperplasia of the Peyer’s patches, changes (Berdanier 2004, Dumont & Robert
or other sites of mucosa associated lymphoid 1980, Pearse 2006b) (Fig. 4.15).
tissue (MALT) in the small and large intestine,
may occur following antigenic stimulation (Faccini
et al 1990, Maekawa et al 1996b, Shackelford &
Elwell 1999). The term is usually reserved for a
prominent increase in lymphoid tissue in an area
where MALT is not normally conspicuous (Shack-
elford & Elwell 1999).
The thymus is larger in young animals compared
to older ones, and reaches maximum size around
the time of sexual maturity. The thymus is usually
larger in females than in males. The normal, age-
associated decrease in the cellularity of the thymus
is termed involution, whereas reductions in cel-
lularity associated with stress or toxicity are
usually termed atrophy (Hardy 1967, Hummel
et al 1966, Pearse 2006b, Wijnands et al 1966).
Involution is characterized by a reduction in size
FIGURE 4.14  Lymphoid hyperplasia of the thymus in an
of the thymus, a loss of cortical lymphocytes, and 18-month-old, male, CD-1 mouse. ×40.
a loss of corticomedullary demarcation (Pearse
2006b), but the thymus does not completely invo-
lute (Faccini et al 1990, Frith & Ward 1988, Frith
et al 1985, 2007, Hardy 1967, Percy & Barthold
2007, Ward et al 1999).
Hassall’s corpuscles are very small or absent in
the mouse thymus (Pearse 2006a, Percy & Bar-
thold 2007, van Rees et al 1996, Ward et al 1999).
Thymic cysts develop from embryonic remnants
arising from the thymopharyngeal duct, or dilata-
tion of thymic tubular structures, and they
increase in number with age (Faccini et al 1990,
Frith & Ward 1988, Frith et al 1985, 2007, Pearse
2006a, b, Ward et al 1999, Wijnands et al 1966).
The cysts are lined by ciliated columnar epithe-
lium and usually contain amorphous, eosinophilic
material (Fig. 4.13).
FIGURE 4.15  Lymphofollicular structure with germinal
centre at the corticomedullary junction in the thymus of
a 20-week-old, female, CD-1 mouse. ×200.

Histopathological evaluation of the bone

FIGURE 4.12  Extramedullary hemopoiesis in the red pulp
of the spleen of a 20-week-old, female, CD-1 mouse.
×200.
Atrophy can affect the red pulp or the white
pulp and may occur spontaneously in older mice
(Faccini et al 1990, Suttie 2006, Ward et al
1999). Pigment accumulation is a common back-
ground lesion in the murine spleen. The pigment,
which is usually hemosiderin, is contained in mac-
rophages in the red pulp, but can also be present
in the white pulp, and can be observed to
FIGURE 4.13  Cyst in the thymus of an 18-month-old,
male, CD-1 mouse. The cyst contains amorphous eosi-
nophilic material. ×100.
marrow in pre-clinical toxicity studies is per-
formed on the marrow presented in conjunction
with the hematoxylin and eosin (H&E)-stained
sections of femur and sternum. A fibro-osseous
lesion involves the partial replacement of bone
marrow with an eosinophilic matrix, and is dis-
cussed in the section relating to bones and joints.
Normal bone marrow is composed of hemat-
opoietic elements including granulocytic, erythro-
cytic, megakaryocytic elements and stem cells.
Each of these may undergo hyperplasia in response
to a physiological need. The most common form
of hyperplasia in murine bone marrow is an
increase in the granulocytic elements in response
to inflammatory lesions elsewhere in the animal

(Faccini et al 1990, Frith et al 1985, 2001) (Fig.
4.16).
FIGURE 4.16  Increased granulopoiesis in the bone
marrow of the sternum of an 18-week-old, male, CD-1
mouse. ×400.
Atrophy of the bone marrow is rare in mice,
but may occur occasionally, and is characterized
by a decrease in number of all elements of the
marrow, and increased adipose tissue replacement
(Faccini et al 1990, Frith & Ward 1988, Frith et al
1985).
Respiratory system
Various reviews of the embryology, anatomy,
structure and physiology of the lungs and the
upper respiratory tract are available (Braun et al
2004, Dixon et al 1999, Harkema et al 2006,
Herbert & Leininger 1999, Kuhn 1985, Pack et al
1981, Pinkerton et al 1996, Renne et al 1992,
Reznik 1990).
The variations in the structure and distribution
of different epithelia within the upper respiratory
tract require consistent sectioning of the nasal
turbinates and larynx. This is important to ensure
consistency in evaluation of the different struc-
tures, which can have different susceptibilities to
pathological changes, particularly as a result of
inhalation exposure to test materials (Herbert &
Leininger 1999, Kittel et al 2004, Renne et al
1992, Ruehl-Fehlert et al 2003). In order to aid
in the identification of the different epithelia
within the nasal turbinates, diagrams of coronal
and sagittal sections of the nasal passages of the
rat and mouse have been produced (Mery et al
1994).
One of the most commonly encountered
changes in the nasal turbinates of the mouse is the
presence of intracytoplasmic, hyaline inclusions
(eosinophilic globules, eosinophilic secretory
inclusions, hyaline droplet accumulation) (Ber-
danier 2004, Braun et al 2004, Dungworth et al
2001, Herbert & Leininger 1999, Leininger et al
1996, Monticello et al 1990, Percy & Barthold
2007, Renne et al 2009, Ward et al 2001). These
inclusions can occur at multiple sites within the
turbinates and can affect all types of epithelium.
The brightly eosinophilic material is observed
within the cytoplasm of the mucosal epithelial
cells and of submucosal glands and ducts and
probably represents proteinaceous secretory
material accumulating in the cells (Fig. 4.17).
Eosinophilic crystals can also occur both within
the epithelium and extracellularly. These changes
occur in many strains of mice, and increase in
severity with age.
FIGURE 4.17  Intracytoplasmic hyaline inclusions in the
olfactory epithelium of the nasal turbinates of a 20-week-
old, male, CD-1 mouse. Cytoplasm of the epithelium is
packed with brightly eosinophilic material. ×200.
Another common change in the nasal turbinates
of mice, which increases in severity with age, is
the accumulation of hyaline material in the nasal
ventral septum (Leininger et al 1996, Monticello
et al 1990, Percy & Barthold). This material has
been identified as amyloid (Herbert & Leininger
1999, Percy & Barthold 2007), but the lack of
positive staining with Congo Red and positive
staining for collagen suggests to some authors that
the material is not amyloid, but a combination of
collagen and an amorphous material produced by
nasal gland epithelial cells (Brayton 2007, Doi
et al 2007, 2009, 2010) (Fig. 4.18). Doi et al
(2010) have reported sex differences in the occur-
rence of eosinophilic substance in the nasal tur-
binates of B6C3F1 mice, with males showing a
greater degree of deposition than females. Inflam-
matory changes in the turbinates occasionally
occur in association with foreign bodies, infection
or injury (Herbert & Leininger 1999) and may
be associated with dysplastic dental changes
encroaching into the nasal passages (Leininger
et al 1996).
FIGURE 4.18  Hyaline material in the nasal septum of a
two-year-old, male, CD-1 mouse. ×100.
Spontaneous, age-related changes in the larynx
are uncommon, but changes can occur commonly
from inhalation of irritant materials (Herbert &
Leininger 1999).
Foreign-body reactions are occasionally observed
in the laryngeal ventral pouch. Foreign material is
Mouse 49
seen in the lumen of the ventral pouch, associated
with an inflammatory reaction and hyperplasia of
the laryngeal epithelium. These changes are not
limited to studies utilizing the inhalation route of
exposure, but can be seen in all types of study
(Fig. 4.19). One lesion described as the only
common change associated with aging in the
larynx of B6C3F1 mice in NTP lifetime studies,
is the dilatation of laryngeal glands with the
accumulation of eosinophilic crystalline material
(Leininger et al 1996).
FIGURE 4.19  Foreign-body reaction in the ventral pouch
of the larynx of a two-year-old, male, CD-1 mouse. A
prominent inflammatory reaction to the foreign material
is seen in the lumen of the laryngeal pouch and in the
laryngeal epithelium and submucosa. Hyperplasia of the
laryngeal epithelium is also evident. ×100.
Spontaneous, age-related changes in the trachea
are uncommon, but changes can occur from inha-
lation of irritant materials (Herbert & Leininger
1999). Intracytoplasmic hyaline inclusions can be
seen in the epithelium of the trachea, bronchi and
bronchioles, with a similar appearance to those
seen in other regions of the respiratory tract
(Dungworth et al 2001, Ward et al 2001, Yang &
Campbell 1964). The incidence and severity of
this change increases with age. Eosinophilic crys-
tals can also be associated with this lesion, and
can accumulate in the lumen of tracheal glands
(Fig. 4.20).
FIGURE 4.20  Trachea: dilated tracheal glands containing
eosinophilic amorphous material and eosinophilic crys-
tals. ×200.
Pulmonary hair emboli are occasionally seen in
the lungs of mice following intravenous injection
into the caudal veins. Hair fragments in the cir­
culation are trapped in the lung vasculature,
50 BACKGROUND LESIONS IN LABORATORY ANIMALS
phagocytosed, and can be seen surrounded by
foreign-body giant cells (Ernst et al 1996, Faccini
et al 1990, Innes et al 1958, Kast 1985).
Osseous metaplasia is occasionally observed in
the lungs of mice, although at a lower incidence
than seen in rats. These small foci are usually
composed of osteoid or bone and may show early
mineralization or calcification. There is usually no
reaction in the surrounding parenchyma (Braun
et al 2004, Dixon et al 1999, Ernst et al 1996,
Faccini et al 1990) (Fig. 4.21).
FIGURE 4.21  Lungs: alveolar osseous metaplasia. Small
focus of mineralized bone in the lungs of a two-year-old,
male, CD-1 mouse. ×200.
Arterial plaque is a lung-specific change occa-
sionally seen in the lungs of mice. Developing
from the tunica media of medium to large pulmo-
nary arteries, there is a focal protrusion into the
lumen of the vessel. The lesion does not occlude
the lumen and is covered by endothelial cells.
There is no associated inflammatory reaction, and
in older lesions mineralization develops within the
plaque (Ernst et al 1996, Rehm et al 1985b) (Fig.
4.22). The walls of the major pulmonary veins of
the lungs in mice contain cardiac muscle (Best &
Heath 1961, Dixon et al 1999, Krinke 2004,
Percy & Barthold 2007) (Fig. 4.23).
FIGURE 4.22  Lungs: arterial plaque. Focal protrusion into
the lumen of a pulmonary vessel with an endothelial
covering in a 20-week-old, female, CD-1 mouse. ×400.
C57BL/6 and 129Sv strains (Guo et al 2000,
Hoenerhoff et al 2006, Ward et al 2001).
Focal accumulations of macrophages (alveolar
histiocytosis) are a common incidental finding in
the lungs (Braun et al 2004, Dixon et al 1999,
Faccini et al 1990, Frith & Ward 1988, Frith et al
2007, Renne et al 2009), and can be seen com-
monly in subpleural areas of aging mice lungs.
Focal hyperplasia of the alveolar epithelium is
occasionally seen in older mice, particularly those
with a high background incidence of bronchioloal-
veolar tumors, and the incidence increases with
age (Braun et al 2004, Dixon et al 1999, Dung-
worth et al 2001, Faccini et al 1990, Frith et al
2007). Accumulations of alveolar macrophages
FIGURE 4.23  Lungs with striated (cardiac) muscle in wall may be associated with the hyperplastic cells. The
of pulmonary vein of a two-year-old, female, CD-1 mouse. affected alveoli are lined by uniform, hypertrophic
×400. epithelium. Cytoplasmic basophilia and enlarged
nuclei are apparent. The cells form a single layer
Eosinophilic crystal accumulation in alveolar that is contiguous throughout the area of hyper-
spaces with associated inflammatory-cell infiltra- plasia and the margins of the lesion are indistinct
tion is seen occasionally in routine toxicity studies (Renne et al 2009) (Fig. 4.25).
(Dixon et al 1999, Ernst et al 1996, Frith et al
2007), and accumulations of eosinophilic material
within alveolar macrophages is associated com-
monly with bronchioloalveolar tumors in carcino-
genicity studies (Dungworth et al 2001, Frith &
Ward 1988, Green 1942). The eosinophilic mate-
rial has been considered to originate from break-
down products of granulocytes or hemoglobin
(Dixon et al 1999, Ernst et al 1996, Marshall et al
1988, Murray & Luz 1990) (Fig. 4.24).
FIGURE 4.25  Focal alveolar epithelial hyperplasia in the
lungs of a two-year-old, male, CD-1 mouse. ×200.
Gastrointestinal system
Reviews of the embryology, anatomy, histology
and physiology of the oral cavity, gastrointestinal
tract, teeth and salivary glands are available
(Berdanier 2004, Botts et al 1999, Leininger et al
1999, Long & Leininger 1999a, Shackelford &
FIGURE 4.24  Accumulations of alveolar macrophages
Elwell 1999, Tucker 2007).
containing eosinophilic material in the lungs of a two-
With the exception of abnormalities of the
year-old, male, CD-1 mouse. Such accumulations are
teeth, there are usually very few spontaneous
often associated with bronchioloalveolar tumors. ×200.
lesions of the oral cavity of the mouse (Leininger
et al 1999). Periodontal disease associated with
Eosinophilic crystalline pneumonia is an idio- the impaction of hair, food or nesting material in
pathic disease affecting certain strains and stocks the gum adjacent to molars and incisors, occurs
of mice; it is characterized by accumulations of occasionally, resulting in inflammatory infiltration
fine needle-like eosinophilic crystals in macro- of the periodontal tissues (Fig. 4.26).
phages and multinucleate giant cells within alveo-
lar and bronchiolar spaces. These are accompanied
by mixed inflammatory cell infiltrates. The disease
can range from a mild, subclinical condition to a
severe and fulminating change resulting in respira-
tory distress and death (Braun et al 2004, Hoen-
erhoff et al 2006, Ward et al 2001).The crystalline
material has been identified as being composed
primarily of Ym1 protein, a chitinase-like protein
associated with neutrophil granule products and
secreted by activated macrophages (Braun et al
2004, Guo et al 2000, Hoenerhoff et al 2006,
Renne et al 2009). This disease occurs with a high
incidence in certain strains of mice, such as the
 Mouse 51

1988, Hummel et al 1996, Krinke 2004, Maekawa

et al 1996a, Ogawa 2003, Percy & Barthold 2007,
Seely 1996b, Tucker 2007). This appears to be
under the control of testosterone, as the differ-
ence in morphology is not apparent in young
males, and castration of male mice results in a
reduced development of granular convoluted
ducts (Smith & Frommer 1975).

FIGURE 4.26  Oral cavity. Periodontal gingivitis involving FIGURE 4.28  Focal atrophy of the submandibular gland
impacted plant material in gum adjacent to tooth of a of a 20-week-old, male, CD-1 mouse. ×100.
two-year-old, male, CD-1 mouse. ×40.
Basophilic hypertrophic foci are seen occasion-
The impacted areas can develop into ulceration, ally in the parotid salivary glands (Berdanier 2004,
abscessation and the formation of periodontal Botts et al 1999, Chiu & Chen 1986, Frith et al
cysts, with disruption of the normal growth of 2007, Krinke 2004, Seely 1996b). These can
the associated dental tissue and bone (Long & occur singly or in multiple locations. There are
Leininger 1999a, Losco 1995, Percy & Barthold sharply demarcated foci of enlarged cells with
FIGURE 4.27a  Sexual dimorphism of the submandibular increased basophilia. There appears to be no asso-
2007, Sakura 1997). Periodontal abscesses can salivary glands. The granular convoluted ducts of the
develop involving the adjacent tissues of the head ciated capsule formation, compression of adjacent
male are larger and contain more prominent eosinophilic tissue or inflammatory involvement. The etiology
and may infiltrate the nasal turbinates (Leininger granules. 18-month-old, male, CD-1 mouse. ×200.
et al 1996, Percy & Barthold 2007). of this change is unclear (Fig. 4.29).
The incisors of rodents grow continuously
throughout their life, and therefore damage to the
teeth can predispose the teeth to abnormal devel-
opment and malformations, either as a result of
damage by repeated clipping of teeth, traumatic
damage, or as a result of inflammation (Long &
Leininger 1999a, Losco 1995, Sakura 1997). This
can result in dental dysplasia with disruption
of the normal growth patterns of odontoblasts
and ameloblasts, and the abnormal deposition
of mineralized dental material (Berdanier 2004,
Leininger et al 1996, Long & Leininger 1999a,
Losco 1995, Maekawa et al 1996a, Sakura 1997,
Weber 2007).
Teeth are not routinely presented for his-
topathological examination in preclinical studies, FIGURE 4.27b  Eighteen-month-old, female, CD-1 mouse FIGURE 4.29  Basophilic hypertrophic focus in the parotid
and are most commonly examined in association with less prominent eosinophilic granules. ×200. salivary gland of a two-year-old, female, CD-1 mouse. A
with sections of the nasal turbinates. As a result,
sharply demarcated focus of enlarged cells with increased
the reported incidence of more subtle changes in
Lymphocytic infiltration of the salivary glands basophilia. ×200.
the teeth of mice may be artificially lower than
is a common finding, and increases in incidence
would occur if sections of teeth were routinely
with age (Berdanier 2004, Botts et al 1999, Spontaneous lesions of the tongue are relatively
examined. Long & Herbert (2002) presented
Faccini et al 1990, Maekawa et al 1996a, Seely rare in mice. Certain strains of inbred mice
information on the occurrence of calcified bodies
1996b). Atrophy occurs occasionally in the sub- develop spontaneous calcification of the muscle
(denticles) within the dental pulp, which they
mandibular and parotid salivary glands; however, layers of the tongue at a very young age (Imaoka
considered could affect the normal growth pattern
it is uncommon in the sublingual glands (Botts et al 1986, Maekawa et al 1996a, Yamate et al
of the teeth and lead to malformations and maloc-
et al 1999, Frith et al 2007, Frith & Ward 1988, 1987), but this is not apparent in CD-1 mice.
clusion of mouse incisors.
Seely 1996b). Atrophy usually affects a single Hemorrhage and degeneration of the muscle can
Three paired salivary glands, the submandibular
lobule with reduction in size of acini and a reduc- be seen occasionally, resulting from blood sam-
(submaxillary), parotid and sublingual glands, are
tion in the diameter and eosinophilic granule pling via the sublingual vein.
closely associated and located in the subcutaneous
content of the granular convoluted ducts. An Few spontaneous lesions occur in the esopha-
tissue of the ventral neck. Salivary glands are also
apparent increase in the number of ductular ele- gus. Gavage accidents may occur rarely, resulting
located at the base of the tongue and may be
ments may be as a result of the reduced size of in rupture of the esophagus, with a consequent
presented in longitudinal sections of the tongue
the acinar component and an apparent crowding associated inflammatory reaction of the muscula-
(Ruehl-Fehlert et al 2003).
of the remaining ductular elements (Fig. 4.28). ris and serosa (Frith & Ward 1988, Leininger et al
There is a striking sexual dimorphism in the
1999) if the animal survives. Hyperkeratosis has
structure of the submandibular salivary glands
been described as the most common lesion in the
which becomes apparent once males reach sexual
esophagus of aging B6C3F1 mice (Leininger et al
maturity (Fig. 4.27a & b). The gland of the male
1999, Maekawa et al 1996a), but occurs rarely
is larger, and the granular convoluted ducts are
in CD-1 mice. Mega-esophagus is occasionally
larger and contain much more prominent eosi-
noted in older mice.
nophilic granules (Berdanier 2004, Botts et al
Minor inflammatory lesions of the forestomach
1999, Brayton 2007, Faccini et al 1990, Frith et al
are occasionally seen, often associated with focal
2007, Frith & Townsend 1985, Frith & Ward
erosions or ulcerations, and squamous hyperplasia
52 BACKGROUND LESIONS IN LABORATORY ANIMALS
and hyperkeratosis (Betton et al 2001, Faccini
et al 1990, Frith et al 2007, Leininger et al 1999,
Maekawa et al 1996a) (Fig. 4.30).
FIGURE 4.30  Focal erosion of the forestomach of a
20-week-old, male, CD-1 mouse. A minimal inflammatory
cell infiltrate is associated with the lesion. ×100.
Hyperplastic lesions of the forestomach are
usually focal in nature, but diffuse hyperplasia and
hyperkeratosis is associated with reduced food
intake in mice (Faccini et al 1990, Leininger et al
1999). The pathologist must be careful to distin-
guish hyperplasia from the thickening seen at the
limiting ridge at the junction of the non-glandular
and glandular stomach.
Developmental anomalies are rare, but ectopic
tissues have been reported in the stomach, such
as hepatocytes in the submucosa/lamina propria
near the limiting ridge, and ectopic pancreatic
tissue in the submucosa (Leininger et al 1990,
1999, Maekawa et al 1996a). Inflammatory lesions
occur less frequently in the glandular stomach.
Adenomatous hyperplasia of the glandular
stomach occurs commonly in aging CD-1 mice,
with a higher incidence in females compared to
males. This change has been variously described
as gastric hyperplasia, glandular hyperplasia,
hypertrophic gastritis, proliferative gastritis, dys-
plasia of gastric epithelium and fundic mucosal
hyperplasia (Betton et al 2001, Faccini et al 1990,
Frith & Ward 1988, Greaves & Boiziau, 1984,
Leininger et al 1999, Maekawa et al 1996a, Rehm
et al 1987). The incidence and severity of this
change is strain- and age-dependent, and is
reported to occur more frequently in densely
housed mice than in singly housed mice (Greaves
& Boiziau, 1984). There is no clinical manifesta-
tion of this change, and it is not associated with
parasitic infection, although an association with
hormonal disturbances and kidney disease has
been suggested (Rehm et al 1987).
The early features of adenomatous hyperplasia
of the glandular stomach are focal hyperplasia of
the gastric mucosa with increased epithelial
basophilia, elongation of the crypts and occasional
cystic glands. As the lesion progresses the mucosa
becomes increasingly thicker with a diffuse epi-
thelial hyperplasia, disorganization of the normal
glandular structure, prominent branching of elon-
gated crypts and cyst formation, and an accompa-
nying inflammatory infiltration in the associated
submucosa. In severe cases there is extension of
disorganized mucosa through the muscularis to
the serosa. This is not, however, considered to be
a sign of malignancy. Within the cystic glands,
hypertrophic cells containing eosinophilic inclu-
sions can be seen (Figs 4.31–4.33). The cause of
the eosinophilic intracytoplasmic inclusions in the
gastric mucosa is unknown.
FIGURE 4.31  Low power view of stomach of an
18-month-old, male, CD-1 mouse showing marked ade-
nomatous hyperplasia of the glandular stomach. There is
a severe thickening of the mucosa, with disruption of the
normal glandular architecture and lymphocytic infiltration
of the submucosa. ×10.
FIGURE 4.34  Ectopic pancreatic tissue in the submucosa
of the duodenum adjacent to the entrance of the pancre-
atic duct. 18-month-old, female, CD-1 mouse. ×100.
FIGURE 4.32  Higher-power view of Fig. 4.31 showing
cystic dilatation of the mucosal glands and extension of FIGURE 4.35  Epidermal inclusion cyst on the serosal
the disorganized mucosa through the muscularis. ×40. surface of the colon of an 18-month-old, male, CD-1
mouse. The cyst is lined by keratinized squamous epithe-
lium and the lumen is filled with keratin. ×100.
Diverticuli occur occasionally in the small and
large intestine (Frith et al 2007). These should be
examined carefully to ensure that they are not a
result of artifactual folding of the section. The
non-proliferative nature of the overlying mucosa
should allow the differentiation between diver-
ticuli and adenocarcinoma (Fig. 4.36).
FIGURE 4.33  High-power view of the disorganized
gastric mucosa, with hypertrophic cells containing eosi-
nophilic, intracytoplasmic inclusions. ×400.
Congenital lesions in the intestine of the mouse
are relatively rare, although ectopic pancreatic
tissue can occasionally be seen in the submucosa
of the duodenum (Shackelford & Elwell 1999)
(Fig. 4.34). Epidermal inclusion cysts occur occa-
sionally in the large intestine (Frith et al 2007).
These cysts occur in the muscularis of the colon
and rectum, are lined by squamous epithelium,
and contain desquamated keratin (Fig. 4.35).
FIGURE 4.36  Diverticulum of the colon of an 18-month-
old, male, CD-1 mouse. Extension of the mucosa through
to the muscularis. ×40.
The Paneth cells of the small intestine of the
mouse contain prominent zymogen granules
(Krinke 2004, Percy & Barthold 2007, Shackelford
& Elwell 1999) (Fig. 4.37). Spontaneous degen­
erative and inflammatory lesions are relatively

uncommon in the mouse intestine, and occur
more frequently in the stomach than in the intes-
tine (Frith et al 2007, Maekawa et al, 1996b,
Shackelford & Elwell 1999).
FIGURE 4.37  Prominent zymogen granules in the Paneth
cells of the ileum of a 20-week-old, male, CD-1 mouse.
×400.
Spontaneous hyperplastic lesions are also rare
in the small and large intestine, but avillous hyper-
plasia of the duodenum is seen in CD-1 mice
(Faccini et al 1990). This change is characterized
by thickening of the duodenal mucosa without
formation of villi, which occur at the first part of
the duodenum adjacent to the pyloric sphincter
(Betton et al 2001, Faccini et al 1990, Rowlatt
et al 1969). These changes are sometimes referred
to as epithelial plaques. Hyperplasia usually
involves all cells of the epithelium, including the
underlying Brunner’s glands. In more severe cases,
dilated mucosal glands can extend towards the
submucosa and be interspersed between dilated
Brunner’s glands. The overlying epithelium lacks
villous projections and can often show erosions or
ulcerations with an associated inflammatory infil-
tration. Submucosal inflammation and edema are
also present (Figs 4.38, 4.39). These changes are
considered to be reactive in nature because of the
inflammatory involvement and the hyperplasia of
more than one type of cell (Betton et al 2001,
Rowlatt et al 1969).
FIGURE 4.38  Avillous hyperplasia of the duodenum of an
18-month-old, female, CD-1 mouse. The section of duo-
denum adjacent to the pyloric sphincter (to the right in
the image) shows thickening, without formation of villi. A
submucosal inflammatory cell infiltrate is evident ×20.
FIGURE 4.39  Higher power view of Fig. 4.38 showing
hyperplastic epithelium with erosion of surface and
inflammatory cell infiltration. ×40.
Intussusception is occasionally seen in the large
and small intestine, and rectal prolapse can occa-
sionally occur. These changes are often associated
with intestinal neoplasms, parasites or other con-
ditions associated with irritation of the intestine
(Betton et al 2001, Frith & Ward 1988, Frith et al
2007, Mahesh Kumar et al 2004, Rowlatt et al
1969, Shackelford & Elwell, 1999). These condi-
tions can lead to intestinal obstruction, inflamma-
tion, necrosis and death. Rectal prolapse is
characterized by eversion of the mucosal surface
of the rectum through the anus (Fig. 4.40).
FIGURE 4.40  Prolapsed rectum of an 18-month-old,
female, CD-1 mouse. ×10.
Pinworms are occasionally seen in the lumen of
the colon (Fig. 4.41). Cross sections of the worms,
usually Syphacia obvelata or Aspiculara tetraptera,
are seen in the lumen. Inflammatory reaction in
the adjacent epithelium is not normally associated
with the presence of the parasites (Faccini et al
1990, Frith et al 2007). Syphacia obvelata infec-
tion is not associated with goblet cell hyperplasia
in the colon of mice (Marillier et al 2008).
Mouse 53
FIGURE 4.41  Adult pinworms (Syphacia obvelata) in the
lumen of the colon of an 18-month-old, male, CD-1
mouse. ×20.
Focal adipose tissue necrosis in the mesentery
is a relatively common finding in mice and rats.
These lesions are grossly visible as yellow nodules.
Histologically, areas of adipose tissue necrosis are
circumscribed by a granulomatous inflammatory
reaction, with fibrous tissue and areas of hemor-
rhage, acute inflammatory infiltration and pigment
deposition. In chronic lesions, areas of mineraliza-
tion may be present. The etiology of these lesions
may be related to focal ischemia (Maekawa et al
1996b, Shackelford & Elwell 1999).
Liver and biliary system
The embryology, anatomy, histology and physiol-
ogy of the liver and gall bladder are reviewed by
Harada et al (1999) and Thoolen et al (2010),
and the functional aspects of liver structure are
reviewed by Malarkey et al (2005).
Congenital lesions in the liver of mice are rela-
tively unusual. Hepatodiaphragmatic nodules,
formed as a result of protrusion of the median
liver lobe through the diaphragm, occur at a much
lower incidence than in rats. Ectopic tissues in the
liver are also rare, but ectopic renal tissue can
occasionally be seen in the mouse liver (Harada
et al 1999) (Fig. 4.42).
FIGURE 4.42  Ectopic renal tissue in the liver of an
18-month-old, male, CD-1 mouse. ×20.
Extramedullary hemopoiesis (EMH) is a normal
feature of the fetal mouse liver, but this function
is lost as the bone marrow takes over this function
in the adult mouse. Extramedullary hemopoiesis
can occur in the adult mouse liver as a response
to functional needs, however, for example in
response to anemia, infectious disease or neoplasia
(Faccini et al 1990, Frith & Ward 1979, 1988,
54 BACKGROUND LESIONS IN LABORATORY ANIMALS
Frith et al 2007, Harada et al 1996, 1999, Jones
1967, Thoolen et al 2010). The nature of the cel-
lular components of EHM can vary depending on
the nature of the initiating factors, but foci may
be found in sinusoids, around central veins or in
periportal areas (Fig. 4.43).
FIGURE 4.43  Small focus of extramedullary hemopoiesis
adjacent to a central vein in the liver of a one-year-old,
male, CD-1 mouse. ×400.
Some of the more striking features of the aging
mouse liver are the presence of cytomegaly, karyo-
megaly, intranuclear and intracytoplasmic inclu-
sions (Berdanier 2004, Faccini et al 1990, Frith &
Ward 1979, 1988, Frith et al 2007, Harada et al
1996, 1999, Jones 1967, Percy & Barthold 2007,
Thoolen et al 2010, Toth & Sugar 1985, Tucker
& Baker 1967, van Zweiten & Hollander 1985).
These changes increase in incidence and severity
with age, and commonly occur within the normal
aging liver. Some of these changes are also evident
in neoplastic hepatocytes.
The relative number of binucleate and multinu-
cleate hepatocytes in the liver increases with age,
either as a result of the failure of dividing cells to
separate during mitosis, or by the fusion of cells
(Wilson & Leduc 1948). The presence of very
large, hepatocellular nuclei in enlarged hepato-
cytes also increases with age, and represents
increased chromosome numbers, as a result of
failure of division of nuclei during mitosis. Poly-
ploid nuclei may contain two, four or eight times
the normal amount of nuclear DNA (Harada et al
1999, Jones 1967) (Fig. 4.44).
FIGURE 4.44  Karyomegaly and multinucleate hepato-
cytes in the liver of a ten-month-old, female, CD-1 mouse.
Variation in the size of the hepatocellular nuclei is evident,
and one hepatocyte has multiple nuclei. ×400.
Intranuclear and intracytoplasmic inclusions
are frequently observed in the aging mouse liver.
Intranuclear, eosinophilic inclusions occur com-
monly in the aging mouse liver (Fig. 4.45). Eosi-
nophilic, homogeneous material forms distinct
spherical inclusions in the nucleus, and these are
considered to be invaginations of cytoplasmic
material (Frith & Ward 1988, Frith et al 2007,
Harada et al 1999, Jones 1967, Thoolen et al
2010, Toth & Sugar 1985).
FIGURE 4.45  Intranuclear, eosinophilic inclusions in the
hepatocytes of an 18-month-old, male, CD-1 mouse. Dis-
tinct spherical inclusions of eosinophilic material are
evident in the nuclei of affected hepatocytes. ×400.
Eosinophilic inclusions are seen occasionally in
the cytoplasm of hepatocytes, the significance of
which is unclear, but they probably represent a
disturbance of protein production by the rough
endoplasmic reticulum (Frith & Ward 1988, Jones
1967, Toth & Sugar 1985). They are observed
frequently in benign hepatocellular tumors, but
they may also occur in normal liver cells (Fig.
4.46).
FIGURE 4.46  Intracytoplasmic, eosinophilic inclusions in
the hepatocytes of an 18-month-old, male, CD-1 mouse.
×400.
Intracytoplasmic accumulation of erythrocytes
is also occasionally observed as a spontaneous
change in the aging mouse liver (Harada et al
1999, Thoolen et al 2010, Tucker & Baker 1967).
The cytoplasm of the enlarged hepatocyte is seen
to contain intact erythrocytes. This is usually a
focal change and is considered to represent eryth-
rophagocytosis by the hepatocyte (Fig. 4.47).
FIGURE 4.47  Intracytoplasmic accumulation of erythro-
cytes in the liver of an 18-month-old, female, CD-1
mouse. Multiple erythrocytes are evident in the cytoplasm
of enlarged hepatocytes. ×400.
A variety of degenerative lesions are seen in the
liver of aging mice. Hepatocellular vacuolation is a
common incidental finding in the liver of aging
mice, and can occur as a focal, zonal or diffuse
change. Vacuolation is more commonly visualized
in males. The vacuoles are the result of lipid
accumulation in the cytoplasm of the hepatocyte,
and appear as clear, distinct, round vacuoles in
the cytoplasm on H&E-stained sections of liver
(Faccini et al 1990, Frith & Ward 1988, Frith et al,
2007, Harada et al 1996, 1999, Percy & Barthold
2007, Thoolen et al 2010, Tucker & Baker 1967).
Macrovesicular vacuolation is characterized by the
presence of a single large vacuole in the cytoplasm,
with displacement of the nucleus. Microvesicular
vacuolation is characterized by the accumulation
of numerous small, round vacuoles in the cyto-
plasm, with no displacement of the nucleus. The
lipid in the vacuoles is lost during routine process-
ing of formalin-fixed tissues, but O Red O staining
of frozen sections can confirm the presence of
lipid (Figs 4.48, 4.49).
FIGURE 4.48  Lipid vacuolation of the liver of an
18-month-old, male, CD-1 mouse. Microvesicular vacu-
olation, characterized by the presence of numerous,
small, round vacuoles in the cytoplasm. ×200.

FIGURE 4.49  Lipid vacuolation of the liver of an
18-month-old, male, CD-1 mouse. Hepatocytes with
macrovesicular and microvesicular vacuolation are
evident in this image. The large vacuoles present in the
cytoplasm of occasional hepatocytes displace the nuclei.
Pigment accumulation is also evident in the majority of
hepatocytes, characterized by the presence of fine,
golden-brown, granular material within the cytoplasm.
×200.
Focal necrosis is a common finding in the liver
of aging mice and can also occur sporadically in
young mice, often with no clear causative agent.
Hepatocyte necrosis can occur in a variety of pat-
terns ranging from individual hepatocytes to
extensive areas of necrosis, sometimes involving
whole liver lobes. The typical morphological fea-
tures consist of coagulative necrosis (Faccini et al
1990, Frith & Ward 1988, Frith et al 2007, Harada
et al 1996, 1999, Jones 1967, Percy & Barthold
2007, Thoolen et al 2010, Tucker & Baker 1967)
(Figs 4.50, 4.51).
FIGURE 4.50  Focal necrosis of the liver of an 18-month-
old, female, CD-1 mouse. The area of necrosis is charac-
terized by increased eosinophilia, loss of normal cellular
structure and infiltration by inflammatory cells. ×100.
FIGURE 4.51  Widespread coagulative necrosis of the
liver of a one-year-old, female, CD-1 mouse. The necrosis
is showing a clear zonal pattern of distribution. ×100.
Pigment accumulation in Kupffer cells and
hepatocytes may be encountered occasionally in
the aging mouse liver. Endogenous pigments such
as hemosiderin, lipofuscin (ceroid) and bile can
accumulate as yellow-brown deposits within the
cytoplasm of hepatocytes and phagocytic cells
(Berdanier 2004, Faccini et al 1990, Frith & Ward
1988, Frith et al 2007, Harada et al 1996, 1999,
Thoolen et al 2010) (Fig. 4.49). Hemosiderin
appears as a golden-brown pigment and can be
identified by special stains such as Perls’ stain.
Lipofuscin is slightly darker brown than hemosi-
derin and can be identified using the Schmorl’s
stain.
Inflammatory lesions are commonly seen in the
liver of mice of all ages, and low background levels
of lymphocytic infiltration in the periportal
regions, or within the sinusoids, are not unusual
(Faccini et al 1990, Harada et al 1999, Thoolen
et al 2010). Small focal granuloma (microgranu-
loma) consisting of collections of macrophages
and lymphocytes are common findings in aging
mice, and appear as individual lesions, or as mul-
tiple scattered foci. (Fig. 4.52) This lesion may be
associated with hepatocyte necrosis, because the
inflammatory areas often contain hepatocellular
debris, or surround single necrotic hepatocytes
(Frith et al 2007, Harada et al 1999). The inflam-
matory areas tend to occur more commonly in
females (Harada et al 1996). Showering of bacte-
ria from the intestine through the bloodstream
has been considered a possible cause (Frith et al
2007).
FIGURE 4.52  Microgranuloma in the liver of a 20-week-
old, male, CD-1 mouse. A focal accumulation of macro-
phages and lymphocytes. ×200.
Mouse 55
Focal angiectasis (peliosis hepatis, telangiecta-
sis) occurs occasionally in the liver of aging mice
(Bannasch et al 1985, Faccini et al 1990, Frith &
Ward 1979, Frith et al 2007, Harada et al 1996,
1999, Thoolen et al 2010). Features of this lesion
are widely dilated sinusoidal spaces containing red
blood cells and lined by normal endothelial cells.
The lesion is focal and is distinguished from sinu-
soidal dilatation by the presence of the lining
endothelium, and from hemangioma by the mor-
phology of the lining endothelium (Bannasch et al
1985, Harada et al 1996) (Fig. 4.6).
Foci of cellular alteration occur at a much lower
incidence in mice compared to rats, but they share
similar morphological characteristics, and are
described in the same terms, based on the size of
the hepatocytes as well as the tinctorial and tex-
tural qualities of the cytoplasm. As with foci of
alteration in the rat, affected hepatocytes merge
with the adjacent hepatocytes without compres-
sion. The different types of foci of alteration
described in mice include basophilic, eosinophilic,
vacuolated, clear cell, amphophilic and mixed
(Deschl et al 2001, Faccini et al 1990, Frith &
Ward 1979, 1988, Frith et al 2007, Harada et al
1996, 1999, Thoolen et al 2010).
Occasional lesions are seen in the intrahepatic
biliary system. Inflammatory infiltration in the
periportal area is relatively common, and often
accompanies biliary proliferation or fibrosis of the
portal area (Deschl et al 2001, Faccini et al 1990,
Frith & Ward 1988, Harada et al 1999).
Simple or multilocular biliary cysts are occa-
sionally seen in the aging mouse liver (Harada et al
1999) (Fig. 4.53). These cysts are lined by a
simple cuboidal or flattened epithelium, with no
evidence of proliferation.
FIGURE 4.53  Multilocular biliary cyst in the liver of an
18-month-old, male, CD-1 mouse. The cyst is lined by a
simple cuboidal and flattened epithelium. ×40.
Hypertrophic and hyperplastic changes of the
intrahepatic bile ducts, associated with accumula-
tion of intracellular eosinophilic material and the
presence of eosinophilic crystals in the lumen are
occasionally seen. (Fig. 4.54) These changes are
variously described as glandular metaplasia (Lewis
1984) or adenomatoid lesion (Harada et al 1999),
and are similar to changes seen in the gall bladder
(Seely 1996a).
56 BACKGROUND LESIONS IN LABORATORY ANIMALS
FIGURE 4.54  Hypertrophic and hyperplastic changes of
the intrahepatic bile duct of an 18-month-old, CD-1
mouse. Vacuolation and accumulation of intracellular
eosinophilic material in the biliary epithelium is present
and eosinophilic crystals are apparent in the lumen of the
duct. A prominent inflammatory cell infiltrate is associ-
ated with this change. ×200.
One of the most common changes described in
the mouse gall bladder is the presence of eosi-
nophilic material within the cytoplasm of the epi-
thelial cells (Fig. 4.55). These changes are often
associated with the presence of eosinophilic crys-
tals in the epithelium and the lumen of the gall
bladder (Berdanier 2004, Deschl et al 2001, Frith
& Ward 1988, Frith et al 2007, Harada et al 1996,
Lewis 1984, Percy & Barthold 2007, Seely 1996a,
Thoolen et al 2010, Yang & Campbell 1964).
These changes are referred to as epithelial hyali-
nosis, eosinophilic cytoplasmic change, glandular
metaplasia or adenomatoid change (Lewis 1984,
Percy & Barthold 2007, Seely 1996a, Thoolen
et al 2010, Ward et al 2001). The etiology of the
crystalline material is obscure, but is considered
to be similar in etiology to that described in the
lungs, and to be the product of the epithelium.
The crystalline material reacts to various staining
techniques in a similar way as the eosinophilic
material within the epithelium (Yang & Campbell
1964). The hyaline material in the epithelium
has been reported to be immunoreactive to
Ym1/Ym2 (Thoolen et al 2010).
FIGURE 4.55  Eosinophilic inclusions in the epithelium of
the gall bladder of a two-year-old, male, CD-1 mouse.
Eosinophilic crystals associated with this change are seen
in the epithelium and the lumen of the gall bladder. ×200.
Other changes recorded in the gall bladder,
including inflammatory infiltration in the sub­
mucosa, which may be associated with mucosal
erosion or the presence of gall stones, are extremely
rare in the mouse (Faccini et al 1990, Harada et al
1999, Seely 1996a, Thoolen et al 2010). Epithelial
hyperplasia may also occur occasionally (Deschl
et al 2001, Harada et al 1996, Lewis 1984) (Fig.
4.56).
FIGURE 4.57  Acinar atrophy of the pancreas of an
18-month-old, male, CD-1 mouse. The acini show a
reduction in size and number and in this example is
accompanied by an inflammatory cell infiltrate. ×100.
FIGURE 4.56  Focal epithelial hyperplasia of the gall
bladder of a two-year-old, female, CD-1 mouse. The epi-
thelium is hypertrophic and thrown into folds. ×200.
Pancreas
The embryology, anatomy, histology and physiol-
ogy of the exocrine pancreas are reviewed by
Eustis & Boorman (1985) and Boorman & Sills
(1999), and the developmental biology of the exo-
crine and endocrine pancreas are reviewed by
Slack (1995). An illustrated review of early pan-
creas development in the mouse, using 3D imaging
techniques, has been presented by Jørgensen
(2007).
Ectopic pancreatic tissue is occasionally seen in
FIGURE 4.58  Severe atrophy of the pancreas of an
the gastro-intestinal tract or abdominal cavity 18-month-old, male, CD-1 mouse. Extensive adipose
(Boorman & Sills 1999, Faccini et al 1990). These
tissue replacement of the acini is evident with normal
foci are composed of normal exocrine pancreatic islets embedded in the adipose tissue stroma. ×40.
tissue (Fig. 4.34).
Atrophic changes are among the most com-
Arteritis is occasionally seen in pancreatic
monly occurring degenerative changes in the aging
vessels, despite an absence of this change in other
mouse pancreas (Berdanier 2004, Boorman & Sills
tissues (Faccini et al 1990). The changes observed
1999, Enomoto et al 1996, Faccini et al 1990,
are typical of those seen in blood vessels through-
Frith & Ward 1988, Frith et al 2007). Atrophy can
out the body, with fibrinoid necrosis of vessel
be focal, lobular or diffuse. There is a reduction
walls and a florid inflammatory reaction (Fig.
in the size and number of acini, occasionally
4.59).
accompanied by an inflammatory infiltrate (Fig.
4.57). In early exocrine atrophy, the reduction in
the cytoplasmic content of the acini, and the more
basophilic appearance, can give the impression of
ductular hyperplasia. As atrophy becomes more
extensive, the acini are replaced with adipose
tissue, and in extensive atrophy, normal islets are
left embedded in an adipose tissue stroma (Fig.
4.58). The etiology of exocrine atrophy is unclear.
Focal, basophilic, tinctorial, cellular change is
occasionally seen in the pancreas (Boorman & Sills
1999, Enomoto et al 1996, Frith et al 2007).
These distinct foci are occasionally seen in long-
term studies.
FIGURE 4.59  Arteritis of a pancreatic vessel of an
18-month-old, male, CD-1 mouse. Inflammatory cell 
infiltration of the adventitia and media is apparent, with
medial hyaline degeneration and intimal proliferation.
×40.

Urinary system
The embryology, anatomy, histology and physiol-
ogy of the kidney are reviewed by Liebelt (1986)
and Seely (1999) and of the lower urinary tract
by Gaillard (1999). The normal development,
growth and aging of the urinary bladder is reviewed
by Kurata & Shibata (1996).
There is a sexual dimorphism in the structure
of the mouse kidney. Male kidneys are generally
larger and heavier than female kidneys, the renal
cortices are larger, the cells of the proximal
tubular epithelium are larger, and there is a greater
percentage of Bowman’s capsules with a cuboidal
parietal layer. In female mice, the parietal layer is
more commonly composed of flattened squamous
epithelium (Brayton 2007, Dunn 1967a, Faccini
et al 1990, Frith & Ward 1988, Frith et al 2007,
Hummel et al 1966, Krinke 2004, Liebelt 1986,
Percy & Barthold 2007, Yabuki et al 1999). These
differences can be strain-specific and are consid-
ered to be under the influence of sex hormones.
The occurrence of cuboidal epithelium is influ-
enced by gonadectomy, and as mice mature, the
number of glomeruli with cuboidal epithelium
increases (Yabuki et al 1999, 2003). In mice older
than two years the incidence decreases (Okada
et al 2005) (Fig. 4.60a & b).
FIGURE 4.60a  Sexual dimorphism in the structure of the
Bowman’s capsule of the mouse kidney. 18-month-old,
male, CD-1 mouse exhibiting a cuboidal parietal epithe-
lium. ×400.
FIGURE 4.60b  Sexual dimorphism in the structure of the
Bowman’s capsule of the mouse kidney. The parietal
layer of this 18-month-old, female, mouse is composed
of a flattened squamous epithelium. ×400.
Congenital lesions are relatively rare in mice,
although spontaneous hydronephrosis and poly-
cystic kidneys are common findings in certain
strains of mice (Dunn 1967a, Frith & Ward 1988,
Frith et al 2007, Goto et al 1984, 1985, Hsu
1986, Percy & Barthold 2007, Seely 1999, Taylor
& Fraser 1973, Wright & Lacy 1988). Hydroneph-
rosis may also occur secondarily to urinary tract
obstruction or inflammatory changes (Ninomiya
et al 1999, Seely 1999) and is characterized by
dilatation of the renal pelvis with associated papil-
lary flattening and in severe cases, compression of
the renal cortex. This change can occur unilater-
ally or bilaterally.
Cysts originating in the cortex, medulla or
papilla are occasionally observed as an isolated
finding, and are often associated with interstitial
inflammatory changes (Faccini et al 1990) (Fig.
4.61).
FIGURE 4.61  Cortical cyst in the kidney of a two-year-
old, male, CD-1 mouse. The cyst is lined by a flattened
epithelium, contains pale proteinaceous material and is
associated with inflammatory cell infiltration. ×20.
Pigment deposition is occasionally encountered
in the tubular epithelium. This pigment usually
represents accumulation of hemosiderin, originat-
ing from the breakdown of hemoglobin, or lipo-
fuscin, a pigment derived from the breakdown of
cellular components (Brown 1986). Lipofuscin is
often referred to as the ‘wear and tear pigment’
and accumulates in a number of tissues with
increasing age. Application of appropriate histo-
chemical techniques (Perls’ for hemosiderin,
Schmorl’s for lipofuscin) will allow differentiation
of these pigments (Fig. 4.62a & b).
FIGURE 4.62a  Pigment accumulation in the cortical
tubules in the kidney of a two-year-old, male, CD-1
mouse. H&E stain. ×200.
Mouse 57
FIGURE 4.62b  Pigment accumulation in the cortical
tubules in the kidney of a two-year-old, male, CD-1
mouse. Schmorl’s positive staining indicating the pres-
ence of lipofuscin. ×200.
Renal mineralization (calcification) occurs occa-
sionally in the cortical, medullary or papillary
regions, either as an incidental finding on its own,
or in association with other inflammatory or
degenerative changes (Frith et al 2007, Morrisey
1986, Seely 1999). There are strain differences
in the occurrence of mineralization and it is
more commonly seen in males than females. The
lesion occurs as basophilic, granular deposits on
H&E stained sections, and can be present within
the epithelium, tubular lumina or the interstitial
spaces.
Perivascular accumulations of lymphocytes or
plasma cells are common in the kidneys of older
mice and deposits of lymphoid cells below the
epithelium of the renal pelvis are not unusual
(Dunn 1967a, Faccini et al 1990). These aggrega-
tions are not usually associated with damage to
the surrounding parenchyma (Fig. 4.63).
FIGURE 4.63  Perivascular accumulation of lymphocytes
in the kidney of a 20-week-old, female, CD-1 mouse.
×100.
More extensive interstitial inflammatory lesions
are commonly associated with obstructive uropa-
thy, and may represent an ascending pyelonephri-
tis. Interstitial inflammatory changes can also be
associated with glomerular, tubular and interstitial
changes. It may be difficult to ascertain whether
the inflammation is a consequence of the tubular
damage or not (Montgomery 1986b). Inflamma-
tory infiltration may affect all areas of the kidney,
the nature of the inflammatory infiltration reflect-
ing the acute or chronic nature of the lesion
(Faccini et al 1990, Montgomery 1986c). Ascend-
ing pyelonephritis is usually associated with
58 BACKGROUND LESIONS IN LABORATORY ANIMALS
inflammatory and proliferative changes in the
lower urinary tract (Seely 1999). Necrosis of the
papilla is often associated with this change, and
cortical scarring can result following progression
to chronic inflammation and fibrosis, or as a
consequence of infarction (Frith et al 2007,
Montgomery 1986a, b & c).
Renal degenerative diseases occur at a relatively
low incidence in CD-1 mice. The spectrum of
changes observed have similarities to those seen
in chronic progressive nephropathy (CPN) with
features as described in aging rats (Frith & Ward
1988, Frith et al 2007, Montgomery 1986b, Percy
& Barthold 2007, Seely 1999, Tucker & Baker
1967, Wolf & Hard 1996), although these changes
tend to occur with a lower incidence and severity
than in rats. The reported incidence of these
changes in individual studies can be variable,
depending on the experience of the study patholo-
gist and their tendency to group individual find-
ings together as syndromes. The threshold applied
by the pathologist for recording CPN can depend
on the background levels of the individual compo-
nents of this change (tubular basophilia, tubular
casts, thickening of basement membrane etc.) and
whether treatment has led to an exacerbation or
reduction of this common, age-related pathologi-
cal lesion.
Glomerulonephritis (glomerulopathy, glomeru-
losclerosis, hyaline glomerulopathy, hyaline
glomerulonephropathy, hyalinization of glomeruli,
membrano-proliferative glomerulonephritis) is a
specific degenerative change, associated with the
glomeruli, which occurs at a relatively low inci-
dence in aging mice. The incidence and severity
of the changes seen are strain-, sex- and age-
dependent, occurring more commonly in females
than in males. (Dunn 1967a, Eaton et al 1980,
Faccini et al 1990, Frith & Ward 1988, Frith
et al 2007, Hoedemaeker et al 1986, Percy &
Barthold 2007, Russell & Meier 1966, Sass 1986,
Seely 1999, Tucker & Baker 1967, Maita et al
1988, Wojcinski et al 1991, Wolf & Hard 1996).
This change is characterized in the initial stages
by mild endothelial and mesangial proliferation
and the deposition of an amorphous, eosinophilic
material in the glomeruli. This material has the
appearance of amyloid on H&E sections, but
is positive with the periodic acid–Schiff (PAS)
stain, and stains negatively with Congo Red. This
material is initially deposited in the mesangium
and basement membrane and progresses to affect
the whole glomerular structure. As the disease
progresses it can be accompanied by tubular and
interstitial changes, with tubular basophilia and
dilatation, cast formation and interstitial inflam-
matory infiltration. The glomeruli become scle-
rotic and dilatation of Bowman’s capsule can
occur. The etiology of this change is complex,
but an immune-mediated mechanism has been
suggested (Hoedemaeker et al 1986, Wojcinski
et al 1991) (Fig. 4.64a).
bladder of the mouse (Frith & Ward 1988, Frith
et al 2007, Gaillard 1999, Krinke 2004). Their
incidence and severity increases with age (Fig.
4.65). Urothelial hyperplasia is an uncommon
spontaneous change, but it can be associated
with calculi or inflammatory infiltration resulting
from obstructive uropathy or ascending infections
(Faccini et al 1990, Frith & Ward 1988, Frith et al
2007, Gaillard 1999) (Fig. 4.66). Calculi are
unusual spontaneous lesions in the bladder of the
mouse (Frith et al 2007, Gaillard 1999).
FIGURE 4.64a  Glomerulosclerosis of the kidney of a ten-
month-old, female, CD-1 mouse. Marked accumulation of
eosinophilic material within the glomeruli is associated
with tubular degenerative changes, interstitial
inflammatory-cell infiltration and the presence of pigment.
The material within the glomeruli stains positively with
PAS, but negatively with Congo Red. ×200.
Although these changes occur at a low inci-
dence, kidney disease is considered to be one of
the main non-neoplastic findings associated with
the early death of mice in long-term studies (Ettlin
et al 1994, Maita et al 1988, Son 2003a, b). Spon- FIGURE 4.65  Submucosal lymphoid infiltrate in the
taneous hyperplastic lesions in the kidney are rare, bladder of a 20-week-old, male, CD-1 mouse. ×200.
but they can be seen in association with inflam-
matory changes in the urinary tract (Hard et al
2001). Occasionally, intranuclear, eosinophilic
inclusion bodies are observed in the renal cortical
epithelium of severe combined immunodeficiency
(SCID) mice (Baze et al. 2006). The tubular
epithelial cells demonstrates large, karyomegalic
nuclei which contain intranuclear inclusions and
marginated chromatin. These cells are randomly
present in the cortex and medulla but may be
more prominent near the corticomedullary junc-
tion (Baze et al. 2006) (Fig. 4.64b).
FIGURE 4.66  Urothelial hyperplasia associated with sub-
mucosal lymphocytic and plasma cell infiltration in the
urinary bladder of a 10-week-old, female, CD-1 mouse.
×100.
Refluxed seminal colloid plugs in the bladder
and urethral plugs (copulatory plugs) occur occa-
sionally in mice and are considered to be an agonal
change (Bendele & Carlton 1986, Gaillard 1999,
Percy & Barthold 2007). Colloid plugs have,
however, been considered to affect urine flow in
FIGURE 4.64b  Large, karyomegalic, renal, cortical, sexually mature males (Taylor 1985) and have
tubular, epithelial nuclei with intranuclear inclusions and been implicated in the pathogenesis of obstructive
marginated chromatin of severe combined immunodefi- uropathy (Bendele & Carlton 1986, Gaillard
ciency (SCID) mouse. ×400. 1999, Maita et al 1988, Ninomiya et al 1999,
Tucker & Baker 1967). The plug is composed of
The ureter is a simple tubular organ lined by eosinophilic proteinaceous material within which
transitional epithelium (Frith et al 1986, Gaillard spermatozoa can be seen (Fig. 4.67).
1999). There are very few lesions which occur
spontaneously in the ureter of the mouse.
Hydroureter may be associated with congenital
hydronephrosis (Seely 1999). Transitional cell
hyperplasia and inflammatory cell infiltration may
be observed in association with similar lesions in
the kidneys or bladder.
Lymphoid follicles in the lamina propria are
a common background change in the urinary

FIGURE 4.67  Refluxed seminal colloid plug in the urinary
bladder of a 20-week-old, male, CD-1 mouse. Amorphous
eosinophilic material in the lumen of the bladder. ×40.
Obstructive uropathy (urologic syndrome) is
considered an important factor in the early deaths
of male mice on long-term studies (Bendele &
Carlton 1986, Maita et al 1988, Son 2003a,b).
This change is associated with gang housing of
male mice in long-term studies and is related to
fighting and genital wounds (Faccini et al 1990,
Gaillard 1999, Seely 1999, Tucker & Baker 1967),
although this change is also seen in singly housed
mice (Bendele & Carlton 1986). The incidence of
this change is also reported to vary with the source
of animals in long-term studies (Engelhardt 1996).
Features of this change include the marked dis-
tension of the urinary bladder, often with colloid
plugs in the neck of the bladder blocking the flow
of urine (Gaillard 1999). Inflammatory, degenera-
tive and proliferative changes of the urinary tract
are associated with obstructive uropathy as is
hydronephrosis of the kidneys.
Endocrine glands
The embryology, anatomy, histology and physiol-
ogy of the pituitary have been reviewed by Capen
(1983a) and Mahler & Elwell (1999). The pitui-
tary gland of the female mouse is reported to be
consistently heavier than that of the male (Chai
& Dickie 1966, Hummel et al 1966).
Cysts are noted commonly in the pars distalis
or at the cleft dividing the pars distalis and pars
intermedia. They occur frequently as an incidental
finding and develop from remnants of the crani-
opharyngeal pouch or Rathke’s cleft, and are
usually lined by ciliated epithelium and contain
eosinophilic material (Capen 1983a, Carlton &
Gries 1983, Faccini et al 1990, Frith & Ward
1988, Frith et al 2007, Hummel et al 1966,
Mahler & Elwell 1999, Morton & Tekeli 1997,
Percy & Barthold 2007, Russfield 1967) (Fig.
4.68a & b).
Mouse 59
FIGURE 4.68a  Multiple developmental cysts in the pars
distalis of an 18-month-old, male, CD-1 mouse. ×40.
FIGURE 4.69a  X zone of the adrenal of an eight-week-
old, female, CD-1 mouse. A distinctive layer of smaller,
basophilic cells between the zona fasciculata and the
medulla is apparent. ×40.
FIGURE 4.68b  High-power view of one of the cysts in Fig.
4.68a showing ciliated lining epithelium and amorphous
eosinophilic material in the lumen. ×400.
Focal hyperplasia may be present occasionally,
particularly in the pars distalis, but at a much
lower incidence than is reported in the rat. The
lesion is focal, but the periphery of the lesion is
poorly demarcated (Berdanier 2004, Capen et al FIGURE 4.69b  Pronounced vacuolation of the X zone of
2001, Faccini et al 1990, Frith & Ward 1988, Frith a 20-week-old, female, CD-1 mouse. ×40.
et al 2007, Mahler & Elwell 1999).
The embryology, anatomy, histology and physi- Accessory cortical tissue is frequently seen in
ology of the adrenal glands have been reviewed by close association with the adrenal capsule and
several authors (Dunn 1970, Frith 1983a, Nyska occurs more commonly in females than males. It
& Maronpot 1999, Rosol et al 2001, Sass 1983a, is composed of normal cortical tissue, in which the
Tischler & Sheldon 1996, Waring 1935, Yarrington zona glomerulosa and fasciculata can often be dis-
1996). tinguished. Medullary tissue is not associated nor-
Sexual dimorphism occurs in the structure of mally with accessory adrenal tissue and the
the mouse adrenal gland. The adrenal glands of accessory cortical tissue can undergo the same
male mice contain less adipocytes, are smaller aging changes as the main body of the adrenal
than female glands, and there are sex differences (Dunn 1970, Faccini et al 1990, Frith & Ward
in the appearance and regression of the transient 1988, Frith et al 2007, Krinke 2004, Nyska &
X zone. There is no discernible zona reticularis in Maronpot 1999, Percy & Barthold 2007, Sass
the mouse adrenal, but in young mice there is an 1983b, Waring & Scott 1937) (Fig. 4.70).
X zone. The X zone is composed of small cells
with distinctly basophilic cytoplasm which devel-
ops postnatally in the inner cortex of mouse
adrenals, being fully formed at weaning. After
weaning it degenerates rapidly in males and disap-
pears by puberty. In pregnant females, the X zone
undergoes vacuolar degeneration during the first
pregnancy. In virgin females, the X zone persists
much longer and increases in size. The X zone
undergoes slow regression and degeneration
during which it develops prominent vacuolation in
females. There are strain differences in the rate of
degeneration of the X zone (Brayton 2007, Chai
& Dickie 1966, Dunn 1970, Faccini et al 1990,
Frith 1983a, Frith & Ward 1988, Frith et al 2007,
Jones 1950, Krinke 2004, Nyska & Maronpot
1999, Percy & Barthold 2007, Rosol et al 2001,
Sass 1983a, Tanaka & Matsuzawa 1995, Waring
1935) (Fig. 4.69a & b).
60 BACKGROUND LESIONS IN LABORATORY ANIMALS
FIGURE 4.70  Accessory cortical tissue of the adrenal of
a 20-week-old, female, CD-1 mouse. The accessory
tissue is clearly demarcated from the adrenal gland and
is exhibiting vacuolation of the X zone. No medullary
tissue is apparent in the accessory tissue. ×100.
Lipogenic pigmentation (brown degeneration,
ceroid pigment, lipofuscin) is an age-related
change which occurs spontaneously in several
stocks and strains of mice, including the CD-1
mouse, and takes the form of pigment deposition
in adrenal cortical cells and macrophages at the
corticomedullary junction. This change is much
more common in females than in males. The cyto-
plasm of the affected cells becomes distended,
brown and foamy and the cells resemble macro-
phages. The pigment in the cells is PAS-positive
(Chai & Dickie 1966, Dunn 1970, Faccini et al
1990, Frith 1983c, Frith & Ward 1988, Frith et al
2007, Hummel et al 1966, Jones 1950, Nyska &
Maronpot 1999, Rosol et al 2001, Yarrington
1996). The appearance of lipogenic pigmentation
is linked to sex hormones (Bernichtein et al 2009)
(Fig. 4.71a & b).
FIGURE 4.71a  Lipogenic pigmentation in the adrenal of
an 18-month-old, female, CD-1 mouse. Distended cells
containing brown, foamy cytoplasm accumulate in the
corticomedullary region. H&E ×200.
FIGURE 4.71b  Periodic acid–Schiff staining of the
adrenal in Fig. 4.71a showing the material in the cells to
be PAS-positive. ×200.
Subcapsular cell hyperplasia (spindle cell hyper-
plasia) is a common, age-related finding in the
cortex of mice of a number of stocks and strains,
including the CD-1 mouse. It occurs rarely in
mice below three months of age. The number of
subcapsular cells increases with age, and occurs at
a greater incidence and severity in females. The
proliferation of subcapsular cells may be focal or
diffuse, and can eventually replace large portions
of the cortex and may ultimately develop into
subcapsular cell tumors. Two types of subcapsular
cells may be identified. Oval to fusiform cells with
scant basophilic cytoplasm (type A cells), and
large, round cells with abundant eosinophilic or
vacuolated cytoplasm (type B cells). Type A cells
predominate in the earlier lesions, but as the
lesions grow larger more type B cells are involved.
The function of the spindle cells is unknown,
but they appear to represent a morphological
variant of epithelial cells in the subcapsular region
(Berdanier 2004, Brayton 2007, Capen et al
2001, Chai & Dickie 1966, Dunn 1970, Faccini
et al 1990, Frith et al 2007, Goodman 1983,
Hummel et al 1966, Kim et al 1997a, 2000,
Krinke 2004, Nyska & Maronpot 1999, Percy &
Barthold 2007, Rosol et al 2001, Yarrington 1996)
(Fig. 4.72a & b).
FIGURE 4.72a  Early subcapsular cell hyperplasia of the
adrenal of a 20-week-old, female, CD-1 mouse. The
basophilic, fusiform (type A) cells extend from the sub-
capsular region towards the medulla. ×200.
FIGURE 4.72b  Subcapsular cell hyperplasia of the
adrenal of an 18-month-old, male, CD-1 mouse. Large,
round cells with abundant vacuolated cytoplasm (type B)
predominate in this lesion. ×200.
There are strain differences in the occurrence
and severity of subcapsular cell hyperplasia, and
also an association with the infiltration of mast
cells. Some authors suggest that mast cells play a
role in the development of subcapsular cell hyper-
plasia (Kim et al 1997a, b, 2000). Sex hormones
play a role in the development of this change, as
gonadectomy of males leads to an increased inci-
dence of subcapsular cell hyperplasia (Bernichtein
et al 2009). Focal hypertrophy/hyperplasia of the
cells of the zona fasciculata is seen occasionally.
The affected cells are enlarged with eosinophilic
cytoplasm (Capen et al 2001, Faccini et al 1990,
Frith et al 2007, Nyska & Maronpot 1999) (Fig.
4.73).
FIGURE 4.73  Focal cortical hypertrophy of the adrenal of
a two-year-old, male, CD-1 mouse. A discrete focus of
enlarged cells with eosinophilic cytoplasm is contained
within the zona fasciculata. ×100.
Proliferative lesions of the adrenal medulla
occur at a much lower incidence than is seen in
rats, but give rise to a continuous spectrum of
lesions from hyperplasia to malignant phaeochro-
mocytoma. Hyperplasia of medullary cells can
occur as focal or diffuse lesions. Focal hyperplasia
can be recognized by an increased basophilia of
the cytoplasm and an increased size of the affected
cells. Diffuse changes involve an increase in the
number and volume of all the medullary cells.
(Capen et al 2001, Faccini et al 1990, Frith
1983b, Frith & Ward 1988, Frith et al 2007,
Nyska & Maronpot 1999, Tischler & Sheldon
1996).
The embryology, anatomy, histology and physi-
ology of the thyroid and parathyroid glands have

been reviewed by several authors (Capen 1983b,
Capen et al 1996, Hardisty & Boorman 1999,
Pour et al 1983a, Thomas & Williams 1996).
Ectopic thyroid tissue can occasionally be found
in the adipose tissue at the base of the heart (Frith
1983d, Frith & Ward 1988), and ectopic parathy-
roid tissue can occasionally be found in the septa
or surface connective tissue of the thymus (Capen
et al 1996, Frith & Fetters 1983, Frith & Ward
1988). Embryonic thymic remnants can give rise
to ectopic thymic tissues in the thyroids or par-
athyroids (Capen et al 1996, Faccini et al 1990,
Frith & Ward 1988, Frith et al 1985, 2007, Krinke
2004, Hardisty & Boorman 1999, Pearse 2006a,
Percy & Barthold 2007, Pour et al 1983b, Ward
et al 1999). This is a common finding in CD-1
mice of all ages (Fig. 4.74).
FIGURE 4.74  Ectopic thymic tissue associated with the
parathyroid gland of a 20-week-old, female, CD-1 mouse.
×100.
The most common, age-related, non-neoplastic
finding in the thyroid of the mouse is cystic dilata-
tion of ultimobranchial bodies (Frith & Ward
1988, Frith et al 2007). In some strains of mouse
this can occur in 90% of animals (Rehm et al
1985a). Cysts occur occasionally in or around the
thyroid and parathyroid. They can be lined by
ciliated, cuboidal epithelium, or squamous epithe-
lial cells, and are considered remnants of the
craniopharyngeal duct or ultimobranchial duct
respectively (Capen 1983b, Capen et al 1996,
Faccini et al 1990, Frith & Ward 1988, Hardisty
& Boorman 1999, Percy & Barthold 2007, Pour
et al 1983b) (Fig. 4.75).
FIGURE 4.75  Ultimobranchial cyst of the parathyroid of a
eight-week-old, male, CD-1 mouse. The cyst contains
eosinophilic flocculent material and is lined by a simple
squamous and ciliated cuboidal epithelium. ×100.
Cystic dilatation of follicles can occasionally be
seen, with individual follicles becoming enlarged
and distended with colloid (Frith et al 2007) (Fig.
4.76). Crystals are occasionally visualized in the
lumen of thyroid follicles, which have been identi-
fied as calcium oxalate in humans (Frith & Ward
1988) (Fig. 4.77a & b). Unlike rats, C-cells are
not prominent in mice, and hyperplasia does not
occur as a common aging change (DeLellis et al
1996, Hardisty & Boorman 1999).
FIGURE 4.76  Cystic dilatation of the follicles of a two-
year-old, male, CD-1 mouse. Individual follicles are  Barthold 2007) (Fig. 4.78a & b).
distended with colloid. ×100.
FIGURE 4.77a  Crystals in the lumen of a thyroid follicle
of an 18-month-old, male, CD-1 mouse. ×400.
FIGURE 4.77b  Polarized image of the follicle in Fig.
4.77a, showing the birefringence of the crystals. ×400.
Hyperplastic changes in the thyroid and par-
athyroid of mice are uncommon (Hardisty &
Boorman 1999). Hyperplasia of the parathyroid
can occur secondarily to chronic renal disease, but
this occurs in mice much less frequently than it is
observed in rats (Capen 1983b, Capen et al 1996,
Mouse 61
Frith & Ward 1988, Hardisty & Boorman 1999,
Capen et al 2001).
The embryology, anatomy, histology and physi-
ology of the exocrine and endocrine pancreas are
reviewed by Boorman & Sills (1999), and the
developmental biology of the exocrine and endo-
crine pancreas are reviewed by Slack (1995). An
illustrated review of early pancreas development
in the mouse using 3D imaging techniques has
been presented by Jørgensen (2007).
The most common spontaneous, non-neoplastic
change noted in the pancreatic islets of the mouse
is islet-cell hyperplasia. There are sex and strain
differences in the occurrence of this change, but
it is seen as an age-related change in many differ-
ent strains and affects males more than females.
The change can involve single or multiple islets,
and the islets are usually rounded, but coalescence
with adjacent islets can result in an irregular
outline. Affected islets are much larger than
normal because of an increased number of cells,
but the cells are morphologically similar to normal
islets. It is not unusual to see a central cystic cavity
in affected islets (Berdanier 2004, Boorman &
Sills 1999, Capen et al 2001, Faccini et al 1990,
Frith & Sheldon 1983, Frith & Ward 1988, Frith
et al 2007, Leiter & Herberg 1996, Percy &
FIGURE 4.78a  Islet cell hyperplasia of the pancreas of an
18-month-old, male, CD-1 mouse. Multiple, enlarged,
coalescing islets, morphologically similar to normal islets.
×40.
FIGURE 4.78b  A hyperplastic islet with a central cystic
cavity. ×100.
Skin and appendages
Various reviews of the embryology, anatomy, his-
tology and physiology of the skin, sebaceous
glands and the mammary gland are available
62 BACKGROUND LESIONS IN LABORATORY ANIMALS

(Peckham & Heider 1999, Seely & Boorman

1999, Sundberg 2004, Sundberg et al 1996,
Thody & Shuster 1989).
The thickness of the skin of the mouse varies
with the area of the body, and also with the stage
of the hair growth cycle (Peckham & Heider 1999,
Sundberg 2004). In the mouse, following the
emergence of the first coat, hair growth occurs in
cycles, with waves of hair growth and follicular
activity occurring in a distinctive pattern along the
body of the animal (Hummel et al 1966, Stenn &
Paus 2001, Sundberg 2004, Sundberg et al 1996).
There are several reviews of hair structure and
growth patterns which allow the pathologist to
easily identify and classify the different stages of FIGURE 4.79  Focal epidermal ulceration of a 20-week-
the hair growth cycle (Hardy 1952, Müller-Röver old, male, CD-1 mouse. Epidermal hyperplasia, hyperk- FIGURE 4.81  Subcutaneous abscess of the submandibu-
et al 2001, Paus et al 1999, Stenn & Paus 2001). eratosis and parakeratosis of the adjacent skin, and a lar region in a 15-month-old, male, CD-1 mouse. Multiple
Loss of hair associated with excessive grooming prominent inflammatory cell infiltration of the ulcerated abscesses displaying Splendore–Hoeppli material are
activity can be seen occasionally. There are sex and area are visible. ×100. evident. ×20.
strain differences in the occurrence of this phe-
nomenon which is often associated with multiple Epidermal inclusion cysts occur occasionally in Congenital lesions are not commonly encoun-
housed animals (Faccini et al 1990, Militzer & mice, although not as commonly as in rats (Faccini tered in the mammary tissue of mice, and the
Wecker 1986). Whisker trimming and barbering et al 1990, Peckham & Heider 1999). These cysts most common age-related lesions in mouse
can also occur as a manifestation of social domi- are lined by squamous epithelium, with desqua- mammary tissues are mammary tumors (Rehm &
nance in some strains of mice (Strozik & Festing mated keratin in the lumen. (Fig. 4.80) They are Leibelt 1996, Seely & Boorman 1999). Other than
1981). considered to be the result of damage to the neoplasms, duct ectasia and hyperplasia of the
Spontaneous degenerative changes are uncom- pilosebaceous unit. Branchial cysts in the superfi- mammary tissue are frequently observed in aging
mon in the skin of mice, and the most common cial tissues of the ventral neck have also been mice (Bruner et al 2001, Faccini et al 1990)
changes observed are related to injuries associated described as an incidental finding in some strains although they occur at a lower frequency than in
with fighting in group-housed, male mice (Faccini of mice. These are lined by cuboidal to columnar, aged rats. Mammary hyperplasia may be focal or
et al 1990, Frith et al 2007, Peckham & Heider non-ciliated epithelium, and are considered to diffuse, and may be lobular or acinar. The change
1999, Tucker & Baker 1967). Similar lesions have develop from embryonic rests (France et al 2000). is characterized by an increased number of normal
also been related to stress, with different strains looking alveoli and ducts with little or no pleomor-
of mice showing different sensitivities (Koopman phism, compression or encapsulation (Fig. 4.82).
et al 1984).
The level of fighting injuries in some mice can
be severe enough to lead to increased mortality in
groups of gang-housed, male mice (Son 2003a, b,
Tucker & Baker 1967). There have been several
studies into the appropriate levels of group
housing and environmental enrichment necessary
to reduce stress and fighting among groups of male
mice, and strain differences in the levels of aggres-
sion shown by male mice have been demonstrated
(Kaliste et al 2006, Van Loo et al 2003, 2004).
Lesions associated with fighting generally occur in
a typical pattern, involving the head, ears, extrem-
ities and perigenital regions (Peckham & Heider FIGURE 4.80  Epidermal inclusion cyst of the skin of the
1999, Son 2003a,b, Tucker & Baker 1967). tail in a 20-week-old, female, CD-1 mouse. The cyst is
The pattern of changes seen histologically lined by a thin, squamous epithelium, and the lumen is FIGURE 4.82  Focal hyperplasia of the mammary glands
reflects the severity and duration of the damage filled with desquamated keratin. ×100. of an 18-month-old, female, CD-1 mouse. ×100.
to the skin and include epidermal erosions or
ulcerations associated with inflammatory cell infil- Occasional mice in long-term studies develop The Zymbal’s gland is a modified sebaceous
tration and squamous hyperplasia of adjacent skin swelling in the submandibular region of such a size gland associated with the auditory meatus. This
(Bruner et al 2001, Frith et al 2007) (Fig. 4.79). that it leads to the early sacrifice of the animal. On gland is rarely examined during routine gross
Inflammatory changes in the skin may also be histopathological examination these swellings are observations and is not commonly presented for
secondary to the presence of mites or immune identified as subcutaneous abscesses (Clarke et al histopathological examination. These glands are
complex-mediated vasculitis (Andrews et al 1978, Son 2003a, b). These can be the result of best examined in transverse sections of the head
1994, Frith & Ward 1988). Similar degenerative opportunistic infection of incisions on the skin, or through the level of the external ears (Ruehl-
and inflammatory changes in the ears have been through the oral mucosa by Staphylococcus aureus. Fehlert et al 2003, Seely & Boorman 1999).
associated with excessive grooming or the pres- The lesions typically contain multiple abscesses Degenerative lesions, other than ductal dilatation
ence of metal ear tags (Bell et al 1970, Kitagaki & demonstrating Splendore–Hoeppli material, with and cyst formation, are uncommon (Seely &
Hirota 2007). the formation of eosinophilic material surrounding Boorman 1999).
the microorganisms in the abscesses (Fig. 4.81).
Muscles, bones and joints
A general discussion on the embryology, anatomy,
histology and physiology of the bones and joints
can be found in Long & Leininger (1999b).
The femur, including the joint and the sternum,
are the two most commonly sampled sites in

routine toxicity testing. The section of long bone
is routinely sectioned longitudinally to include the
femur, knee joint and tibia. This section will allow
evaluation of the bone, bone marrow, growth
plates and articular surfaces. The sternum is rou-
tinely sectioned longitudinally, allowing examina-
tion of two or three sternebrae and the associated
bone marrow (Morawietz et al 2004).
One of the more common degenerative changes
of the cartilage is chondromucinous degeneration
of the cartilage of the sternum (Fig. 4.83). This
has been described as a common finding in certain
strains of rat (Long et al 1996), but occurs at a
relatively high incidence in older mice (Long &
Leininger 1999b). This change is characterized by
increased eosinophilia, loss of chondrocytes and,
ultimately, necrosis of the cartilage and the forma-
tion of cystic cavities within the sternebral joints.
This change increases in incidence and severity
with age and occurs commonly in both sexes.
FIGURE 4.83  Chondromucinous degeneration of the
sternum of a 20-month-old, male, CD-1 mouse. Loss of
normal chondrocytes in sternebral joint with central area
of necrosis. ×100.
Degenerative joint disease, also described as
osteoarthropathy, osteoarthritis, and osteoarthro-
sis, is a non-inflammatory, progressive, degenera-
tive disorder affecting the joints which shows an
increase in incidence and severity with age (Faccini
et al 1990, Long & Leininger 1999b, Russell &
Meier 1966, Sokoloff 1967, Wancket et al 2008,
Yamamoto & Iwase 1998, Yamamoto et al 1999).
There are strain differences in the occurrence of
this disorder and also sex differences, with males
being more susceptible than females (Long &
Leininger 1999b, Sokoloff 1967, Walton 1978,
Wancket et al 2008, Yamamoto & Iwase 1998,
Yamamoto et al 1999). The knee joint is most
commonly affected, indicating that mechanical
trauma may be a factor in the etiology (Russell &
Meier 1966), although an age-related loss of joint
innervation is also considered to play a role in the
development of this disorder (Salo et al 2002).
Features of this lesion include destruction of the
articular cartilage with cartilage regeneration and
hyperplasia, formation of subchondral cysts and
ossification of the joint capsule in more severe
cases (Fig. 4.84).
FIGURE 4.84  Femur, knee joint and tibia. Degenerative
joint disease in a two-year-old, female, CD-1 mouse.
Disruption of the normal joint structure, involving cartilage
hyperplasia, remodelling of the articular surfaces, forma-
tion of bony bridges between the bones of the joint, and
formation of cyst-like structures at the surface of the
joints. A fibro-osseous lesion can also be seen in the
marrow space of the tibia, characterized by replacement
of the marrow space with eosinophilic material and
spindle-shaped cells. ×20.
Fibro-osseous lesion (myelofibrosis, osteofibro-
sis, osteosclerosis) occurs as a spontaneous and
induced change in the marrow cavity of sternebrae
and long bones (Albassam & Courtney 1996,
Berdanier 2004, Faccini 1990, Frith & Ward 1988,
Frith et al 2007, Percy & Barthold 2007, Travlos
2006b). There are strain and sex differences in the
occurrence of fibro-osseous lesion, with females
showing a higher incidence and severity (Albassam
et al 1991, Long & Leininger 1999b, Ritting-
hausen et al 1997, Wancket et al 2008) suggesting
that sex hormones play a role in the etiology.
In certain strains this lesion is accompanied
by reproductive tract lesions, including ovarian
atrophy and uterine cervical dysplasia (Wancket
et al 2008). This change is characterized by the
replacement of the marrow by an eosinophilic
matrix, including fibroblast-like cells and osteo-
clasts (Frith & Ward 1988, Long & Leininger
1999b) (Fig. 4.84).
This lesion should be distinguished from fibrous
osteodystrophy, a metabolic bone disease associ-
ated with disturbances of calcium homeostasis.
Fibrous osteodystrophy involves extensive remod-
elling of bones resulting from excessive secretion
of parathyroid hormone. Commonly seen in end-
stage renal disease, this lesion is usually accompa-
nied by hypertrophy of the parathyroids and
metastatic mineralization in the soft tissues and
vasculature throughout the body. This change is
more commonly seen in rats than mice (Long et al
1996, Long & Leininger 1999b, Travlos 2006b).
Hypertrophy of the bone (hyperostosis) is occa-
sionally seen in bones and joints of mice, charac-
terized by increased bone mass affecting the
periosteum, endosteum or trabecular bone (Long
& Leininger 1999b) (Fig. 4.85).
Mouse 63
FIGURE 4.85  Hyperostosis of the epiphyseal region of
bone from a two-year-old, female, CD-1 mouse. ×40.
Synovial hyperplasia of the joints also occurs
sporadically, usually accompanying inflammatory
or degenerative joint changes (Long & Leininger
1999b) (Fig. 4.86a).
FIGURE 4.86a  Synovial hyperplasia of the knee joint of a
two-year-old, female, CD-1 mouse. Proliferation of the
synovium associated with inflammatory cell infiltration in
the joint and periosteum. ×100.
A review of the embryology, anatomy, histology
and physiology of skeletal muscle can be found in
Leininger (1999).
Spontaneous changes in the skeletal muscle of
mice are rare. There are certain inbred strains of
mice which show spontaneous degenerative
changes or calcification at an early age (Leininger
1999, Sokoloff 1967), but these are not routinely
used in toxicity testing. Congenital lesions of the
muscle in CD-1 mice are rare, although in chronic
studies, small foci of inflammation, degeneration,
necrosis and regeneration may occasionally occur
spontaneously (Novilla & Smith 1996) and dys-
trophic or metastatic mineralization may occa-
sionally be seen (Leininger 1999).
Inflammatory processes in the skin or other
tissues may sometimes involve the adjacent
muscle (Faccini et al 1990), and degenerative and
inflammatory changes are a common response to
the injection of saline or vehicle in studies involv-
ing intramuscular administration of test materials
(Leininger 1999, Thuilliez et al 2009). Atrophic
changes may also occur secondary to denervation
as a result of traumatic damage to the associated
nerve fibers (Leininger 1999, Van Vleet & Ferrans
1991b). The muscle spindle is a normal structure
in skeletal muscle, responsible for proprioception
(Fig. 4.86b).
64 BACKGROUND LESIONS IN LABORATORY ANIMALS
FIGURE 4.86b  The nuclear bag fiber of the mouse
muscle spindle. ×200.
Brain and nervous system
A review of the embryology, anatomy, histology
and physiology of the central and peripheral
nervous system can be found in Radovsky &
Mahler (1999).
Due to the small size of the mouse brain, and
the relatively few sections examined in routine
toxicity studies, standardization of the trimming
and sectioning of the brain is important to ensure
that homologous sections are examined for all
animals in a particular study. The mouse brain is
also very susceptible to handling artifacts and
should be processed with care (Morawietz et al
2004, Radovsky & Mahler 1999). The brain is a
highly complex structure and a relatively small
proportion of the tissue is examined in the three
or four coronal sections prepared for routine his-
topathological examination in pre-clinical studies.
If the reader is required to investigate potential
changes in particular structures within the central
nervous system, the use of a brain atlas is recom-
mended to identify the position and extent of the
different regions of the mouse brain. Such maps
are now available on-line and give the pathologist
the opportunity to compare the histological sec-
tions for individual animals with high-resolution
images of step-serial sections through the mouse
brain.
One such source of information is the High
Resolution Mouse Brain Atlas (Sidman et al,
available electronically at http://www.hms.
harvard.edu/research/brain/index.html) which
provides step-serial, coronal sections stained alter-
natively with myelin and Nissl substance stains,
and labelled with all the major grey and white
matter structures.
An alternative source of images is provided by
BrainMaps.org at: (http://brainmaps.org/index.
php?p=speciesdata&species=mus-musculus), a
site which presents images of the brains of various
species, including the mouse. Histochemical and
immunohistochemically-stained coronal, sagittal
and horizontal sections of the mouse brain are
available.
Developmental cysts are relatively common
congenital lesions, variously identified as epider-
moid cysts, epithelial cysts, squamous epithelial
cysts or epithelial inclusion cysts (Faccini et al
1990, Frith et al 2007, Frith & Ward 1988, Garner
et al 1967, Krinke et al 2000, Kulwich 1994,
Percy & Barthold 2007, Radovsky & Mahler 1999,
Satoh & Furuhama 2001). These occur in the
brain and the spinal cord, usually occurring at the
midline in the brain and peripherally located
beneath the pia of the spinal cord. They usually
consist of cystic structures lined by stratified squa-
mous epithelium and containing desquamated
keratin (Fig. 4.87). Occasional cysts in the brain
are lined by cuboidal, ciliated epithelium and
contain flocculent material, similar to develop-
mental cysts seen in the pituitary (Fig. 4.88).
Developmental cysts are considered to be embry-
ological remnants, are often seen in young mice,
may increase in size with age, and are not usually
associated with clinical signs.
FIGURE 4.87  Epidermoid cyst in the cervical spinal cord
of an 18-week-old, male, CD-1 mouse. The cyst is lined
by a stratified squamous epithelium and contains desq-
uamated keratin. ×200.
FIGURE 4.88  Developmental cyst in the brain of an
18-month-old, male, CD-1 mouse. A multilocular cyst
lined by a squamous epithelium, occasionally cuboidal
and ciliated. The cyst contains faintly basophilic, floccu-
lent material, and is located in the hypothalamus, adja-
cent to the third ventricle. ×100.
Lipomatous hamartoma is the accumulation of
normal adipocytes, predominantly located in the
midline or ventricles of the brain (Fig. 4.89).
Lipomatous hamartoma have been reported as
lipomas, but are considered to result from faulty
development of the meninges or choroid plexus.
They do not grow rapidly or progress to malignant
tumors, and are not usually associated with
clinical signs (Frith & Ward 1988, Frith et al
2007, Krinke et al 2000, 2001a, Morgan &
Sheldon 1988, Percy & Barthold 2007). Adkison
& Sundberg (1991) reported lipomatous hamarto-
mas in the brain of inbred strains of mice associ-
ated with the growth of normal adipose tissue
in the subcutis above the dorsal midline of the
cranium. These lesions were distinct from the
lipomatous hamartomas reported here in that they
represented an extension of an extracranial growth
through the cranial sutures of the skull into the
brain of affected mice and did not represent a
disturbance in the embryological development of
the brain.
FIGURE 4.89  Lipomatous hamartoma of the brain of an
18-month-old, male, CD-1 mouse. A well demarcated
cluster of mature adipose cells is present in the midline
of the brain, occupying the third ventricle, and distorting
the normal architecture of the hippocampus. ×40.
Cerebral mineralization consists of small depos-
its of basophilic concretions, often with concen-
tric laminations, occurring in the thalamic region
of the brain in aging mice and is usually associated
with blood vessels (Fig. 4.90). The mineralized
material occurs bilaterally, and they are common
in CD-1 and B6C3F1 mice. Their incidence can
be dependent on the level of brain sectioned
(Faccini et al 1990, Elwell & Mahler 1999, Frith
& Ward 1988, Frith et al 2007, Percy & Barthold
2007, Radovsky & Mahler 1999).
FIGURE 4.90  Cerebral mineralization of the brain of an
18-month-old, male, CD-1 mouse. The foci of basophilic,
rounded, irregular bodies are associated with blood
vessels in the thalamus. ×200.
Degenerative changes are occasionally seen in
the spinal cord and peripheral nerves of aging
mice, and are characterized by the swelling of
axons and vacuolation of the associated myelin
sheaths with cellular debris and the presence of
macrophages (Fig. 4.91). These changes may be
observed in the sciatic nerves and all levels of the
spinal cord, but generally occur at a much lower
incidence than is seen in aging rats (Berdanier
2004, Engelhardt 1996, Faccini et al 1990, Krinke
1996).

FIGURE 4.91  Focal degeneration in the thoracic spinal
cord of an 18-week-old, female, CD-1 mouse. Focal area
of degenerate fibers with vacuolation and accumulation
of macrophages. ×200.
Eye and Ear
A review of the embryology, anatomy, histology
and physiology of the eye and associated glands
can be found in Geiss & Yoshitomi (1999) and
Botts et al (1999). Payne (1994) presents an
extensive review of the structure and function of
the Harderian gland.
The causes of retinal atrophy (retinal degenera-
tion, retinal dystrophy) are diverse. A genetic pre-
disposition to retinal degeneration is prevalent in
several strains of mice and results in the degenera-
tion and loss of the outer nuclear layers of the
retina within the first month of life (Frame &
Slone 1996, Frith et al 2007, Fuller & Wimer
1966, Percy & Barthold 2007, Saunders 1967,
Serfilippi et al 2004, Stirling et al 1983). In those
mice without a genetic predisposition to degen-
eration in the first weeks of life, the incidence of
retinal atrophy varies with stock, strain, age and
housing conditions. Light-induced retinal atrophy
is more prevalent in albino than in pigmented
mice (Faccini et al 1990, Frame & Slone 1996,
Frith & Ward 1988, Frith et al 2007, Geiss &
Yoshitomi 1999, Serfilippi et al 2004). Atrophy
may be unilateral or bilateral, and may be partial.
It is characterized by loss of the rods and the outer
plexiform and nuclear layers (Fig. 4.92).
FIGURE 4.92  Retinal atrophy in the eye of an 18-week-
old, male, CD-1 mouse showing loss of the outer nuclear
layer of the retina. ×200.
Degenerative changes in the cornea (corneal
degeneration) can include mineralization, inflam-
mation, ulceration, epithelial hyperplasia and
keratinization, usually resulting from traumatic
damage to the cornea (Frame & Slone 1996,
Geiss & Yoshitomi 1999, Percy & Barthold 2007)
(Fig. 4.93). Spontaneous corneal degeneration and
dystrophic mineralization without an associated
inflammatory response, can occur in some stocks
and strains of mice, including the CD-1 strain
(Faccini et al 1990, Frame & Slone 1996, Percy &
Barthold 2007, Yamate et al 1987), with females
being more susceptible than males (Fig. 4.94).
FIGURE 4.93  Corneal degeneration of the eye of an
18-month-old, male, CD-1 mouse. Acute inflammatory
infiltration of the cornea (keratitis) is apparent, with asso-
ciated epithelial hyperplasia and keratinization of the
surface epithelium. Anterior synechia (attachment of iris
to Descemet’s membrane) is also evident. ×40.
FIGURE 4.94  Corneal mineralization in an 18-month-old,
male, CD-1 mouse. Basophilic accumulations are appar-
ent at the junction of the corneal epithelium and the
stroma. ×100.
Mineralization of the iris occurs at a low inci-
dence in CD-1 mice (Faccini et al 1990). The
etiology of this change is unknown, but it occurs
occasionally as a unilateral or bilateral change in
older CD-1 mice, usually with no apparent inflam-
matory reaction (Fig. 4.95).
Mouse 65
FIGURE 4.95  Mineralization of the iris of an 18-month-
old, female, CD-1 mouse. A focus of basophilic material
is visible within the iris. These depositions can be rounded
or irregular in shape and usually elicit no inflammatory
response. ×200.
Histologically, all degenerative changes in the
lens, capsule, lens epithelium or fibers which
result in reduced permeability of the lens to light
are termed cataracts (Frith et al 2007, Geiss &
Yoshitomi 1999). Early lesions consisting of foci
of swollen or degenerate lens fibers are relatively
common in mice (Faccini et al 1990, Saunders
1967). Multiple, globoid, circumscribed bodies,
and the development of vacuoles within the lens,
are seen in the later stages of the disease (Frith
& Ward 1988, Saunders 1967). Fibrosis and
mineralization are features of late-stage, lenticular
degeneration (Saunders 1967). There are strain
differences in the occurrence and severity of these
changes (Berdanier 2004, Frame & Slone 1996,
Geiss & Yoshitomi 1999). Cataracts may occur
unilaterally or bilaterally and although early
degenerative changes are relatively common,
severe changes are an unusual occurrence in mice
(Fig. 4.96).
FIGURE 4.96  Lenticular degeneration in a two-year-old,
male, CD-1 mouse. Swollen and degenerate lens fibers
and globoid bodies are apparent and a large vacuole is
present within the lens. ×200.
There is sexual dimorphism in the function of
the Harderian gland of mice, with females pro-
ducing more porphyrin than males (Krinke 2004,
Krinke et al 1996, Payne 1994). The number of
spontaneous changes found in the Harderian gland
are limited, and the most common finding is the
presence of lymphoid aggregates within the gland
of aging mice. Duct ectasia and focal hyperplasia
may occasionally be seen and the incidence of
atrophy also increases with age (Botts et al 1999,
66 BACKGROUND LESIONS IN LABORATORY ANIMALS
Faccini et al 1990, Frith & Ward 1988, Frith et al
2007, Krinke et al 1996, 2001b).
Focal or widespread necrosis of the Harderian
gland may also be observed occasionally as a result
of damage to the gland during retro-orbital blood
sampling procedures. There is usually no risk of
mistaking these findings as being related to the
administration of a test material given the unilat-
eral occurrence and association with the blood
sampling procedure (Fig. 4.97).
FIGURE 4.97  Focal necrosis of the Harderian gland of a
20-week-old, male, CD-1 mouse. Loss of the normal epi-
thelial structure of the acini with associated inflammatory-
cell infiltration. ×40.
Foci of lymphocytic infiltration are the most
commonly observed finding in the lacrimal glands
of mice (Botts et al 1999, Krinke et al 1996).
As mice age, the incidence of ectopic Harderian
gland within the lacrimal gland (Harderianization)
increases, although this change is more common
in females (Frith & Ward 1988, Krinke et al 1996)
(Fig. 4.98). Atrophy of the gland also occurs with
an increased incidence in older mice (Botts et al
1999). An unusual, apparently proliferative, lesion
of the mouse lacrimal gland occurs occasionally in
older CD-1 mice. Foci of degenerate acinar cells,
with replacement by spindle-shaped cells (often
resembling cholesterol clefts), and areas of calci-
fication, occur at a low incidence in male mice
over 1 year old. This finding is usually termed
mesenchymal proliferation and the incidence
varies from 0 to 8% in control animals (Fig. 4.99).
The etiology of this change is unclear, although a
hormonal influence may be involved given that it
does not appear to occur in females.
FIGURE 4.98  Harderianization of the lacrimal gland of
a two-year-old, male, CD-1 mouse. Harderian acini are
present within an area of atrophic, lacrimal acini. An
island of normal lacrimal acini is present at the bottom
right of the image and a focus of lymphocytic cell infiltra-
tion is present at the top right. ×100.
appearance in H&E stained sections. Amyloid
stains positively with Congo Red and under polar-
ized light exhibits an apple-green birefringence
(HogenEsch et al 1996, Jakob 1971, Percy &
Barthold 2007, Sass 1983c).
Primary amyloidosis is considered to be geneti-
cally determined, and the term secondary amy-
loidosis is used for amyloid associated with chronic
inflammation (Dunn 1967b). Amyloidosis is also
associated with skin lesions related to fighting
in male mice (Brayton 2007, Frith et al 2007,
HogenEsch et al 1996, Page & Glenner 1972,
Tucker & Baker 1967).
The kidney is a predilection site for the deposi-
tion of amyloid, where it accumulates in the
FIGURE 4.99  Mesenchymal proliferation of the lacrimal glomeruli (Chandra & Frith 1994, Dunn 1967b).
gland of an 18-month-old, male, CD-1 mouse. Prolifera- Amyloid deposits in the kidney are associated
tion of spindle-shaped cells within the gland, associated with degenerative kidney disease and papillary
with atrophic acini and inflammatory-cell infiltration. necrosis in susceptible strains of mice (Cornelius
×100. 1970, Dunn 1967b, Russell & Meier 1966), and
renal amyloidosis is often considered the main
factor contributing to the death of susceptible
Systemic disease strains of mice (Chai 1978, Frith et al 2007).
In systemic amyloidosis, amyloid deposition can
There are frequent references to amyloidosis as a occur in various organs, and the pattern varies
significant disease of the CD-1 mouse in the sci- with the strain, but particular sites of deposition
entific literature relating to mouse pathology include the gastrointestinal tract, with marked
(Brayton 2007, Elmore 2006, Engelhardt 1996, deposition in the lamina propria of the small intes-
Frith & Chandra 1991, Frith et al 2007, Glaister tine, the adrenal cortex, salivary glands, the alveo-
1986, Gruys et al 1996, Maita et al 1988, Percy lar septa of the lungs, the spleen (where it can
& Barthold 2007, Suttie 2006, Thoolen et al involve the entire red pulp in extreme cases),
2010). Our current experience with CD-1 mice lymph nodes, and the liver (periportal deposits)
from Charles River UK, however, is that amyloid (Dunn 1967b, Elmore 2006, Jakob 1971, Percy &
occurs at a very low incidence in control animals Barthold 2007, Sass 1983c, Suttie 2006) (Figs
in long-term studies, and usually manifests only as 4.100, 4.101a & b).
an incidental finding in a single tissue in an occa-
sional animal. The occurrence of systemic amy-
loidosis as a significant finding, and a cause of
death in control animals from long-term studies,
has reduced to zero in the last 20 years in the
author’s experience. This experience is reflected
in some reports in the literature. Significant dif-
ferences in the incidence of amyloidosis in Charles
River CD-1 mice from three different sources in
the USA have been reported (Engelhardt 1996,
Engelhardt et al 1993). In males, the incidence of
amyloidosis ranged from 2% to 45%, and in
females from 0.7% to 30.7%. A similar report on
the causes of death in long-term studies using
CD-1 mice from Charles River Japan reported an
abrupt decrease in the incidence of systemic amy-
loidosis in Charles River CD-1 mice born after FIGURE 4.100  Amyloid accumulation in the lamina
1980 (Maita et al 1988). The factors contributing propria of the jejunum of a two-year-old, male, CD-1
to death in control CD-1 mice from 20 long-term mouse. ×100.
studies performed between 1990 and 2002 were
reviewed (Son 2003a, b). All of the animals in
these studies were supplied by Charles River UK,
and systemic amyloidosis did not feature as a
factor contributing to death in any of the decedent
animals in these studies.
Notwithstanding these changes in the occur-
rence of this finding in CD-1 mice, amyloidosis is
still reported as a significant finding in a number
of strains and stocks of mice (Brayton 2007, Percy
& Barthold 2007, Frith et al 2007), and the toxi-
cological pathologist should be aware of the fea-
tures of this important systemic disease.
The structure, distribution, incidence and
pathogenesis of senile amyloidosis is reviewed by
HogenEsch et al (1996). The disease is character-
ized by the extracellular deposition of fibrillar
protein, which has a characteristic eosinophilic

FIGURE 4.101a  Perivascular accumulation of amyloid in
the liver of a two-year-old, male, CD-1 mouse. Congo Red
staining. ×200.
FIGURE 4.101b  Image from Fig. 4.101a under polarized
light showing apple green birefringence of the amyloid.
×200.
References
Adkison, D.L., Sundberg, J.P., 1991. ‘Lipomatous’
hamartomas and choristomas in inbred laboratory
mice. Vet. Pathol. 28, 305–312.
Albassam, M.A., Wojcinski, Z.W., Barsoum, N.J., et al.,
1991. Spontaneous fibro-osseous proliferative lesions
in the sternums and femurs of B6C3F1 mice. Vet.
Pathol. 28, 381–388.
Albassam, M.A., Courtney, C.L., 1996. Nonneoplastic
and neoplastic lesions of the bone. In: Mohr, U.,
Dungworth, D.L., Capen, C.C., Carlton, W.W.,
Sundberg, J.P., Ward, J.M. (Eds.), Pathobiology of
the aging mouse, vol. 2. ILSI Press, Washington,
pp. 425–437.
Andrews, A.G., Dysko, R.C., Spilman, S.C., et al.,
1994. Immune complex vasculitis with secondary
ulcerative dermatitis in aged C57BL/6NNia mice.
Vet. Pathol. 31, 293–300.
Anagnostopoulos, A.V., Mobraaten, L.E., Sharp, J.J.,
et al., 2001. Transgenic and knockout databases:
behavioral profiles of mouse mutants. Physiol. Behav.
73, 675–689.
Bannasch, P., Wayss, K., Zerban, H., 1985. Peliosis
hepatis, rodents. In: Jones, T.C., Mohr, U., Hunt,
R.D. (Eds.), Digestive system, ILSI monographs on
pathology of laboratory animals. Springer-Verlag,
Berlin, pp. 110–115.
Baze, W.B., Steinbach, T.J., Fleetwood, M.L.,
Blanchard, T.W., Barnhart, K.F., McArthur, M.J.,
2006. Karyomegaly and intranuclear inclusions in
the renal tubules of sentinel ICR mice (Mus
musculus). Comp. Med. 56, 435–438.
Bell, J.F., Moore, G.J., Clifford, C.M., et al., 1970.
Dry gangrene of the ear in white mice. Lab. Anim.
4, 245–254.
Bendele, A.M., Carlton, W.W., 1986. Urologic
syndrome, mouse. In: Jones, T.C., Mohr, U., Hunt,
R.D. (Eds.), Urinary system, ILSI monographs on
pathology of laboratory animals. Springer-Verlag,
Berlin, pp. 369–375.
Berdanier, C.D., 2004. Gastrointestinal system and
metabolism. In: Hedrich, H.J., Bullock, G. (Eds.),
The handbook of experimental animals: the
laboratory mouse. Elsevier, San Diego, pp. 245–259.
Bernichtein, S., Peltoketo, H., Huhtaniemi, I., 2009.
Adrenal hyperplasia and tumors in mice in
connection with aberrant pituitary-gonadal function.
Mol. Cell. Endocrinol. 300, 164–168.
Best, P.V., Heath, D., 1961. Interpretation of the
appearances of the small pulmonary blood vessels in
animals. Circ. Res. 9, 288–294.
Betton, G.R., Whiteley, L.O., Anver, M.R., et al.,
2001. Gastrointestial tract. In: Mohr, U. (Ed.),
International classification of rodent tumors, the
mouse. Springer, Berlin, pp. 23–58.
Bolon, B., 2006. Internet resources for phenotyping
engineered rodents. ILAR. J. 47, 163–171.
Bolon, B., 2007. Genetically engineered animals. In:
Kubinyi, H. (Ed.), Comprehensive medicinal
chemistry, second ed. Drug discovery technologies,
vol. 3. Elsevier, Oxford, pp. 151–170.
Boorman, G.A., Sills, R.C., 1999. Exocrine and
endocrine pancreas. In: Maronpot, R.R., Boorman,
G.A., Gaul, B.W. (Eds.), Pathology of the mouse,
reference and atlas. Cache River Press, Vienna, ch 8,
pp. 185–205.
Botts, S., Jokinen, M.M., Gaillard, E.T., et al., 1999.
Salivary, harderian and lacrimal glands. In:
Maronpot, R.R., Boorman, G.A., Gaul, B.W. (Eds.),
Pathology of the mouse, reference and atlas. Cache
River Press, Vienna, ch 5, pp. 49–79.
BrainMaps.org – digital atlases of mouse brain available
at: http://brainmaps.org/index.php?p=speciesdata&
species=mus-musculus, 08 July 2011.
Braun, A., Ernst, H., Hoymann, H.G., et al., 2004.
Respiratory tract. In: Hedrich, H.J., Bullock, G.
(Eds.), The handbook of experimental animals:
the laboratory mouse. Elsevier, San Diego,
pp. 225–243.
Brayton, C., 2007. Spontaneous diseases in commonly
used inbred mouse strains. In: Fox, J.G., Barthold,
S.W., Davisson, M.T., et al (Eds.), The mouse in
biomedical research, second ed, vol. 3. Academic
Press, Burlington, ch 25, pp. 623–717.
Brayton, C., Justice, M., Montgomery, C.A., 2001.
Evaluating mutant mice: anatomic pathology. Vet.
Pathol. 38, 1–19.
Brown, R., 1986. Pigment deposition, kidney, mouse.
In: Jones, T.C., Mohr, U., Hunt, R.D. (Eds.),
Urinary system, ILSI monographs on pathology of
laboratory animals. Springer-Verlag, Berlin, pp.
244–245.
Bruner, R., Kuttler, K., Bader, R., et al., 2001.
Integumentary system. In: Mohr, U. (Ed.),
International classification of rodent tumors,
the mouse. Springer, Berlin, pp. 1–22.
Capen, C.C., 1983a. Functional and pathologic
interrelationships of the pituitary gland and the
hypothalamus. In: Jones, T.C., Mohr, U., Hunt, R.D.
(Eds.), Endocrine system, monographs on pathology
of laboratory animals. Springer-Verlag, Berlin,
pp. 101–120.
Capen, C.C., 1983b. Structural and biochemical
aspects of parathyroid gland function in animals. In:
Jones, T.C., Mohr, U., Hunt, R.D. (Eds.), Endocrine
system, monographs on pathology of laboratory
animals. Springer-Verlag, Berlin, pp. 217–247.
Capen, C.C., Gröne, A., Bucci, T.J., et al., 1996.
Changes in structure and function of the parathyroid
gland. In: Mohr, U., Dungworth, D.L., Capen, C.C.,
Carlton, W.W., Sundberg, J.P., Ward, J.M. (Eds.),
Pathobiology of the aging mouse, vol. 1. ILSI Press,
Washington, pp. 109–123.
Mouse 67
Capen, C.C., Karbe, E., Deschl, U., et al., 2001.
Endocrine system. In: Mohr, U. (Ed.), International
classification of rodent tumors, the mouse. Springer,
Berlin, pp. 269–322.
Carlton, W.W., Engelhardt, J.A., 1991. Atrial
thrombosis, rat, mouse, and hamster. In: Jones, T.C.,
Mohr, U., Hunt, R.D. (Eds.), Cardiovascular and
musculoskeletal system, ILSI monographs on
pathology of laboratory animals. Springer-Verlag,
Berlin, pp. 37–41.
Carlton, W.W., Gries, C.L., 1983 Cysts, pituitary: rat,
mouse, and hamster. In: Jones, T.C., Mohr, U.,
Hunt, R.D. (Eds.), Endocrine system, monographs
on pathology of laboratory animals. Springer-Verlag,
Berlin, pp. 161–163.
Cesta, M.F., 2006a. Normal structure, function, and
histology of the spleen. Toxicol. Pathol. 34,
455–465.
Cesta, M.F., 2006b. Normal structure, function, and
histology of mucosa-associated lymphoid tissue.
Toxicol. Pathol. 34, 599–608.
Chai, C.K., 1978. Spontaneous amyloidosis in LLC
mice. Am. J. Pathol. 90, 381–398.
Chai, C.K., Dickie, M.M., 1966. Endocrine variations.
In: Green, E.L. (Ed.), Biology of the laboratory
mouse. McGraw-Hill, New York, ch 20,
pp. 387–403.
Chandra, M., Frith, C.H., 1994. Spontaneous renal
lesions in CD-1 and B6C3F1 mice. Exp. Toxicol.
Pathol. 46, 189–198.
Charles River website: http://www.criver.com/, 08 July
2011.
Chiu, T., Chen, H.C., 1986. Spontaneous basophilic
hypertrophic foci of the parotid glands in rats and
mice. Vet. Pathol. 23, 606–609.
Clarke, M.C., Taylor, R.J., Hall, G.A., et al., 1978. The
occurrence in mice of facial and mandibular
abscesses associated with Staphylococcus aureus.
Lab. Anim. 12, 121–123.
Cook, M.J., 1965. The anatomy of the laboratory
mouse. Academic Press, London, available
electronically at http://www.informatics.jax.org/
cookbook/.
Cornelius, E.A., 1970. Amyloidosis and renal papillary
necrosis in male hybrid mice. Am. J. Pathol. 59,
317–326.
Cotchin, E., Roe, F.J.C. (Eds.), 1967. Pathology of
laboratory rats and mice. Blackwell, Oxford.
DeLellis, R.A., Sheldon, W.G., Bucci, T.J., 1996.
Changes in thyroid c cells. In: Mohr, U.,
Dungworth, D.L., Capen, C.C., Carlton, W.W.,
Sundberg, J.P., Ward, J.M. (Eds.), Pathobiology of
the aging mouse, vol. 1. ILSI Press, Washington,
pp. 103–107.
Deschl, U., Cattley, R.C., Harada, T., et al., 2001.
Liver, gall bladder, and exocrine pancreas. In: Mohr,
U., (Ed.), International classification of rodent
tumors, the mouse. Springer, Berlin, pp. 59–86.
Dixon, D., Herbert, R.A., Sills, R.C., et al., 1999.
Lungs, pleura, and mediastinum. In: Maronpot, R.R.,
Boorman, G.A., Gaul, B.W. (Eds.), Pathology of the
mouse, reference and atlas. Cache River Press,
Vienna, ch 12, pp. 293–332.
Doi, T., Kotani, Y., Kokoshima, H., et al., 2007.
Eosinophilic substance is ‘not amyloid’ in the mouse
nasal septum. Vet. Pathol. 44, 796–802.
Doi, T., Kotani, Y., Kokoshima, H., et al., 2009.
Deposition process of eosinophilic substance in
mouse nasal septum. J. Vet. Med. Sci. 71, 931–935.
Doi, T., Kokoshima, H., Kanno, T., et al., 2010. New
findings concerning eosinophilic substance deposition
in mouse nasal septum: sex difference and no
increase in seniles. Toxicol. Pathol. 38, 631–636.
Donnelly, K.B., 2008. Cardiac valvular pathology:
comparative pathology and animal models of
acquired cardiac valvular diseases. Toxicol. Pathol.
36, 204–217.
68 BACKGROUND LESIONS IN LABORATORY ANIMALS
Dumont, F., Robert, F., 1980. Age- and sex-dependent
thymic abnormalities in NZB X SJL F1 hybrid mice.
Clin. Exp. Immunol. 41, 63–72.
Dungworth, D.L., Rittinghausen, S., Schwartz, L.,
et al., 2001. Respiratory system and mesothelium.
In: Mohr, U. (Ed.), International classification
of rodent tumors, the mouse. Springer, Berlin,
pp. 87–137.
Dunn, T.B., 1967a. Renal disease of the mouse. In:
Cotchin, E., Roe, F.J.C. (Eds.), Pathology of
laboratory rats and mice. Blackwell, Oxford, ch 6,
pp. 149–179.
Dunn, T.B., 1967b. Amyloidosis in mice. In: Cotchin,
E., Roe, F.J.C. (Eds.), Pathology of laboratory rats
and mice. Blackwell, Oxford, ch 7, pp. 181–212.
Dunn, T.B., 1970. Normal and pathologic anatomy of
the adrenal gland of the mouse, including neoplasms.
Natl. Cancer. Inst. 44, 1323–1389.
Eaton, G.J., Custer, R.P., Johnson, F.N., et al., 1978.
Dystrophic cardiac calcinosis in mice: genetic,
hormonal, and dietary influences. Am. J. Pathol. 90,
173–186.
Eaton, G.J., Johnson, F.N., Custer, R.P., et al., 1980.
The Icr:Ha(ICR) mouse: a current account of
breeding, mutations, diseases and mortality. Lab.
Anim. 14, 17–24.
Elangbam, C.S., Colman, K.A., Lightfoot, R.M., et al.,
2002. Endocardial myxomatous change in Harlan
Sprague-Dawley rats (Hsd:S-D) and CD-1 mice: its
microscopic resemblance to drug-induced
valvulopathy in humans. Toxicol. Pathol. 30,
483–491.
Elmore, S.A., 2006. Histopathology of the lymph
nodes. Toxicol. Pathol. 34, 425–454.
Elwell, M.R., Mahler, J.F., 1999. Heart, blood and
lymphatic vessels. In: Maronpot, R.R., Boorman
G.A., Gaul, B.W. (Eds.), Pathology of the mouse,
reference and atlas. Cache River Press, Vienna,
ch 14, pp. 361–380.
Engelhardt, J.A., 1996. Variability in the occurrence of
spontaneous lesions in Crl:CD-1(r)(ICR) mice
related to source. In: Mohr, U., Dungworth, D.L.,
Capen, C.C., Carlton, W.W., Sundberg, J.P., Ward,
J.M. (Eds.), Pathobiology of the aging mouse. 1,
vol. 1. ILSI Press, Washington, pp. 45–49.
Engelhardt, J.A., Gries, C.L., Long, G.G., 1993.
Incidence of spontaneous neoplastic and
nonneoplastic lesions in Charles River CD-1 mice
varies with breeding origin. Toxicol. Pathol. 21,
538–541.
Enomoto, M., Hirouchi, Y., Tsutumi, M., 1996.
Pancreas. In: Mohr, U., Dungworth, D.L., Capen,
C.C., Carlton, W.W., Sundberg, J.P., Ward, J.M.
(Eds.), Pathobiology of the aging mouse, vol. 2. ILSI
Press, Washington, pp. 251–260.
Ernst, H., Dungworth, D.L., Kamino, K., et al., 1996.
Nonneoplastic lesions in the lungs. In: Mohr, U.,
Dungworth, D.L., Capen, C.C., Carlton, W.W.,
Sundberg, J.P., Ward, J.M. (Eds.), Pathobiology of
the aging mouse, vol. 1. ILSI Press, Washington,
pp. 281–300.
Ettlin, R.A., Stirnimann, P., Prentice, D.E., 1994.
Causes of death in rodent toxicity and
carcinogenicity studies. Toxicol. Pathol. 22,
165–178.
Eustis, S.L., Boorman, G.A., 1985. Embryology,
histology, and ultrastructure of the exocrine
pancreas. In: Jones, T.C., Mohr, U., Hunt, R.D.
(Eds.), Digestive system. ILSI monographs on
pathology of laboratory animals. Springer-Verlag,
Berlin, pp. 209–218.
Faccini, J.M., Abbott, D.P., Paulus, G.J.J. (Eds.), 1990.
Mouse histopathology. Elsevier, Amsterdam.
Frame, S.R., Slone, T.W., 1996. Nonneoplastic and
neoplastic changes in the eye. In: Mohr, U.,
Dungworth, D.L., Capen, C.C., Carlton, W.W.,
Sundberg, J.P., Ward, J.M. (Eds.), Pathobiology of
the aging mouse, vol. 2. ILSI Press, Washington,
pp. 97–103.
France, M.P., Sundberg, J.P., Martinic, G., 2000.
Branchial cysts in laboratory mice. J. Comp. Pathol.
123, 55–58.
Frith, C.H., 1983a. Histology, adrenal gland, mouse.
In: Jones, T.C., Mohr, U., Hunt, R.D. (Eds.),
Endocrine system, monographs on pathology of
laboratory animals. Springer-Verlag, Berlin, pp. 8–12.
Frith, C.H., 1983b. Pheochromocytoma, adrenal
medulla, mouse. In: Jones, T.C., Mohr, U., Hunt,
R.D. (Eds.), Endocrine system, monographs on
pathology of laboratory animals. Springer-Verlag,
Berlin, pp. 27–30.
Frith, C.H., 1983c. Lipogenic pigmentation, adrenal
cortex, mouse. In: Jones, T.C., Mohr, U., Hunt,
R.D. (Eds.), Endocrine system, monographs on
pathology of laboratory animals. Springer-Verlag,
Berlin, pp. 60–64.
Frith, C.H., 1983d. Ectopic thyroid, mouse. In: Jones,
T.C., Mohr, U., Hunt, R.D. (Eds.), Endocrine
system, monographs on pathology of laboratory
animals. Springer-Verlag, Berlin, pp. 171–172.
Frith, C.H., Chandra, M., 1991. Incidence,
distribution, and morphology of amyloidosis in
Charles Rivers CD-1 mice. Toxicol. Pathol. 19,
123–127.
Frith, C.H., Fetters, J., 1983. Ectopic parathyroid,
mouse. In: Jones, T.C., Mohr, U., Hunt, R.D.
(Eds.), Endocrine system, monographs on pathology
of laboratory animals. Springer-Verlag, Berlin, pp.
263–264.
Frith, C.H., Sheldon, W.D., 1983. Hyperplasia,
adenoma and carcinoma of pancreatic islets, mouse.
In: Jones, T.C., Mohr, U., Hunt, R.D. (Eds.),
Endocrine system, monographs on pathology of
laboratory animals. Springer-Verlag, Berlin, pp.
297–303.
Frith, C.H., Townsend, J.W., 1985. Histology and
ultratructure, salivary glands, mouse. In: Jones, T.C.,
Mohr U., Hunt, R.D. (Eds.), Digestive system. ILSI
monographs on pathology of laboratory animals.
Springer-Verlag, Berlin, pp. 177–184.
Frith, C.H., Ward, J.M., 1979. A morphologic
classification of proliferative and neoplastic hepatic
lesions in mice. J. Environ. Pathol. Toxicol. 3,
329–351.
Frith, C.H., Ward, J.M., 1988. Color atlas of neoplastic
and non-neoplastic lesions in aging mice. Elsevier,
Amsterdam.
Frith, C.H., Pattengale, P.K., Ward, J.M., 1985. A color
atlas of hematopoietic pathology of mice. Toxicology
Pathology Associates, Little Rock.
Frith, C.H., Townsend, J.W., Ayres, P.H., 1986.
Histology, ultrastructure, urinary tract, mouse. In:
Jones, T.C., Mohr, U., Hunt, R.D. (Eds.), Urinary
system. ILSI monographs on pathology of laboratory
animals. Springer-Verlag, Berlin, pp. 281–284.
Frith, C.H., Ward, J.M., Harleman, J.H., et al., 2001.
Hematopoietic system. In: Mohr, U. (Ed.),
International classification of rodent tumors, the
mouse. Springer, Berlin, pp. 417–451.
Frith, C.H., Goodman, D.G., Boysen, B.G., 2007. The
mouse, pathology. In: Gad, S.C. (Ed.), Animal
models in toxicology, second ed. Taylor & Francis,
Boca Raton, pp. 72–122.
Fuller, J.L., Wimer, R.E., 1966. Neural, sensory, and
motor functions. In: Green, E.L. (Ed.), Biology of
the laboratory mouse. McGraw-Hill, New York, ch
32, pp. 609–628.
Gaillard, E.T., 1999. Ureter, urinary bladder, and
urethra. In: Maronpot, R.R., Boorman, G.A., Gaul,
B.W. (Eds.), Pathology of the mouse, reference and
atlas. Cache River Press, Vienna, ch 10, pp. 235–258.
Gad, S.C. (Ed.), 2007. Animal models in toxicology,
second ed. Taylor & Francis, Boca Raton, pp. 72–122.
Garner, F.M., Innes, J.R.M., Nelson, D.H., 1967.
Murine neuropathology. In: Cotchin, E, Roe, F.J.C.
(Eds.), Pathology of laboratory rats and mice.
Blackwell, Oxford, ch 11, pp. 295–348.
Geiss, V., Yoshitomi, K., 1999. The eyes. In: Maronpot,
R.R., Boorman, G.A., Gaul, B.W. (Eds.), Pathology
of the mouse, reference and atlas. Cache River
Press, Vienna, ch 18, pp. 471–489.
Glaister, J.R., 1986. The ageing mouse. In: Principles
of toxicological pathology. Taylor & Francis, London,
pp. 182–203.
Goodman, D.G., 1983. Subcapsular-cell hyperplasia,
adrenal, mouse. In: Jones, T.C., Mohr, U., Hunt,
R.D. (Eds.), Endocrine system, monographs on
pathology of laboratory animals. Springer-Verlag,
Berlin, pp. 66–68.
Goto, N., Nakajima, Y., Onodera, T., et al., 1984.
Inheritance of hydronephrosis in the inbred mouse
strain DDD. Lab. Anim. 18, 22–25.
Goto, N., Nakajima, Y., Imamura, K., et al., 1985.
Influence of testosterone on hydronephrosis in the
inbred mouse strain DDD. Lab. Anim. 19, 85–88.
Greaves, P., Boiziau, J.L., 1984. Altered patterns of
mucin secretion in gastric hyperplasia in mice. Vet.
Pathol. 21, 224–228.
Green, E.L. (Ed.), 1966. Biology of the laboratory
mouse. McGraw-Hill, New York.
Green, E.U., 1942. On the occurrence of crystalline
material in the lungs of normal and cancerous Swiss
mice. Cancer Res. 2, 210–217.
Gruys, E., Tooten, P.C., Kuijpers, M.H., 1996. Lung,
ileum and heart are predilection sites for AApoAII
amyloid deposition in CD-1 Swiss mice used for
toxicity studies. Pulmonary amyloid indicates
AApoAII. Lab. Anim. 30, 28–34.
Guo, L., Johnson, R.S., Schuh, J.C., 2000. Biochemical
characterization of endogenously formed eosinophilic
crystals in the lungs of mice. J. Biol. Chem. 275
(17), 8032–8037.
Hagiwara, A., Tamano, S., Hirose, M., 1996. Changes
in the heart. In: Mohr, U., Dungworth, D.L., Capen,
C.C., Carlton, W.W., Sundberg, J.P., Ward, J.M.
(Eds.), Pathobiology of the aging mouse, vol. 1. ILSI
Press, Washington, pp. 361–371.
Haines, D.C., Chattopadhyay, S., Ward, J.M., 2001.
Pathology of aging B6;129 mice. Toxicol. Pathol. 29,
653–661.
Harada, T., Maronpot, R.R., Enomoto, A., et al., 1996.
Changes in the liver and gallbladder. In: Mohr, U.,
Dungworth, D.L., Capen, C.C., Carlton, W.W.,
Sundberg J.P., Ward, J.M. (Eds.), Pathobiology of
the aging mouse, vol. 2. ILSI Press, Washington,
pp. 207–241.
Harada, T., Enomoto, A., Boorman, G.A., et al., 1999.
Liver and gallbladder. In: Maronpot, R.R., Boorman,
G.A., Gaul, B.W. (Eds.), Pathology of the mouse,
reference and atlas. Cache River Press, Vienna, ch 7,
pp. 119–183.
Hard, G.C., Durchfeld-Meyer, B., Short, B., et al.,
2001. Urinary system. In: Mohr, U. (Ed.),
International classification of rodent tumors, the
mouse. Springer, Berlin, pp. 139–162.
Hardisty, J.F., Boorman, G.A., 1999. The thyroid and
parathyroid glands. In: Maronpot, R.R., Boorman,
G.A., Gaul, B.W. (Eds.), Pathology of the mouse,
reference and atlas. Cache River Press, Vienna, ch
21, pp. 537–554.
Hardy, M.H., 1952. The histochemistry of hair follicles
in the mouse. Am. J. Anat. 90, 285–337.
Hardy, J., 1967. Haematology of rats and mice. In:
Cotchin, E., Roe, F.J.C. (Eds.), Pathology of
laboratory rats and mice. Blackwell, Oxford, ch 16,
pp. 501–536.
Harkema, J.R., Carey, S.A., Wagner, J.G., 2006. The
nose revisited: a brief review of the comparative
structure, function, and toxicologic pathology of the
nasal epithelium. Toxicol. Pathol. 34, 252–269.
Harlan Laboratories web site: http://www.harlan.com/,
08 July 2011.
Hedrich, H.J., Bullock, G. (Eds.), 2004. The handbook
of experimental animals: the laboratory mouse.
Elsevier, San Diego.

Herbert, R.A., Leininger, J.R., 1999. Nose, larynx, and
trachea. In: Maronpot, R.R., Boorman, G.A., Gaul,
B.W. (Eds.), Pathology of the mouse, reference and
atlas. Cache River Press, Vienna, ch 11, pp. 259–292.
Hoedemaeker, P.J., Fleuren, G.J., Weening, J.J., 1986.
Immune mechanisms in injury to glomeruli and
tubulo-interstitial tissue. In: Jones, T.C., Mohr, U.,
Hunt, R.D. (Eds.), Urinary system, ILSI
monographs on pathology of laboratory animals.
Springer-Verlag, Berlin, pp. 153–174.
Hoenerhoff, M.J., Starost, M.F., Ward, J.M., 2006.
Eosinophilic crystalline pneumonia as a major cause
of death in 129S4/SvJae mice. Vet. Pathol. 43,
682–688.
HogenEsch, H., Gruys, E., Higuchi, K., 1996. Senile
amyloidosis. In: Mohr, U., Dungworth, D.L., Capen,
C.C., Carlton, W.W., Sundberg, J.P., Ward, J.M.
(Eds.), Pathobiology of the aging mouse, vol. 1. ILSI
Press, Washington, pp. 237–244.
Hsu, H.H., 1986. Hereditary hydronephrosis, mouse.
In: Jones, T.C., Mohr, U., Hunt, R.D. (Eds.),
Urinary system, ILSI monographs on pathology
of laboratory animals. Springer-Verlag, Berlin,
pp. 273–275.
Hummel, K.P., Richardson, F.L., Fekete, E., 1966.
Anatomy. In: Green, E.L. (Ed.), Biology of the
laboratory mouse. McGraw-Hill, New York, ch 13,
pp. 247–307.
Imaoka, K., Honjo, K., Doi, K., et al., 1986.
Development of spontaneous tongue calcification
and polypoid lesions in DBA/2NCrj mice. Lab.
Anim. 20, 1–4.
Innes, J.R.M., Donati, E.J.,Yevich, P.P., 1958.
Pulmonary lesions in mice due to fragments of hair,
epidermis and extraneous matter accidentally
injected in toxicity experiments. Am. J. Pathol. 34,
161–167.
Jakob, W., 1971. Spontaneous amyloidosis of
mammals. Vet. Pathol. 8, 292–306.
Jones, I.C., 1950. The effect of hypophysectomy on
the adrenal cortex of the immature mouse. Am. J.
Anat. 86, 371–403.
Jones, T.C., 1967. Pathology of the liver of rats and
mice. In: Cotchin, E., Roe, F.J.C. (Eds.), Pathology
of laboratory rats and Mice. Blackwell, Oxford, ch
1, pp. 1–23.
Jones, T.C., Mohr, U., Hunt, R.D. (Eds.), 1983.
Endocrine system, monographs on pathology of
laboratory animals. Springer-Verlag, Berlin.
Jones, T.C., Mohr, U., Hunt, R.D. (Eds.), 1985a.
Respiratory system. ILSI monographs on pathology
of laboratory animals. Springer-Verlag, Berlin.
Jones, T.C., Mohr, U., Hunt, R.D. (Eds.), 1985b.
Digestive system. ILSI monographs on pathology of
laboratory animals. Springer-Verlag, Berlin.
Jones, T.C., Mohr, U., Hunt, R.D. (Eds.), 1986.
Urinary system. ILSI monographs on pathology of
laboratory animals. Springer-Verlag, Berlin.
Jones, T.C., Mohr, U., Hunt, R.D. (Eds.), 1988.
Nervous system. ILSI monographs on pathology of
laboratory animals. Springer-Verlag, Berlin.
Jones, T.C., Mohr U., Hunt R.D. (Eds.), 1991
Cardiovascular and musculoskeletal system. ILSI
monographs on pathology of laboratory animals.
Springer-Verlag, Berlin.
Jørgensen, M.C., Ahnfelt-Rønne, J., Hald, J., et al.,
2007. An illustrated review of early pancreas
development in the mouse. Endocr. Rev. 28,
685–705.
Kaliste, E.K., Mering, S.M., Huuskonen, H.K., 2006.
Environmental modification and agonistic behavior in
NIH/S male mice: nesting material enhances fighting
but shelters prevent it. Comp. Med. 56, 202–208.
Kast, A., 1985. Pulmonary hair embolism rat. In:
Jones, T.C., Mohr, U., Hunt, R.D. (Eds.),
Respiratory system. ILSI monographs on pathology
of laboratory animals. Springer-Verlag, Berlin,
pp. 186–194.
Kim, J.S., Kubota, H., Doi, K., et al., 1997a.
Correlation of mast cells with spindle cell
hyperplasia in the adrenal cortex of IQI/Jic mice.
Exp. Anim. 46, 103–109.
Kim, J.S., Kubota, H., Kiuchi, Y., et al., 1997b.
Subcapsular cell hyperplasia and mast cell infiltration
in the adrenal cortex of mice: comparative study in
7 inbred strains. Exp. Anim. 46, 303–306.
Kim, J.S., Kubota, H., Nam, S.Y., et al., 2000.
Expression of cytokines and proteases in mast cells
in the lesion of subcapsular cell hyperplasia in mouse
adrenal glands. Toxicol. Pathol. 28, 297–303.
Kitagaki, M., Hirota, M., 2007. Auricular chondritis
caused by metal ear tagging in C57BL/6 mice. Vet.
Pathol. 44, 458–466.
Kittel, B., Ruehl-Fehlert, C., Morawietz, G., et al.,
2004. Revised guides for organ sampling and
trimming in rats and mice-Part 2. A joint publication
of the RITA and NACAD groups. Exp. Toxicol.
Pathol. 55, 413–431.
Komárek, V., 2004. Gross anatomy. In: Hedrich, H.J.,
Bullock, G. (Eds.), The handbook of experimental
animals: the laboratory mouse. Elsevier, San Diego,
pp. 117–132.
Koopman, J.P., Van der Logt, J.T., Mullink, J.W., et al.,
1984. Tail lesions in C3H/He mice. Lab. Anim. 18,
106–109.
Krinke, G.J., 1996. Nonneoplastic and neoplastic
changes in the peripheral nervous system. In: Mohr,
U., Dungworth, D.L., Capen, C.C., Carlton, W.W.,
Sundberg, J.P., Ward, J.M. (Eds.), Pathobiology of
the aging mouse, vol. 2. ILSI Press, Washington,
pp. 83–93.
Krinke, G.J., 2004. Normative histology of organs. In:
Hedrich, H.J., Bullock, G. (Eds.), The handbook of
experimental animals: the laboratory mouse.
Elsevier, San Diego, pp. 133–166.
Krinke, G.J., Schaetti, P.R., Krinke, A.-L., 1996.
Nonneoplastic and neoplastic changes in the
harderian and lacrimal glands. In: Mohr, U.,
Dungworth, D.L., Capen, C.C., Carlton, W.W.,
Sundberg, J.P., Ward, J.M. (Eds.), Pathobiology of
the aging mouse, vol. 2. ILSI Press, Washington,
pp. 139–152.
Krinke, G.J., Kaufmann, W., Mahrous, A.T., et al.,
2000. Morphologic characterization of spontaneous
nervous system tumors in mice and rats. Toxicol.
Pathol. 28, 178–192.
Krinke, G., Fix, A., Kaufmann, W., et al., 2001a.
Central nervous system. In: Mohr, U. (Ed.),
International classification of rodent tumors, the
mouse. Springer, Berlin, pp. 323–345.
Krinke, G., Fix, A., Jacobs, M., et al., 2001b. Eye and
harderian gland. In: Mohr U (Ed.), International
classification of rodent tumors, the mouse. Springer,
Berlin, pp. 347–359.
Kuhn, III, C., 1985. Structure and function of the
lung. In: Jones, T.C., Mohr, U., Hunt, R.D. (Eds.),
Respiratory system. ILSI monographs on pathology
of laboratory animals. Springer-Verlag, Berlin,
pp. 89–98.
Kulwich, B.A., 1994. Epidermoid cysts in the central
nervous system of rats and mice: an incidental
finding in toxicity/oncogenicity studies. Vet Pathol
31, 475–478.
Kurata, Y., Shibata, M.-A., 1996. Aging changes in the
urinary bladder. In: Mohr, U., Dungworth, D.L.,
Capen, C.C., Carlton, W.W., Sundberg, J.P., Ward,
J.M. (Eds.), Pathobiology of the aging mouse, vol. 1.
ILSI Press, Washington, pp. 345–357.
Leininger, J.R., 1999. Skeletal muscle. In: Maronpot,
R.R., Boorman, G.A., Gaul, B.W. (Eds.), Pathology
of the mouse, reference and atlas. Cache River
Press, Vienna, ch 24, pp. 637–643.
Leininger, J.R., McDonald, M.M., Abbott, D.P., 1990.
Hepatocytes in the mouse stomach. Toxicol. Pathol.
18, 678–686.
Leininger, J.R., Herbert, R.A., Morgan, K.T., 1996.
Aging changes in the upper respiratory tract. In:
Mouse 69
Mohr, U., Dungworth, D.L., Capen, C.C., Carlton,
W.W., Sundberg, J.P., Ward, J.M. (Eds.),
Pathobiology of the Aging Mouse, vol. 1. ILSI Press,
Washington, pp. 247–260.
Leininger, J.R., Jokinen, M.P., Dangler, C.A., et al.,
1999. Oral cavity, esophagus and stomach. In:
Maronpot, R.R., Boorman, G.A., Gaul, B.W. (Eds.),
Pathology of the mouse, reference and atlas. Cache
River Press, Vienna, ch 4, pp. 29–48.
Leiter, E.H., Herberg, L., 1996. Aging, pancreatic
islets, and glucose homeostasis in inbred mice. In:
Mohr, U., Dungworth, D.L., Capen, C.C., Carlton,
W.W., Sundberg, J.P., Ward, J.M. (eds), Pathobiology
of the aging mouse, vol. 1. ILSI Press, Washington,
pp. 153–170.
Lewis, D.J., 1984. Spontaneous lesions of the mouse
biliary tract. J. Comp. Pathol. 94, 263–271.
Liebelt, A.G., 1986. Unique features of anatomy,
histology, and ultrastructure, kidney, mouse. In:
Jones. T.C., Mohr, U., Hunt, R.D. (Eds.), Urinary
system. ILSI monographs on pathology of laboratory
animals. Springer-Verlag, Berlin, pp. 24–44.
Linder, C.C., 2003. Mouse nomenclature and
maintenance of genetically engineered mice. Comp.
Med. 53, 119–125.
Linder, C.C., 2006. Genetic variables that influence
phenotype. ILAR. J. 47, 132–140.
Linder, C.C., Davisson, M.T., 2004. Strains, stocks,
and mutant mice. In: Hedrich, H.J., Bullock, G.
(Eds.), The handbook of experimental animals: the
laboratory mouse. Elsevier, San Diego, pp. 25–46.
Long, P.H., Herbert, R.A., 2002. Epithelial-induced
intrapulpal denticles in B6C3F1 mice. Toxicol.
Pathol. 30, 744–748.
Long, P.H., Leininger, J.R., 1999a. The teeth. In:
Maronpot, R.R., Boorman, G.A., Gaul, B.W. (Eds.),
Pathology of the mouse, reference and atlas. Cache
River Press, Vienna, ch 3, pp. 13–28.
Long, P.H., Leininger, J.R., 1999b. Bones, joints, and
synovia. In: Maronpot, R.R., Boorman, G.A., Gaul,
B.W. (Eds.), Pathology of the mouse, reference and
atlas. Cache River Press, Vienna, ch 25, pp. 645–678.
Long, P.H., Leininger, J.R., Ernst, H., 1996.
Proliferative lesions of bone, cartilage, tooth and
synovium in rats, MST-2. In: Guides for toxicologic
pathology. STP/ARP/AFIP, Washington.
Losco, P.E., 1995. Dental dysplasia in rats and mice.
Toxicol. Pathol. 23, 677–688.
Maeda, N., Doi, K., Mitsuoka, T., 1986. Development
of heart and aortic lesions in DBA/2NCrj mice. Lab.
Anim. 20, 5–8.
Maekawa, A., Enomoto, M., Hirouchi, Y., et al., 1996a.
Changes in the upper digestive tract and stomach.
In: Mohr, U., Dungworth, D.L., Capen, C.C.,
Carlton, W.W., Sundberg, J.P., Ward, J.M. (Eds.),
Pathobiology of the aging mouse, vol. 2. ILSI Press,
Washington, pp. 267–286.
Maekawa, A., Enomoto, M., Iwata, H., 1996b.
Changes in the intestine and peritoneum. In: Mohr,
U., Dungworth, D.L., Capen, C.C., Carlton, W.W.,
Sundberg, J.P., Ward, J.M. (Eds.), Pathobiology of
the aging mouse, vol. 2. ILSI Press, Washington,
pp. 287–294.
Mahesh Kumar, M.J., Nagarajan, P., Venkatesan, R.,
et al., 2004. Case report and short communication:
Rectal prolapse associated with an unusual
combination of pinworms and citrobacter species
infection in FVB mice colony. Scand. J. Lab. Anim.
Sci. 31, 221–223.
Mahler, J.F., Elwell, M.R., 1999. The pituitary gland.
In: Maronpot, R.R., Boorman G.A., Gaul, B.W.,
(Eds.), Pathology of the mouse, reference and atlas.
Cache River Press, Vienna, ch 19, pp. 491–507.
Mahler, J.F., Stokes, W., Mann, P.C., et al., 1996.
Spontaneous lesions in aging FVB/N mice. Toxicol.
Pathol. 24, 710–716.
Maita, K., Hirano, M., Harada, T., et al., 1988.
Mortality, major cause of moribundity, and
70 BACKGROUND LESIONS IN LABORATORY ANIMALS
spontaneous tumors in CD-1 mice. Toxicol. Pathol.
16, 340–349.
Malarkey, D.E., Johnson, K., Ryan, L., et al., 2005.
New insights into functional aspects of liver
morphology. Toxicol. Pathol. 33, 27–34.
Marillier, R.G., Michels, C., Smith, E.M., et al., 2008.
IL-4/IL-13 independent goblet cell hyperplasia in
experimental helminth infections. BMC. Immunol.
9, 11 (http://www.biomedcentral.
com/1471-2172/9/11).
Maronpot, R.R., 2006. A monograph on
histomorphologic evaluation of lymphoid organs.
Toxicol. Pathol. 34, 407–408.
Maronpot, R.R., Boorman, G.A., Gaul, B.W. (Eds.),
1999. Pathology of the mouse, reference and atlas.
Cache River Press, Vienna.
Marshall, G.E., Shortland, J.R., Hudson, G., 1988.
Crystalloid material in cells of the murine
mononuclear phagocyte system. J. Anat. 157,
217–228.
Meier, H., Hoag, W.G., 1966. Blood coagulation. In:
Green, E.L. (Ed.), Biology of the laboratory mouse.
McGraw-Hill, New York, ch 18, pp. 373–376.
Mery, S., Gross, E.A., Joyner, D.R., et al., 1994. Nasal
diagrams: a tool for recording the distribution of
nasal lesions in rats and mice. Toxicol. Pathol. 22,
353–372.
Michael, L.H., Taffet, G.E., Frangogiannis, N.G., et al.,
2004. The cardiovascular system. In: Hedrich, H.J.,
Bullock, G. (Eds.), The handbook of experimental
animals: the laboratory mouse. Elsevier, San Diego,
pp 207–224.
Militzer, K., Wecker, E., 1986. Behaviour-associated
alopecia areata in mice. Lab. Anim. 20, 9–13.
Mohr, U. (Ed.), 2001. International classification of
rodent tumors, the mouse. Springer, Berlin.
Mohr, U., Dungworth, D.L., Capen, C.C., et al (Eds.),
1996a. Pathobiology of the aging mouse, vol. 1. ILSI
Press, Washington.
Mohr, U., Dungworth, D.L., Capen, C.C., et al (Eds.),
1996b. Pathobiology of the aging mouse, vol. 2. ILSI
Press, Washington.
Montgomery, Jr., C.A., 1986a. Infarction, kidney, rat,
mouse. In: Jones, T.C., Mohr, U., Hunt, R.D.
(Eds.), Urinary system. ILSI monographs on
pathology of laboratory animals. Springer-Verlag,
Berlin, pp 179–184.
Montgomery, Jr., C.A., 1986b. Interstitial nephritis,
mouse. In: Jones, T.C., Mohr, U., Hunt, R.D.
(Eds.), Urinary system. ILSI monographs on
pathology of laboratory animals. Springer-Verlag,
Berlin, pp. 210–215.
Montgomery, Jr., C.A., 1986c. Suppurative nephritis,
pyelonephritis, mouse. In: Jones, T.C., Mohr, U.,
Hunt, R.D. (Eds.), Urinary system. ILSI
monographs on pathology of laboratory animals.
Springer-Verlag, Berlin, pp. 215–219.
Monticello, T.M., Morgan, K.T., Uraih, L., 1990.
Nonneoplastic nasal lesions in rats and mice.
Environ. Health. Perspect. 85, 249–274.
Morawietz, G., Ruehl-Fehlert, C., Kittel, B., et al.,
2004. Revised guides for organ sampling and
trimming in rats and mice.Part 3. A joint publication
of the RITA and NACAD groups. Exp. Toxicol.
Pathol. 55, 433–449.
Morgan, K.T., Sheldon, W.G., 1988. Lipoma, brain,
mouse. In: Jones, T.C., Mohr, U., Hunt, R.D.
(Eds.), Nervous system. ILSI monographs on
pathology of laboratory animals. Springer-Verlag,
Berlin, pp 130–134.
Morrisey, R., 1986. Renal calcifications, mouse.
In: Jones, T.C., Mohr, U., Hunt, R.D. (Eds.),
Urinary system. ILSI monographs on pathology
of laboratory animals. Springer-Verlag, Berlin,
pp. 361–364.
Morton, D., Tekeli, S., 1997. ‘Have you seen this?’
Pituitary cysts in a mouse. Toxicol. Pathol. 25, 333.
Mouse Genome Informatics website: http://
www.informatics.jax.org/, 08 July 2011.
Müller-Röver, S., Handjiski, B., van der Veen, C.,
et al., 2001. A comprehensive guide for the accurate
classification of murine hair follicles in distinct hair
cycle stages. J. Invest. Dermatol. 117, 3–15.
Mullink, J.W., Haneveld, G.T., 1979. Polyarteritis in
mice due to spontaneous hypertension. J. Comp.
Pathol. 89, 99–106.
Murray, A.B., Luz, A., 1990. Acidophilic macrophage
pneumonia in laboratory mice. Vet. Pathol. 27,
274–281.
National Toxicology Program, Histrocial Controls
Database: http://ntp.niehs.nih.gov/, 08 July 2011.
Ninomiya, H., Inomata, T., Ogihara, K., 1999.
Obstructive uropathy and hydronephrosis in male
KK-Ay mice: a report of cases. J. Vet. Med. Sci. 61,
53–57.
Novilla, M.N., Smith, C.K., 1996. Changes in skeletal
muscle. In: Mohr, U., Dungworth, D.L., Capen,
C.C., Carlton, W.W., Sundberg, J.P., Ward, J.M.
(Eds.), Pathobiology of the aging mouse, vol. 2. ILSI
Press, Washington, pp. 401–413.
Nyska, A., Maronpot, R.R., 1999. The adrenal gland.
In: Maronpot, R.R., Boorman, G.A., Gaul, B.W.,
(Eds.), Pathology of the mouse, reference and atlas.
Cache River Press, Vienna, ch 20, pp. 509–536.
Ogawa, Y., 2003. Immunocytochemistry of
myoepithelial cells in the salivary glands. Prog.
Histochem. Cytochem. 38, 343–426.
Okada, A., Yabuki, A., Matsumoto, M., et al., 2005.
Development of gender differences in DBA/2Cr
mouse kidney morphology during maturation. J. Vet.
Med. Sci. 67, 877–882.
Pack, R.J., Al-Ugaily, L.H., Morris, G., 1981. The cells
of the tracheobronchial epithelium of the mouse: a
quantitative light and electron microscope study. J.
Anat. 132, 71–84.
Page, D.L., Glenner, G.G., 1972. Social interaction
and wounding in the genesis of ‘spontaneous’ murine
amyloidosis. Am. J. Pathol. 67, 555–567.
Paus, R., Müller-Röver, S., Van Der Veen, C., et al.,
1999. A comprehensive guide for the recognition
and classification of distinct stages of hair follicle
morphogenesis. J. Invest. Dermatol. 113, 523–532.
Payne, A.P., 1994. The harderian gland: a tercentennial
review. J. Anat. 185, 1–49.
Pearse, G., 2006a. Normal structure, function and
histology of the thymus. Toxicol. Pathol. 34,
504–514.
Pearse, G., 2006b. Histopathology of the thymus.
Toxicol. Pathol. 34, 515–547.
Peckham, J.C., Heider, K., 1999. Skin and subcutis. In:
Maronpot, R.R., Boorman, G.A., Gaul, B.W. (Eds.),
Pathology of the mouse, reference and atlas. Cache
River Press, Vienna, ch 22, pp. 555–612.
Percy, D.H., Barthold, S.W., 2007. Pathology of
laboratory rodents and rabbits, third ed. Blackwell
Publishing, Ames, ch 1, pp. 3–124.
Pinkerton, K.E., Cowin, L.L., Witschi, H., 1996.
Development, growth, and aging of the lungs. In:
Mohr, U., Dungworth, D.L., Capen, C.C., Carlton,
W.W., Sundberg, J.P., Ward, J.M. (Eds.),
Pathobiology of the aging mouse, vol. 1. ILSI Press,
Washington, pp. 261–272.
Plendi, J., Kölle, S., Sinowatz, F., et al., 1996.
Nonneoplastic lesions of blood vessels. In: Mohr, U.,
Dungworth, D.L., Capen, C.C., Carlton, W.W.,
Sundberg, J.P., Ward, J.M. (Eds.), Pathobiology of
the aging mouse, vol. 1. ILSI Press, Washington,
pp. 385–391.
Pour, P.M., Qureshi, S.R., Salmasi, S., 1983a. Anatomy,
histology, ultrastructure, parathyroid, mouse. In:
Jones, T.C., Mohr, U., Hunt, R.D. (Eds.), Endocrine
system, monographs on pathology of laboratory
animals. Springer-Verlag, Berlin, pp. 252–257.
Pour, P.M., Wilson, J.T., Qureshi, S.R., et al., 1983b.
Cysts, parathyroid, hamster, rat, mouse. In: Jones,
T.C., Mohr, U., Hunt, R.D. (Eds.), Endocrine
system, monographs on pathology of laboratory
animals. Springer-Verlag, Berlin, pp. 288–294.
Radovsky, A., Mahler, J.F., 1999. The nervous system.
In: Maronpot, R.R., Boorman, G.A., Gaul, B.W.
(Eds.), Pathology of the mouse, reference and atlas.
Cache River Press, Vienna, ch 17, pp. 445–470.
Ramot, Y., Manno, R.A., Okazaki, Y., et al., 2009.
Spontaneous aortitis in the Balb/c mouse. Toxicol.
Pathol. 37, 667–671.
Rao, G.N., Boorman, G.A., 1999. History of the
B6C3F1 mouse. In: Maronpot, R.R., Boorman, G.A.,
Gaul, B.W. (Eds.), Pathology of the mouse,
reference and atlas. Cache River Press, Vienna, ch 1,
pp. 1–6.
Rehm, S., Liebelt, A.G., 1996. Nonneoplastic and
neoplastic lesions of the mammary gland. In: Mohr,
U., Dungworth, D.L., Capen, C.C., Carlton, W.W.,
Sundberg, J.P., Ward, J.M. (Eds.), Pathobiology of
the aging mouse, vol. 2. ILSI Press, Washington,
pp. 381–398.
Rehm, S., Nitsche, B., Deerberg, F., 1985a. Non-
neoplastic lesions of female virgin Han:NMRI mice,
incidence and influence of food restriction
throughout life span. I: Thyroid. Lab. Anim. 19,
214–223.
Rehm, S., Wcislo, A., Deerberg, F., 1985b. Non-
neoplastic lesions of female virgin Han:NMRI mice,
incidence and influence of food restriction
throughout life span. II: Respiratory tract. Lab.
Anim. 19, 224–235.
Rehm, S., Sommer, R., Deerberg, F., 1987.
Spontaneous nonneoplastic gastric lesions in female
Han:NMRI mice, and influence of food restriction
throughout life. Vet. Pathol. 24, 216–225.
Renne, R.A., Gideon, K.M., Miller, R.A., et al., 1992.
Histologic methods and interspecies variations in the
laryngeal histology of F344/N rats and B6C3F1
mice. Toxicol. Pathol. 20, 44–51.
Renne, R., Brix, A., Harkema, J., et al., 2009.
Proliferative and nonproliferative lesions of the rat
and mouse respiratory tract. Toxicol. Pathol. 37,
5S–73S.
Reznik, G.K., 1990. Comparative anatomy, physiology,
and function of the upper respiratory tract. Environ.
Health. Perspect. 85, 171–176.
Rittinghausen, S., Kohler, M., Kamino, K., et al., 1997.
Spontaneous myelofibrosis in castrated and
ovariectomized NMRI mice. Exp. Toxicol. Pathol.
49, 351–353.
Rosol, T.J., Yarrington, J.T., Latendresse, J., et al.,
2001. Adrenal gland: structure, function, and
mechanisms of toxicity. Toxicol. Pathol. 29, 41–48.
Rowlatt, C., Franks, L.M., Sheriff, M.U., et al., 1969.
Naturally occurring tumors and other lesions of the
digestive tract in untreated C57BL mice. J. Natl.
Cancer. Inst. 43, 1353–1364.
Ruehl-Fehlert, C., Kittel, B., Morawietz, G., et al.,
2003. Revised guides for organ sampling and
trimming in rats and mice–part 1. Exp. Toxicol.
Pathol. 55, 91–106.
Russell, E.S., 1966. Lifespan and aging patterns. In:
Green, E.L. (Ed.), Biology of the laboratory mouse.
McGraw-Hill, New York, ch 26, pp. 511–519.
Russell, E.S., Meier, H., 1966. Constitutional diseases.
In: Green, E.L. (Ed.), Biology of the laboratory
mouse. McGraw-Hill, New York, ch 29, pp.
571–587.
Russfield, A.B., 1967. Pathology of the endocrine
glands, ovary and testis of rats and mice. In:
Cotchin, E., Roe, F.J.C. (Eds.), Pathology of
laboratory rats and mice. Blackwell, Oxford, ch 14,
pp. 391–467.
Sakura, Y., 1997. Periodontal inflammatory and cystlike
lesions in BDF1 and B6C3F1 mice. Vet. Pathol. 34,
460–463.
Salo, P.T., Seeratten, R.A., Erwin, W.M., et al., 2002.
Evidence for a neuropathic contribution to the

development of spontaneous knee osteoarthrosis in a
mouse model. Acta. Orthop. Scand. 73, 77–84.
Sass, B., 1983a. Embryology, adrenal gland, mouse. In:
Jones, T.C., Mohr, U., Hunt, R.D. (Eds.), Endocrine
system, monographs on pathology of laboratory
animals. Springer-Verlag, Berlin, pp. 3–7.
Sass, B., 1983b. Accessory adrenocortical tissue,
mouse. In: Jones, T.C., Mohr, U., Hunt, R.D.
(Eds.), Endocrine system, monographs on pathology
of laboratory animals. Springer-Verlag, Berlin,
pp. 12–15.
Sass, B., 1983c. Amyloidosis, adrenal, mouse. In:
Jones, T.C., Mohr, U., Hunt, R.D. (Eds.), Endocrine
system, monographs on pathology of laboratory
animals. Springer-Verlag, Berlin, pp. 57–59.
Sass, B., 1986. Glomerulonephritis, mouse. In: Jones,
T.C., Mohr, U., Hunt, R.D. (Eds.), Urinary system.
ILSI monographs on pathology of laboratory animals.
Springer-Verlag, Berlin, pp. 192–210.
Satoh, H., Furuhama, K., 2001. ‘Have you seen this?
An epidermoid cyst with rosette-like structures of
the cerebrum in a male BALB/c Mouse. Toxicol.
Pathol. 29, 498–500.
Saunders, L.Z., 1967. Ophthalmic pathology in rats
and mice. In: Cotchin, E., Roe, F.J.C. (Eds.),
Pathology of laboratory rats and mice. Blackwell,
Oxford, ch 12, pp. 349–371.
Seely, J.C., 1996a. Gallbladder. In: Mohr, U.,
Dungworth, D.L., Capen, C.C., Carlton, W.W.,
Sundberg, J.P., Ward, J.M. (Eds.), Pathobiology of
the aging mouse, vol. 2. ILSI Press, Washington,
pp. 243–249.
Seely, J.C., 1996b. Salivary glands. In: Mohr, U.,
Dungworth, D.L., Capen, C.C., Carlton, W.W.,
Sundberg, J.P., Ward, J.M. (Eds.), Pathobiology of
the aging mouse, vol. 2. ILSI Press, Washington,
pp. 261–265.
Seely, J.C., 1999. Kidney. In: Maronpot, R.R.,
Boorman, G.A., Gaul, B.W. (Eds.), Pathology of the
mouse, reference and atlas. Cache River Press,
Vienna, ch 9, pp. 207–234.
Seely, J.C., Boorman, G.A., 1999. Mammary gland and
specialized sebaceous glands (zymbal, preputial,
clitoral, anal). In: Maronpot, R.R., Boorman, G.A.,
Gaul, B.W. (Eds.), Pathology of the mouse,
reference and atlas. Cache River Press, Vienna,
ch 23, pp. 613–635.
Serfilippi, L.M., Pallman, D.R., Gruebbel, M.M., et al.,
2004. Assessment of retinal degeneration in outbred
albino mice. Comp. Med. 54, 69–76.
Shackelford, C.C., Elwell, M.R., 1999. Small and large
intestine, and mesentery. In: Maronpot, R.R.,
Boorman, G.A., Gaul, B.W. (Eds.), Pathology of the
mouse, reference and atlas. Cache River Press,
Vienna, ch 6, pp. 81–118.
Sidman, R.L., et al., High-resolution mouse brain atlas,
available electronically at: http://www.hms.harvard.
edu/research/brain/index.html, 08 July 2011.
Slack, J.M., 1995. Developmental biology of the
pancreas. Development. 121, 1569–1580.
Smith, R.J., Frommer, J., 1975. Quantitative
morphology and carbohydrate histochemistry of the
mouse submandibular gland following prepubertal
castration. Am. J. Anat. 144, 137–147.
Sokoloff, L., 1967. Articular and musculoskeletal
lesions of rats and mice. In: Cotchin, E., Roe, F.J.C.
(Eds.), Pathology of laboratory rats and mice.
Blackwell, Oxford, ch 13, pp. 373–390.
Son, W.C., 2003a. Factors contributory to early death
of young CD-1 mice in carcinogenicity studies.
Toxicol. Lett. 145, 88–98.
Son, W.C., 2003b. Analysis of fighting-associated
wounds causing death of young male CD-1 mice in
carcinogenicity studies. Scand. J. Lab. Anim. Sci. 30,
101–111.
Staats, J., 1966. The laboratory mouse. In: Green, E.L.
(Ed.), Biology of the laboratory mouse. McGraw-
Hill, New York, pp. 1–9.
Stenn, K.S., Paus, R., 2001. Controls of hair follicle
cycling. Physiol. Rev. 81, 449–494.
Stirling, P., Tullo, A.B., Blyth, W.A., et al., 1983.
Retinal degeneration in NIH (inbred) mice. Exp.
Eye. Res. 36, 761–763.
Strozik, E., Festing, M.F., 1981. Whisker trimming in
mice. Lab. Anim. 15, 309–312.
Sundberg, J.P., 2004. Skin and adnexa of the laboratory
mouse. In: Hedrich, H.J., Bullock, G. (eds.), The
handbook of experimental animals: the laboratory
mouse. Elsevier, San Diego, pp. 195–206.
Sundberg, J.P., Hogan, M.E., King, Jr., L.E., 1996.
Normal biology and aging changes of skin and hair.
In: Mohr, U., Dungworth, D.L., Capen, C.C.,
Carlton, W.W., Sundberg, J.P., Ward, J.M. (Eds.),
Pathobiology of the aging mouse, vol. 2. ILSI Press,
Washington, pp. 303–323.
Suttie, A.W., 2006. Histopathology of the spleen.
Toxicol. Pathol. 34, 466–503.
Tanaka, S., Matsuzawa, A., 1995. Comparison of
adrenocortical zonation in C57BL/6J and DDD
mice. Exp. Anim. 44, 285–291.
Taylor, D.M., Fraser, H., 1973. Hydronephrosis in
inbred strains of mice with particular reference to
the BRVR strain. Lab. Anim. 7, 229–236.
Taylor, D.M., 1985. Urethral plugs and urine retention
in male mice. Lab. Anim. 19, 189–191.
Tew, J.G., Thorbecke, G.J., Steinman, R.M., 1982.
Dendritic cells in the immune response:
Characterisitics and recommended nomenclature
(A report from the Reticuloendothelial Society
Committee on Nomenclature). J. Reticuloendothel.
Soc. 31, 371–380.
Thody, A.J., Shuster, S., 1989. Control and function of
sebaceous glands. Physiol. Rev. 69, 383–416.
Thomas, G.A., WilliamsWilliams, E.D., 1996. Changes
in the structure and function of the thyroid
follicular cell. In: Mohr, U., Dungworth, D.L.,
Capen, C.C., Carlton, W.W., Sundberg, J.P., Ward,
J.M. (Eds.), Pathobiology of the aging mouse, vol. 1.
ILSI Press, Washington, pp. 87–101.
Thoolen, B., Maronpot, R.R., Harada, T., et al., 2010.
Proliferative and nonproliferative lesions of the rat
and mouse hepatobiliary system. Toxicol. Pathol. 38,
5S–81S.
Thuilliez, C., Dorso, L., Howroyd, P., et al., 2009.
Histopathological lesions following intramuscular
administration of saline in laboratory rodents and
rabbits. Exp. Toxicol. Pathol. 61, 13–21.
Tischler, A.S., Sheldon, W., 1996. Adrenal medulla. In:
Mohr, U., Dungworth, D.L., Capen, C.C., Carlton,
W.W., Sundberg, J.P., Ward, J.M. (Eds.),
Pathobiology of the aging mouse, vol. 1. ILSI Press,
Washington, pp. 135–151.
Toth, K., Sugar, J., 1985. Intranuclear and
intracytoplasmic inclusions in normal and neoplastic
hepatocytes, mouse. In: Jones, T.C., Mohr, U.,
Hunt, R.D. (Eds.), Digestive system, ILSI
monographs on pathology of laboratory animals.
Springer-Verlag, Berlin, pp. 92–100.
Travlos, G.S., 2006a. Normal structure, function, and
histology of the bone marrow. Toxicol. Pathol. 34,
548–565.
Travlos, G.S., 2006b. Histopathology of bone marrow.
Toxicol. Pathol. 34, 566–598.
Tucker, A.S., 2007. Salivary gland development. Semin.
Cell. Dev. Biol. 18, 237–244.
Tucker, M.J., Baker, S.B.D.e.C., 1967. Diseases if
specific pathogen-free mice. In: Cotchin, E., Roe,
F.J.C. (Eds.), Pathology of laboratory rats and mice.
Blackwell, Oxford, ch 24, pp. 787–824.
Van den Broeck, W., Derore, A., Simoens, P., 2006.
Anatomy and nomenclature of murine lymph nodes:
Descriptive study and nomenclatory standardization
in BALB/cAnNCrl mice. J. Immunol. Methods. 312,
12–19.
van der Heijden, A., van Dijk, J.E., Lemmens, A.G.,
et al., 1995. Spleen pigmentation in young C57BL
Mouse 71
mice is caused by accumulation of melanin. Lab.
Anim. 29, 459–463.
Van Loo, P.L., Van Zutphen, L.F., Baumans, V.,
2003. Male management: Coping with aggression
problems in male laboratory mice. Lab. Anim. 37,
300–313.
Van Loo, P.L., Van der Meer, E., Kruitwagen, C.L.,
et al., 2004. Long-term effects of husbandry
procedures on stress-related parameters in male
mice of two strains. Lab. Anim. 38, 169–177.
van Rees, E.P., Sminia, T., Dukstra, C.D., 1996.
Structure and development of the lymphoid organs.
In: Mohr, U., Dungworth, D.L., Capen, C.C.,
Carlton, W.W., Sundberg, J.P., Ward, J.M. (Eds.),
Pathobiology of the aging mouse, vol. 1. ILSI Press,
Washington, pp. 173–187.
Van Vleet, J.F., Ferrans, V.J., 1986. Myocardial diseases
of animals. Am. J. Pathol. 124, 98–178.
Van Vleet, J.F., Ferrans, V.J., 1991a. Inherited
dystrophic cardiac calcinosis, mouse. In: Jones, T.C.,
Mohr, U., Hunt, R.D. (Eds.), Cardiovascular and
musculoskeletal system. ILSI monographs on
pathology of laboratory animals. Springer-Verlag,
Berlin, pp. 9–14.
Van Vleet, J.F., Ferrans, V.J., 1991b. Pathologic
reactions of skeletal muscle to injury. In: Jones, T.C.,
Mohr, U., Hunt, R.D. (Eds.), Cardiovascular and
musculoskeletal system. ILSI monographs on
pathology of laboratory animals. Springer-Verlag,
Berlin, pp. 109–126.
van Zweiten, M.J., Hollander, C.F., 1985. Polyploidy,
liver, rat. In: Jones, T.C., Mohr, U., Hunt, R.D.
(Eds.), Digestive system. ILSI monographs on
pathology of laboratory animals. Springer-Verlag,
Berlin, pp. 83–86.
Walton, M., 1978. A spontaneous ankle deformity
in an inbred strain of mouse. J. Pathol. 124,
189–194.
Wancket, L.M., Devor-Henneman, D., Ward, J.M.,
2008. Fibro-osseous (FOL) and degenerative joint
lesions in female outbred NIH black Swiss mice.
Toxicol. Pathol. 36, 362–365.
Ward, J.M., Mann, P.C., Morishima, H., et al., 1999.
Thymus, spleen, and lymph nodes. In: Maronpot,
R.R., Boorman, G.A., Gaul, B.W. (Eds.), Pathology
of the mouse, reference and atlas. Cache River
Press, Vienna, ch 13, pp. 333–360.
Ward, J.M., Anver, M.R., Mahler, J.F., et al., 2000.
Pathology of mice commonly used in genetic
engineering (C57BL/6, 129, B6,129, and FVB/N).
In: Ward, J.M., Mahler, J.F., Maronpot, R.R., et al.
(Eds.), Pathology of genetically engineered mice.
Iowa State University Press, Ames, ch 13, pp.
161–179.
Ward, J.M., Yoon, M., Anver, M.R., et al., 2001.
Hyalinosis and Ym1/Ym2 gene expression in the
stomach and respiratory tract of 129S4/SvJae and
wild-type and CYP1A2-null B6, 129 mice. Am. J.
Pathol. 158, 323–332.
Waring, H., 1935. Memoirs: The development of the
adrenal gland of the mouse. Q. J. Microscop. Sci.
78, 329–366.
Waring, H., Scott, E., 1937. Some abnormalities of the
adrenal gland of the mouse with a discussion on
cortical homology. J. Anat. 71, 299–314.
Weber, K., 2007. Induced and spontaneous lesions in
teeth of laboratory animals. J. Toxicol. Pathol. 20,
203–213.
Wijnands, M.V.W., Kuper, C.F., Schuurman, H.-J.,
et al., 1996. Nonneoplastic lesions of the
hematopoietic system. In: Mohr, U., Dungworth,
D.L., Capen, C.C., Carlton, W.W., Sundberg, J.P.,
Ward, J.M. (Eds.), Pathobiology of the aging mouse,
vol. 1. ILSI Press, Washington, pp. 205–217.
Willard-Mack, C.L., 2006. Normal structure, function,
and histology of lymph nodes. Toxicol. Pathol. 34,
409–424.
Wilson, J.W., Leduc, E.H., 1948. The occurrence and
formation of binucleate and multinucleate cells and
72 BACKGROUND LESIONS IN LABORATORY ANIMALS
polyploid nuclei in the mouse liver. Am. J. Anat. 82,
353–391.
Wojcinski, Z.W., Albassam, M.A., Smith, G.S., 1991.
Hyaline glomerulopathy in B6C3F1 mice. Toxicol.
Pathol. 19, 224–229.
Wolf, D.C., Hard, G.C., 1996. Pathology of the
kidneys. In: Mohr, U., Dungworth, D.L., Capen,
C.C., Carlton, W.W., Sundberg, J.P., Ward, J.M.
(Eds.), Pathobiology of the aging mouse, vol. 1. ILSI
Press, Washington, pp. 331–344.
Wright, J.R., Lacy, P.E., 1988. Spontaneous
hydronephrosis in C57BL/KsJ mice. J. Comp.
Pathol. 99, 449–454.
Yabuki, A., Suzuki, S., Matsumoto, M., et al., 1999.
Morphometrical analysis of sex and strain
differences in the mouse nephron. J. Vet. Med. Sci.
61, 891–896.
Yabuki, A., Matsumoto, M., Nishinakagawa, H., et al.,
2003. Age-related morphological changes in kidneys
of SPF C57BL/6Cr mice maintained under
controlled conditions. J. Vet. Med. Sci. 65,
845–851.
Yamamoto, H., Iwase, N., 1998. Spontaneous
osteoarthritic lesions in a new mutant strain of the
mouse. Exp. Anim. 47, 131–135.
Yamamoto, H., Iwase, N., Kohno, M., 1999.
Histopathological characterization of spontaneously
developing osteoarthropathy in the BCBC/Y mouse
established newly from B6C3F1 mice. Exp. Toxicol.
Pathol. 51, 15–20.
Yamate, J., Tajima, M., Maruyama, Y., et al., 1987.
Observations on soft tissue calcification in
DBA/2NCrj mice in comparison with CRJ:CD-1
mice. Lab. Anim. 21, 289–298.
Yang, Y.H., Campbell, J.S., 1964. Crystalline
excrements in bronchitis and cholecystitis of mice.
Am. J. Pathol. 45, 337–345.
Yarrington, J.T., 1996. Adrenal cortex. In: Mohr, U.,
Dungworth, D.L., Capen, C.C., et al. (Eds.),
Pathobiology of the aging mouse, vol. 1. ILSI Press,
Washington, pp. 125–133.
Yoshiki, A., Moriwaki, K., 2006. Mouse phenome
research: implications of genetic background. ILAR.
J. 47, 94–102.

HAMSTERS
Introduction
The total numbers of Syrian hamsters (Mesocrice-
tus auratus) used in biomedical research is much
less than the numbers of rats and mice used,
however, the Syrian hamster is an important
research subject. Hamsters are used in specialized
research situations where the use of their cheek
pouch for tumor induction and transplantation has
contributed significantly to carcinogenesis research
(Strandberg, 1987). Hamsters are also used in
contract research laboratories and pharmaceutical
companies for occasional studies. Information
about the incidence of spontaneously occurring
lesions is essential if the Syrian hamster is to be
used in acute and chronic toxicity studies (Pour
et al 1976a). The interpretation of tumor inci-
dences in hamster carcinogenicity studies depends
on a good knowledge of the background neoplastic
and non-neoplastic lesions of the species involved
(Pour et al 1976a).
Hamsters are occasionally chosen as the candi-
date species for carcinogenicity studies in preclini-
cal studies. Reasons include the fact that hamsters
are less sensitive in developing olfactory mucosa
toxicity (Toth et al 1961) and hamsters have a
low incidence of spontaneous neoplastic lesions
(Kamino et al 2001a).
Cardiovascular system
Atrial thrombosis, secondary to heart failure, is
seen commonly in older hamsters (Gad 2007a).
This lesion generally occurs in the left atrium and
auricle and may cause dilatation of the left ventri-
cle (Fig. 5.1). Female hamsters are more com-
monly affected than male hamsters and the
syndrome may be associated with amyloidosis
(Percy & Barthold 2007). Atrial thrombosis may
be associated with myocardial degeneration and
fibrosis as well as medial degeneration and calcifi-
cation of the coronary arteries (Percy & Barthold
2007). In addition, vegetative endocarditis or
aortic valve thrombosis has also been observed in
older hamsters (Fig. 5.2). Vascular calcinosis has
been noted in the aorta and coronary and renal
arteries; however, this lesion is thought to be
related to diet (Pour et al 1976d). Fibrinoid
degeneration of arterioles may also be observed in
aging hamsters (Percy & Barthold 2007).
FIGURE 5.1  Left atrial thrombosis and dilatation of left
ventricle in hamster. ×40.
FIGURE 5.2  Aortic valve thrombosis of hamster heart.
×100.
Hemolymphoreticular system
In common with other laboratory animals, ham-
sters display atrophy and involution of the thymus
as they reach adulthood (Fig. 5.3).
FIGURE 5.3  Severe atrophy of the thymus in a hamster.
×100.
Respiratory system
The severe cardiac insufficiency caused by atrial
thromboses or aortic valve thromboses causes
chronic pulmonary congestion and this results in
Hamsters and guinea pigs 73
CHAPTER 5 
Elizabeth F McInnes
Hamsters and guinea pigs
an increase in hemosiderin-laden, alveolar macro-
phages within the alveoli of the lung (Fig. 5.4).
Neuroendocrine cell hyperplasia occurs in small
foci in the bronchioles and within the trachea
(Fig. 5.5). The presence of spontaneous C-cell
or neuroendocrine cell hyperplasia or benign or
malignant tumors of these cells in the larynx
(Pour et al 1985) or the trachea (Ernst et al 1995)
may be unique to hamsters. The frequency varies
considerably between colonies of hamsters (Mohr
et al 1996). Pour and co-workers (1985) reported
an incidence of 9% in the larynx of Syrian ham-
sters. Ernst and co-workers have confirmed the
neuroendocrine origin of these cells and stated
that they occur in the larynx and upper trachea.
The proliferation of neuroendocrine cells in the
larynx and upper trachea is first observed at 15
months of age in Syrian hamsters and it is dif-
ficult to differentiate between hyperplasia, benign
and malignant tumors involving these cells (Mohr
et al 1996). The cytoplasm of the neuroendocrine
cells tends to be clear with large, central nuclei
(Mohr et al 1996). Mitotic figures are rare (Mohr
et al 1996). Ernst and coworkers 1995 demon-
strated that the neuroendocrine cells in these
laryngeal and tracheal locations are positive for
calcitonin, calcitonin gene related peptide, neu-
ron-specific enolase (NSE), and serotonin using
immunohistochemistry. Benign tumors of neu-
roendocrine cells do not display invasion while
malignant tumors of these cells demonstrate sub-
mucosal and lumenal invasion and metastasis to
the lung (Ernst et al 1995). The term C-cell
hyperplasia rather than neuroendocrine cell
hyperplasia is now favored since the origin of
these cells is thought to be the C-cells from the
thyroid. In addition, alveolar histiocytosis may be
observed in the lungs of older hamsters (Percy &
Barthold 2007).
FIGURE 5.4  Severe alveolar histiocytosis and congestion
due to chronic passive congestion due to atrial thrombo-
sis. ×200.
74 BACKGROUND LESIONS IN LABORATORY ANIMALS
FIGURE 5.5  Neuroendocrine cell hyperplasia in the
hamster trachea. ×100. Turusov, V.S., Mohr, U. (eds),
1996. Pathology of Tumors in Laboratory Animals, Vol. III,
Tumors of the Hamster, 2nd edition. IARC Scientific Pub-
lications No. 126, 189–222, IARC (International Agency
for Research on Cancer), Oxford University Press.
Gastrointestinal system
The Syrian hamster has a long duodenum, long
jejunum, short ileum, large cecum and long colon.
The hamster cecum is divided into apical and basal
portions separated by a semi-lunar valve and a
series of four valves in the ileocaecocolic area. The
hamster is a pregastric fermenter. The hamster
stomach has a distinct constriction between the
forestomach and glandular stomach and there is
almost no lesser curvature, resulting in two blind-
ending sacs.
Cyst formation and mineralization in the hard
palate of a hamster is occasionally observed (Fig.
5.6). Mineralization in the tongue of a hamster
is also noted (Fig. 5.7). Mineralization of the
gastric mucosa is observed commonly in hamsters
(Fig. 5.8). Ulcers and erosions are observed com-
monly in the hamster forestomach as well as in
the glandular region (Pour et al 1976a & b) (Fig.
5.9). Herniation of gastric glands into the tunica
muscularis occurs in the hamster stomach (Fig.
5.10). Tumors of granular cells and areas of pro-
liferation composed of granular cells in the intes-
tinal walls of white hamsters have been described
by Pour and co-workers (1973). This finding may
be observed in the serosa of the small and large
intestine.
FIGURE 5.6  Cyst formation and mineralization in the hard
palate of a hamster. ×100.
FIGURE 5.7  Mineralization in the tongue of a hamster.
×100.
FIGURE 5.8  Mineralization in gastric mucosa of hamster
stomach. ×200.
FIGURE 5.9  Focal erosion in hamster stomach. ×100.
FIGURE 5.10  Herniation of gastric glands into tunica
muscularis in hamster stomach. ×100.
Papillary proliferation of pancreatic common
duct epithelium with goblet cell metaplasia is an
age-related change in the hamster.
Liver and biliary system
In the liver, intranuclear inclusions are a non-
specific background lesion and the cause is not
known; however, the inclusions are thought to
be formed by invagination of nuclear mem-
brane with incorporation of some cell cytoplasm.
The inclusions are weakly eosinophilic with
H&E, PAS-negative, 5–8 micrometers in diame-
ter, homogeneous or granular in appearance and
usually eccentrically located within the nucleus.
The nuclei containing the inclusions are frequently
enlarged and have an irregular wrinkled profile
(Fig. 5.11).
FIGURE 5.11  Intranuclear inclusions in hepatocytes in
the hamster liver. ×400.
Intracytoplasmic, non-glycogenic vacuolation is
present in the liver under normal circumstances,
but is more numerous if there is hepatic damage.
The vacuolation is eosinophilic with H&E stain,
strongly PAS-positive before and after diastase
digestion, thus indicating its non-glycogenic char-
acter, and it stains intensely with Sudan Black,
suggesting the presence of bound lipids. The vacu-
olation is also acid-fast with Ziehl–Neelsen stain
and is usually 2–30 µm in diameter.
Liver cysts are common in the hamster and are
thought to derive from bile ducts (Gad 2007a)
(Fig. 5.12). Polycystic liver disease may occur in
hamsters and is characterized by multiple hepatic
cysts observed in older animals (Percy & Barthold
2007). Cysts are most common in the liver and
epididymis as well as in the seminal vesicles and
pancreas (Percy & Barthold 2007).
FIGURE 5.12  Multiple biliary cysts in hamster liver. ×100.

Atrophy of the adjacent hepatocytes, hemosi-
derin deposition, bile duct hyperplasia and peri-
portal lymphocytes may be observed in conjunction
with the liver cysts (Percy & Barthold 2007).
Clear-cell foci are a normal background change in
the hamster liver (Kamino et al 2001b) (Fig.
5.13). In common with rats and mice, bile-duct
hyperplasia is observed in the hamster liver (Fig.
5.14) (Percy & Barthold 2001, 2007). Bile-duct
proliferation forms part of the hepatic cirrhosis
complex in hamsters, which includes periportal
fibrosis, bile duct proliferation, nodular hepato­
cellular proliferation, degeneration, necrosis and
a mixed inflammatory-cell infiltrate (Percy &
Barthold 2007).
FIGURE 5.13  Clear-cell focus in a hamster liver. ×100.
FIGURE 5.14  Bile duct hyperplasia and dilatation in the
hamster liver. ×100.
Amyloidosis occurs frequently in hamsters, par-
ticularly in older animals and in female animals.
Testosterone administration will inhibit the occur-
rence of amyloidosis in female hamsters (Percy &
Barthold 2007). The liver, kidneys, adrenals and
gonads are the most commonly affected organs.
In the liver, amyloid deposition is observed around
the portal areas, within blood vessel walls and
within the sinusoids (Fig. 5.15). Amyloidosis is
an important cause of renal failure in hamsters
(Percy & Barthold 2007). Gall bladder cholelithi-
asis and submucosal lymphocyte aggregates are
occasionally observed in the hamster gall bladder
(Fig. 5.16).
Hamsters and guinea pigs 75
FIGURE 5.17  Hyaline casts and tubular dilatation in the
hamster kidney forming part of the glomerulonephropathy
complex. ×200.
FIGURE 5.15  Severe diffuse amyloidosis of the hamster
liver. ×40. Arteriolar nephrosclerosis, with subsequent
glomerulosclerosis and interstitial nephritis, is a
disease which is seen commonly in hamsters (Gad
2007a). In addition, papillary mineralization is
also a common background change in the hamster
kidney (Gad 2007a).
Endocrine glands
Cortical cysts and cortical hyperplasia of the
adrenal are common background lesions in the
hamster adrenal (Pour et al 1976c). In addition,
the male hamster adrenal is very large due to the
presence of a large adrenal cortex (Fig. 5.18).
FIGURE 5.16  Hamster gall bladder cholelithiasis and
submucosal lymphocyte aggregates. ×100.
Urinary system
Amyloidosis is a disease that occurs in the kidneys
of aging hamsters. Although it is a disease and not
a spontaneous background change, it is mentioned FIGURE 5.18  Large adrenal cortex in male hamster.
here due to the high incidence of this phenome- ×200.
non, which can be as high as 88% (Gad 2007a).
Chronic glomerulonephropathy is also observed
In the hamster pancreas, islet-cell hyperplasia,
in aging hamsters (Fig. 5.17). The disease is char-
similar to that seen in mice, is observed (Pour et al
acterized by glomerulosclerosis, interstitial inflam-
1976c) (Fig. 5.19). Ductal mucus epithelial meta-
mation and fibrosis, hyaline casts, dilatation and
plasia may be observed in the pancreas of a
atrophy of tubules and amyloid deposition in the
hamster (Fig. 5.20).
glomeruli. This degenerative renal disease causes
significant mortality in older hamsters (Percy &
Barthold 2007). The disease occurs more com-
monly in female animals and the cause is not
known, although high protein diets are thought to
play a role (Percy & Barthold 2007).
FIGURE 5.19  Minimal islet cell hyperplasia in a hamster
pancreas. ×100.
76 BACKGROUND LESIONS IN LABORATORY ANIMALS

Reproductive system
Trophoblastic giant cells are derived from the fetal
placenta, specifically from the trophoblast, i.e. the
epithelial cell layer covering the blastocyst. The
blastocyst erodes the uterine mucosa to establish
the hemochorial placenta seen in hamsters, guinea
pigs, other rodents, and primates. The placenta in
the hamster is of the labyrinthine hemotrichorial
type (Percy & Barthold 2007).
Trophoblastic giant cells are in direct contact
with the maternal bloodstream. These cells
exhibit migratory activity and are frequently
found inside mesometrial uterine arteries and
FIGURE 5.20  Ductal mucus epithelial metaplasia in the ovarian arteries. Trophoblast giant cells apparently
pancreas of a hamster. ×200. only migrate toward arterial blood and can be seen FIGURE 5.24  Granular cell accumulation (hyperplasia) in
at three weeks postpartum. A decidual reaction the uterus of a hamster. ×100.
Hyperplasia of the parathyroid is observed may be noted in the uterus of hamster (Fig. 5.22).
occasionally in Syrian hamsters (Pour et al 1976c). Sparse copora lutea are noted in the hamster ovary
(Fig. 5.23).
Skin and appendages
Cheek pouches in the hamster are well developed
and are highly distensible evaginations of lateral
buccal walls. Cheek pouches are used to store
and transport food and can be easily evaginated
under anaesthesia to use as a common site for
experimental tumor implantation and vascular
physiology studies. The hamster cheek pouch is
an immunologically privileged site.
Hamsters have small pigmented spots and the
costovertebral spot or flank organ (Ghadially and
Ghadially 1996). The small pigmented spot is a
term used to describe pigmented spots in the FIGURE 5.25  Granular cell hyperplasia on the serosal
hamster skin which are made up of melanocytes surface of the large intestine. ×100.
and melanophages and which are surrounded by FIGURE 5.22  Decidual reaction in uterus of hamster.
pilosebaceous units. The costovertebral spot or ×100.
flank organ is a paired, black organ on the flank of
the hamster comprising large sebaceous glands
discharging into large hair follicles. Large numbers
of melanocytes are visible around the neck of
these hair follicles. Hip or flank glands in hamsters
secrete during sexual arousal in both sexes and are
used for olfactory marking of territory (Fig. 5.21).
Ulceration may occur over the site of the male
flank gland.

FIGURE 5.26  Granular cell hyperplasia adjacent to

FIGURE 5.21  Male flank organ of the hamster with focal
ulceration. ×100.
FIGURE 5.23  Sparse copora lutea in hamster ovary.
×100.
Granular cell foci are visible in the female
hamster uterus (Kamino et al 2001b) (Fig. 5.24).
In addition, granular cell hyperplasia has been
observed on the serosal surface of the large intes-
tine (Fig. 5.25) and adjacent to the hamster
kidney (Fig. 5.26). Cysts in the ovary are common
in female hamsters (Pour et al 1976c).
hamster kidney. ×200.
In male hamsters, tubular atrophy of the testis
is often noted as a background lesion (Kamino
et al 2001b) (Fig. 5.27) as well as epithelial hyper-
plasia of the hamster epididymis (Fig. 5.28). Pre-
pucial abscesses are common in the hamster (Fig.
5.29). Epithelial atrophy and mineralization of a
hamster prostate are also observed in male ham-
sters (Fig. 5.30).

Hamsters can develop a bedding-associated der-

matitis (Percy & Barthold 2007). The lesion is
characterized by a granulomatous inflammatory
response in the dermis and subcutis of the digits.
A subcutaneous abscess in the skin of a Syrian
hamster has been reported (Kondo et al 2008).
 Hamsters and guinea pigs 77

FIGURE 5.27  Atrophic seminiferous tubules in the

hamster testis. ×100. FIGURE 5.31  Dysplasia of the hamster sternum. ×100. FIGURE 5.34  Mineralization in the brain of a hamster.
×100.

FIGURE 5.28  Epithelial hyperplasia of the hamster epidi-

dymis. ×100. FIGURE 5.32  Cartilage degeneration in the sternum of a
hamster. ×200.
FIGURE 5.35  Chronic suppurative inflammation in the ear
of the hamster. ×40.
Eye and ear
Multifocal retinal dysplasia manifesting as rosette-
like structures making up of the photoreceptor
layer, the outer-membrane layer and the outer
nuclear layer are noted sometimes in the hamster
eye (Gad 2007a). Degeneration of lens fibres, i.e.
cataract, may be observed in the eye of the
hamster (Fig. 5.33).

FIGURE 5.29  Prepuce abscess in a hamster. ×100.

FIGURE 5.36  Middle ear fibrosis in the ear of the hamster.

×40.

FIGURE 5.30  Epithelial atrophy and mineralization of a
hamster prostate. ×100.
Muscle, bones and joints
Cartilage dysplasia (Fig. 5.31) or chondromu­
cinous degeneration (Fig. 5.32) is often observed
in the sternum of the hamster (Kamino et al
2001b).
FIGURE 5.33  Degeneration of lens fibres in the eye of a
hamster. ×200.
In common with other laboratory animals,
mineralization in the brain of a hamster is encoun-
tered (Fig. 5.34). In addition, age-related vacuo­
lation of the brain is also reported (Gerhauser
et al 2012). Chronic suppurative inflammation in
the ear of the hamster (Fig. 5.35) is occasionally
observed and may lead to middle ear fibrosis of
the middle ear (Fig. 5.36).
GUINEA PIGS
Introduction
The Hartley guinea pig strain is most commonly
used in research environments (Percy & Barthold
2007). Heterophils are the equivalent of the
neutrophil in guinea pigs. Guinea pigs (Cavia por-
cellus) originate in South America and are used in
research studies on immunology, audiology and
infectious diseases (Gad et al 2007b). Guinea pigs
are not used in great numbers as they are expen-
sive and do not have readily accessible peripheral
veins for collection of blood. (Gad et al 2007b).
78 BACKGROUND LESIONS IN LABORATORY ANIMALS
Cardiovascular system
Rhabdomyomatosis (nodular glycogen infiltration)
is an incidental finding in guinea pigs of various
ages (Hueper 1941, Rooney 1961, Takahashi et al
1985, Vink 1969). The condition is thought to be
a congenital disease caused by a disorder of glyco-
gen metabolism (Percy & Barthold 2007). The
lesion is present in the heart muscle, usually in the
left ventricle, but also in the atria, interventricular
septum and papillary muscle. If the lesion is large,
it is generally visible at necropsy as pink to yellow
foci (Fig. 5.37) or streaks (Percy & Barthold
2007); however, the lesion may be too small to be
visible macroscopically.
FIGURE 5.37  Rhabdomyomatosis in the heart of a guinea
pig.
Upon histopathological examination, the lesion
is visible as a network of vacuolated myofibres
composed of finely fibrillar to granular, eosi-
nophilic cytoplasm. The vacuoles contain large
quantities of glycogen which will be washed out
during routine fixation and processing (Percy &
Barthold 2007). The glycogen may be demon-
strated in alcohol-fixed specimens stained with
PAS (Percy & Barthold 2007).
There may be displacement and flattening of
myocyte nuclei in some of the affected fibres
(Percy & Barthold 2007). Myofibres with centrally-
located nuclei and radiating fibrillar processes are
called ‘spider cells’ (Percy & Barthold 2007).
Poorly differentiated cardiac myofibres may be
present with identifiable cross-striations. The
lesion does not appear to compromise cardiac
function and is considered to be an incidental
finding.
Hemolymphoreticular system
Degenerate lymphocytes are observed in associa-
tion with Hassall’s corpuscles (Percy & Barthold
2007) in the guinea pig thymus. Thymic cysts in
guinea pigs are common (Percy & Barthold 2007).
Respiratory system
Moderate to marked pulmonary arterial hyperpla-
sia is observed in the guinea pig lung (Percy &
Barthold 2007). Perivascular lymphoid aggregates
are often present in the guinea pig lung and this
lesion may be visible as white, subpleural foci at
necropsy (Percy & Barthold 2007). These lesions
may be caused by antigenic stimuli, but no viral
cause has been isolated (Percy & Barthold 2007).
In common with other laboratory animals, osseous
metaplasia is occasionally noted in the guinea
pig lung. Guinea pigs are prone to foreign-body
pneumonitis (Percy & Barthold 2007). The lesion
is characterized by granulomas made up of gener-
ally plant fibres, surrounded by an inflammatory
cell infiltrate made up predominantly of neu-
trophils, lymphocytes and foreign-body giant
cells. Pseudo­gland formation associated with
diffuse or nodular basal cell hyperplasia of the
olfactory epithelium of nasal tract of young
hamster has been reported.
Gastrointestinal system
Intestinal hemosiderin deposition in the lamina
propria of the large intestine is common in guinea
pigs (Percy & Barthold 2007). The hemosiderin is
contained within macrophages within the lamina
propria. Adipocyte infiltration into the pancreas
is observed in the older guinea pigs (Percy &
Barthold 2007). The areas of adipose tissue may
become quite large and adipocytes may also be
noted within the islets of Langerhans (Percy &
Barthold 2007).
Liver and biliary system
Focal areas of hepatic necrosis with a minimal
inflammatory response are observed in guinea pig
livers (Percy & Barthold 2007). In common with
other laboratory animals, guinea pigs also display
chronic, idiopathic cholangiohepatitis (Percy &
Barthold 2007). The lesion is characterized by
the presence of hepatocyte degeneration, bile-
duct hyperplasia and portal fibrosis (Percy &
Barthold 2007). The cause of the lesion is not
known. Occasionally, focal tears of the liver
capsule with hemorrhage into the peritoneum
are observed at necropsy in guinea pigs (Percy &
Barthold 2007). The cause of this lesion is thought
to be trauma.
Urinary system
Segmental nephrosclerosis is an aging change
observed in guinea pigs older than one year (Percy
& Barthold 2007). The cause of the renal lesions
is not known, however a high protein diet has
been noted to exacerbate the renal lesions (Percy
& Barthold 2007). Segmental nephrosclerosis is
characterized by the presence of pitted, granular,
pale white kidneys at necropsy, and the
histopathological lesion consists of interstitial
fibrosis, tubular dilatation, hyaline casts, inter­
stitial lymphocyte infiltrates, glomerular fibrosis
and medial hypertrophy of arterioles (Percy &
Barthold 2007). Osseous metaplasia has been
observed in the kidney of the guinea pig.
Reproductive system
Kurloff cells are unique mononuclear cells found
in certain sites in guinea pigs (Percy & Barthold
2007), particularly adult, female guinea pigs.
Kurloff cells are encountered routinely in the
spleen, bone marrow and thymus. The cells
increase and decrease in number based on the
estrus cycle of the guinea pig. Increased numbers
of these cells are noted during pregnancy (Percy
& Barthold 2007) and large numbers of them
are present in the placenta where they are
thought to play a role in preventing maternal
rejection of the fetus (Percy & Barthold 2007).
The cells demonstrate granular cytoplasm with
displacement of the nucleus (Percy & Barthold
2007). Kurloff cells are thought to be lymphoid
cells, possibly large granular lymphocytes, and the
intracytoplasmic material is PAS positive. Kurloff
cells may also play a role in tumor rejection (Percy
& Barthold 2007).
Multiple cystic areas may be noted in the
ovaries of female guinea pigs, particularly guinea
pigs older than one year. (Percy & Barthold 2007).
The cysts are lined with cuboidal to columnar
epithelium and the cysts may cause atrophy of the
surrounding ovarian tissue (Percy & Barthold
2007). Large ovarian cysts are thought to derive
from the rete ovarii (Percy & Barthold 2007).
Cystic endometrial hyperplasia, mucometra,
endometritis and endometrial polyps are lesions
noted in conjunction with ovarian cysts (Percy &
Barthold 2007).
Muscles, bones and joints
Myocardial and skeletal muscle degeneration
occurs in guinea-pig muscle. The lesions may also
demonstrate mineralization (Percy & Barthold
2007). The muscles of the hind limbs are particu-
larly prone to this syndrome, although the affected
animal may not demonstrate clinical signs. A
mononuclear cell infiltrate and later fibrosis,
accompanies the degenerative lesions in both
skeletal muscle and cardiac muscle.
Metastatic mineralization occurs in guinea pigs
and results in mineral deposition in the soft tissues
around the elbows and ribs (Percy & Barthold
2007). Mineralization of other tissues such as
lung, trachea, heart, aorta, liver, kidney, stomach,
uterus and eye are also observed. The syndrome
is thought to be caused by dietary mineral imbal-
ances (Percy & Barthold 2007).
Brain and nervous system
Otitis media may occur in guinea pigs. The disease
is caused by various bacteria (such as Streptococcus
pneumoniae, Streptococcus zooepidemicus, Borde-
tella and Pseudomonas sp.) (Percy & Barthold
2007), and may cause otosclerosis of the tympanic
bullae. Bacterial conjunctivitis is also common in
guinea pigs and may be caused by coliforms, Strep-
tococcus zooepidemicus, Staphylococcus aureus
and Pasteurella multocida (Percy & Barthold
2007).
References
Ernst, H., Heinrichs, M., Bargsten, G., et al., 1995.
Neuroendocrine hyperplasias and tumours of the
larynx and trachea in the Syrian hamster. In: Jones,
T.C., Mohr, U., Hunt, R.D., (Eds.), Respiratory
system, second ed. Monographs on Pathology
of Laboratory Animals. Springer-Verlag, Berlin,
pp. 107–116.
Gad, S.C., Hess, F.G., Gad, S.C., 2007a. The
Hamster. In: Gad, S.C., (Ed.), Animal models in
toxicology, second ed. Taylor & Francis, Boca Raton,
pp. 277–312.
Gad, S.C., Peckham, J.C., Gad, S.C., 2007b. The
Guinea pig. In: Gad, S.C., (Ed.), Animal models in
toxicology, second ed. Taylor & Francis, Boca Raton,
pp. 333–400.
Gerhauser, I., Wohlsein, P., Ernst, H., Germann, P.G.,
Baumgartner, W., 2012. Vacuolation and mineralisa-
tion as dominant age-related findings in hamster
brains. Exp. Toxicol. Pathol. (In press).

Ghadially, F.N., Ghadially, R., 1996. Tumours of the
skin. In: Turusov, V.S., Mohr, U., (Eds.), Pathology
of tumours in laboratory animals. Vol 3. Tumours of
the hamster. IARC Scientific Publications no 126,
Lyon IARC, pp. 1–44.
Hueper, W.C., 1941. Rhabdomyomatosis of the
heart in a guinea pig. Am. J. Pathol. 17,
121–124.
Kamino, K., Tillman, T., Mohr, U., 2001a. Spectrum
and age-related incidence of spontaneous tumours
in a colony of Han: AURA hamsters. Exp. Toxic.
Pathol. 52, 539–544.
Kamino, K., Tillmann, T., Boschmann, E., Mohr, U.,
2001b. Age-related incidence of spontaneous
non-neoplastic lesions in a colony of Han:
AURA hamsters. Exp. Toxicol. Pathol. 53,
157–164.
Kondo, H., Onuma, M., Shibuya, H., et al., 2008.
Spontaneous tumors in domestic hamsters. Vet.
Pathol. 45, 674–680.
Mohr, U., Emura, M., Dungworth, D.L., Ernst, H.,
1996. Tumours of the lower respiratory tract. In:
Turusov, V.S., Mohr, U., (Eds.), Pathology of
tumours in laboratory animals. Vol 3. Tumours of
the hamster. IARC Scientific Publications no 126,
Lyon IARC, pp. 189–222.
Percy, D.H., Barthold, S.W., 2007. Pathology
of laboratory rodents and rabbits. third ed.
Blackwell Publishing, Ames, ch 1, pp. 168–196
and 209–247.
Pour, P.M., 1985. Clear cell carcinoma in the larynx of
the Syrian Hamster. In: Jones, T.C., Mohr, U.,
Hunt, R.D., (Eds.), Respiratory system (monographs
on pathology of laboratory animals). Springer-Verlag,
Berlin, pp. 75–77.
Pour, P., Althoff, J., Cardesa, A., 1973. Granular cells
in tumours and in non-tumourous tissues. Arch.
Pathol. 95, 135–138.
Pour, P., Kmoch, N., Greiser, E., et al., 1976a.
Spontaneous tumors and common diseases in two
colonies of Syrian hamsters. I. Incidence and sites.
J. Natl. Cancer Inst. 56, 931–935.
Pour, P., Mohr, U., Cardesa, A., et al., 1976b.
Spontaneous tumors and common diseases in two
colonies of Syrian hamsters. II. Respiratory tract
and digestive system. J. Natl. Cancer Inst. 56,
937–948.
Pour, P., Mohr, U., Althoff, J., et al., 1976c.
Spontaneous tumors and common diseases in two
colonies of Syrian hamsters. III. Urogenital system
and endocrine glands. J. Natl. Cancer Inst. 56,
949–961.
Hamsters and guinea pigs 79
Pour, P., Mohr, U., Altoff, J., et al., 1976d.
Spontaneous tumors and common diseases in two
colonies of Syrian hamsters. IV. Vascular and
lymphatic systems and lesions of other sites. J. Natl.
Cancer Inst. 56 (5), 963–974.
Rooney, J.R., 1961. Rhabdomyomatosis in the heart of
the guinea pig. Cornell Vet. 51, 388–394.
Sandberg, J.D., 1987. Neoplastic diseases. In: Hoosier,
G.L., McPherson, C.W., (Eds.), Laboratory Hamsters.
Academic Press Inc, London, pp. 157–167.
Takahashi, M., Iwata, S., Matsuzawa, H., Fujiwara, H.,
1985. Pathological findings of cardiac
rhabdomyomatosis in the guinea pig. Jikken
Dobutsu. 34 (4), 417–424.
Toth, B., Tomatis, L., Shubik, P., 1961. Multipotential
carcinogenesis with urethan in the Syrian golden
hamster. Cancer Res. 21, 1537–1541.
Turusov, V.S., Mohr, U., (Eds.), 1996. Pathology of
Tumours in Laboratory Animals, Vol. III, Tumours of
the Hamster, second edition. IARC Scientific
Publications No. 126, IARC (International Agency
for Research on Cancer), Oxford University Press,
pp. 189–222.
Vink, H.H., 1969. Rhabdomyomatosis (nodular
glycogenic infiltration) of the heart in guinea-pigs.
J. Pathol. 97 (2), 331–334.
 Minipigs 81
CHAPTER 6 
Elizabeth F McInnes

Minipigs

cells are sometimes encountered within the

Introduction minipig aorta. Occasionally focal arteritis may be
Minipigs are now used in greater numbers in con- observed in the minipig kidney.
tract research organizations as well as in the phar-
maceutical industry. They provide significant
advantages over dogs due to their greater toler-
ance of non-steroidal anti-inflammatory drugs,
antihypertensive agents and sympathicomimetic
drugs (Dincer 2007). In addition, minipigs are
advantageous models for human drugs since their
digestion is similar to that of humans, and their
smaller size makes them more manageable labora-
tory animals. The minipig offers many advantages
as a model in dermal toxicity because of the simi-
larity between porcine and human skin (Mortensen
et al 1998, Lavker et al 1991), including similari- FIGURE 6.3  Lymphoid infiltrate into the cerebrum. ×200.
ties in skin thickness, permeability, pigmentation,
allergic reaction and reaction to burning.
In addition, the gastrointestinal system, diges-
FIGURE 6.1  Arteritis and periarteritis of artery in the
tion and metabolism, as well as the renal and
coronary groove. ×100.
immune systems of minipigs are similar to those
in man. There are also similarities to humans in
reproductive tract histology, physiology, and
in cervical and vaginal secretions and vaginal Hemolymphoreticular system
pH (minipigs are used for intravaginal studies)
The most common histopathological background
(Jørgensen et al 1998). Finally, cardiovascular
lesion noted in minipigs is the focal accumulation
physiology and anatomy, ventricular performance,
of mononuclear inflammatory cells in various
electrophysiology and coronary artery distribution
organs. These cells are predominantly lym-
are similar to those in humans.
phocytes, macrophages and plasma cells (Madsen
Most contract research organizations or phar-
et al 1998), and are generally noted in perivascu-
maceutical companies use the Göttingen minipig
lar and interstitial locations (Madsen et al 1998).
which displays a number of important background
The foci may be observed in the adrenal glands
lesions. Minipigs are derived from the Eurasian FIGURE 6.4  Renal cortical interstitial lymphoid infiltrate.
(Fig. 6.2), cerebrum (Fig. 6.3), epididymis,
wild boar (Sus scrofa). In total, some 40 strains ×200.
oesophagus, kidneys (Fig. 6.4), liver, lung, man-
exist, and the Sinclair, Yucatan and Göttingen
dibular salivary gland, meninges, parotid gland,
strains are the most common. Maturity occurs at Ellipsoids are pale, eosinophilic structures
rectum, stomach, testes, tongue and vagina
four to six months of age. The estrous cycle is arranged concentrically around capillaries or small
(Madsen et al 1998).
approximately 20 days and gestation is approxi- arterioles in the spleen. The ellipsoids display vari-
mately 114 days. able prominence; they consist of phagocytic cells
Advantages to the use of minipigs include their and reticular fibres and have been referred to in
ready availability and their small size compared the literature as Schweigger–Seidel sheaths
with conventional pigs. Extensive data on anatomy, (Charles 1996) (Fig. 6.5).
reproductive physiology, clinical parameters and
histopathology are available for the Göttingen
minipig. Minipigs are also easier to house, handle,
dose and sample than non-human primates and
there are no zoonoses (in contrast to non-human
primates). There are fewer ethical or conservation
concerns, sexually mature animals are readily
available and global transportation of minipigs is
easier.
Disadvantages include the fact that the minipig
is larger than the Beagle dog or non-human
primate, a greater quantity of test material is FIGURE 6.2  Lymphoid infiltrate into the adrenal cortex.
required and minipigs are extremely vocal! ×200.

Cardiovascular system
Arteritis and periarteritis are commonly observed FIGURE 6.5  Schweiggel–Siedel sheaths in the minipig
in a variety of organs, including the epididymides, spleen. ×100.
heart, intestine, kidney, lung, spleen, stomach and
urinary bladder (Fig. 6.1). In the heart, small foci
of lymphocytes and occasional areas of myofibre
necrosis are observed. Mononuclear inflammatory
82 BACKGROUND LESIONS IN LABORATORY ANIMALS
In common with other pig breeds, minipigs
display a reversed cortex and medulla in the
lymph nodes, i.e. the medulla is present on the
outside of the node and the cortex with germinal
centres is visible in the middle of the node. In the
mesenteric lymph node the presence of a perime-
senteric plexus may sometimes be confusing to
pathologists (Fig. 6.6). The vascular supply to the
lymph nodes differs from that of other mammals.
Major nodal arteries subdivide into branches
which envelop the node as a capsular network
from which further branches penetrate the paren-
chyma (Charles 1996).
FIGURE 6.6  Perimesenteric plexus adjacent to the
mesenteric lymph node in the minipig. ×100.
Iron deposits are often visible within macro-
phages in the sinuses of local lymph nodes
(Madsen et al 1998). The widespread iron depos-
its visible in the minipig are thought to be due to
the intramuscular injections of iron administered
to neonatal piglets to prevent anemia (Svendsen
et al 1998). In addition, iron deposits may also be
encountered in the kidney, liver and mandibular
lymph node.
Small lymph node abscesses are common in the
mandibular node (Dincer 2007) (Fig. 6.7) and are
thought to be secondary to local irritation or
inflammation. Sinus histiocytosis is also com-
monly observed in the minipig lymph node. An
increase in granulocytic cells such as eosinophils
can sometimes be observed in the sinuses of the
mesenteric lymph nodes (Dincer 2007) (Fig. 6.8).
FIGURE 6.7  Mandibular lymph node abscess with central
mineralization in the minipig. ×100.
Gastrointestinal system
Interstitial and periductular lymphocytes and
foci of mineralization are commonly seen in the
salivary glands of minipigs (Fig. 6.11), and perig-
landular edema adjacent to the mandibular sali-
vary glands is common. In addition, foci of
squamous epithelial metaplasia may be noted in
the cuboidal epithelium of the salivary-gland
ducts (Fig. 6.12). In the tongue, focal chondro-
cytes are occasionally observed, unassociated with
inflammation (Fig. 6.13). The origin of the carti-
lage cells is thought to be ectopic. Erosive and
ulcerative changes and inflammatory cell infiltra-
tion are occasionally evident in the glandular and
FIGURE 6.8  Eosinophils in the subcapsular sinus. ×100. non-glandular stomach (Dincer 2007) (Fig. 6.14).
Respiratory system
Multifocal aggregates of alveolar macrophages
may be observed in the minipig lung (Fig. 6.9).
Most lesions are focal and mild, and interstitial or
perivascular inflammatory cell infiltrates are often
seen, usually involving lymphocytes and macro-
phages. In addition, mineralized material within
the alveoli (Dincer 2007) (Fig. 6.10) as well as
focal areas of interstitial or purulent bronchopneu-
monia are commonly seen (Dincer 2007). The
pulmonary mineralization is often associated with
macrophages. The pneumonic lesions may be due
to mycoplasma or other bacterial infections
(Dincer 2007). Foreign-body granulomata, often
FIGURE 6.11  Focal area of mineralization in the salivary
surrounding hair or food particles, are also com-
gland of the minipig. ×200.
monly observed in the minipig lung (Dincer
2007). Occasionally pleural thickening or pleuritis
is observed (Dincer 2007).
FIGURE 6.12  Squamous metaplasia of duct epithelium in
sublingual salivary gland. ×10.
FIGURE 6.9  Alveolar macrophage aggregates in the
minipig lung. ×200.
FIGURE 6.13  Foci of cartilage present in minipig tongue.
×10.
FIGURE 6.10  Foci of mineralization in the lung of a
minipig. ×100.
 Minipigs 83

(Svendsen et al 1998). The lesions are generally

Liver and biliary system small and focal, and the basophilic tubules are
Chronic necrotic cholecystitis is occasionally often related to areas of tubular degeneration and
observed in the minipig gall bladder (Svendsen regeneration. Foci of mineralization, particularly
1998) (Fig. 6.16). This lesion is generally observed in the papilla, as well as cortical cysts and occa-
at necropsy where the necrotic gall bladder sionally hyaline casts are also observed in the
appears thickened. Histopathologically the lesion minipig kidney (Dincer 2007) (Fig. 6.19).
is characterized by necrosis of the mucosa with
large numbers of inflammatory cells extending
into the submucosa and smooth muscle layers
(Dincer 2007). Cholecystitis is not associated
with clinical signs or changes in clinical pathology
parameters. Hypoplasia or aplasia of the gall
bladder are also observed as background findings.
FIGURE 6.14  Focal, minimal gastric erosion in minipig
stomach. ×100.

Minimal to moderate hemorrhage and inflam-

mation of the lamina propria and submucosa of
the small and large intestine is observed in mini-
pigs. Moderate numbers of eosinophils may be
noted in the lamina propria of the small intestine
(Fig. 6.15a). Occasional erosive changes are also FIGURE 6.18  Basophilic tubules and interstitial lym-
visible in the intestinal mucosa. Hyperkeratosis is phocytes in the minipig kidney. ×100.
occasionally noted in the stomach due to the
feeding of finely ground rations and pelleted food
(Madsen et al 1998). Herniated large intestinal
glands are often noted within the underlying sub-
mucosal lymphoid tissue in the large intestine of
the minipig (Fig. 6.15b). FIGURE 6.16  Necrotic cholecystitis in the minipig gall
bladder. ×200.

The minipig liver can often display small multi-

focal areas of inflammatory cells, and occasional
foci of single-cell necrosis or necrosis can be seen
throughout the liver (Dincer 2007) (Fig. 6.17). In
addition, fibrous tissue is noted in the interlobular
and subcapsular areas (Dincer 2007). Clear-cell
foci can occasionally be observed in the minipig
liver, and the clear hepatocyte cytoplasm is
thought to be due to glycogen accumulation FIGURE 6.19  Mineralization in papilla of minipig kidney.
(Dincer 2007). Iron deposits are often visible ×200.
within Kupffer cells in the liver.
Multifocal inflammatory-cell infiltration is occa-
sionally observed in the urinary bladder mucosa
FIGURE 6.15a  Moderate numbers of eosinophils in the and submucosa of minipigs (Dincer 2007). Inflam-
lamina propria of the jejunum of a minipig. ×200. mation, made up predominantly of eosinophilic
granulocytes, is occasionally observed in the
ureter of the minipig. Thromboses or areas of
mineralization may be noted in the umbilical
blood vessels within the serosa of the urinary
bladder (Fig. 6.20).

FIGURE 6.17  Focal hepatocyte necrosis in the minipig

liver. ×200.

FIGURE 6.15b  Herniated colonic glands in submucosal

Urinary system
lymphoid tissue. ×100. The minipig has a multipapillate kidney, similar
in structure to the human kidney, including the
presence of true calices. Multifocal interstitial
lymphoid infiltrates and fibrosis, as well as multi-
focal cortical basophilic tubules, are observed FIGURE 6.20  Thrombosed umbilical blood vessel in
commonly in the minipig kidney (Fig. 6.18) serosa of the urinary bladder of minipig. ×100.
84 BACKGROUND LESIONS IN LABORATORY ANIMALS
Endocrine glands
Accessory adrenal cortical tissue is sometimes
observed. Ultimobranchial cysts lined by squa-
mous epithelium and filled with keratin may be
observed in minipigs in the thyroid. The pituitary
gland may sometimes display cysts lined by cili-
ated epithelium as well as mineralization of single
cells (Dincer 2007).
The parathyroid glands in the minipig are dif-
ficult to locate and are generally found within the
thymus. The thyroids are on the ventral surface
of the trachea at the thoracic inlet and are also
difficult to locate. The thyroids are often damaged
by venipuncture procedures (Fig. 6.21), and this
can affect thyroid hormone levels. The parathy-
roids are not attached to the thyroids in the
minipig, but are located close to the carotid artery
bifurcation, within the thymus (Fig. 6.22), adipose
or connective tissue. The thymus is located in the
neck and cranial thorax, lying adjacent to the
trachea. Occasionally, parathyroid glands display
cysts (Dincer 2007).
FIGURE 6.21  Peritracheal hemorrhage and inflammation
due to venipuncture procedures in the minipig. ×100.
FIGURE 6.22  The minipig parathyroid within the thymus.
×20.
Skin and appendages
The skin of the minipig demonstrates multifocal
areas of dermal inflammatory cells and acanthosis
of the epidermis (Dincer 2007). Focal areas of
epidermal ulceration are also common (Fig. 6.23).
FIGURE 6.23  Severe focal epidermal ulceration and FIGURE 6.25  Serous atrophy of adipose tissue in the
dermal inflammatory-cell infiltration in the minipig skin. minipig bone marrow. ×200.
×100.
Brain and nervous system
Reproductive system Mineralization is often observed in the meninges
of the minipig brain (Fig. 6.26), and occasional
In the testes, uni- or bilateral seminiferous tubular areas of perivascular lymphocyte infiltration are
hypoplasia or atrophy is commonly observed and noted in the brain. In addition, meningeal
is characterized by the presence of vacuolation in inflammatory cell infiltrates are sometimes seen.
Sertoli cells and multinucleate cells (Dincer
2007). In the epididymis reduced numbers of
spermatids can be observed, generally related to
tubular atrophy in the testes. Interstitial lymphoid
infiltration is commonly observed in the prostate
of the minipig (Dincer 2007).
Muscles, bones and joints
Multifocal areas of necrosis and inflammatory cell
infiltration are noted in the skeletal muscle of
minipigs. This can be confused with treatment-
related effects. The lesion is observed in skeletal
muscle at many sites, and there is generally
minimal evidence of myofibre regeneration (Fig.
6.24).
FIGURE 6.26  Focal mineralization of the meninges of the
minipig brain. ×200.
Eye and ear
Focal retinal dysplasia, lymphocytic keratitis, and
superficial keratitis have been observed occasion-
ally in the eyes of minipigs (Madsen et al 1998).
Periocular hyperkeratosis can be observed in
minipigs, and yeasts (Candida spp.) are occasion-
ally found within the debris. In addition, cupping
of the optic disk is often noted as a background
finding in the minipig eye (Fig. 6.27).
FIGURE 6.24  Myodegeneration and inflammation of the
skeletal muscle of the minipig. ×200.
Serous atrophy of the adipose tissue within
bone marrow is observed in the minipig (Svendsen
et al 1998). The lesion occurs in the epiphysis and
metaphysis and may be related to nutritional
imbalances (Bollen & Skydsgaard 2006) (Fig.
6.25). It occurs in both sexes at any age, possibly
more frequently in male animals, and it occurs
under normal physiological conditions. The lesion
is of no pathological significance and may be con-
sidered as a normal variation in the bone marrow FIGURE 6.27  Cupping of the optic disk in the minipig eye.
cell population. ×40.
 Minipigs 85

References Jørgensen, K.D., Ellegaard, L., Klastrup, S., et al.,

1998. Haematological and clinical chemical values in
microbiologically defined Gottingen minipigs. Scand.
J. Lab. Anim. Sci. 25, 159–166.
pregnant and juvenile Göttingen minipigs. Scand. J. Mortensen, J.T., Brinck, P., Lichtenberg, J., 1998.
Bollen, P., Skydsgaard, M., 2006. Restricted feeding
Lab. Anim. Sci. 25, 181–190. The minipig in dermal toxicology. A literature
may induce serous fat atrophy in male Göttingen
minipigs. Exp. Toxicol. Pathol. 57, 347–349. Lavker, R.M., Dong, G., Zheng, P.S., et al., 1991. review. Scand. J. Lab. Anim. Sci. 35 (Suppl. 1),
Hairless micropig skin. A novel model for studies 77–83.
Charles, J.A., 1996. Lymph nodes and thymus. In:
of cutaneous biology. Am. J. Pathol. 138, Svendsen, O., Skydsgaard, M., Aarup, V., et al., 1998.
Sims, L.D., Glastonbury, J.R.W. (Eds.), Pathology of
687–697. Spontaneously occurring microscopic lesions in
the pig. Industry House, Australia, pp. 185–210.
Madsen, L.W., Jensen, A.L., Larsen, S., 1998. selected organs of the Gottingen minipig. Scand. J.
Dincer, Z., 2007. The minipig. In: Gad, S.C. (Ed.),
Spontaneous lesions in clinically healthy, Lab. Anim. Sci. 25 (Suppl. 1), 231–234.
Animal models in toxicology. CRC Press, Boca
Raton, pp. 739–759.

Introduction
Rabbits are classified in the order Lagomorpha.
They differ from rodents in that they possess an
additional pair of incisor teeth directly behind the
large incisors of the upper jaw. There are over 100
different breeds of rabbit that are descendants of
the European wild rabbit, Oryctolagus cuniculus.
New Zealand White rabbits are used extensively
in toxicological studies; however, the design of the
majority of study protocols are such that full his-
topathology of all organ systems seldom occurs.
The animals in toxicology studies are euthanized
at the end of the study period whilst still relatively
young and so pre-neoplastic and neoplastic dis-
eases are rarely seen. The range and extent of
neoplastic and hyperplastic lesions that occur in
older rabbits have been well described in the
standard veterinary medicine and pathology texts
on laboratory animals (Hrapkiewicz et al 1998,
Saunders & Rees-Davies 2005, Percy & Barthold
2007). This chapter will focus on lesions encoun-
tered as spontaneous background findings in young
rabbits which are between six and 24 months old.
Tissue reactions are generally conserved
between species, and very few spontaneous back-
ground conditions occur in the rabbit that are
species-specific. The majority of toxicological
studies for which rabbits are used are immunologi-
cal, teratological (reproductive toxicity), dermal,
ocular, implant and vaccine safety assessments.
The latter often involve histopathological sam-
pling of eyes and injection sites only. The rabbit
eye is particularly sensitive to the deposition of
immune complexes.
Cardiovascular system
The heart is relatively small compared with other
species of similar size. The chambers of the right
side of the rabbit heart are thin and the right
ventricle often contains clotted blood with no evi-
dence of contraction (Percy & Barthold 2007).
The rabbit right atrioventricular valve is bicuspid
rather than tricuspid.
Occasionally evidence of the closure of the
ductus arteriosus may be seen as foci of mineraliza-
tion in the media of major vessels of young rabbits
in toxicology studies, depending on the plane of
section. Mononuclear inflammatory-cell infiltrates
are recorded infrequently in the myocardium
(Lehr 1965). The foci are usually at the base of
the interventricular septum. These are inert cel-
lular infiltrates with no accompanying myocardial
necrosis or fibrosis associated with the lesion.
Other findings recorded regularly at a low inci-
dence in the rabbit heart include mineralization of
the left atrial appendage, pericardial lipomatosis
(Fig. 7.1) and pericardial fibrosis. Calcification
of the aorta and large veins (Fig. 7.2) (arterioscle-
rosis) may be seen at necropsy in older animals
(e.g. ex-breeding colonies). Foci of myofibre
degeneration and fibrosis with a mononuclear-cell
infiltrate in the heart may be associated with
administration of ketamine/xylazine combinations
or detomidine in Dutch Belted rabbits (Percy &
Barthold 2007).
FIGURE 7.1  Rabbit heart with adipose tissue infiltration
in the right ventricle (base). ×100.
FIGURE 7.2  Medial mineralization of the aorta of the
rabbit. ×100.
The Watanabe rabbit is used extensively as an
animal model of natural endogenous atherosclero-
sis (Garibaldi & Pecquet 1988). Atherosclerosis
may also be observed in the aorta of New Zealand
White rabbits (Salamon et al 2007).
Hemolymphoreticular system
In the rabbit polychromasia of the erythrocytes
is a normal finding. Heterophils are the counter-
part of the neutrophil and have distinct acido-
philic granules. Hyposegmented neutrophils may
be observed in a rabbit blood smear. This is known
as the Pelger–Huet syndrome, and this anomaly is
inherited as a partial dominant trait in rabbits
(Salamon et al 2007). Basophils in the rabbit may
be relatively numerous and occasionally represent
up to 30% of circulating leukocytes (Percy &
Barthold 2007).
Large eosinophilic histiocytes are commonly
seen within the thymus, lymph nodes and follicles
of the epithelium-associated lymphoid tissue
(e.g. bronchiolar and gut-associated lymphoid
tissue) (Fig. 7.3). These histiocytes may have
particulate debris in the cytoplasmic vacuoles.
In the thymus they are usually situated in the
medullary area of the peripheral lobules. Other
New Zealand White rabbit 87
CHAPTER 7 
Alys E Bradley
New Zealand White rabbit
findings recorded regularly at a low incidence in
the hematopoietic tissues of rabbits include:
ectopic parathyroid in the thymus, increased
extramedullary hematopoiesis in the spleen,
accessory spleen (Weisbroth et al 1976), eryth-
rophagocytosis, sinusoidal dilation, and pigmented
macrophages in the lymph nodes. In addition,
lymphocyte infiltrations may be noted in the liver,
heart and lungs.
FIGURE 7.3  Macrophage aggregates, often containing
debris, present in the rabbit GALT (gut associated lym­
phoid tissue). ×100.
Respiratory system
The mucosa lining the ventral surface of the nose
contains some hair follicles, and so the presence
of these should not be misdiagnosed as hamar-
toma. The submucosa of the nasal cavity contains
individual or small aggregates of lymphocytes, but
no lymphoid follicles are present. There are no
respiratory bronchioles in rabbits. Alveolar macro-
phage accumulations are often seen in the lungs.
The alveolar macrophage aggregates have no par-
ticular distribution pattern and can be peripher-
ally placed just beneath the pleura, or at the
bronchoalveolar junction. The macrophages are
hypertrophic with pale pink cytoplasm, but
usually there is no associated inflammatory cell
response. Other findings recorded regularly in the
rabbit lung include: perivascular mononuclear or
eosinophilic inflammatory cell infiltrations, ectopic
bone and bronchus-associated lymphoid tissue
(BALT) hyperplasia in older rabbits. Submucosal
mononuclear inflammatory cell infiltrations are
seen occasionally in the trachea and larynx. Bar-
biturate euthanasia can result in petechiae on the
surface of the lung which disappear during
processing (Salamon et al 2007). In addition, the
process of euthanasia may cause alveolar edema
and congestion in the rabbit lung (Fig. 7.4). Occa-
sional thrombi with arteritis and periarteritis may
be noted in the rabbit lung (Fig. 7.5).
88 BACKGROUND LESIONS IN LABORATORY ANIMALS
FIGURE 7.4  Agonal alveolar edema present in rabbit
lung. ×100.
FIGURE 7.5  Rabbit lung with pulmonary thrombus, arteri­
tis and periarteritis. ×100.
Gastrointestinal system
Malocclusion of the teeth is reasonably common
in rabbits due to congenital mandibular prognathia
or secondary to inadequate wear. Ulcerations of
the tongue may be caused by lower molar maloc-
clusion. Upper molar malocclusion causes buccal
ulcerations of the cheek. Malocclusion of teeth is
controlled in contract research organizations or
pharmaceutical industries by the clipping of over-
grown incisors. A large pair of circumvallate papil-
lae on the dorsal surface of the tongue are
occasionally mistaken for papillomatous growths.
Vomiting is not possible in the rabbit, and food
and cecal pellets are always present in the stomach.
Hairballs or trichobezoars in the stomach or
pylorus are encountered commonly, but are gener-
ally incidental findings at necropsy (Haugh 1987).
The hairballs are masses of hair and ingesta that
result from excessive self-grooming and boredom
(Percy & Barthold 2007). Predisposing factors to
hairball formation may include low-fibre diets,
experimental manipulation and stress. Heter-
ophils and lymphocytes are common in the sub-
mucosal area of the stomach. In contrast to other
laboratory animals, Brunner’s glands are present
throughout the length of the rabbit duodenum
(Percy & Barthold 2007).
Rabbits are hindgut fermenters with a large and
complex digestive system. They practise cecotro-
phy, which is the ingestion of mucus-coated night
faeces. Cecotrophy is controlled by the adrenal
glands and may be altered during periods of stress.
The large intestines consist of a spiral cecum, a
sacculated colon and the rectum. The sacculus
rotundus is a round, thick-walled enlargement at
the ileocecal junction (Percy & Barthold 2007). At
the ileocecal junction adjacent to the sacculus Other findings regularly recorded at low inci-
rotundus, the cecum has a large mass of lymphoid dences in the gastrointestinal tract of the rabbit
tissue called the cecal tonsil. This may be referred include dilated glands in the fundus of the
to as the ileocecal tonsil, or ampulla ilei in older stomach, mineralized foci in the fundus of the
anatomy texts. The walls of this area are thick- stomach, and micro-abscesses in the lamina
ened by aggregations of organized lymphoid tissue propria of the cecum.
and macrophages and this area should not to be Rabbits may occasionally be affected by enteric
mistaken for lymphoid hyperplasia or increased infectious diseases; however, breakdowns in test
cellularity. The grossly thickened walls of the sac- facility closed systems are rare and usually result
culus rotundus, caecal tonsil and vermiform in parasitic disease only. Other than coccidiosis,
appendix are due to aggregates of lymphoid tissue which may cause clinical disease that could
in the lamina propria and submucosa. At the junc- compromise a toxicology study, parasitic lesions
tion between the transverse and descending colon are generally only incidental interesting back-
is the fusus coli, which is unique to rabbits and is ground findings. Adult nematodes of Passalurus
made up of ganglion cells which regulate the flow ambiguus (pinworm) (Fig. 7.6), may occasionally
of ingesta into the descending colon (Salamon be observed in the cecum and colon. Larval forms
et al 2007, Cruise and Brewer, 1994). The rectum may be seen in the mucosa of the small intestine
often contains submucosal lymphocytes rather or cecum. Evidence of parasitic transit through
than areas of gut-associated lymphoid tissue the intestine may manifest as mineralized parasitic
(GALT). Marked plasma-cell infiltrations into the granulomata. Cysticercus larvae of the tapeworm
intestinal tract have been described (Salamon et al Taenia pisiformis are occasionally seen hanging
2007). In the small intestine, mucosal erosions, from the mesentery or embedded in the liver
dilatation of lacteals and blunting of overlying parenchyma (Soulsby 1986), though this would
villi occur in conjunction with a severe plasma- be a rare finding in purpose-bred laboratory
cell infiltration into the lamina propria (Percy & rabbits.
Barthold 2007).
Peyer’s patches are normally very large in the
rabbit and should not be misdiagnosed as lym-
phoid hyperplasia. Lymphoid tissues are large and
prominent in the small and large intestines of the
rabbit. The gut associated lymphoid tissue (GALT)
of the rabbit represents more than 50% of the
total mass of lymphoid tissue in the body and
accounts for the relatively small spleen size in the
rabbit.
Rabbits are peculiar among laboratory species in
that their main reaction to external stressors of
any type is an episode of diarrhea. This can some-
times occur days after a stressful event. Any
change in gut microflora, pH or motility will lead
to diarrhea. Rabbits react negatively to changes of FIGURE 7.6  Passalurus ambiguus ova in faecal flotation
environment (moving to a research facility, new taken from a rabbit. ×400.
technicians, visitors to the animal rooms) and any
such event may trigger an episode of diarrhea
two to three days later. This is a behavioural trait Liver and biliary system
of rabbits and has to be taken into account when
Rabbit livers are often pale yellow at necropsy due
assessing clinical signs. Rabbits are hindgut fer-
to the amount of stored glycogen in hepatocytes.
menters adapted to digest a low-quality, high-fibre
This cytoplasmic vacuolation (Fig. 7.7), associated
diet. Stress events slow peristalsis and impede
with glycogen accumulation, varies between sexes
caecotrophy or coprophagia. In addition, caecal
and also varies diurnally. Vacuolation is also influ-
pH becomes more alkaline due to the slowed
enced by standard diets and fasting before eutha-
passage of food material, and Escherichia coli can
nasia (Percy & Barthold 2007). Generally rabbits
then dominate the caecal environment.
not starved before euthanasia show high levels of
Mucoid enteropathy (epizootic rabbit entero­
glycogen accumulation. Therefore animals eutha-
pathy) is a disease of unknown etiology that
nized in the morning will have more glycogen in
occurs in young rabbits. It is a common cause of
their livers than those euthanized in the after-
death in commercial and laboratory rabbit colo-
noon. Pathologists should be aware of this diurnal
nies, but is rare in pet hutch and housed rabbits.
change and interpret any changes in glycogen
Affected animals may recover if switched to a
vacuolation with the knowledge of whether the
high-fibre, low-energy diet. Upon histopatholo­
animal was starved prior to euthanasia, for how
gical examination, there is marked goblet-cell
long, and at what time of day the necropsy
hypertrophy and hyperplasia, mainly affecting the
occurred (Wells et al 1988; Weisbroth et al 1990).
small intestine and colon, with only occasional
Vacuolation of hepatocytes is more common in
cecal changes. Small intestinal sections may show
female rabbits. Other findings regularly recorded
atrophy and fusion of villi. There may be minimal
at a low incidence in the liver include extramedul-
lymphocytic/plasmacytic inflammatory cell infil-
lary hematopoiesis, periportal hepatocyte vacuola-
trates in some animals, but in general an inflam-
tion, multifocal hepatic mineralization and tension
matory reaction is not usually associated with this
lipidosis.
condition. The proposed bacterial proliferation
(E. coli and/or Clostridium spiroforme) is not
always evident on tissue sections.

FIGURE 7.7  Vacuolation of hepatocytes in the rabbit liver.
×100.
The most frequently recorded background
lesions in the livers of rabbits consist of lym-
phocytic cell infiltration into the periportal area
of the liver. Lymphocytic infiltration increases
with age and is more common in female rabbits.
Torsion of the caudate lobe occasionally occurs
and may result in fatal hepatic rupture (Salamon
et al 2007, Weisbroth 1975). Duplication of the
gall bladder (Fig. 7.8) and cholecystitis (Fig. 7.9)
have been reported in rabbits (Gupta 1975).
Serosal hemorrhages have been reported at
necropsy in rabbit gall bladder (Salamon et al
2007).
FIGURE 7.8  Duplication of rabbit gall bladder. ×100.
FIGURE 7.9  Necrotic cholecystitis in the rabbit gall
bladder. ×100.
Hepatic coccidiosis is a disease of young rabbits
caused by Eimeria stiedae and is a common cause
of condemnation of rabbit livers in slaughter-
houses. Recently coccidiosis has increased in com-
mercial rabbit establishments and caused a paucity
in the availability of rabbits for research. This
infection is of great concern due to the difficulty
in eradication. The extent of the lesions and clini-
cal signs associated with the disease may invali-
date a toxicity study. Developmental stages are
usually seen in biliary epithelium, proximal to the
cell nucleus, five to six days after infection, but
may be seen after three days in heavy infections
(Fig. 7.10). Oocysts are shed around day 23 and
patency lasts until day 37 post infection (Soulsby
1986). Mineralized foci in the biliary epithelium
may be the only sign of previous infection in a
clinically normal rabbit. In some cases oocysts
may be present in calcified nodules, and have been
misdiagnosed/misreported as the eggs of and
infection with the nematode Capillaria hepatica.
Histologically, the liver shows well circumscribed
focal granulo­matous lesions surrounding the bile
ducts, with schizonts and zygotes of E. stiedae in
the lumen.
FIGURE 7.10  Coccidiosis in the gall bladder of the rabbit
liver. ×100.
Pancreas
The most common finding in the exocrine pan-
creas is the presence of an accessory spleen (Fig.
7.11) (Weisbroth et al 1976). These nodules are
usually fairly large and may be described as raised,
dark foci or masses at necropsy. The architecture
of the splenic tissue resembles that of normal
spleen with clearly discernible follicles. Acinar cell
degranulation is also occasionally recorded in
control rabbits. This lesion is usually associated
with fasting, and is generally associated with
animals euthanized mid afternoon and correlates
with decreased glycogen accumulation in the liver.
Individual acinar necrosis may also be reported in
rabbits subjected to prior fasting (Salamon et al
2007). Small lymphocytic infiltrations into the
pancreas are common.
FIGURE 7.11  Pancreas with accessory spleen in the
rabbit. ×50.
New Zealand White rabbit 89
Urinary system
The background data in the literature on non-
infectious, spontaneous kidney lesions in labora-
tory rabbits are limited. Vacuolation of the
proximal convoluted tubule epithelium is a regular
finding in young, non-pregnant does. These vacu-
oles stain positively with Oil red O stain for
neutral lipids.
Calcium absorption and metabolism in the
rabbit is poorly understood. Rabbits normally have
a higher blood calcium range than other laboratory
animal species and are predisposed to cystic, ure-
thral, ureteral and renal calculi. Rabbits excrete
excess calcium though the urine as calcium car-
bonate. Mineralized foci are commonly seen in the
collecting ducts and medulla of rabbit kidneys.
The mineralized foci correlate with the normal
alkaline, cloudy to pigmented urine of rabbits
caused by the presence of albumin, fine calcium
carbonate and ammonium magnesium phosphate
crystals. Rabbit urine varies in colour from creamy
yellow to dark red depending on the presence
of porphyrin pigments derived from the diet
(Percy & Barthold 2007), urobilin or xenobiotics
(e.g. antibiotics). Care must be taken to differen-
tiate normal rabbit urine from red urine caused by
hematuria. Mineralized foci in the tubules or
interstitial areas of the cortex are present in large
numbers of rabbits (Fig. 7.12). The lesions are
distributed evenly between males and females,
and have been previously reported in the litera-
ture with similar incidences (Ritskes-Hoitinga
et al 2004). Focal mineralization is also occasion-
ally recorded in the urothelium of the urinary
bladder.
FIGURE 7.12  Mineralization in the interstitium of the
rabbit kidney. ×100.
Aging glomerular changes are observed at about
one year of age in the rabbit kidney (Salamon
et al 2007). These changes consist of mesangial
proliferation with a multifocal distribution. Scars
on the surface of the kidneys are commonly seen
at the necropsy of clinically normal, apparently
healthy rabbits. These are the most common find-
ings in rabbit organs at slaughterhouses. The
lesions are seen in animals from colonies free from
Encephalitozoon cuniculi and are the result of a
spontaneous nephropathy syndrome. There are no
clinical signs accompanying these lesions and no
evidence of progression or greater severity of the
findings on longer term studies. The lesion is not
thought to be progressive, unlike the nephropathy
of rats. The histological findings are generally
recorded separately as basophilic tubules (Fig.
90 BACKGROUND LESIONS IN LABORATORY ANIMALS
7.13), dilated or cystic tubules, pigmented tubules
and interstitial inflammatory cell infiltration.
Spontaneous findings of mineralization, tubular
basophilia and dilatation have been reported pre-
viously in young laboratory rabbits under one year
of age (Burek et al 1988, Hinton 1981), but the
incidence of this lesion seems to be increasing in
the New Zealand White rabbit population. The
abundance of spontaneous renal findings may
mask nephrotoxic effects. Nephrotoxins can
induce or exacerbate chronic progressive neph-
ropathy in the rat after single or acute exposure
(Khan & Alden 2002). Although the pathogenesis
of the lesion in rabbits is unclear and does not
appear to be progressive, this masking effect may
occur and so renal evaluation should be approached
with care. It is always good practice to record
and grade each finding separately. Any increased
incidences will be apparent once the study is
complete and findings can be merged in incidence
tables as ‘nephropathy’ if appropriate.
FIGURE 7.13  Focal area of basophilic tubules in cortical
interstitium of rabbit. ×100.
Other findings recorded regularly at low inci-
dences in the rabbit kidney include focal fibrosis,
cortical cyst and acute pyelitis in the kidney. Sub-
mucosal lymphoid hyperplasia (Fig. 7.14) and
papillary hyperplasia of the urothelium in the
bladder are background changes associated with
the high crystalline content of normal rabbit urine.
FIGURE 7.14  Submucosal lymphoid hyperplasia noted in
the rabbit urinary bladder. ×100.
Endocrine system
The rabbit adrenal often contains vacuolated
cortical areas and extracapsular cortical tissue is
also a feature. Hyperplastic areas in the rabbit
adrenal cortex have been reported during spring
(Greene 1965).
Lymphocytic infiltration is commonly recorded
in the thyroid gland of the rabbit. There is usually
no active inflammatory component to the lesion
and it has not been reported to be associated with
immune-mediated changes or the presence of
autoantibodies. In some animals lymphoid folli-
cles form in the thyroid and it is considered that
these lymphocytic infiltrates reflect the high inci-
dence of extra lymphoid tissue in the rabbit com-
pared with other species. Other findings regularly
recorded at low incidences in the rabbit endocrine
system include ectopic thymus, dilated or cystic
follicles and ultimobranchial cysts in the thyroid FIGURE 7.15  Focal squamous metaplasia with hyper­
and focal mineralization in the adenohypophysis keratosis in the rabbit prostate. ×100.
of the pituitary. The rabbit pituitary often displays
endothelium-lined spaces at the junction between The rabbit uterus has two horns and two sepa-
the pars distalis and intermedia. rate cervices, and the placentation is hemochorial.
Does are induced ovulators. Although tumors of
Skin and appendages the uterus are common in older animals (i.e. adeno-
carcinomas, deciduomas), findings in the female
Lesions caused by fighting are common, particu- reproductive tract of young animals on routine
larly in aggressive males. Hair chewing and bar­ toxicology studies are uncommon. The most fre-
bering are occasionally observed among young quent findings recorded at low incidences include
group-housed rabbits. The lesion is characterized follicular cysts, hemorrhagic follicles and mineral-
by alopecia without dermatitis on the face and ized atretic follicles in the ovary and mesonephric
back. Boredom and low-roughage diets are consid- duct cysts in the oviduct. Pigmented macrophages,
ered to be predisposing factors (Percy & Barthold endometrial edema, dilated endometrial glands
2007). Ulcerative dermatitis or pododermatitis is and endometrial cysts in the uterus and subepi-
observed in older, heavier rabbits kept in wire thelial cysts and distended veins on the surface of
cages (Percy & Barthold 2007). the vagina are also recorded. Endometrial venous
Moist dermatitis occurs due to the contact aneurysms are considered to be congenital defects
wetting of the fur. Multifocal aggregates of lym- characterized by multiple, blood-filled endome-
phocytes can be observed surrounding hair folli- trial varices that are composed of dilated, thin-
cles or in the deep dermis. The cause is unknown. walled veins that rupture and bleed periodically in
Exfoliative dermatitis and sebaceous adenitis have the uterine lumen (Bray et al 1992).
been reported in older rabbits (Salamon et al Abundant interstitial tissue in the female rabbit
2007). ovary is referred to as the ovarian interstitial gland
(Mori & Matsumoto 1973) (Fig. 7.16).
Reproductive system
The male rabbit has patent inguinal canals which
connect to the abdominal cavity and which often
accumulate white to brown secretions produced
by scent glands in the inguinal canal wall (Salamon
et al 2007). Tubular atrophy is an infrequent
finding in the male rabbit testis. This may be
unilateral or bilateral, but is generally localized,
affecting only a few tubules. Periarteritis and
perivascular lymphoid infiltration are common
lesions observed in the rabbit testis. There is an
accompanying oligospermia and sloughing of sem-
iniferous epithelium in the epididymes. Clinically
silent, focal, acute inflammation is occasionally
recorded in the bulbourethral gland and prostate
of the rabbit. Spontaneous, focal, keratinized FIGURE 7.16  The interstitial gland in the rabbit ovary.
squamous metaplasia of the prostate (Fig. 7.15) ×10.
and seminal vesicle epithelium is an infrequent
lesion peculiar to the rabbit. It is important to
recognize this as a spontaneous lesion when evalu-
Mammary gland
ating test articles with androgenic or oestrogenic Mammary gland dysplasia and cystic mammary
actions. This lesion is well described by Zwicker glands are associated with pituitary adenomas and
and co-workers (1985). uterine adenocarcinoma in older rabbits. Cystic
mammary gland hyperplasia is associated with a
condition called cystic mastitis. This lesion occurs
in one or more glands of non-breeding does over the
age of three years and appears to be a preneoplastic
condition. The hyperplastic teats may contain a
brown, serosanguinous fluid and masses or fluid-
filled cysts may be palpable in the mammary tissues
(Richardson 2000).

Muscle, bones and joints
The preferred location for intramuscular injec-
tions in the rabbit is the dorsal lumbar muscle
group. Alum granulomas, indicative of previous
vaccination, or minimal focal myofibre degenera-
tion may be seen at this site. The skeleton of the
rabbit comprises only six to eight percentage of
total body weight (Percy & Barthold 2007). The
bones of rabbits are fragile and fractures occur
readily, especially with improper handling. Verte-
bral fracture is caused by improper handling
leading to sudden, unsupported movement of the
forelimbs which causes fracture or vertebral luxa-
tion. Most fractures occur in the lumbosacral
region and cause spinal cord damage and paralysis.
Vertebral spondylosis is common in rabbits
(Richardson 2000).
Brain and nervous system
Occasionally ventricular dilation of the rabbit cer-
ebrum is recorded at necropsy. There is no under-
lying pathology in the brain tissue and the lesion
is presumed to be congenital and related to the
domed shape of the brachycephalic rabbit skull.
Focal meningeal aggregates of lymphocytes and
minimal cortical focal gliosis, are often observed
in the right brain and spinal cord of unhealthy
rabbits. Similar aggregates have also been observed
in the optic nerve (Salamon et al 2007).
Eye and ear
Inherited buphthalmia or congenital glaucoma
occurs in New Zealand White rabbits as an auto-
somal recessive trait. One or both eyes may be
affected (Burrows et al 1995). The globe is
enlarged due to increased intraocular pressure as
a result of the absence or underdevelopment of
the aqueous humour outflow channels with
incomplete lavage of the iridocorneal angles
(Burrows et al 1995). This manifests in clinically
normal rabbits as foci of inflammatory cells in the
cornea of the eye. Occasionally these lesions may
progress to conjunctivitis and panophthalmitis,
with involvement of the nictitating membrane and
eyelids.
Prolapse of the deep gland of the third eyelid
has been recently reported and appears histolo­
gically as a mass composed of bilobed glands
arranged into an alveolar-like pattern without
inflammation. The cause is proposed to be abnor-
mal laxity of the supporting connective tissue
(Janssens et al 1999).
Metabolic diseases
Pregnancy toxaemia occurs in obese rabbits (Percy
& Barthold 2007). At necropsy, obesity is evident,
and the liver and kidneys are pale yellow due to
mobilized fat stores. Pregnant does may have
uterine hemorrhages and dead fetuses in the
uterine horns.
References
Bray, M.V., Weir, E.C., Brownstein, D.G., et al., 1992.
Endometrial venous aneurysms in three New
Zealand white rabbits. Lab. Anim. Sci. 42,
360–362.
Burek, J.D., Duprat, P., Owen, R., et al., 1988.
Spontaneous renal disease in laboratory animals. Int.
Rev. Exp. Pathol. 30, 231–319.
Burrows, A.M., Smith, T.D., Atkinson, C.S., et al.,
1995. Development of ocular hypertension in
congenitally buphthalmic rabbits. Lab. Anim. Sci.
45, 443–444.
Cruise, L.J., Brewer, N.R., 1994. Anatomy. In:
Manning, P.J., Ringler, D.H., Newcomer, C.E.,
(Eds.), The biology of the laboratory rabbit, second
ed. Academic Press, pp. 47–61.
Garibaldi, B.A., Pecquet Goad, M.E., 1988. Lipid
keratopathy in the Watanabe rabbit. Vet. Path. 25,
173–174.
Greene, H.S.N., 1965. Lesions of the spontaneous
diseases of the rabbit. In: Ribelin, W.E., McCoy, J.R.
(Eds.) Pathology of laboratory animals. Springfield,
IL, pp. 330–350.
Gupta, B.N., 1975. Duplication of the gall bladder in a
rabbit. Lab. Anim. Sci. 25, 646.
Haugh, P.G., 1987. Hairballs in rabbits: alternative
treatment. Can. Vet. J. 28, 280.
Hinton, M., 1981. Kidney disease in the rabbit: a
histological survey. Lab. Anim. 15, 263–265.
Hrapkiewicz, K., Medina, L., Holmes, D.D., 1998.
Clinical medicine of small mammals and primates,
second edn. Manson Publishing, London, UK.
New Zealand White rabbit 91
Janssens, G., Simoens, P., Muylle, S., et al., 1999.
Bilateral prolapse of the deep gland of the third
eyelid in a rabbit: diagnosis and treatment. Lab.
Anim. Sci. 49, 105–109.
Khan, K.N.M., Alden, C.L., 2002. Kidney. In:
Hashek-Hock, WM, Rousseaux, C.G. (Eds.),
Handbook of toxicologic pathology, vol. 2, second
ed. Academic Press, San Diego, pp. 255–336.
Lehr, D., 1965. Lesions of the cardiovascular system.
In: Ribelin, W.E., McCoy, J.R., (Eds.), The pathology
of laboratory animals. Thomas, pp. 124–159.
Mori, H., Matsumoto, K., 1973. Development of the
secondary interstitial gland in the rabbit ovary.
J. Anat. 116, 417–430.
Percy, D.H., Barthold, S.W., 2007. Pathology of
laboratory rodents and rabbits, third ed. Blackwell,
Ames, IA, USA.
Richardson, V.C.G., 2000. Rabbits: health, husbandry
and diseases. Blackwell Science, Malden MA.
Ritskes-Hoitinga, M., Skott, O., Uhrenholt, T.R., 2004.
Nephrocalcinosis in rabbits – a case study. Scand. J.
Lab. Anim. Sci. 31, 143–148.
Salamon, C.M., Mackenzie, K.M., Peckham, J.C.,
et al., 2007. The rabbit. In: Gad, S.C., (Ed.),
Animal Models in Toxicology, second ed. Taylor &
Francis, Boca Raton, pp. 421–492.
Saunders, R.A., Rees-Davies, R., 2005. Notes on rabbit
internal medicine. Blackwell, Ames, IA, USA.
Soulsby, E.J.L., 1986. Helminths, arthropods and
protozoa of domesticated animals, seventh ed.
Baillière Tindall, England.
Weisbroth, S.H., 1975. Torsion of the caudate lobe of
the liver in the domestic rabbit (Oryctolagus). Vet.
Pathol. 12, 13–15.
Weisbroth, S.H., Fox, R.R., Scher, S., et al., 1976.
Accessory spleens in domestic rabbits (Oryctolagus
suniculus). II. Increased frequency in hematological
diseases and experimental induction with
phenylhydrazine. Teratology 13, 253–262.
Weisbroth, S.H., Mauer, J.K., Bennett, F.B., et al.,
1990. Hepatocellular vacuolization in rabbits: Effects
of feed restriction, orchidectomy and ovariectomy.
Toxicol. Pathol. 18, 56–60.
Wells, M.Y., Weisbrode, S.H., Maurer, J.K., et al.,
1988. Variable hepatocellular vacuolization
associated with glycogen in rabbits. Toxicol. Pathol.
16, 360–365.
Zwicker, G.M., Killinger, J.M., McConnell, R.F., 1985.
Spontaneous vesicular and prostatic gland epithelial
squamous metaplasia, hyperplasia and keratinised
nodule formation in rabbits. Toxicol. Pathol. 13,
222–228.

Introduction
What one may see upon microscopic examination
of sections of animal tissues is not always related
to the normal histology or pathology of the tissue
in question. Defects or abnormalities in tissue sec-
tions may result from the faulty processing of the
tissue specimens. These defects are referred to as
artifacts. Some artifacts are easily distinguished
from normal or pathological tissue components,
and some are difficult to distinguish from such
entities (McInnes 2005). This chapter will endeav-
our to illustrate some of the many hundreds of
artifacts that are most frequently encountered in
the preparation of microscopic tissue sections.
Artifacts are defined as being any structure or
feature that has been produced by the processing
of a tissue or has been introduced artificially. Since
artifacts are generally produced at the different
stages, this chapter will follow the chronological
sequence that is routinely employed in the collec-
tion, fixation and processing of tissue specimens.
Each artifact will be described as well as its cause
or causes and methods for its prevention and/or
correction, if these exist. Artifacts occur at each
of the following stages in the processing of tissue
sections: before death, at postmortem or necropsy,
during the fixation, dehydration, clearing, impreg-
nation and embedding with paraffin wax and
microtomy of tissues and during the mounting of
tissue sections onto glass slides, during staining
procedures and cover slipping.
Before death/ante mortem
Several artifacts that are encountered in micro-
scopic tissue sections are not the fault of histology
technicians but have, in fact, been caused by clini-
cal procedures performed ante mortem or by ante
mortem environmental factors. Examples include
the inclusion of suture material and carbon parti-
cles into tissues (Thompson & Luna 1978) as well
as thermal and chemical dehydration. Thermal
dehydration results from tissues removed by
cautery, which causes coagulation of proteins and
chemical dehydration results from the use of
caustic chemicals to sterilize the surgical instru-
ments. Carbon particles result from the inhalation
of dust and are ingested by alveolar macrophages
within the lungs. The condition is called arthraco-
sis and is common in dog, non-human primate and
human lungs.
Although the hemorrhage and gliosis present
within the optic nerve, caused by retro-orbital
bleeding, represent a genuine pathological process,
this lesion may still be considered to be an ante
mortem artifact because it occurs regularly and
can be confusing to an inexperienced pathologist
(Fig. 8.1). In larger animals, euthanasia is generally
achieved with intravenous barbiturate injection.
Occasionally, the inability to find a suitable vein
results in intracardiac barbiturate injection in
order to cause rapid euthanasia. Barbiturate is
highly caustic and when injected into the cardiac
muscle it causes the intense eosinophilia of the
cardiac myocytes as well as separation of shrunken
eosinophilic fibres by a pink-staining amorphous
material. The changes are considered to be arti-
factual and a result of barbiturate lysis (Darke
et al 1996). This change may, however, be con-
fused with severe degeneration or necrosis of the
cardiac myocytes. The absence of concomitant
inflammatory cells in the areas of eosinophilia
militate against this diagnosis (Fig. 8.2). A similar
lesion may be observed in the liver of monkeys
euthanased with barbiturates. Here, the centri-
lobular hepatocytes demonstrate dilatation of the
sinusoids and rarefaction of the hepatocytes (Fig.
8.3). Bone marrow biopsy before death under ter-
minal anesthesia can result in bone marrow emboli
lodging in the blood vessels of the lung (Fig. 8.4).
FIGURE 8.1  Hemorrhage and gliosis in optic nerve
caused by retro-orbital bleeding in the rat. ×100.
FIGURE 8.2  Barbiturate intracardiac injection causing
eosinophilia of cardiomyocytes. ×100.
Artifacts in histopathology 93
CHAPTER 8 
Elizabeth F McInnes
Artifacts in histopathology
FIGURE 8.3  Dilatation of centrilobular sinusoids and rar-
efaction of hepatocytes in cynomolgus monkey liver after
barbiturate injection. ×100.
FIGURE 8.4  Bone marrow thrombus in dog lung caused
by ante mortem bone marrow collection. ×100.
Postmortem/necropsy
Numerous artifacts that are observed in micro-
scopic tissue sections occur during the necropsy
procedure. Examples of such artifacts include
talcum powder inclusion, pressure effects, plant
material, hair contaminants, bone fragment con-
tamination and freezing artifacts (Thompson &
Luna 1978). Talcum powder may become incor-
porated into the tissues at necropsy when powder
contamination of the carcass occurs with the use
of powdered latex gloves (Thompson & Luna
1978). Talcum powder is made up of starch and
hydrous magnesium silicate. The powder appears
as grey crystals in the processed tissues. Hair and
grass may contaminate tissues at necropsy and
may be observed in cross section in the processed
slide (Fig. 8.5). In addition, it is important to note
that autolysis (which is an artifact in itself) will
start to occur within three minutes of death in the
small intestine, resulting in the swelling of the
villus tips and epithelial denudation of a few villi
(Pearson & Logan 1978).
94 BACKGROUND LESIONS IN LABORATORY ANIMALS
FIGURE 8.5  Plant material in brain tissue due to contami-
nation at necropsy. ×100.
Tissues may be deliberately or accidentally
frozen after death. Freezing artifacts (Fig. 8.6) are
characterized by vacuolation and cracks caused by
formation of ice crystals intracellularly and extra-
cellularly due to freezing of the cadaver prior to
necropsy examination. The ice crystals melt when
the tissue is placed in fixative and this causes
cell rupture and the formation of spaces within
the interstitium (Thompson & Luna 1978). Softer
tissues containing greater amounts of fluid are
more vulnerable to freezing artifacts than are
harder tissues such as smooth muscle (Thompson
& Luna 1978). Freezing artifacts may also occur
when tissues are frozen after placement into fixa-
tive, usually when transported in freezing weather
with inadequate insulation (Thompson & Luna
1978).
FIGURE 8.6  Cracks and open spaces produced in the
liver due to freezing of the tissue. ×100.
Perfusion of the lungs with buffered formalin
at necropsy is an accepted technique to improve
the histology of the pulmonary architecture
(Hausmann et al 2004, Braber et al 2010). This
may result in a lesion that resembles perivascular
edema (Fig. 8.7). Failure to perfuse lung tissue at
necropsy results in collapse of the interstitial,
alveolar tissue and the lung may appear to dem-
onstrate pneumonia. Pressure exerted on tissues
by forceps (Fig. 8.8) at necropsy may result in
‘pinch’ defects which do not resolve during
processing (Thompson & Luna 1978). Such arti-
facts cannot be corrected during processing of the
tissue specimen; however, they may be prevented
by solicitous handling of fresh tissue specimens
prior to fixation. Fragments of bone are inserted
commonly deep into the brain tissue (Fig. 8.9),
due to carelessness on the part of the prosector,
during the opening of the cranium to remove the
brain. Such fragments of non-decalcified bone are
extremely hard in comparison with the surround-
ing soft brain tissue. In addition to nicking the
edge of the microtome knife, the bone fragments
are moved by the microtome knife-edge during
cutting and this causes shattering and distortion
of the tissue section (Thompson & Luna 1978).
FIGURE 8.7  Perivascular expansion resembling edema
in lung caused by perfusion of lung tissue with formalin.
×100.
FIGURE 8.8  Pinch artifact caused by forceps handling of
lung tissue at necropsy. ×100.
FIGURE 8.9  Bone and skeletal muscle contamination of
brain tissue during brain removal. ×100.
Tissue may become contaminated with frag-
ments of the animal’s hair (Fig. 8.10) at the time
of necropsy and prior to fixation. Such surface
contaminants are often not removed by washing
the tissue specimens subsequent to fixation and
prior to further processing, although this is the
only method of removal. In some cases, depending
on the orientation of the shaft of the hair, the
knife can push the hair or bone further into the
tissue and produce shattering of softer tissue
(Thompson & Luna 1978). Suture material may
also become embedded in tissues, and a similar
effect is caused when suture material is pushed
ahead of the microtome knife and causes shatter-
ing of the tissue adjacent to the site of localization
(Thompson & Luna 1978).
FIGURE 8.10  A fragment of hair incorporated into brain
tissue. ×100.
Fixation
Fixation of tissue will always prove to be a com-
promise between the need to preserve the cellular
detail of the cells and the technical constraints of
tissue processing.
Many of the artifacts observed in microscopic
tissue sections are the result of incorrect fixation
procedures. Examples of fixation artifacts include
the use of an incorrect fixative for a particular
tissue specimen and the formation of acid forma-
lin hematin pigment. In addition, autolytic changes
due to specimen adherence to the inner surface
of the fixative container or the use of inadequate
quantities of fixative may also occur. The practice
of placing tissues in a container and then adding
the fixative often results in the adherence of the
tissue to the sides of the container, with conse-
quent autolysis of the adherent surface of the
tissue. Thick tissue specimens, or insufficient time
spent in fixative, may result in a similar artifact
with a focal area of autolysis visible in the middle
of the tissue, generally pink (Fig. 8.11). Cracking
in the centre of tissues may occur due to inade-
quate fixation in formalin. When the tissue is
processed, the central unfixed tissue is effectively
fixed with ethanol, and shrinks, and thus forms
more cracks than the peripheral formalin-fixed
tissue during subsequent processing (Fig. 8.12).
FIGURE 8.11  Central areas of pink autolytic tissue in a
liver specimen due to inadequate penetration of fixative.

FIGURE 8.12  Cracking in centre of liver caused by inad-
equate fixation. ×100.
Biopsy specimens are often not fixed for a suf-
ficient length of time. Fixation requires at least 48
hours and a change of fixative at 24 hours is rec-
ommended. Formalin penetrates tissues at a rate
of 16 to 28 mm per 24 hours. If the tissues are
thicker than 6 mm, then longer fixation is recom-
mended. The maximum thickness of a specimen
should not exceed 6 mm, and for each volume of
tissue, 20 volumes of fixative should be used
(Thompson & Luna 1978). Saprophytic bacteria
may occur in areas of autolysis. Slaoui & Fiette
(2011) review the precautions that should be
taken when fixing tissues in 10% neutral buffered
formalin. The saprophytic bacteria are basophilic
and are not accompanied by inflammatory cells
when the tissue is examined under the micro-
scope. In addition, gas bubbles, due to the pres-
ence of clostridial bacteria, may also be observed
in autolysed tissues.
Black, amorphous crystals may result from
fixing in mercuric chloride–formaldehyde (this
artifact is not very common today due to the toxic-
ity of mercuric chloride and its subsequent disuse).
The mercuric chloride artifact may be removed
with iodine and sodium thiosulphate. Contamina-
tion of the specimen with rust (formed as a result
of oxidation of the metal cap covering the formalin
container) is another artifact which may occur
in histopathological tissues (Thompson & Luna
1978).
Acid formalin hematin pigment is a dark
brown, anisotropic, microcrystalline, iron-negative
pigment (Fig. 8.13). It is produced by the reaction
of formic acid from unbuffered formalin with the
heme of hemoglobin at an acid pH, and it can be
removed with a saturated alcoholic picric acid
solution or by fixation in phenol–formalin (Thomp-
son & Luna 1978). This pigment results in a brown
formalin pigment in, on, or around erythrocytes
within tissue sections. Consequently the pigment
may be encountered in a wide variety of sites
within tissue specimens because these sites may
contain heme. In tissue sections, a black to brown,
amorphous to microcrystalline pigment is observed
particularly over blood vessels and it is often not
in the plane of section. In the kidney, formalin
pigment tends to accumulate in glomeruli.
FIGURE 8.13  Acid hematin, formalin pigment present in
a blood vessel in the lung. ×100.
Contaminants may also be embedded in the
wax block such as bone adjacent to the tissue and
may nick the knife during sectioning or cause the
specimen to crumble and fall out of the paraffin
wax block as it is cut (Thompson & Luna 1978).
Fixation in 10% buffered formalin for longer than
48 hours is not recommended if immunohisto-
chemistry is to be performed on the tissues;
however, the success of various antibodies depends
largely on the ability of the histologist to retrieve
antigen using citrate buffer and microwave tech-
niques. Conventional fixing of liver in neutral
buffered 10% formalin causes the glycogen in
hepatocytes to migrate to one side of the cell
(Thompson & Luna 1978). Alcoholic formalin
fixatives give better and more consistent preserva-
tion of the hepatic glycogen (Thompson & Luna
1978). Tissues fixed in Zenker’s fixative for
greater than eight hours, demonstrate eosinophilia
and a loss of basophilia with indistinct nuclei
(Thompson & Luna 1978). In addition, crystalliza-
tion of erythrocytes may also be encountered with
the use of Zenker’s fixative.
Occasionally, tissues are fixed in a fixative
which is not recommended for that particular
type of tissue. This may occur with testes, where
fixation in modified Davidson’s fixative is recom-
mended over Bouin’s fixative (Lanning et al 2002;
Latendresse et al 2002) or over fixation in forma-
lin (Fig. 8.14). Modified Davidson’s fixative is
recommended over Bouin’s fixative because the
morphological detail is good and there is minimal
shrinkage of central tubules. Over-decalcification
may occur if bone tissue is placed in formic acid
for a long period of time (Fig. 8.15). No nuclear
or chromatin details are observed in the cells
because the decalcification in formic acid will
hydrolyse the nucleic acids in the cell. Slow freez-
ing of tissues (for immunohistochemistry) can
result in shrinkage of tissue as well as unfrozen
areas, which appear as holes in the sections.
Artifacts in histopathology 95
FIGURE 8.14  Distortion of testicular tissue after fixation
in formalin. ×100.
FIGURE 8.15  Loss of cellular detail in bone which has
been decalcified in formic acid for 14 days. ×100.
Processing
The ability to cut thin sections from a tissue block
depends on the consistent hardness of the tissues.
This is achieved by embedding the tissue in a
paraffin wax block in the series of steps that
involve dehydration, clearing, impregnation and
embedding with paraffin wax. Some of the arti-
facts that result from the inefficient processing of
tissues include the presence of crystals of fixative
within the tissue due to imperfect removal of
processing fluids and shrinkage of bone marrow
during processing and paraffin infiltration. The net
effect of poor fixation and clearing in chloroform
or xylene is excessive shrinkage of the tissue com-
ponents. Very little shrinkage will occur upon
clearing in chloroform and xylene if the tissue
has been adequately fixed in formalin previously.
Problems with paraffin wax sectioning include wax
that is too hard, which causes vibration marks and
wax that is too soft, which causes compression of
the tissue and sections of varying thickness.
Artifacts occur either during the trimming of
the gross tissue specimen for processing or within
the tissue processor (Figs 8.16, 8.17). During
macroscopic sectioning of the fixed, gross speci-
mens prior to processing, care should be taken to
keep the surface of the cutting block free of tissue
debris. If contamination occurs at this stage, the
fragments of tissue debris should be in the same
focal plane as the adjacent tissue. If the artifact is
not in the same focal plane as the surrounding
tissues, then it is logical to assume that the artifact
may have occurred during the flotation of the
tissue section in the water bath during the mount-
ing of the section (Thompson & Luna 1978).
Tissue specimens with detachable components
such as exposed, epithelium-lined surfaces should
96 BACKGROUND LESIONS IN LABORATORY ANIMALS
be processed in separate biopsy bags and tissue
cassettes to avoid the problem of tissue detach-
ment. Tissue processors are used to carry fixed
tissue specimens automatically through the differ-
ent processing fluids. If an automatic processor is
improperly adjusted, or a power failure occurs,
the basket of cassettes may remain elevated, and
the tissue specimens may become dehydrated by
exposure to air. Inadequately-filled solution con-
tainers will pro­duce a similar artifact which will
result in the tissue gaining a dry, homogenous
appearance upon staining.
FIGURE 8.16  Bone marrow tissue incorporated into the
brain during trimming resulting in an apparent inflamma-
tory focus. ×100.
FIGURE 8.17  Pieces of small intestinal tissues appear
adjacent to the brain due to incorporation during trimming
or processing. ×100.
Inadequate dehydration occurs during tissue
processing when the water is removed from the
specimen prior to paraffin wax infiltration. Water
will remain trapped within the tissue if the tissue
is inadequately dehydrated. The reasons for poor
dehydration include failure to cover the processing
solutions and failure to change the processing
solutions frequently. The alcohols used in pro­
cessing lose their effectiveness as dehydrating
agents as they become diluted by moisture from
the atmosphere and tissues. In the stained section,
inadequately dehydrated tissue will be partially
unstained (Fig. 8.18). In this situation, the tissue
block may be melted down and the tissue may be
reprocessed in the case of a valuable specimen.
FIGURE 8.18  Pale areas in the white matter of the spinal
cord due to inadequate dehydration during processing.
×100.
If a tissue is inadequately infiltrated with paraffin
wax during processing it will spread out rapidly
when floated on the water bath. This causes the
tissues (particularly the sclera of the eye) to
become pulled apart on the glass microscope slide.
This is a common artifact in ophthalmic pathology.
Inadequate paraffin wax infiltration may occur as a
result of inadequate fixation, dehydration, clearing
or insufficient time in molten wax. Inadequate
infiltration with paraffin wax can also result in
wrinkles that run in all directions of the tissue.
Utilization of a vacuum oven during paraffin infil-
tration, for difficult tissues such as bone, will help
to alleviate this problem (Thompson & Luna 1978).
Shrinkage of bone marrow in bone sections (Fig.
8.19) is also caused by inadequate infiltration of
paraffin wax and cannot be prevented if paraffin-
embedding techniques are being used. Shrinkage
of bone marrow can be prevented by the use of
the time-consuming, colloidin-embedding tech-
nique. Boonstra and co-workers (1983) have dem-
onstrated that in, morphometric studies, a total
shrinkage of about 15% of the original dimensions
has to be taken into consideration. Examples of
faulty paraffin wax embedding procedures include
entrapped air around specimens and multiple
embedding of tissues of varied consistencies. Air
may be trapped around the specimen within the
paraffin wax block. This allows the tissue to fall
out or to vibrate during the cutting procedure.
Cramming of tissue into a cassette before process-
ing results in empty spaces in the tissue section
(Fig. 8.20).
FIGURE 8.19  Shrinkage of bone marrow due to formalin
fixation. ×100.
FIGURE 8.20  Empty spaces caused in tissue due to
cramming of tissue into the cassette before processing.
×100.
Cutting
The rotary microtome is now the most popular
microtome used in histology today because it is
strong and can cut semi-thin and thin sections for
light microscopy. Histologists are advised to cut
tissues at a steady rate as cutting too fast can cause
loss of tissue in the block with resultant holes in
the tissue section. The slow cutting of tissue sec-
tions results in tissue sections of varying thickness
as the tissue can warm up and expand between
cutting strokes. Vibration of the tissue specimen
within a wax block results in an artifact that resem-
bles a venetian blind, with compressed zones of
tissue separated by open spaces (chatter) (Fig.
8.21). This effect can also be produced by a loose
screw on the knife or chuck of the microtome.
FIGURE 8.21  Vibration artifact in lymphoma tissue
caused by vibration of the blade when sectioning. ×100.
Knife artifacts include chatter and compression
caused by a dull blade, nicks in the blade edge
which cause splits and nicks in the tissue section
(Fig. 8.22), incorrect clearance angle of the knife
which causes sections to stick to the block and
to roll up and to be of varying thickness. A loose
blade in a holder causes chatter marks in the tissue
section and debris on the knife edge causes tissue
sections to stick to the block. Incorrect orienta-
tion of the tissue in the block causes incomplete
sections with missing areas. Hard, calcified mate-
rial in the paraffin wax causes slits and nicks in the
tissue section.

FIGURE 8.22  Scoring of tissue due to nick in the knife’s
cutting surface. ×100.
The causes of wrinkles in the tissue include a
dull knife or inadequate infiltration of the tissue
with embedding medium. In addition, wrinkles
may also be caused by different tissue compo-
nents within the paraffin wax block expanding at
different rates within the hot water bath. Traces
of processing or clearing fluids in the embedded
block of tissue may result in a variety of defects
in tissue sections. These residues are hydrophilic;
however, the tissue section, when placed in the
water bath, is confined by the presence of hydro-
phobic wax. The net result is that the tissue
section becomes wrinkled, and it retains these
wrinkles, even after being flattened in the water
bath or on the microscope slide. These wrinkles
usually extend the length of the section and stain
more intensely because the stain has access to
both tissue surfaces of the wrinkle. This problem
cannot be corrected, and the discarding of the
poorly processed wax blocks is recommended
(Thompson & Luna 1978). Folds and wrinkles,
due to thin sections that become folded as
they are being cut, often cannot be completely
smoothed out. In addition, wrinkles may occur if
the temperature of the water bath is too hot or
too cold: the tissue will not spread out adequately
(cold) or will spread out excessively (hot) and will
produce cracks (Fig. 8.23). Contamination of the
cutting edge of the microtome knife with pig-
ments or bacteria taken from one block and placed
onto another tissue, may occur if the knife edge
is not cleaned regularly.
FIGURE 8.23  Cracks in lymphoma tissue due to exces-
sive heat of water bath. ×100.
If the microtome knife is set at too acute an
angle, the knife compresses the tissue specimen
as it is being cut. The compression tends to
occur at normal points of weakness. The firm
components, such as arteries, are pushed ahead as
they are cut and compress the adjacent paren-
chyma. A dull or blunt knife results in numerous
compression artifacts, streaks and cracks, which
are parallel to the blunt edge of the microtome
knife and which can render the tissue inadequate
for microscopic examination. Epithelial cells are
frequently rubbed off from the technician’s fingers
during microtomy or are shed on the knife surface
or unmounted slide as dandruff. These epithelial
cells are then stained and become part of the
microscope slide. The cells are highly eosinophilic
and have large nuclei and resemble skin epidermal
cells. Splitting and disruption of lens fibres in the
eye is a common artifact when sectioning eyes
(Fig. 8.24). The solution is to soften the lens in
the wax block with a softening agent.
FIGURE 8.24  Cracks in lens fibre material in the eye
which occur during sectioning. ×100.
Mounting onto glass slides
Examples of artifacts which occur during mount-
ing of the paraffin wax ribbons of tissue onto the
glass slide, include failure to remove tissue debris
from the tissue flotation water bath, contamina-
tion of the water bath with fungi (Fig. 8.25),
wrinkling of tissue sections, air bubbles beneath
tissue sections and improper drying of mounted
tissue sections. Contaminants such as fungi may
be present in the water bath or may have been on
the glass slide before mounting due to inadequate
storage of the glass slides. Every effort should be
made to eliminate water bath contaminants such
as bacteria and fungi because they may be con-
fused with pathological bacteria or fungi within
the tissue. In addition to the fungal and bacterial
contaminants, a dirty water bath may also contain
remnants of previous tissues, which may become
incorporated into the present tissue section such
as the liver tissue present within the lung (Fig.
8.26). As the tissue sections are flattened in the
water bath, a bubble of air may become trapped
beneath them. If the bubble is not removed, it
breaks and the overlying tissue is shattered
(Thompson & Luna 1978). The tissue overlying
the bubble will stain more intensely because the
stain has access to both surfaces of the tissue at
the site of the bubble.
Artifacts in histopathology 97
FIGURE 8.25  The presence of fungi on a kidney tissue
section due to a contaminated water bath. Note the fact
that no inflammatory cells are present. ×100.
FIGURE 8.26  Incorporation of liver tissue into lung due to
a contaminated water bath. ×100.
Staining
Unstained tissues are clear and transparent and
no cellular detail is visible under the microscope.
The commonly-used stain hematoxylin and eosin
(H&E) allows nuclear and cytoplasmic detail to be
visualized under the microscope and most tissue
sections are thus stained with hematoxylin and
eosin. Residual paraffin wax will occur in a tissue
when the deparaffinization in xylene has not been
complete and the xylene has become saturated.
Alcohol and water droplets may be retained
within the tissue section on the slide after rehy-
dration through graded alcohols to water. These
bubble artifacts obscure the morphological details
of the underlying tissue sections.
Examples of staining artifacts include faulty
clearing techniques resulting in inadequate stain-
ing and stain deposits. Alum hematoxylin oxidizes
during use and sections stained at the end of the
week may be paler than those stained at the start
of the week. When a staining dish containing
a working solution of hematoxylin is allowed to
stand at room temperature for long periods of
time, a metallic-appearing scum accumulates on
the surface of the solution. This scum consists of
oxidized mordant (used to increase the stability
of the stain) and hematein, and this may contami-
nate tissue sections that are stained in the old dye
solution. Eosin flakes, seen above the focal plane
of the tissue section, are precipitated dye derived
from an unfiltered stock solution. Eosin stain
deposit and eggs from an unknown organism in
the bronchiole of the lung may be seen in Fig.
8.27. The use of either hard or soft water for
98 BACKGROUND LESIONS IN LABORATORY ANIMALS
differentiation during the staining process will
have a marked effect on the appearance of the
stained slides. Occasionally splashes of acid onto
stained tissues (without a cover slip), will result
in pale areas scattered throughout the tissue (Fig.
8.28).
FIGURE 8.27  Deposition of an eosin flake during staining
and deposition of unidentified eggs and lint in bronchiole
of lung tissue. ×100.
FIGURE 8.28  Pale area within stained tissue caused by
acid splashing of the tissue before cover slipping. ×100.
Artifacts may also occur during the immunohis-
tochemistry process; these include bubbles, drying
out of tissue at the edges and blunt-blade artifacts
caused by a dull knife during the cryostat section-
ing of frozen tissues.
Cover slipping
Cover slipping of slides with the use of a mount-
ant is recommended to prevent the tissue from
drying out, to prevent surface damage to the
tissue, to allow long-term storage of tissues and to
improve the transparency of the tissue. Examples
of cover slipping artifacts include contamination
of the slides with pollen grains (Fig. 8.29), spores,
eggs, dust, dirt, cotton fibres, contamination of
the mounting medium, use of cover slips that are
too small, faulty positioning of the cover slip,
insufficient mountant, which leads to drying back,
entrapment of air during the cover slipping and
the use of too much mounting medium. Lint fibres
and other debris can be deposited on stained sec-
tions at the time of cover slipping and the lint of
linen fibres may be mistaken for a nematode (Fig.
8.30). A trapped mite, present on the surface of
adipose tissue, is illustrated in Fig. 8.31.
FIGURE 8.29  Pollen deposited onto the surface of a
tissue section before cover slipping. ×100.
FIGURE 8.30  Lint fibre deposited on the surface of the
spleen before cover slipping. ×100.
FIGURE 8.31  A trapped mite deposited on the surface of
adipose tissue before cover slipping. ×200.
Artifacts in the central
nervous system (CNS) and
special senses
The central nervous system (CNS) is susceptible
to some artifacts that affect all tissues, but it also
has artifacts that affect only the CNS. Garman
(2003) and Fix & Garman (2000) have described
different techniques that may be used to produce
optimal sections of central nervous system tissue.
Artifactual alterations to the grey matter may look
similar to the effects of trauma and hypoxia and
the vacuolar changes seen in myelin may appear
similar to myelin edema or Wallerian degenera-
tion. The most useful action, when examining a
CNS lesion under the microscope, to determine
whether the change is an artifact or a genuine
lesion, is to look for evidence of response to
putative injury. This includes vascular endothelial
swelling, the infiltration of neutrophils into areas
of degeneration and the presence of macrophages
within myelin digestion chambers (Summers et al
1995).
Postmortem myelin vacuolation is a common
artifact observed in the CNS. It generally produces
widespread, uniform, fine, vacuolation of the tissue
(Wells & Wells 1989). Prolonged holding of fixed
CNS tissue in 70% alcohol within an enclosed-
system automatic tissue processor (which may
happen over a weekend) may produce vacuolation
of the white matter (Fig. 8.32). The effect is con-
sistently reproduced in calf brains but not in pig
brains, suggesting a possible species difference in
tissue susceptibility (Wells & Wells 1989). In
addition, the degree of vacuolation depends on
undetermined processor factors additional to pro-
longed immersion in 70% alcohol (Wells & Wells
1989). The artifact resembles forms of intra­
myelinic edema and can be avoided by holding
tissue in primary fixative (Wells & Wells 1989).
FIGURE 8.32  Vacuolation of the white matter of the cer-
ebrum as a result of a suspected extended period in
alcohol during processing. ×100.
Retraction spaces around capillaries and scat-
tered oligodendrocytes are artifacts associated
with CNS tissue which is not perfusion-fixed and
which may have been improperly handled at
necropsy (Garman 1990). Cerebellar, cortical,
Purkinje cells (Fig. 8.33) often appear ischemic
(i.e. deeply eosinophilic and shrunken); however,
in most cases this is an artifact. If the lesion were
genuine, the Purkinje cells would be condensed,
elongated and deeply eosinophilic, and the nucleus
would be pyknotic. Mucocytes (Buscaino bodies)
are a rare artifact in the white matter that resemble
pale, blue to grey, amorphous bodies. Mucocytes
are PAS-positive and metachromatic and appear
to arise from the solubilization and subsequent
precipitation by the fixative of some component
of myelin (Garman 1990, Summers et al 1995).
Excessive handling of the brain at necropsy pro-
duces artifactual, dark-staining, basophilic neurons
(Fix & Garman 2000, Garman 2006, Summers
et al 1995). These neurons may appear to be
ischemic, but they are found in most areas (whereas
ischemic neurons generally occur at the base of the
sulci). Autolysis of the rat lens produces artifactual
change similar to cataract formation in the eye (Fig.
8.34). Many of the above artifacts may be avoided
by perfusion fixation (Summers et al 1995) fol-
lowed by a delay of several hours before the brain
is removed from the cranial vault (Garman 2003).

FIGURE 8.33  Eosinophilic Purkinje cells in the cerebel-
lum. ×100.
FIGURE 8.34  Autolysis of the rat lens appears similar to
cataract formation. ×200.
Acknowledgments
The figures and small excerpts of the text from
this chapter have been published with kind per-
mission from Springer Science+Business Media:
Comparative Clinical Pathology, Artefacts in
histopathology, 13, 2005, 100–108, Elizabeth F
McInnes.
References
Braber, S., Verheijden, K.A., Henricks, P.A., et al.,
2010. A comparison of fixation methods on lung
morphology in a murine model of emphysema. Am J
Physiol Lung Cell Mol Physiol. 299, L843–851.
Boonstra, H., Oosterhuis, J.W., Oosterhuis, A.M.,
et al., 1983. Cervical tissue shrinkage by
formaldehyde fixation, paraffin wax embedding,
section cutting and mounting. Virchows Arch. A.
Pathol. Anat. Histopathol. 402 (2), 195–201.
Darke, P.G.G., Donagura, J.D., Kelly, D.F., 1996.
Color atlas of veterinary cardiology. Mosby, St Louis.
Fix, A.S., Garman, R.H., 2000. Practical aspects of
neuropathology: a technical guide for working with
the nervous system. Toxicol. Pathol. 28, 122–131.
Garman, R.H., 1990. Artifacts in routinely immersion
fixed nervous tissue. Toxicol. Pathol. 18 (1 pt 2),
149–153.
Garman, R.H., 2003. Evaluation of large-sized brains
for neurotoxic endpoints. Toxicol. Pathol. 31,
32–43.
Garman, R.H., 2006. The return of the dark neuron.
A histological artifact complicating contemporary
Artifacts in histopathology 99
neurotoxicologic evaluation. Neurotoxicology 27 (6),
1126.
Hausmann, R., Bock, H., Biermann, T., et al, 2004.
Influence of lung fixation technique on the state of
alveolar expansion—a histomorphometrical study.
Leg. Med. (Tokyo) 6, 61–65.
Lanning, L.L., Creasy, D.M., Chapin, R.E., et al., 2002.
Recommended approaches for the evaluation of
testicular and epididymal toxicity. Toxicol. Pathol.
30, 507–520.
Latendresse, J.R., Warbrittion, A.R., Janssen, H., et al.,
2002. Fixation of testes and eyes using a modified
Davidson’s fluid: comparison with Bouin’s fluid and
conventional Davidson’s fluid. Toxicol. Pathol. 30,
524–533.
McInnes, E.F., 2005. Artefacts in histopathology.
Comp. Clin. Pathol. 13, 100–108.
Pearson, G.R., Logan, E.F., 1978. The rate of
development of postmortem artefact in the small
intestine of neonatal calves. Br. J. Exp. Pathol. 59,
178–182.
Slaoui, M., Fiette, L., 2011. Histopathology
procedures: from tissue sampling to histopathological
evaluation. Methods Mol Bioi 691, 69–82.
Summers, B.A., Cummings, J.F., De Lahunta, A.,
1995. Veterinary neuropathology. Mosby Year Book,
pp. 34–35.
Thompson, S.W., Luna L.G., 1978. An atlas of artifacts
encountered in the preparation of microscopic tissue
sections. Charles Louis Davis DVM Foundation,
Springfield, IL.
Wells, G.A., Wells, M., 1989. Neuropil vacuolation in
brain: a reproducible histological processing artefact.
J. Comp. Pathol. 101 (4), 355–362.

Reproduction of the rat, mouse, dog, non-human primate and minipig
Introduction
The reproductive system has been assigned a sepa-
rate chapter for a number of reasons. Most of these
reasons relate to the significant morphological vari-
ability that is seen in the reproductive organs of
all species as a result of normal physiological altera-
tions: for example, changes associated with estrus
or menstrual cyclicity in the female reproductive
tissues, changes associated with sexual maturation
and reproductive senescence in both sexes, regres-
sion and recrudescence of reproductive tissues in
seasonal breeding animals such as the hamster and
the rhesus monkey. It is essential that the patholo-
gist is familiar with these normal variations in
morphology so that these findings are not mistaken
for pathological changes and so that the true range
of ‘normal’ is appreciated.
In addition to physiological endocrine-mediated
changes, the reproductive tissues show a range of
background pathology which is sometimes species-
specific, but more often similar across species. The
chapter is divided into separate sections on the
male and female reproductive background changes,
and the most common species (rat, mouse, dog,
non-human primate and minipig) are covered in
each section.
REPRODUCTION: MALE
Male reproductive tract
Recognizing normal
spermatogenesis in the testis
In the male animal, recognizing the different cel-
lular associations that make up the spermatogenic
cycle within seminiferous tubules is essential to
be able to identify when cell populations are
missing or, in the case of spermatid retention,
when they are inappropriately present. An addi-
tional challenge in the male animal is the problem
of recognizing immaturity and distinguishing it
from degenerative conditions. This is a particular
problem in the dog and non-human primate.
Spermatogenesis staging is aided by the use
of the correct fixative to preserve the testes
(Latendresse et al 2002, Lanning et al 2002).
Spermatogenesis is essentially the same in all
mammalian species. It consists of continuous
division and maturation of precursor stem cells
(spermatogonia), which then enter meiosis (sper-
matocytes) where they undergo DNA replication
and reduction division to form the haploid germ
cells (spermatids). These cells undergo a complex
morphogenesis, changing from normal appearing
cells (round spermatids) to thin whip-like cells
(elongating spermatids) that possess a head, body
and tail. The mature spermatid is released from
the seminiferous epithelium (spermiation) and
transported in seminiferous tubule fluid to the
collecting reservoir of the rete testis and on into
the epididymis via the efferent ducts. The overall
process is the same between species, but the
Reproduction of the rat, mouse, dog, non-human primate and minipig
detailed characteristics of the different cell types,
the kinetics of the process, and the regulatory
pathways vary slightly.
The different types of germ cell (spermatogo-
nia, spermatocytes and spermatids) are arranged
in a very regular, layered pattern within the sem-
iniferous tubules (Figs 9.1, 9.2). They are sup-
ported both structurally and metabolically by the
somatic Sertoli cell, which entirely surrounds each
germ cell with cytoplasmic processes not readily
visible by light microscopy. The true extent of the
Sertoli cell becomes more apparent when germ
cells are lost from the tubule, leaving only Sertoli
cells lining the tubules (Fig. 9.4).
Elongating spermatid Round spermatid
Pachytene
spermatocyte
Spermatogonium
Sertoli cell
FIGURE 9.1  Rat seminiferous tubule stage V. Tubules in
the first half of the cycle (stages I–VIII) contain four layers
of germ cells. There are two generations of spermatids
(round and elongating spermatids) and only one genera-
tion of spermatocytes (pachytene spermatocytes). There
is a basal layer of spermatogonia interspersed by Sertoli
cell nuclei. ×200.
Pachytene Elongating
spermatocyte spermatid
Prepachytene
spermatocyte
Spermatogonium
Sertoli cell
FIGURE 9.2  Rat seminiferous tubule stage XII. Tubules in
the second half of the cycle (stages IX–XIV) also contain
four layers of germ cells. There is only one generation of
spermatids (elongating spermatids) but there are two
generations of spermatocytes (pachytene and prepach-
ytene). There is also a layer of spermatogonia inter-
spersed by Sertoli cell nuclei. ×200.
The germ cells proceed through spermatogen-
esis in a highly controlled manner, and in any given
tubule there are always four generations of germ
cells developing in complete synchrony with one
101
CHAPTER 9 
Dianne Creasy
another. This gives rise to the concept of stages
of the spermatogenic cycle, where each stage
can be identified by the precise morphological
characteristics of the individual germ cells from
the four synchronously developing generations.
The detailed description of the cell associations
is beyond the scope of this chapter but is com­
prehensively covered in a number of reviews
(Creasy 1997, Creasy & Foster 2002, Russell et al
1990). It is important that the toxicological
pathologist be familiar with the concept of the
spermatogenic cycle and the different appear-
ances of tubules, at least at the beginning, middle
and end of the cycle (Figs 9.1, 9.2) (Foley 2001).
Rat and mouse
Immaturity, peripuberty, maturity
Rats start releasing sperm from the testis at about
six to seven weeks of age, but sperm output does
not reach its full potential until about 10–11 weeks
of age (for review see Marty et al 2003). The first
cycle of spermatogenesis is relatively inefficient,
with reduced numbers of maturation-phase sper-
matids and numerous degenerating germ cells
(multinucleate and apoptotic cells) in the seminif-
erous tubules. The cauda epididymis is only par-
tially expanded and contains variable numbers of
sperm as well as cell debris and degenerating
sloughed germ cells from the testes (Fig. 9.3a &
b). Not all rats will mature at the same rate, and
so the appearance can easily be mistaken for tes-
ticular toxicity, especially if the test-article-treated
animals are slightly less sexually mature due to
decreased food intake or decreased body weight.
FIGURE 9.3a  Epididymis from a peripubertal eight-week-
old rat. The initial segment and caput epididymis are
expanded and filled with sperm. ×100.
102 BACKGROUND LESIONS IN LABORATORY ANIMALS
FIGURE 9.3b  Epididymis from a peripubertal eight-week-
old rat. The ducts of the distal corpus and cauda epidi-
dymis are still contracted and contain few sperm and cell
debris. ×40.
Puberty occurs earlier in the mouse, with sperm
being present in the epididymis at approximately
five weeks of age. As with the rat, degenerating
germ cells and low epididymal sperm numbers
accompanied by cell debris will be present until
spermatogenesis reaches full efficiency at around
eight to nine weeks of age.
Congenital or developmental lesions
Absence or incomplete development of the testis
(aplasia or hypoplasia) is occasionally seen but is
relatively uncommon. Hypoplasia is characterized
by a small testis containing reduced numbers of
seminiferous tubules. Cryptorchidism, caused by
delayed descent of the testis into the scrotal sac,
is occasionally seen and results in tubular degen-
eration and atrophy. This is often difficult to iden-
tify at necropsy because rodents can readily
retract the testes into the abdominal cavity. His-
tologically, the cryptorchid testis displays seminif-
erous tubular atrophy and the tubules are lined
solely by Sertoli cells with few spermatogonia.
Lesions seen in
young animals: testes
In general, the incidence of background degenera-
tive findings in rodent testes is low, making them
a very sensitive species for detecting sperma-
togenic abnormalities.
Tubular degeneration/atrophy
Occasional tubules (five tubular profiles/testis)
with partial or complete depletion of germ cells is
a common finding in young adult rats (Fig. 9.4).
Tubuli recti, which are the transitional tubules
that connect the seminiferous tubules with the
rete testis, should not be mistaken for atrophic
tubules. Tubuli recti are generally seen immedi-
ately adjacent to the main rete testis lumen that
is situated at the cranial pole of the testis (Fig.
9.5). The tubuli may also be present in a more
lateral subcapsular position, because fingerlike
extensions of the rete radiate out from the main
sac-like structure.
FIGURE 9.4  Occasional (1–5) atrophic tubules, lined only FIGURE 9.6a  Diffuse tubular atrophy in the rat. Seminif-
by Sertoli cells, are commonly seen as a background erous tubules lined only by Sertoli cells with all germ cells
finding in rat testes. The finding probably represents a missing. This can often be seen as a unilateral or bilateral
loss of germ cells from a short segment of one convoluted lesion and may be present in young rats and mice. The
seminiferous tubule. ×100. tubules may have a dilated tubular lumen and be sur-
rounded by interstitial edema. ×100.
*
FIGURE 9.5  Rete testis (rat) lined by cuboidal epithelium
with a transitional tubule (tubulus rectus) emptying into FIGURE 9.6b  Diffuse tubular atrophy in the rat. The
it. It is common to see tubules lined only by Sertoli cells tubules may be contracted with luminal closure (b). This
(*) close to the rete testis. These are normal profiles of can also be seen as an end-stage, chemically induced
the tubuli recti and should not be mistaken for atrophic lesion (unilateral or bilateral) and so it is important to
tubules. They can often be seen in a subcapsular position know the historical background incidence of this finding
somewhat distant from the visible rete because the in the appropriate strain of mouse or rat. ×100.
finger-like projections of the rete testis radiate some dis-
tance from the main sac. ×100. Other findings that can occasionally be seen
include tubular dilatation (unilateral or bilateral)
More extensive tubular atrophy, often involving (Fig. 9.7) which may be associated with a dilated
complete loss of all germ cells from all tubules in
rete. Tubular degeneration characterized by
one or both testes, can sometimes be seen as a varying numbers of degenerating germ cells,
background finding in young adult rats (Fig. 9.6a).
including multinucleate giant cells and/or disor-
If the atrophy has been present for more than a ganization of germ cell layers, is sometimes seen
few weeks the corresponding epididymis will as a bilateral background finding (Fig. 9.8).
probably be depleted of sperm and/or germ cells.
This can be a useful indication of whether the
atrophy was pre-existing, prior to study start, and
therefore a background change. Moderate to
severe tubular atrophy is often accompanied by
increased interstitial fluid (sometimes termed
edema) (Fig. 9.6b). Care should be taken when
using this diagnosis because fluid accumulation
can occur as a fixation artifact, particularly in
Bouins-fixed testes (Fig. 9.6b).

FIGURE 9.7  Unilateral or bilateral tubular luminal dilata-
tion is occasionally seen in young adult rats. The affected
testis or testes are enlarged and generally show increased
weight. The lesion may be associated with a dilated rete
and likely reflects fluid build-up due to obstruction of the
efferent ducts. The lesion generally progresses rapidly to
complete tubular atrophy (as in FIGURE 9.6a) due to pres-
sure atrophy, but the tubular lumina often remain dilated.
×100.
FIGURE 9.8  Tubular degeneration (rat) can be seen as an
incidental background finding in young rats and mice. It
is generally bilateral, and varying numbers of germ cells
may show degeneration. The germ cells may form multi-
nucleate or syncytial bodies due to coalescence of germ
cells that are normally joined by cytoplasmic bridges.
Round spermatids often develop chromatin margination
of their nuclei as they degenerate. The degenerate cells
are either sloughed into the lumen or phagocytozed by
the Sertoli cells. As more cells die, the regular layering of
the germ cell layers is lost. ×200.
Abnormal residual bodies (Fig. 9.9) are most
commonly seen in the mouse and are character-
ized by the presence of abnormally large and
dense residual bodies that are usually located on
the luminal surface of the germinal epithelium in
tubules at various stages of the spermatogenic
cycle. In contrast, normal residual bodies are
much smaller and should only be present at the
luminal surface of stage VIII–IX tubules, follow-
ing which they rapidly descend to the basal region
of stage X–XI tubules where they are phagocy-
tosed. Abnormal residual bodies are often seen in
tubules in other stages of the cycle.
Reproduction of the rat, mouse, dog, non-human primate and minipig
FIGURE 9.9  Mice testes occasionally contain abnormal-
appearing residual bodies (arrowed) as a background
lesion. They are larger than normal residual bodies and
often occur in tubular stages not normally associated with
residual body formation. They may also be increased as
a chemically-induced lesion. ×200.
Lesions seen in young
animals: epididymides
In the rat and mouse epididymis, sperm is nor-
mally present and abundant from the initial
segment through to the cauda. In the adult rat,
there are generally negligible numbers of sloughed
germ cells mixed in with the sperm and so the
presence of sloughed cells or debris in the epidi-
dymis (particularly in the head) is a very sensitive
indicator of testicular injury. Sloughed germ cells
are slightly more frequent in the mouse. Sperma-
togenic disturbances in the testes will generally be
reflected by decreased sperm and cell debris in
the epididymis.
Sperm granulomas are one of the most common
changes seen in young rat epididymides. They may
be unilateral or bilateral and can occur anywhere
in the epididymis, but are more commonly noted
in the cauda (Fig. 9.10). Occasional lymphoid
aggregates in the interstitium of the epididymis are
a common finding, but are generally few in number.
FIGURE 9.10  Epididymal sperm granuloma is a common
background finding in young adult rats. The lesion repre-
sents a granulomatous response to sperm (which are
antigenically foreign) that have ruptured through the
epididymal epithelial lining. Sperm granulomas are more
common in the cauda than the caput epididymis. ×100.
Cribriform change, characterized by intra-
epithelial microcysts, is a common finding at the
junction of the corpus/cauda (Fig. 9.11).
103
FIGURE 9.11  Epithelial cystic vacuolation (rat), some-
times referred to as cribriform change or pseudo-gland
formation, is often seen at the junction between the distal
corpus and cauda epididymis. ×100.
Age-related lesions:
testes and epididymides
Increased germ cell degeneration or increased
numbers of tubules with germ cell depletion
occur in the testis with increasing age, but gener-
ally spermatogenesis remains relatively efficient
throughout the entire life of the rat and mouse.
Minimal, diffuse hyperplasia may accompany the
decline in spermatogenesis as a secondary hormo-
nal response.
The most common age-related testicular find-
ings include degenerative and inflammatory lesions
of the vasculature, including fibrinoid necrosis,
hypertrophy and inflammation of the vascular wall
as part of the more generalized condition of pol-
yarteritis nodosa (Fig. 9.12a). Lymphoid aggre-
gates within the interstitium of the rat epididymis
are common (Fig. 9.12b). The testis is also a
common site for deposition of amyloid in systemic
amyloidosis of the aging mouse (Fig. 9.13). Focal
proliferative changes in the Leydig cell population
(hyperplasia and tumors) (Fig. 9.14) are very
common in the F344 rat, and are also present at
a lower incidence in other strains of rats and mice.
Another common proliferative lesion in the mouse
testis is hyperplasia of the rete epithelium (Fig.
9.15). Lipofuscin pigmentation is associated with
aging and is observed in the interstitial macro-
phages and Leydig cells.
FIGURE 9.12a  Fibrinoid necrosis of arteriolar walls and
periarteritis in the interstitial vasculature of the testis of
an aging rat. ×100.
104 BACKGROUND LESIONS IN LABORATORY ANIMALS

FIGURE 9.12b  Lymphoid aggregates within in the inter- FIGURE 9.15  Hyperplasia of the rete epithelium in the FIGURE 9.17a  Purulent inflammation with regenerative
stitium of the rat epididymis are common. ×100. mouse. The normal low cuboidal epithelium has been epithelial hyperplasia of the ventral prostate in an aging
replaced by a tall columnar hyperplastic epithelium. rat with a urogenital infection. ×100.
×100.

In the aging rat, the segment of the epididymis

at the junction between the caput and corpus
commonly develops a foamy, basophilic vacuola-
tion within the epithelium (Fig. 9.16). This prob-
ably represents the normal glycoprotein secretory
product that has become modified in the aging
rat. If the cystic rete testis is filled with sperma-
tozoa, the lesion is referred to as a spermatocoele.
Spermatocoeles are also common at the junction
between the epididymis and vas deferens.

FIGURE 9.13  Amyloid deposition in the interstitial com-

partment of a mouse testis. The testis is a common site FIGURE 9.17b  Epithelial hyperplasia in the acinus of the
for amyloidosis. ×100. rat prostate. ×200.

FIGURE 9.16  Basophilic vacuolation is a common age-

related change in the epididymal epithelium of rats. It only
occurs in one region of the epididymis that is at the junc-
tion of the caput and corpus. ×200.

FIGURE 9.14  Focal Leydig cell hyperplasia and Leydig
cell tumors are very common in some strains of rat
(F344), but are also seen at a low incidence in other
strains of rats and mice. ×100.
Age-related lesions: accessory sex
organs and penis
Urogenital infections affecting the penis, prepu-
tial glands, prostate, coagulating glands and
seminal vesicles are common in aging rodents, par-
ticularly in mice (Suwa et al 2001). Acute or
chronic active inflammation with abscess forma-
tion (Figs 9.17a, 9.18) may be present throughout
the genital tract (Fig. 9.17a). Focal epithelial
hyperplasia is observed in the aging rat prostate
(Fig. 9.17b).
FIGURE 9.18  Chronic inflammation of the rat preputial
gland. ×100.
In the accessory sex organs, common age-
related lesions include distension of the seminal
vesicles with degradation of the seminal fluid (Fig.
9.19) or contraction of the vesicles with atrophy
of the epithelium. The prostate also shows pro-
gressive acinar atrophy and mineralized concre-
tions in the acinar lumina (Fig. 9.20). Cystic
dilatation of the preputial gland ducts with secre-
tion or cystic atrophy of the acini are common
findings in both rats and mice (Figs 9.21, 9.22).
Similar cystic dilation can also be seen in the
bulbourethral gland (Fig. 9.23).

FIGURE 9.19  Grossly distended seminal vesicle of a
mouse with degradation of the secretory product. ×100.
FIGURE 9.20  Mineralized concretions and progressive
acinar atrophy of the prostate are common age-related
findings in the rat. ×100.
FIGURE 9.21  Rat preputial gland with cystic dilatation of
the duct with secretion. This often results in gross
enlargement of the gland. ×100.
FIGURE 9.22  Mouse preputial gland with cystic atrophy
of the acini. ×100.
Reproduction of the rat, mouse, dog, non-human primate and minipig 105
FIGURE 9.23  Cystic dilatation of the bulbourethral gland FIGURE 9.24  Totally quiescent testis from a dog,
of a rat, which is expanded with secretion. ×200. five to six months of age. Tubules are lined by Sertoli cells
interspersed with occasional gonocytes. There is no
lumen within the tubules because fluid is not yet being
Dog secreted. ×100.
Background changes in Beagle dog testes are
common and may obfuscate chemically-induced
toxicity. Spermatogenesis in the dog appears to
be much less efficient than that in the rodent or
non-human primate and seminiferous tubules fre-
quently show incomplete spermatogenesis (hypo-
spermatogenesis). One of the greatest problems
identifying chemically-induced toxicity is caused
by using dogs that are still sexually immature or
peripubertal at the time of examination. It is
therefore important that the pathologist under-
stands the characteristics of immature, peripuber-
tal and mature reproductive tissues so that the
degenerative changes that characterize this period
of morphological development are not mistaken
for toxicologically-induced disruption of sperma-
togenesis. The normal appearance of sperma- FIGURE 9.25  Testis from a dog, five to six months old,
togenesis in the dog testis and the cell associations where the Sertoli cells are just beginning to secrete fluid
of the stages of the spermatogenic cycle have which is forming vacuoles within the Sertoli cell lining and
been described in detail by Russell and coworkers producing a lumen in the gradually expanding tubules.
(1990). The gonocytes are proliferating to form spermatogonia,
some of which are apoptotic. ×100.
Immaturity, peripuberty, maturity
Dogs mature over a relatively wide age range
(seven to twelve months of age), with most reach-
ing sexual maturity by ten months of age. The age
of sexual maturity is also influenced by the sup-
plier. Dorso and coworkers (2008) reported that
90% of dogs 31–40 weeks of age supplied by
Harlan, France, were sexually mature compared
with only 10% of dogs of the same age supplied
by Marshall Farms, USA. Dogs less than ten
months of age usually show a range of degenera-
tive morphologies reflecting ongoing maturation.
Dogs on the borderline of maturity will have
varying numbers of degenerating, missing and
sloughed germ cells in the testis and the epididy-
mal lumina and some tubules may be vacuolated,
FIGURE 9.26  Testis from a dog, six to seven months old.
contracted or dilated. The appearance of the
The tubules are progressively expanding due to the fluid
testes and epididymal contents can be indistin-
secreted by the Sertoli cells and there are numerous
guishable from chemically-induced toxicity, and
fluid-filled vacuoles within the Sertoli cell lining. Sperma-
evaluating the relationship of such changes to
togonia and early spermatocytes can be seen in the
treatment is made more difficult by the small
tubules, many of which are undergoing apoptosis. ×100.
number of animals in each group. The confusion
can be avoided by ensuring that dogs are at least
ten months and preferably 12 months of age at
the end of the study. The morphological charac-
teristics of the developing dog testis and epidi-
dymis are illustrated in Figs 9.24–9.29 and Figs
9.30–9.34 respectively.
106 BACKGROUND LESIONS IN LABORATORY ANIMALS
FIGURE 9.27  Testis from a dog, six to nine months old.
The tubules are variously populated with germ cells
extending up to round spermatids. Spermatogenesis is
still patchy in most tubules, and many of the newly devel-
oped germ cells are degenerating or being sloughed into
the lumen. ×20.
FIGURE 9.28  Testis from a dog, six to nine months
of age. Most tubules have layers of germ cells up to
pachytene spermatocytes or round spermatids. There are
also occasional elongating spermatids. Degenerating and
sloughed germ cells are fewer but still quite frequent.
×200.
FIGURE 9.29  Testis from a 12-month-old dog. All tubules
have complete spermatogenesis and have a full comple-
ment of elongating spermatids the tails of which fill the
lumen. Occasional degenerate cells are still present but
they are infrequent. ×200.
FIGURE 9.30  Cauda epididymis from a dog, five to six
months of age. The ductal diameter is small due to lack
of any fluid secretion by the testis. There are no sloughed
cells or cell debris.
FIGURE 9.31  Cauda epididymis from a dog, six to eight
months of age. The ductal lumen is beginning to expand
and contains a small amount of fluid and sloughed germ
cells from the testis.
FIGURE 9.32  Cauda epididymis from a dog, six to eight
months of age. The luminal diameter is gradually increas-
ing and contains fluid and cells.
FIGURE 9.33  Cauda epididymis from a dog, 10–12
months of age. The ductal lumen is fully expanded with
sperm and there are only occasional sloughed germ cells.
FIGURE 9.34  Caput epididymis from a dog, six to eight
months of age. The ducts are filled with sloughed, apop-
totic and degenerating germ cells from the developing
testis but there are no sperm. Sloughed cells are more
obvious in the head than the tail of the epididymis. ×200.
Unlike the rodent, the dog goes through a
developmentally quiescent period with respect to
testicular development. During this period, the
seminiferous tubules are lined by Sertoli cells and
small numbers of gonocytes (pre-spermatogonia)
which do not begin development until around five
months of age (Fig. 9.24). At that time the gono-
cytes begin to proliferate and produce spermato-
gonia which will populate the basal layer of the
seminiferous tubule and initiate the process of
spermatogenesis. At the same time, the Sertoli
cell resumes its maturation and begins to secrete
seminiferous tubule fluid. This often takes the
form of fluid-filled vacuoles within the layer of
Sertoli cells and the gradual formation of a tubular
lumen and expansion of the overall tubular diam-
eter (Figs 9.25, 9.26). Spermatogonia begin to
proliferate and produce spermatocytes, which in
turn divide and produce round spermatids. The
round spermatids gradually elongate and mature
into the final, whip-like mature spermatids, which
are released into the tubular lumina. From there
they are transported on into the epididymis. Sper-
matogenesis does not develop at the same rate in
all tubules, and so different tubules will be more
or less advanced than others and because the
Sertoli and germ cells are not yet up to full func-
tional efficiency, there will be considerable degen-
eration and sloughing of germ cells in this first
cycle of spermatogenesis (Fig. 9.27). The gradual
increase in seminiferous tubule fluid production
and the high rate of germ cell attrition is reflected

by the luminal contents of the epididymis (Figs
9.30–34). Small-diameter, empty ducts will grad-
ually expand with fluid and contain degenerate
sloughed germ cells through to puberty (Fig.
9.34), to be replaced by expanded ducts filled
with mature sperm and relatively few degenerate
cells in the fully mature adult (Fig. 9.33).
Identifying the status of sexual maturation in
the dog testis should be done on the basis of the
presence of sperm in the corpus and cauda epidi-
dymis (the caput does not concentrate sperm suf-
ficiently to be able to visualize it) and the presence
and number of elongating spermatids in the testis.
The size and secretory activity of the prostate is
not very useful for identifying maturity because
the prostate matures at a different rate.
Lesions seen in young
mature animals: testes
Changes that can easily be confused with imma-
turity, but which are background degenerative
changes of the sexually mature dog, are seen com-
monly in dogs that are more than 10 months
of age. The changes have been described by
Rehm and coworkers (2000) and by Goedken
and coworkers (2008). The most common find-
ings are tubular hypospermatogenesis and tubular
hypoplasia/atrophy, both of which have an approx-
imately 30% incidence in normal dogs. Hyposper-
matogenesis is characterized by the absence of
one or more generations of germ cells from the
seminiferous epithelium (Figs 9.35, 9.36). Hypo-
spermatogenesis may be distinguished from
immaturity by the fact that many of the tubules
will have a layer of mature, elongate spermatids
at the luminal surface, but with the underlying
layer of round spermatids and/or layer of pach-
ytene spermatocytes missing or partially depleted.
This is indicative of transient (cyclical) inhibition
of spermatogonial division and is different from
the pattern of germ cell degeneration and patchy
germ cell depletion typical of immaturity (com­
pare with Fig. 9.27). Hypospermatogenesis can
range in severity from minimal (<10% tubules
affected) to severe (>75% tubules affected). Most
testes will have a few tubules affected by
hypospermatogenesis.
* adult dog testis include swollen spermatocytes.
FIGURE 9.35  Stage VI tubule with hypospermatogenesis
(*). This tubule is in the same stage of spermatogenesis
as the other two tubules (identifiable by the shape of the
elongating spermatids). Although this tubule contains a
normal population of elongating spermatids, it lacks the
underlying layers of pachytene and prepachytene sper-
matocytes that are present in the other two tubules. ×200.
Reproduction of the rat, mouse, dog, non-human primate and minipig 107
*
FIGURE 9.36  Stage V tubule with hypospermatogenesis FIGURE 9.38  Tubules containing large, ‘swollen’, densely
(*). The tubule on the left is at the same stage. Although staining spermatocytes (arrowed). Most testes will have
the elongating spermatid population is normal in the a few tubules that contain these cells. They probably
hypospermatogenic tubule, it is almost entirely missing represent secondary spermatocytes which have failed to
the underlying layers of round spermatids, pachytene complete their second meiotic division and have become
spermatocytes and spermatogonia that are present in the arrested in their development while the rest of the cells
normal stage V tubule. ×200. in the tubule continue maturation. ×200.
Segmental hypoplasia is typically seen as a
discrete group of contracted tubules, often in a
wedge shape and situated in a subcapsular loca-
tion, which are completely devoid of germ cells
(Fig. 9.37). These tubules give the appearance of
never having been populated by germ cells, hence
the term hypoplasia. The lobular distribution sug-
gests that the group of tubular profiles belong to
a single, coiled, seminiferous tubule.
FIGURE 9.39  A few multinucleate giant cells, which rep-
resent degenerating spermatids or spermatocytes, may
be found in most mature dog testes. ×200.
Orchitis, characterized by a lymphocytic or
mixed inflammatory cell infiltrate in the intersti-
tium of the testis, may occasionally be seen in
the dog testes (Fig. 9.40) (Fritz et al 1976). The
inflammation may also extend into the tubular
epithelium, resulting in destruction of the affected
FIGURE 9.37  Tubular hypoplasia typically presents as a
tubules. The lesion is generally accompanied by a
triangular wedge of contracted tubules lined only by
lymphocytic inflammation in the epididymis (Fig.
Sertoli cells. The surrounding tubules show normal sper-
9.41) and in some cases is genetically associated
matogenesis. The affected tubules have the appearance
with lymphocytic thyroiditis (Fritz et al 1976).
of never having been populated with germ cells. ×100.
The lesion is thought to have an autoimmune
etiology.
Other common background findings in the
These are probably secondary spermatocytes
which have failed to complete their second
meiotic division and remain sequestered in the
germinal epithelium, arrested in meiosis, while
the surrounding cells continue their development
into round spermatids (Fig. 9.38). Occasional
multinucleate germ cells, which represent degen-
erating spermatocytes or spermatids, are also a
common feature of dog testes (Fig. 9.39). Both
findings can usually be seen in a few tubules from
most dog testes.
108 BACKGROUND LESIONS IN LABORATORY ANIMALS
FIGURE 9.40  Lymphocytic orchitis is an uncommon
background finding in the dog. The infiltrating lym-
phocytes and macrophages often penetrate and destroy
seminiferous tubules in the affected areas. ×100.
FIGURE 9.41  Lymphocytic epididymitis often accompa-
nies lymphocytic orchitis. ×100.
Lesions observed in young
mature animals: testes
The sperm content of the dog epididymis varies
depending on location. Sperm is generally not seen
in the efferent ducts or the head of the epididymis
because it is very dilute. Sperm becomes visible
in the distal cauda and appears dense throughout
the corpus. Sperm content within the large-
diameter ducts of the cauda can be variable and
may often be missing due to loss during processing.
The sperm content of the corpus and proximal
cauda should be used to evaluate normal sperm
content and maturity status.
Sperm stasis, sometimes accompanied by
inflammation, is a common finding in the efferent
ducts situated in the head of the epididymis, and
probably represents impaction of sperm in blind-
ending efferent ducts (Fig. 9.42) (Foley et al 1995).
True sperm granulomas, similar to those seen in
rodents, also occur throughout the epididymis.
FIGURE 9.42  The efferent ducts are embedded in the
head of the dog epididymis and some are blind-ending.
This may result in sperm stasis and dilation. ×100.
Cribriform change, characterized by infolding
of the ductal epithelium with the formation of
pseudoglandular cysts (Fig. 9.43), is often observed
at the corpus/caudal junction. This is similar to
the lesion in rodents. Intranuclear eosinophilic
inclusion bodies are a feature of the canine epidi-
dymis (Fig. 9.44), particularly in the caput,
although their significance is unknown.
FIGURE 9.43  Cribriform change or ‘pseudoglandular’
cysts are frequently present at the junction between the
distal corpus and cauda epididymis of the dog. ×100.
FIGURE 9.44  Intranuclear eosinophilic inclusions are
commonly seen in the epithelial cells of the caput epidi-
dymis of the dog. Their significance is unknown. ×200.
Interstitial lymphocytic infiltrates are common
and may also be accompanied by arteritis or
periarteritis either in the interstitium or in the
surrounding adipose tissue of the dog epididymis.
This is a common site for arteritis associated with
the more generalized condition of ‘beagle pain
syndrome’. Epithelial hyperplasia of the ductus
epididymis is a normal feature of the aged beagle
dog epididymis (James & Heywood 1979).
Lesions seen in young
mature animals: prostate
The features of the maturing dog prostate and
some of the more common background lesions
have been reviewed by Dorso and coworkers
(2008). The prostatic acini of the dog gradually
expand as secretory activity increases in the epi-
thelium. The prostate of the immature dog is
small, the acini are contracted and lined by a low,
basophilic, cuboidal or squamous epithelium. As
the gland becomes functionally active under the
regulation of testosterone and dihydrotestoster-
one, the epithelium becomes tall and columnar
and filled with intensely eosinophilic secretion. In
contrast to the rat, in the dog the secretory fluid
produced by the prostate is stored in the acinar
epithelium and not in the acinar lumen. Although
there is generally good correlation between the
timing of testis and prostate maturity, they are not
always in synchrony, and one can be mature while
the other is immature.
The most common background lesions of the
dog prostate include focal or lobular cystic acini
(Figs 9.45, 9.46). The acini may be lined by low
cuboidal ‘atrophic’ appearing epithelium or by
columnar secretory epithelium. Minor inflamma-
tory infiltrates are also often noted in the prostate,
ranging from focal or periurethral interstitial lym-
phoid infiltrates to subacute or chronic inflamma-
tion affecting the prostatic acini (Fig 9.47). The
inflammatory response is generally accompanied
by atrophy of the affected acinar epithelium.
There are age-related variations in the testes and
prostate of beagle dogs (James & Heywood 1979)
and benign prostate enlargement is a normal
feature of the aged male Beagle.
FIGURE 9.45  Focal, cystic dilation of prostatic acini is a
common finding in dogs. ×100.

FIGURE 9.46  Focal, fluid-filled, cystic acini and an area
of acinar atrophy in the dog prostate. ×100.
FIGURE 9.47  Area of prostatic acinar atrophy with inter-
stitial chronic inflammatory infiltrate in the dog. ×100.
Non-human primate
Cynomolgus monkeys (Macaca fascicularis) are
the most common non-human primates used in
preclinical studies. The geographical origin of the
animal influences the background pathology that
is observed. Many non-human primates are cur-
rently imported from Indo-China and are in most
cases captive-bred. Cynomolgus males become
sexually mature at around four to five years of age,
but it is common for studies to be conducted with
animals that range from totally immature, through
pubertal maturation, to totally mature. This
makes evaluation of any testicular toxicity almost
impossible. It is important for the pathologist to
be aware of the characteristics of the maturing
reproductive system so that degenerative changes
associated with puberty are not confused with
chemically induced degenerative changes.
Characteristics of the prepubertal, peripubertal
and adult cynomolgus testis are illustrated in Figs
9.48–9.50. A detailed description of the normal
cynomolgus testis and the cell associations of the
spermatogenic cycle have been described by
Dreef and coworkers (2007). As with other
species, degenerating and sloughed germ cells are
present in significant numbers in the testis and in
the epididymal lumina of pre- and peripubertal
monkeys (Fig. 9.49). Sometimes, a discrete focus
of tubules may develop complete spermatogenesis
in an otherwise immature testis (Fig. 9.51). The
immature or pubertal testis will also be associated
with absence or low numbers of sperm in the
epididymis. The secretory activity and size of the
seminal vesicles and prostate are also dependent
on the maturational status of the monkey.
Reproduction of the rat, mouse, dog, non-human primate and minipig
FIGURE 9.48  The cynomolgus monkey testis undergoes
a quiescent period until around three and three-and-a-half
years of age. Prior to this, the seminiferous tubules are
contracted with no tubular lumen and lined by Sertoli cells
and gonocytes. ×100.
FIGURE 9.49  Testis from a prepubertal monkey. Sperma-
togenesis has only proceeded to the level of pachytene
spermatocytes and a few round spermatids, many of
which are degenerating and being sloughed into the
tubular lumen and will end up in the epididymis. ×100.
FIGURE 9.50  Testis from a young adult monkey. The
tubules are expanded with a patent tubular lumen and all
germ cell types are present, including a small number of
mature sperm at the luminal surface. ×100.
109
FIGURE 9.51  Occasionally a focal area of tubules in an
otherwise immature testis will develop complete sperma-
togenesis ahead of all the other tubules. ×100.
Seasonal changes
Rhesus monkeys (Macaca mulatta) are seasonal
breeders and spermatogenesis will undergo cycli-
cal regression and recrudescence depending on the
time of year. The changes occur in response to
decreased levels of testosterone, so the entire
reproductive tract will show varying degrees of
degeneration and atrophy in a cyclical manner.
The appearance of the testes and epididymides
are identical to those of prepubertal and peripu-
bertal cynomolgus monkeys (as seen in Fig. 9.49)
Incidental findings
In the sexually mature testis, focal tubular dilation
with thinning of the epithelium, sperm stasis and/
or decreased numbers of germ cells occurs quite
frequently (Fig. 9.52). Germ cell degeneration
and germ cell depletion (tubular degeneration/
atrophy) are rarely seen as diffuse changes in the
mature non-human primate, but one or several
tubular profiles within a lobule (probably repre-
senting a single tubule) may show degenerative
changes including vacuolation, and partial germ
cell degeneration and depletion (Fig. 9.53).
FIGURE 9.52  Focal tubular dilation, which is often
accompanied by sperm stasis within the dilated lumen (as
here), is a relatively common finding. It may be associated
with partial depletion of germ cells as well as thinning of
the germinal epithelium. ×100.
110 BACKGROUND LESIONS IN LABORATORY ANIMALS
FIGURE 9.53  Focal tubular degeneration and atrophy is
sometimes present as an incidental background lesion.
Tubules may be totally or partially depleted of germ cells,
with some tubules lined only by Sertoli cells. The change
is often focal or lobular in distribution. ×100.
Fibrous hypoplasia is a lesion that has become
more prevalent in recent years and appears to be
associated with the importation of cynomolgus
monkeys from Indo-China. The change, which is
assumed to be a congenital condition, is character-
ized by large tracts of mature collagen connective
tissue replacing the normal lobular distribution of
seminiferous tubules (Fig. 9.54). Sperm granulo-
mas are rarely seen in the epididymis of cynomol-
gus monkeys. The prostate of the cynomolgus
monkey frequently contains mineralized concre-
tions (Fig. 9.55).
FIGURE 9.54  Fibrous hypoplasia of the monkey testis.
This is a relatively common finding in cynomolgus
monkeys originating from Indo-China. It is presumed  REPRODUCTION: FEMALE
to be a congenital condition and is generally present
bilaterally. ×20.
FIGURE 9.55  Mineralized concretions in the prostate of
a cynomolgus monkey. ×100.
Minipig
Immaturity, peripuberty and maturity
A characteristic of the mature pig testis is a large
number of interstitial (Leydig) cells. This should
not be mistaken for Leydig cell hyperplasia. The
interstitial cells begin increasing in number around
three to four months of age as the animals reach
puberty, but carry on increasing in number over
the next few months. This may mean that there
is significant variation in Leydig cell numbers in
animals that are three to six months of age. As
with other species, increased numbers of degen-
erating germ cells, reduced numbers of sperm in
the epididymis, and incomplete spermatogenesis FIGURE 9.57  Severe tubular hypoplasia or atrophy with
may be present in animals that are prepubertal or contracted tubules lined only by Sertoli cells. There is also
peripubertal. moderate Leydig cell hyperplasia. ×100.
The normal cell associations of the pig sperma-
togenic cycle have been described by Jorgensen
et al (1998). The overall pattern of spermatogen-
esis is similar to that in other species.
Incidental findings in mature animals
Congenital aplasia and cryptorchidism of one or
both testes have been described in minipigs, with
a 1.5% incidence of the latter (Jorgensen et al
1998).
Tubular hypoplasia or tubular atrophy is a rela-
tively common finding in mature pig testes
(Dincer & Svendsen 2006, Jorgensen et al 1998).
The affected tubules are often contracted and the
interstitial space is generally packed with Leydig
cells, which may be increased in number. The
severity of the finding can range from a few to the FIGURE 9.58  Reduced or absent sperm may be present
majority of tubules affected (Figs 9.56, 9.57). in the epididymides associated with testes that have sig-
When the finding is more severe, the epididymis nificant tubular hypoplasia/atrophy. ×100.
shows reduced or absent sperm (Fig. 9.58). In
some cases the hypoplasia and atrophy may be In the epididymides, aggregates of lymphocytes
associated with cryptorchidism. may be present in the interstitial tissue. In the
ductal contents, a few sloughed testicular germ
cells may be admixed with the spermatozoa, but
these are generally few in number. Sperm granu-
lomas are only rarely seen in the pig epididymis.
Chronic inflammatory cell infiltrate and inter-
stitial mineralized concretions may occasionally
be observed in the prostate (Dincer & Svendsen
2006).
Female reproductive tract
The morphological appearance of the female
reproductive tissues is dependent on hormonal
balance. This includes the hormonal interactions
FIGURE 9.56  Minimal tubular hypoplasia or tubular of the hypothalamic–pituitary–gonad axis as well
atrophy in a mature pig. Note the large number of Leydig as the balance of hormones secreted by the ovary
cells in the interstitial space, which is a normal feature of that regulate uterine and vaginal morphology.
the pig testis. ×100. Hormonal balance and therefore morphology
will change in response to cyclicity (estrus or
menstrual), maturational status and reproductive
senescence. It is on the background of this con-
stantly changing profile of morphological charac-
teristics that the toxicological pathologist must
evaluate tissues for evidence of toxicity.
With the exception of rabbits, which are
induced ovulators, all other species used for toxic-
ity testing undergo an estrus cycle, or in the
case of non-human primates, a menstrual cycle.
Both cycles are driven by a complex interplay of
 Reproduction of the rat, mouse, dog, non-human primate and minipig 111

hormones which produce cyclical morphological continues over the subsequent four or five cycles.
changes in the ovary, uterus, cervix, vagina, and in The continued presence of corpora lutea over
some species the mammary gland. In the female several cycles can make detection of failed ovula-
rodent, the ovaries, uterus and vagina all change tion difficult in shorter-term studies unless the
their morphology in rapid succession during the detailed morphology of the corpora lutea and fol-
constantly repeating four to five-day estrus cycle. licles is examined.
In contrast, the Beagle bitch comes into estrus
only once or twice a year and spends most of
the intervening time in the resting, anestrus
phase. In addition to the morphological changes
in the reproductive tract, the appearance of the
mammary gland shows dramatic cyclical altera-
tions in the dog, whereas there are negligible
changes in the mammary gland of most other
species. The mammary gland of the male rat
differs histologically from that of the female rat. FIGURE 9.61  During estrus (rat) the vagina is lined by a
In addition to the physiological changes in thick keratinized squamous epithelium. The cornified
hormone status due to maturation, cyclicity and layer is progressively shed as the cycle moves into 
senescence, the reproductive tissues show a metestrus. ×100.
variety of background lesions. The following
species-specific sections are organized to summa-
rize the main morphological features associated
with these categories of change. FIGURE 9.59  Ovary of a young adult rat contains primary,
secondary and tertiary follicles as well as corpora lutea
from the most recent ovulation and regressing corpora
Rat and mouse lutea from previous cycles. ×40.
With some exceptions (which are noted in the
text), the response of the female reproductive The cyclical changes in the vaginal epithelium
tract to hormonal changes and the range of back- are the easiest and most obvious morphological
ground findings are broadly similar between rat changes for the pathologist to use when staging
and mouse. the estrus cycle. However, these must be used in
conjunction with the changing morphology of the
Immaturity and peripuberty uterus and, to a lesser extent, that of the ovary to
identify the stage of estrus and to ensure that the
Rats and mice reach puberty and begin cycling at
various parts of the reproductive tract are in syn-
the same time. This occurs at such a young age
chrony with one another. The most obvious cycli- FIGURE 9.62  During metestrus (rat), the non-keratinized
(approximately four to five weeks of age) that
cal changes in vaginal morphology are illustrated squamous epithelium of the vagina becomes progres-
morphological changes associated with immatu-
in Figs 9.60–9.63, but the full spectrum of changes sively thinner and infiltrated with leucocytes. The epithe-
rity or peripuberty are not seen in general toxicity
and their correlation with uterine and ovarian lium is thinnest as the cycle enters diestrus. ×100.
studies and do not present a problem for inter-
morphology are best illustrated in the excellent
preting test-article-related changes.
reviews by Li & Davis (2007) and Westwood
(2008).
Changes associated with estrus
cyclicity in young rodents
The average duration of the rodent estrus cycle is
four to five days. The cycle comprises pro-estrus
(12–14 hours), estrus (25–27 hours), metestrus
(six to eight hours) and diestrus (55–57 hours),
based on vaginal smears. There are a number of
comprehensive reviews and illustrations of the
morphological changes in the ovary, uterus and
vagina in the different stages of the estrus cycle
of the rat (Westwood 2008, Yuan & Foley 2002).
A brief summary is provided here.
The short duration of the estrus cycle in the
FIGURE 9.63  At the beginning of diestrus (rat), the vagina
rodent means that most stages of follicular and
has a thin squamous epithelium, which is only a few cells
corpora luteal development and regression are
thick. As diestrus continues towards pro-estrus the epi-
present in the ovary, regardless of the stage of
FIGURE 9.60  Vagina of a rat during proestrus with a thelium becomes progressively thicker. ×100.
the cycle (Fig. 9.59). The corpus luteum is the
main structure that can be used to recognize prominent layer of mucified cells above a layer of corni-
fication. This will be shed as the cycle moves into estrus. Cyclical changes in the uterine endometrial epi-
cyclical changes. Immediately following ovulation
×100. thelium and stroma are less obvious than those in
(estrus), the corpus luteum is composed of small
the vagina, but they can sometimes be mistaken
basophilic cells and contains small amounts of
by the inexperienced pathologist as abnormal find-
hemorrhage and may sometimes also contain a
ings. These include dilation of the uterine lumen
central cyst. As it matures, the luteal cells enlarge
in pro-estrus and estrus (Glaister 1986) (Fig.
and become filled with lipid vacuoles (containing
9.64), pro­minent apoptosis of the epithelium and
steroid and cholesterol) and are intensely eosi-
glands during metestrus (Fig. 9.65), mitotic activ-
nophilic (metestrus). The structure is highly vas-
ity in the epithelium and glands during diestrus,
cularized in the early stages and then begins to
edema of the stroma during pro-estrus, and
regress as leucocytes, macrophages and fibroblasts
varying degrees of leucocytic infiltration of the
infiltrate the corpus luteum (diestrus). Regression
endometrium and cervical mucosa throughout the
112 BACKGROUND LESIONS IN LABORATORY ANIMALS
cycle (Fig. 9.66). These characteristics should be
viewed in conjunction with the vaginal and ovarian
morphology to determine normality of the repro-
ductive tract.
FIGURE 9.64  During late pro-estrus and early estrus (rat),
the uterine lumen becomes dilated with fluid. The horns
are often noted to be grossly dilated at necropsy. ×40.
FIGURE 9.65  There is prominent apoptosis of the surface
epithelium and glandular epithelium during metestrus
(rat). ×200.
FIGURE 9.66  Leucocytic infiltration, particularly by eosi-
nophils, is a common feature of the estrus cycle (rat).
×100.
Background pathology
in young rodents
Apart from the hormone-mediated cyclical
changes of the estrus cycle (described above)
and the early changes of reproductive senescence
(described below), relatively few background
changes occur in the reproductive tract of the
young female rodent. The most common are
occasional cysts (bursal, follicular, luteal or rete
ovarii) in the ovary (Figs 9.67, 9.68), and cysts in
the cervix and vagina, which may sometimes be
filled with keratin (Figs 9.69, 9.70a). Dilated
endometrial glands begin to develop in the young
rat as it approaches reproductive senescence;
these are the early stages of cystic endometrial
hyperplasia which is a very common end-stage
lesion in the aged rodent uterus and is described
in more detail below. Foci of prostatic tissue can
be encountered in female rats adjacent to the
urethra and vagina (Fig. 9.70b). Wollfian duct
remnants are made up of small foci of tubular
structures in the cervix of the uterus, generally
below the serosa or in the outer wall.
FIGURE 9.67  Follicular cysts in a young adult mouse
ovary. ×40.
FIGURE 9.68  Rete ovarii cyst in a mouse. ×100.
FIGURE 9.69  Keratin cyst in the cervix of a rat. Hyper-
plasia of the cervical epithelium and stromal thickening
are also present. ×40.
FIGURE 9.70a  Epithelium-lined cyst in the vagina of a
mouse. ×100.
FIGURE 9.70b  Prostatic tissue adjacent to the vagina.
×10.
Reproductive senescence
in aging rodents
Reproductive senescence can be a confounding
factor for interpreting toxicity studies in rodents.
Disturbances in hormone balance related to early
senescence are commonly seen in rats and mice
at the end of a 13-week toxicity study. Over 10%
of Sprague–Dawley rats show persistent estrus
by 20–21 weeks of age (Eldridge et al 1999).
The morphological profile of changes is dependent
on the nature of the hormone imbalance and
whether there is unopposed estrogen or unop-
posed progesterone at the time of examination.
Increased prolactin secretion from pituitary
hyperplasia or pituitary tumors can also be a major
determinant.
In the early stages of reproductive senescence,
the estrus cycle loses its normal four to five-day
cyclicity and becomes longer, generally due to
longer periods spent in estrus (persistent estrus),
which is due to an excess of estrogen compared
with progesterone. This is usually accompanied by
the presence of cystic follicles in the ovary (Fig.
9.71) due to failed ovulation but an otherwise rela-
tively normal (estrus) morphology of the vagina
and uterus. However, prolonged estrogen stimula-
tion will result in hyperplasia and/or squamous
metaplasia of the uterine epithelium (Fig. 9.72).
Periods of persistent estrus are generally inter-
spersed with periods of pseudopregnancy, where
progesterone is secreted in excess of estrogen. The
morphological characteristics of this hormonal
profile are persistent corpora lutea (maintained
by a concomitant increase in prolactin secretion)
 Reproduction of the rat, mouse, dog, non-human primate and minipig 113

(Fig. 9.73), an atrophic uterus with a folded epi-

thelium (Fig. 9.74), and most strikingly a vagina
lined by a mucified epithelium, which may appear
atrophic or hyperplastic (Fig. 9.75). Persistent
anestrus is the end stage of reproductive senes-
cence in rodents and is characterized by an ovary Int
that is devoid of follicles and corpora lutea. These
are generally replaced by an excess of interstitial
cells (Figs 9.76a, 9.77), which are formed by
thecal cells from atretic follicles and which secrete
androgens. The remains of the oocytes from the
atretic follicles can also be recognized (Fig. 9.77).
The uterus and vagina show an atrophic morphol-
ogy, similar to that seen in diestrus. These changes
(inactive ovary, uterus and vagina) are often also
associated with non-specific stress, which results FIGURE 9.73  Ovary of a rat in pseudopregnancy. FIGURE 9.76a  The end stage of ovarian atrophy in the
in decreased secretion of gonadotropin releasing Increased numbers of large eosinophilic corpora lutea and rat is characterized by interstitial gland and follicular
hormone. absence of tertiary follicles. ×40. cysts. Profiles of tubular structures lined by Sertoli-like
cells (arrowed) are often present. ×100.

FIGURE 9.71  Cystic follicles increase and corpora lutea

decrease as reproductive senescence progresses. This FIGURE 9.76b  Rat ovotestis with seminiferous tubules
reflects failed ovulation and is generally accompanied by and ovarian tissue. ×10.
features of persistent estrus in the uterus and vagina (rat).
×100.
FIGURE 9.74  Uterus of a rat in pseudopregnancy. The
uterus is atrophic and typically has a lumen lined by
folded endometrium. ×40.

*
*
*
FIGURE 9.77  Atrophic ovary from an aging mouse. The
tissue contains large amounts of interstitial gland, atretic
FIGURE 9.72  Squamous metaplasia of the endometrial follicles (arrow) and the remains of degenerating oocytes
glands in the uterus is a feature of prolonged estrogen (*). ×100.
stimulation (rat). ×100.
FIGURE 9.75  Vagina of a rat in pseudopregnancy. Here
the epithelium is atrophic and mucified, but it can also be Background pathology
hypertrophic and mucified. ×100. in aging rodents
Most of the background changes seen in the female
reproductive tissues of old mice and rats are a
mixture of degenerative and proliferative lesions
and are the result of chronic hormone imbalance
between estradiol and progestogens. The high
incidence of prolactin-secreting pituitary tumors,
particularly in the Sprague–Dawley rat, has a
major influence on the pathology of the gonadal
tissues and the mammary gland. Although the
overall changes in the various tissues are broadly
114 BACKGROUND LESIONS IN LABORATORY ANIMALS
similar between rat and mouse, there are species-
and strain-specific differences in the incidence and
severity of the changes. This is partly a reflection
of the differing species and strain-specific profiles
of hormonal imbalance. The general changes are
illustrated here, but the detailed characteristics of
the changes can be found in the literature (Alison
et al 1990, Davis et al 1999, Leininger et al 1990).
Ovaries
End-stage ‘atrophy’ of the ovary generally involves
the presence of numerous cysts and proliferation
of interstitial glands. The ovary may be partially
or entirely replaced by cysts (Figs 9.71, 9.78)
or by interstitial gland proliferation and fibrous
stroma (Figs 9.76a, 9.77). The atrophic rodent
ovary is generally smaller in size.
FIGURE 9.78  In the aged mouse, the ovary is often
replaced entirely by cysts that are filled with proteina-
ceous fluid and erythrocytes. ×100.
Ovarian cysts come in many forms. The most
common are derived from follicles that fail to
ovulate. These may be empty, fluid-filled, or
blood-filled (hematocysts) (Fig. 9.78). In the
mouse, hematocysts are often associated with vas-
cular ectasia, which may be difficult to distinguish
from hemangioma. Rupture of large hematocysts
can be a cause of death in aging mice. Other cysts
include luteal cysts lined by multiple layers of
luteinized cells, bursal cysts, which are commonly
detected at necropsy but which may rupture
during processing, rete cysts arising from the rete
ovarii (Fig. 9.68), paraovarian cysts in the meso-
varium lined by ciliated epithelium and containing
smooth muscle in their walls, epidermoid cysts
lined by squamous epithelium and filled with
keratin and epithelial inclusion cysts. Epithelial
inclusion cysts are lined by columnar epithelium
which may form papillary projections into the
lumen of the cyst. It has been suggested that in
the mouse many of the cysts, including rete,
paraovarian, and epithelial inclusion cysts, arise
from mesonephric duct remnants (Long 2002).
Brown pigment reflecting hemosiderin and/or
hematoidin from breakdown of hematocysts, vas-
cular thrombi and hemorrhage, lipofuschin or
ceroid pigment from lipid degradation, dystrophic
mineralization, fibrosis and chronic inflammatory
infiltrates are all common sequelae of the degen-
erative processes involved in age-related ovarian
atrophy. These lesions are commonly seen, par-
ticularly in mouse ovaries. Adipose tissue degen-
eration or adipose tissue metaplasia can often be
seen in the stromal tissue of atrophic ovaries. It is
characterized by areas of unremarkable adipose
tissue in the ovarian stroma. In the mouse, the
ovary is also a common site for polyarteritis
nodosa, and many small arterioles may be noted
with fibrinoid necrosis and perivascular inflamma-
tion. The ovary is also a common site for systemic
amyloidosis in the mouse (Fig. 9.79).
FIGURE 9.79  Mouse ovary partially replaced by eosi-
nophilic amyloid. Remaining tissue shows tubulostromal
hyperplasia. ×100.
In addition to the hyperplasia of interstitial cells
in the atrophic ovary, other common hyperplastic
lesions in the ovary include epithelial (or tubulos-
tromal) hyperplasia, characterized by cords of
epithelial cells forming tubular structures which
appear to originate from the layer of germinal
epithelium surrounding the ovary (Figs 9.79,
9.80). This change is often diffuse and extensive
in the mouse. Sertoli cell or Sertoliform hyperpla-
sia is also common in some strains of rat and
mouse and takes the form of tubular structures
lined by cells with the characteristics of testicular
Sertoli cells (Fig. 9.76a), with large, pale white
cytoplasm, and large, pale basal nuclei. Less fre-
quently, granulosa cell and luteal cell hyperplasia
can occur. Ovotestis or hermaphroditism are con-
genital syndromes characterized by the combina-
tion of male and female reproductive organs. The
ovotestis is made up of seminiferous tubules and
ovarian tissue (Fig. 9.76b).
FIGURE 9.80  Mouse ovary with tubulostromal hyperpla-
sia. Tubular structures appear to arise as down growths
from the surface germinal epithelium. The change is often
diffuse and particularly prominent at the periphery of the
ovary. ×100.
The most common neoplasms of the ovary
include cyst adenoma, which is a neoplastic pro-
gression of the hyperplastic epithelial inclusion
cyst, tubulostromal adenomas and adenocarcino-
mas, which appear to arise from, and may be
difficult to distinguish from, diffuse epithelial
(tubular) hyperplasia, and Sertoli cell tumors,
which progress from Sertoliform hyperplasia.
Hemangiomas and hemangiosarcomas commonly
occur in the mouse ovary and may be tissue-
specific or part of a multicentric distribution.
They can be difficult to distinguish from vascular
ectasia and hematocysts. Tumors of the granulosa,
thecal and luteal cells occur, but are less common.
Germ cell tumors (dysgerminoma, teratoma, cho-
riocarcinomas and yolk sac tumors) are considered
rare in both rat and mouse.
Uterus and cervix
Ovarian follicular cysts that develop in the aging
ovary commonly secrete an abundance of estro-
gen, which maintain the animal in persistent
estrus and are also associated with endometrial
epithelial hyperplasia, cystic endometrial glands
and inflammatory lesions in the uterus. Cystic
endometrial hyperplasia is the most common, age-
related change in the uterus of rats and mice (Fig.
9.81). The change is characterized by increased
numbers of dilated or cystic endometrial glands,
which often contain proteinaceous fluid and
inflammatory cells. The irregular, cystic glands can
grossly distort the anatomy of the endometrium
and replace much of the endometrial stroma and
the normal lumen (Fig. 9.81). Hyperplasia of the
endometrial epithelium can be extensive and
polypoid in nature, with vascular ectasia and squa-
mous metaplasia often present in areas of the
endometrium (Figs 9.82, 9.83).
FIGURE 9.81  Cystic endometrial hyperplasia in the rat.
This is the most common age-related change in the
uterus of rats and mice. ×100.
FIGURE 9.82  Mouse uterus with extensive cystic
endometrial hyperplasia forming a polypoid structure that
extends into the cervical lumen. ×40.

FIGURE 9.83  Cystic endometrial hyperplasia in a mouse
associated with prominent vascular ectasia. ×100.
Atrophy of the uterus due to age-related,
decreased gonadotropin release is also a common
change and reveals itself as persistent anestrus.
The uterus is contracted and lined by a low cuboi-
dal epithelium with few endometrial glands and a
collagenous stroma.
Vascular lesions, including vascular thrombi,
hemorrhage, ectasia (of the endometrium or myo-
metrium), commonly accompany cystic endome-
trial hyperplasia (Fig. 9.83). Acute or purulent
inflammation of the uterus (pyometra) may also
be seen in animals with cystic endometrial hyper-
plasia. High estradiol and progesterone levels con-
tribute to inflammatory lesions and infection in
the reproductive tract.
Squamous epithelial hyperplasia of the cervix
(Fig. 9.69) accompanies prolonged periods of per-
sistent estrus. Stromal hyperplasia or hypertrophy
is occasionally noted, particularly in the cervix of
rats at the junction with the vagina. This is the
region containing the portio vaginalis uteri, which
project down from the cervix into the vaginal
opening and become hypertrophic in the aging
rat uterus (Leininger et al 1990). Cysts lined by
squamous epithelium and sometimes containing
keratin may be seen in the cervix (Fig. 9.69).
In the mouse and occasionally in the rat, adeno-
myosis may be seen as a background change in the
uterus. This is characterized by the downgrowths
of endometrial glands into the myometrium
(Fig. 9.84a). Sometimes these may extend onto
the serosal surface, so it is important that they
are not mistaken for invasive endometrial
adenocarcinoma.
FIGURE 9.84a  Adenomyosis in the mouse characterized
by down growths of endometrial glands deep into the
myometrium. This should not be mistaken for adenocar-
cinoma. ×100.
Reproduction of the rat, mouse, dog, non-human primate and minipig
Decidual alteration is an unusual lesion that is
occasionally seen in the aging rat. It generally
appears as a poorly defined nodule within the
uterine wall or a polypoid mass projecting into the
uterine lumen. The ‘mass’ is made up of cells with
varying morphology, including large epithelioid
(histiocytic-like) cells with eosinophilic cytoplasm
intermixed with spindle- or fibroblastic-like cells
with wide intercellular spaces and, characteristi-
cally, small cells with intensely eosinophilic gran-
ules, which are also PAS-positive. A lesion with
similar cellular characteristics, but forming a more
discrete and organized nodule, may be observed
more rarely in the young adult rat, where it is
termed a deciduoma. For additional details on this
lesion see Leininger and coworkers (1990) and
Karbe and coworkers (1998). The characteristics
of the change resemble those of the decidual
response of the pregnant, implanted uterus, which
involves development of the metrial gland (Picut
et al 2009) (Fig. 9.84b).
FIGURE 9.84b  Deciduoma in the uterus of a female rat.
×200.
The most common neoplasm in the rat and
mouse uterus is the endometrial stromal polyp,
which is composed of loosely organized stromal
cells interspersed by blood vessels and occasional
trapped endometrial glands (Fig. 9.85). The polyp
can be edematous, congested, and sometimes
infarcted, especially when it extends down into
the cervix or vagina. Polyps may be solitary or
multifocal.
FIGURE 9.85  Benign stromal polyp in the uterine horn of
a rat. This is the most common tumor of the reproductive
tract in rats and mice. The polyp is composed of loose
fibrous tissue with few endometrial glands. ×40.
Other tumors that are relatively common in
the uterus and cervix of some strains are endome-
trial adenoma and carcinoma, stromal sarcoma,
115
leiomyoma and leiomyosarcoma, shwannoma,
hemangioma and hemangiosarcoma. The uterus
and cervix are also common sites for granular cell
tumors, which comprise sheets or nests of large
cells with abundant pale cytoplasm containing
fine, PAS-positive granules and small hyperchro-
matic nuclei (Markovits et al 2000, Veit et al
2008). The possibility that this ‘tumor’ represents
a non-neoplastic proliferation of the metrial gland,
as seen in decidual alteration, has been suggested
by Picut et al (2009). Less common tumors
include squamous cell carcinoma, fibroma and
fibrosarcoma.
Vagina
Squamous cell hyperplasia and inflammation of the
vagina may be seen with prolonged periods of per-
sistent estrus (high estrogen, low progestogen).
Conversely, mucinous atrophy is also commonly
seen in response to high progestogen : estrogen
ratio.
Solitary cysts are common in the vaginal mucosa
(Fig. 9.69). They may occasionally contain keratin.
Tumors of the vagina are relatively uncommon.
The most common tumors include granular cell
tumor, schwannoma, and occasionally squamous
cell papilloma or carcinoma.
Mammary gland
In most strains of rats and mice, reproductive
senescence is accompanied by increased secretion
of prolactin, which is generally the result if hyper-
plasia and neoplasia occur in the pituitary gland.
Raised prolactin results in ductal and alveolar
hyperplasia of the mammary gland and increased
secretory product (Fig. 9.86). An increase in
brown-pigmented cells within the acini is noted in
older rat mammary tissue. Prolonged stimulation
of the mammary gland by prolactin also results in
a high incidence of mammary gland tumors, the
most common being fibroadenoma (Fig. 9.87).
Adenomas and adenocarcinomas of the mammary
gland are also common.
FIGURE 9.86  Alveolar and ductal hyperplasia with
increased secretory product (rat) is generally associated
with age-related increased prolactin secretion and pitui-
tary gland hyperplasia or neoplasia. ×100.
116 BACKGROUND LESIONS IN LABORATORY ANIMALS
FIGURE 9.87  Fibroadenoma is the most common neo-
plasm of the mammary gland in rats. The ratio of fibrous
and glandular components can vary significantly between
tumors and within areas of the same tumor. ×100.
Clitoral gland
Cystic atrophy (Fig. 9.88) or ductal ectasia filled
with secretion (Fig. 9.89) occur in the clitoral
gland of aging rats and mice. These lesions are
often sampled as gross inguinal ‘masses’ at
necropsy.
FIGURE 9.88  Clitoral glands of a mouse with cystic
ectasia of the ducts and atrophy of the acinar glands. ×40.
FIGURE 9.89  Rat clitoral gland with cystic ducts dis-
tended with secretory product. ×40.
Dog
The morphological appearance of the female dog
reproductive tract varies significantly with matu-
rational status and the stage of the estrus cycle.
Relatively few background changes occur in the
age of dogs routinely used in toxicity studies and
reproductive senescence is not an issue in the
young dogs used in routine studies.
Immaturity and peripuberty
In the beagle dog, the first estrus occurs between
10 and 14 months of age, and therefore a high
proportion of dogs that are routinely used in toxic-
ity studies will be immature or peripubertal.
Chandra and Adler (2008) examined a cohort of
young adult dogs more than 11 months of age and
recorded the percentage of dogs in the different
stages of the estrus cycle, or that were immature.
They reported five dogs that were immature, with
ages in the range 12.5–15.5 months.
The main characteristic of the prepubertal
ovary is the lack of current or regressing corpora
lutea (Fig. 9.90). This contrasts with the ovary of
a mature female that has entered anestrus, which FIGURE 9.91  Mammary gland from a prepubertal dog.
always contains remnants of corpora lutea (Fig. The absence of glandular development identifies that the
9.92). In very young females, the ovary will only animal has not reached puberty. ×40.
contain small primary and primordial follicles. As
the dog approaches puberty, antral follicles will
begin to develop, but all will undergo atresia until Changes associated with
the onset of the first cycle. At that point, a sub- estrus cyclicity in dogs
population of antral follicles will develop into ter-
tiary and Graafian follicles and continue through The detailed morphology of the cycle-dependent
to ovulation and become luteinized to form changes in the ovary, uterus, vagina and mammary
corpora lutea. gland are beyond the scope of this chapter, and
the reader is referred to the excellent reviews by
Rehm et al (2007) and Chandra & Adler (2008).
The major changes are briefly summarized and
illustrated in this section.
Following first estrus, subsequent cycles begin
approximately every seven to eight months. The
cycle comprises pro-estrus and estrus, each of
which lasts one to two weeks followed by a long
but variable period of diestrus lasting approxi-
mately two to three months, followed by an inac-
tive anestrus phase of variable duration (three to
five months) (Rehm et al 2007). The latter part
of the estrus phase is sometimes termed met­
estrus, but it is very short and generally consid-
ered as part of estrus (Rehm et al 2007). As would
be expected from the durations of these phases,
FIGURE 9.90  Ovary from a prepubertal dog. The ovary the majority of sexually mature dogs will be in
anestrus or diestrus. Chandra & Adler (2008) con-
only contains primary and secondary follicles with large
amounts of stroma. There are no remnants of previous ducted a retrospective analysis of stage of estrus
corpora lutea. Also note follicles containing multiple in dogs, 11–22.5 months old, used in previous
oocytes (arrowed). ×100. toxicity studies and found 54% of dogs in anestrus
and 27% of dogs in diestrus.
In the case of the dog, the mammary gland as
The immature uterus is small, with a narrow
well as the ovary, uterus and vagina show marked
lumen, a dense basophilic stroma containing few,
cyclical changes depending on the stage of the
contracted and inactive glands lined by a thin epi-
cycle.
thelial layer. The myometrium is made up of
small, basophilic, smooth muscle cells. The vaginal
Anestrus
mucosa is thin and lined with a non-keratinizing,
stratified squamous epithelium. During anestrus all the tissues are in a resting
One of the easiest features that can be used to phase. The ovary is largely populated with small
identify a prepubertal animal is the glandular primary follicles and a few atretic antral follicles,
development of the mammary gland. This only and there will be remnants of regressing corpora
occurs once the animal has undergone its first lutea with highly vacuolated cells from previous
estrus but is maintained throughout the rest of cycles (Fig. 9.92).
the cycle, including anestrus. Thus the lack of
mammary gland tissue (Fig. 9.91) and absence
of corpora luteal remnants (Fig. 9.90) indicates
immaturity. A direct comparison of the morpho-
logical features of the reproductive tissues in an
immature female and a female in anestrus is pro-
vided by Chandra & Adler (2008).

FIGURE 9.92  Ovary from a dog in anestrus containing
primary and secondary follicles as well as the vacuolated
and shrunken remnants of a previous corpus luteum.
×100.
The uterus is small with an atrophic endo­
metrium containing small endometrial glands
surrounded by a compact ‘atrophic’ stroma (Fig.
9.93) and a basophilic myometrium made up of
small, spindled, smooth muscle cells.
FIGURE 9.93  Uterus from a dog in anestrus with low
cuboidal epithelium, few and small endometrial glands
and prominent dense stroma. ×100.
The vaginal epithelium is cuboidal and only one
or two cell layers thick (Fig. 9.94).
FIGURE 9.94  Vagina from a dog in anestrus with low
cuboidal epithelium only one or two cells thick. ×100.
The mammary gland is made up of relatively
few, atrophic-appearing alveoli and ducts, which
may contain secretory fluid. This is similar to the
appearance of the gland in pro-estrus (Fig. 9.95).
Reproduction of the rat, mouse, dog, non-human primate and minipig
FIGURE 9.95  Mammary gland from a dog in anestrus.
The alveoli are contracted and atrophic in appearance but
there is still occasional residual secretion in the ducts.
×100.
Pro-estrus
During pro-estrus, a number of tertiary follicles
with antral spaces develop (Fig. 9.96). They are
lined by multiple layers of basophilic granulosa
cells without folds.
FIGURE 9.96  Ovary in pro-estrus containing tertiary fol-
licles with large antral space lined by a flattened layer of
granulosa cells. ×100.
The uterus becomes slightly edematous and the
endometrial surface epithelium proliferates and
forms crypts into the underlying stroma while the
basal endometrial glands also increase in number
and start to dilate (Fig. 9.97). The myometrial
smooth muscle cells also enlarge and become
more eosinophilic.
FIGURE 9.97  Uterus in pro-estrus showing slight edema
of the endometrial stroma and crypt formation of the
surface endometrial epithelium along with proliferation,
dilation and branching of the basal endometrial glands.
×100.
117
The squamous epithelium of the vagina increases
in thickness, but is not yet keratinized (Fig. 9.98).
FIGURE 9.98  Pro-estrus vagina with thickened non-
keratinized epithelium. ×100.
The mammary gland of a dog in the pro-estrus
of its first cycle will have no glandular develop-
ment, but if the animal has been through at least
one previous cycle, it will have the same morphol-
ogy as that associated with anestrus, including
atrophic alveoli and ducts containing secretion
(Fig. 9.95). Some alveolar proliferation may be
identified at the end of pro-estrus.
Estrus
The developing follicles in the ovary become large
tertiary follicles lined by a thin layer of granulosa
cells that form projections or folds into the antral
lumen. Following ovulation, the granulosa cells
rapidly luteinize (Fig. 9.99), and although this is
technically metestrus, it lasts a relatively short
time and may be referred to as the ‘metestrus
phase of estrus’ (Rehm et al 2007).
FIGURE 9.99  Recently ovulated follicle in the ‘metestrus’
phase of estrus. The granulosa cells surround a large
central cavity that has just expelled the oocyte. These
cells undergo rapid luteinization. ×40.
The uterus in early estrus resembles that in
pro-estrus, but during late estrus the stroma
becomes more collagen-rich and contains an
increasing number of hypertrophic glands through-
out the endometrium. The myometrium contin-
ues to expand with hypertrophy of the smooth
muscle cells.
During estrus the vaginal epithelium reaches its
maximum thickness and develops a cornified layer
of keratin (Fig. 9.100) which is sloughed towards
the end of estrus. The epithelium is thrown into
ridges and folds and the underlying connective
118 BACKGROUND LESIONS IN LABORATORY ANIMALS
tissue is increased in thickness with dense collagen
fibres.
FIGURE 9.100  Vagina in estrus is lined by a keratinized
epithelium which starts to shed its superficial layers into
the lumen. ×100.
The mammary gland shows alveolar and ductal
proliferation with prominent edema surrounding
the islands of glandular tissue and stromal connec-
tive tissue proliferation (Fig. 9.101). The edema
is accompanied by increased vascularisation, hem-
orrhage and inflammatory infiltrate.
FIGURE 9.101  Estrus mammary gland shows edema
around the proliferating alveoli and stromal proliferation.
There may also be hemorrhage and an inflammatory cell
infiltrate. ×100.
Diestrus
This is a long stage of the cycle and the morphol-
ogy varies from early to late diestrus. The main
change in the ovary is the formation, maturation
and regression of the corpora lutea. At the start
of diestrus the luteal cells proliferate around
a fluid and hemorrhagic central cavity. This is
replaced by a homogenous structure made up of
plump eosinophilic cells, and the corpora lutea
take up most of the ovary (Fig. 9.102). As the
corpora lutea regress, the cells become apoptotic
and increasingly vacuolated and begin to shrink.
FIGURE 9.102  Ovary in diestrus containing large eosi-
nophilic corpora lutea. The luteinized cells are gradually
filling the cavity left following ovulation. ×100.
The uterus demonstrates a striking morphology
during diestrus. The endometrium and myo-
metrium are at their greatest thickness, but the
endometrium is divided into two different zones,
with the basophilic, basal endometrial glands
forming one layer and the surface epithelium with
its crypts lined by a pale-staining, secretory and
vacuolated tall columnar epithelium, forming the
other layer (Fig. 9.103).
FIGURE 9.103  Uterus in diestrus with two different zones
in the endometrium. The surface epithelium and crypts
are lined by pale pink, columnar secretory cells, whereas
the basal glands are more basophilic and coiled. ×100.
The vaginal epithelium decreases in thickness
during diestrus and contains mucus-secreting cells
and variable numbers of inflammatory cells (Fig.
9.104).
FIGURE 9.104  The vagina in diestrus becomes reduced
in thickness and infiltrated by leucocytes. ×100.
During diestrus the alveoli and ducts of the
mammary gland proliferate and gradually become
filled with eosinophilic secretion (Fig. 9.105).
Regressive changes, including apoptosis, macro-
phage infiltration and alveolar contraction, occur
in the gland during late diestrus.
FIGURE 9.105  Mammary gland in diestrus displaying
marked proliferation of the alveoli which are filled with
secretory fluid. ×100.
Background changes
in the young adult dog
There are relatively few changes in the reproduc-
tive tissues of young female dogs.
Pseudopregnancy or segmental cystic endo­
metrial hyperplasia (Schulman & Bolton 1997)
(Fig. 9.106) occurs occasionally, and the uterine
changes can be quite dramatic and mistaken for
real pregnancy. The ovary contains prominent and
persistent corpora lutea. The uterine surface
endometrium is replaced by cavernous spaces
lined by hyperplastic, columnar epithelium with
pale pink, secretory cytoplasm, similar to that
seen in the diestrus uterus, and the lower
endometrium contains dilated basal glands.
Necrotic tissue derived from the endometrial
surface is generally present in the uterine lumen.
FIGURE 9.106  Uterus from dog with segmental, cystic
endometrial hyperplasia. The superficial glands are
expanded into cavernous spaces filled with mucinous
secretion and lined by pale pink, secretory cells. The basal
glands form a totally distinct zone and are also dilated.
The lumen is filled by necrotic debris from the surface
endometrium. ×100.
The canine ovary often contains bi- or multinu-
cleate oocytes as a normal feature (Fig. 9.59).
Follicular or luteal cysts are occasionally present
in the ovary.

Non-human primates
In non-human primates, puberty occurs at approx-
imately two-and-a-half to four years of age which
leads to significant variation in the pubertal status
of monkeys that are nominally the same age. Sea-
sonality varies with species; rhesus are highly sea-
sonal, with sexual activity occurring primarily
between October and March, whereas cynomolo-
gus monkeys show very few seasonal changes at
different times of the year.
The normal physiology, morphology and pathol-
ogy of the female reproductive system of the
macaque have been comprehensively described in
a monograph by Buse and co-authors (2008). The
following section provides a summary of the major
morphological features associated with matura-
tion and cyclicity, as well as some common back-
ground findings.
Immaturity and puberty
The reproductive tissues of the immature non-
human primate are similar to those of the dog. If
the animal is very young, the ovary is small with
only primordial or primary follicles and stroma. If
the non-human primate is approaching puberty,
follicles may begin to develop antral cavities, but
most will show evidence of atresia. The uterus is
contracted with a basophilic, inactive-appearing
endometrium and myometrium. The vagina is also
small in diameter and lined by a thin, cuboidal
epithelium. The immature non-human primate
mammary gland has a rudimentary ductal tree
which begins to develop just prior to the first
menstrual cycle. There is elongation and branch-
ing of the major ducts accompanied by develop-
ment of terminal end buds surrounded by loose,
myxoid, periglandular stroma (Cline & Wood
2008). There is extensive lobular development in
the peripubertal non-human primate connective
tissue (Fig. 9.107). There is also significant inter-
animal variation in the rate of this growth. It is
important for the pathologist to be aware of this
when comparing individuals and pathologists
should not mistake normal pubertal glandular
development for hyperplasia or neoplasia (Cline
& Wood 2008).
FIGURE 9.107  This is the normal appearance of the
mammary gland from a cycling cynomolgus monkey.
There are abundant alveoli with secretion, but there is
significant inter-animal variation in the volume of glandu-
lar tissue. ×100.
Reproduction of the rat, mouse, dog, non-human primate and minipig 119
Changes associated
with female cyclicity
The non-human primate undergoes a menstrual
cycle that is approximately 30 days in duration,
but this is often very irregular, particularly in
the period adjacent to menarche, and intervals
of three months between the start of consecutive
cycles are not uncommon. The features of the
menstrual cycle and the changing morphology of
the reproductive tissues have been comprehen-
sively reviewed by Weinbauer and coworkers
(2008) and Van Esch and coworkers (2008). A
brief summary is provided here.
The cycle can be divided into three phases on
the basis of the ovarian changes; these are the FIGURE 9.110  Ovary in late luteal phase with a corpus
follicular phase, the luteal phase and the men- luteum undergoing regression. Also note mineralized
strual phase. The follicular phase of 12–14 days is deposits in the cortex; these probably represent mineral-
characterized by growth and maturation of the ized atretic oocytes or primordial follicles. ×40.
ovarian follicles (Fig. 9.108) and ovulation, where
the oocyte is released from a single ovulating fol- The uterus undergoes cyclic morphological
licle. The luteal phase lasts 14–16 days and changes which coincide with and are regulated by
the ovarian phases. These are the proliferative
includes establishment of the corpus luteum (Fig.
(follicular) phase (Figs 9.111, 9.112), the secre-
9.109), and then gradual regression of the corpus
tory (luteal) phase (Figs 9.113, 9.114) and the
luteum during the subsequent menstrual period
menstrual phase (Figs 9.115, 9.116). Van Esch
(Fig. 9.110). Remnants of regressing corpora lutea
and coworkers (2008) also describe a fourth,
will remain through multiple, successive cycles.
regenerative phase which follows the menstrual
phase and has different characteristics from the
proliferative phase.
FIGURE 9.108  Ovary in follicular phase with a number of
tertiary antral follicles developing. Also present are the
remnants of a regressing corpus luteum. ×100.
FIGURE 9.111  Uterus in proliferative phase. The endome-
trial glands are relatively sparse and have a straight
profile. ×100.
FIGURE 9.109  Ovary in the luteal phase with a newly
formed single corpus luteum. ×100.
FIGURE 9.112  Uterus in proliferative phase showing
mitotic activity (arrowed) in the glandular epithelium and
straight profile of the glands. ×200.
120 BACKGROUND LESIONS IN LABORATORY ANIMALS
FIGURE 9.113  Uterus in secretory phase with densely
packed, tortuous-appearing glands. Note prominent spiral
arteries surrounded by connective tissue (circled). ×100.
FIGURE 9.114  Endometrial glands from secretory phase
with saccular profile. The epithelium is tall and secretory  a wider lumen, particularly in the basal zone of
and contains sub- and supra-nuclear, glycogen-filled
vacuoles. ×100.
FIGURE 9.115  Uterus during menstruation with hemor-
rhage, necrosis and sloughing of the surface endometrium
into the lumen. ×100.
FIGURE 9.116  Menstrual uterus with hemorrhage and
sloughing of surface layers. ×100.
The proliferative phase is characterized by pro-
gressive increases in mitotic activity of the
endometrial glandular epithelium and stromal
fibroblasts (Fig. 9.112). During this phase, the
endometrial glands are relatively sparse, straight
glands, lined by pseudostratified epithelium with
a narrow lumen. The endometrial blood vessels
are not very prominent (Fig. 9.111).
Following ovulation, the uterus enters the
secretory phase, which corresponds with the
ovarian luteal phase. Mitotic activity in the
endometrium decreases, and the endometrial
glands are lined by medium to tall columnar epi-
thelium with sub- and supra-nuclear, glycogen-
filled vacuoles (Fig. 9.114). The glands are more
prominent than in the proliferative phase and
become more tortuous, with a saccular profile and
the endometrium, as the phase progresses (Fig.
9.113). During the luteal phase, the endometrium
becomes infiltrated by large numbers of lym-
phocytes, many of which contain granules. In
addition, the spiral arteries of the vasculature
become very prominent in the endometrium, with
areas containing cross sections of spiral arteries
surrounded by prominent connective tissue (Fig.
9.113).
During the menstrual phase, the spiral arteries
in the upper parts of the endometrium constrict
and cause ischemia and sloughing of the surface
layers. Tissue necrosis, hemorrhage, vascular
thrombi and leucocytic infiltration are the hall-
marks of menstruation. During this phase, blood
and necrotic tissue can be found in the cervix and
vagina (Figs 9.115, 9.116).
As the menstrual phase comes to an end, the
remaining glands in the lower endometrium that
have escaped necrosis begin to show proliferative
activity and the epithelium grows out to form a
confluent layer over the denuded stroma. This
signals the start of the regenerative phase. During
this stage, the underlying stroma of the surface
endometrium is composed of small, compact,
basophilic cells. The spiral arteries remain promi-
nent in the basal region but are not visible in the
newly forming surface endometrium while the
basal endometrial glands become quiescent in
appearance.
Although there are minor changes in the prolif-
erative activity of the ductal and alveolar tissue of
the mammary gland during the menstrual cycle,
the changes are not of sufficient magnitude to be
readily visible (Cline & Wood 2008).
Background changes in
the non-human primate
reproductive tract
Common and uncommon background changes
in the female reproductive tissues of non-human
primates have been reviewed by Cooper &
Gabrielson (2007) and Cline and coworkers
(2008). The more common changes that are seen
in young macaques used in toxicity studies are
mentioned and illustrated here. In general, back-
ground pathology is uncommon in the pubertal
and young adult non-human primates used in tox-
icity studies. The main changes that can be recog-
nized are the result of minor hormonal disturbances
that are associated with social or environmental
stress and hormonal imbalances associated with
puberty. These are recognized by inactivity of the
endometrium and inappropriate shedding of the
surface epithelium (irregular bleeding) due to
early regression of the corpus luteum.
Ovarian cysts are quite common and include
cysts that arise from structures in the paraovarian
tissue, e.g., embryonic mesonephric remnants,
dilated rete ovarii and cystic follicles or corpora
lutea (Fig. 9.117).
FIGURE 9.117  Follicular cyst compressing surrounding
tissue. These are quite common in the cynomolgus ovary.
×40.
Cortical mineral deposits are commonly seen in
the young adult non-human primate ovary and
probably represent dystrophic mineralization of
atretic follicles (Fig. 9.110). Follicles containing
multiple oocytes are also commonly seen as a back-
ground change, as is hyperplasia of the ovarian
surface epithelium.
Hemorrhage of the uterine surface epithelium
is a characteristic of the menstrual phase of the
uterine endometrium, but it often occurs in the
peripubertal female that is not yet ovulating or
cycling in a regular manner and represents irregu-
lar bleeding. Irregular endometrial hemorrhage
can be distinguished from true menstruation by
the fact that it occurs in an endometrium that is
in the proliferative phase, whereas normal men-
struation occurs in an endometrium that is in the
secretory phase of the cycle. In addition, men-
struation involves full endometrial shedding,
whereas irregular endometrial hemorrhage is char-
acterized by superficial hemorrhage and very little
endometrial shedding or necrosis. Examination of
the ovary should also provide clues on the basis of
presence or absence of regressing corpora lutea
(present in the menstruating female and absent in
the prepubertal female). Inactive (non-cycling)

uterine morphology can often be seen in non-
human primates that are postpubertal (i.e. ovaries
contain corpora lutea remnants) and is the result
of anovulatory cycles. This is often seen in females
that have relatively recently attained puberty or
may be a reflection of non-specific stress, e.g.,
social stress or stress associated with test article
administration in toxicity studies.
Adenomyosis, characterized by the presence of
occasional endometrial glands in the myometrium,
is a relatively common background finding in the
non-human primate uterus.
The myometrial vasculature of non-human
primates that have previously given birth show
marked expansion of the tunica media by eosi-
nophilic hyaline material that resembles amyloid.
The affected vessels often have obliteration of the
lumina. The change is a consequence of the tro-
phoblast invasion of the uterus during pregnancy.
Nulliparous females do not show these changes.
This lesion is also noted in rodents which have had
previous litters of pups.
Lymphoid aggregates and follicles as well as
varying numbers of infiltrating inflammatory cells
are commonly present in the mucosa of the vagina
and cervix (Figs 9.118–9.120). Mucus-filled cysts
are also commonly seen in the superficial cervical
mucosa.
FIGURE 9.118  Lymphocytic inflammatory infiltrates of
the cervix and focal squamous metaplasia of the normal
mucus secreting epithelium (arrow). ×100.
FIGURE 9.119  Prominent lymphoid follicles and lymphoid
infiltrate of the lamina propria of the vagina. ×100.
Reproduction of the rat, mouse, dog, non-human primate and minipig
FIGURE 9.120  Normal cornification of the vaginal epithe-
lium with diffuse subepithelial inflammatory cell infiltrate.
×200.
Squamous metaplasia of the endocervical glands
is a common finding in the pre- or peripubertal
female non-human primates due to the prevailing
hormonal status (oestrogen excess) and should not
be considered to be an abnormal finding.
Minipig
The female pig reaches sexual maturity at four
to five months of age and the estrus cycle is
approximately 20 days in duration.
Apart from occasional follicular cysts in the
ovary and minimal hemorrhage in the uterine
myometrium, no background changes have been
reported in the female reproductive tract of young
adult Gottingen pigs (Dincer & Svendsen 2006).
References
Alison, R.H., Morgan, K.T., Montgomery, C.A., 1990.
In: Boorman, G.A., Eustis, S.L., Elwell, M.R.,
Montgomery, C.A., MacKenzie, W.F. (Eds.),
Pathology of the Fischer rat: reference and atlas.
Academic Press, San Diego, ch 26, pp. 429–442.
Buse, E., Cline, J.M., de Rijk, E.P.C.T., et al., 2008. A
monograph on female reproductive pathophysiology
in macaques. Toxicol. Pathol. 36 (suppl), 5S–172S.
Chandra, S.A., Adler R.R., 2008. Frequency of
different estrous stages in purpose-bred beagles: a
retrospective study. Toxicol. Pathol. 36, 944–949.
Cline, J.M., Wood, C.E., 2008. The mammary gland of
macaques. Toxicol. Pathol. 36 (suppl), 130S-141S.
Cline, J.M., Wood, C.E., Vidal, J.D., et al., 2008.
Selected background findings and interpretation of
common background lesions in the female
reproductive system in macaques. Toxicol. Pathol.
36 (suppl), 142S-163S.
Cooper, T.K., Gabrielson, K.L., 2007. Spontaneous
lesions in the reproductive tract and mammary gland
of female non-human primates. Birth Def. Res. (Part
B) 80, 149–170.
Creasy D.M., 1997. Evaluation of testicular toxicity in
safety evaluation studies: the appropriate use of
spermatogenic staging. Toxicol. Pathol. 25:119–131.
Creasy, D.M., Foster P.M.D., 2002. Male reproductive
system. In: Haschek, W.M., Rousseaux, C.G.,
Wallig, M.A. (Eds.), Handbook of toxicologic
pathology, vol 2. second ed. Academic Press, San
Diego, pp. 785–846.
Davis, B.J., Dixon, D., Herbert R.A., 1999. Ovary,
oviduct, uterus, cervix and vagina. In: Maronpot,
R.R., Boorman, G.A., Gaul, B.W. (Eds.), Pathology of
the mouse. Cache River Press, Vienna, pp. 409–444.
Dincer, Z., Svendsen, O., 2006. The minipig: pathology.
In: Gad, S. (Ed.), Animal models in toxicology. CRC
Press, Boca Raton, FL, pp. 739–760.
121
Dorso, L., Chanut, F., Howroyd, P., Burnett, R., 2008.
Variability in weight and histological appearance of
the prostate of beagle dogs used in toxicology
studies. Toxicol. Pathol. 36, 917–925.
Dreef, H.C., van Esch, E., De Rijk, E.P.C.T., 2007.
Spermatogenesis in the cynomolgus monkey
(Macaca fascicularis): a practical guide for routine
morphological staging. Toxicol. Pathol. 35, 395–404.
Eldridge, J.C., Wetzel, L.T., Tyrey, L., 1999. Estrous
cycle patterns of Sprague–Dawley rats during acute
and chronic atrazine administration. Reprod. Toxicol.
13, 491–499.
Foley G.L., 2001. Overview of male reproductive
pathology. Toxicol. Pathol. 29(1):49–63.
Foley, G.L., Bassily, N., Hess, R.A., 1995. Intratubular
spermatic granulomas of the canine efferent
ductules. Toxicol. Pathol. 23, 731–734.
Fritz, T.E., Lombard, S.A., Tyler, S.A., Norris W.P.,
1976. Pathology and familial incidence of orchitis
and its relation to thyroiditis in a closed beagle
colony. Exp. Mol. Pathol. 24, 142–158.
Glaister, J.R., 1986. The young rat. In: Principles of
toxicological pathology. Taylor & Francis, London,
pp. 132–145.
Goedken, M.J., Kerlin, R.L., Morton, D., 2008.
Spontaneous and age related testicular findings in
beagle dogs. Toxicol. Pathol. 36, 465–471.
James R.W., Heywood R., 1979. Age-related variations
in the testes and prostate of beagle dogs. Toxicology
12:273–279.
Jorgensen, K.D., Kledal, T.S.A., Svendsen, O.,
Skakkeboek, N.E., 1998. The Gottingen minipig as
a model for studying effects on male fertility. Scand.
J. Lab. Anim. Sci. 25 (suppl. 1), 161–169.
Karbe, E., Hartmann, E., George, C., et al., 1998.
Similarities between the uterine decidual reaction
and the ‘mesenchymal lesion’ of the urinary bladder
in aging mice. Exp. Toxicol. Pathol. 50, 330–340.
Latendresse J.R., Warbrittion A.R., Jonassen H., et al.,
2002. Fixation of testes and eyes using a modified
Davidson’s fluid: comparison with Bouin’s fluid and
conventional Davidson’s fluid. Toxicol. Pathol. 30:
524–533.
Lanning L.L., Creasy D.M., Chapin R.E., et al., 2002.
Recommended approaches for the evaluation of
testicular and epididymal toxicity. Toxicol. Pathol.
30:507–520.
Leininger, J.R., Jokinen, M.P., 1990. Oviduct, uterus
and vagina. In: Boorman, G.A., Eustis, S.L., Elwell,
M.R., Montgomery, C.A., MacKenzie, W.F. (Eds.),
Pathology of the Fischer rat: reference and atlas.
Academic Press, San Diego, ch 27, pp. 443–460.
Li, S., Davis, B., 2007. Evaluating rodent vaginal and
uterine histology in toxicity studies. Birth Defects
Res. B. Dev. Reprod. Toxicol. 80, 246–252.
Long, G.G., 2002. Apparent mesonephric duct (rete
anlage) origin for cysts and proliferative epithelial
lesions in the mouse ovary. Toxicol. Pathol. 30,
592–598.
Markovits, J.E., Sahota P.S., 2000. Granular cell lesions
in the distal female reproductive tract of aged
Sprague–Dawley rats. Vet. Pathol. 37, 439–448.
Marty, M.S., Chapin, R.E., Parks, L.G., Thorsrud B.A.,
2003. Development and maturation of the male
reproductive system. Birth Defects Res. B. Dev.
Reprod. Toxicol. 68, 125–136.
Picut, C.A., Swanson, C.L., Oarker, R.F., Scully, K.L.,
Parker G.A., 2009. The metrial gland in the rat and
its similarities to granular cell tumors. Toxicol.
Pathol. 37, 474–480.
Rehm, S., 2000. Spontaneous testicular lesions in
purpose bred beagle dogs. Toxicol. Pathol. 28,
782–787.
Rehm, S., Stanislaus, D.J., Williams, A.M., 2007.
Estrous cycle-dependent histology and review of sex
steroid receptor expression in dog reproductive
tissues and mammary gland and associated hormone
122 BACKGROUND LESIONS IN LABORATORY ANIMALS
levels. Birth Defects Res. B. Dev. Reprod. Toxicol.
80, 233–245.
Russell, L.D., Ettlin, R.A., Sinha Hikim, A.P.,
CleggClegg, E.D., 1990. Histological and
histopathological evaluation of the testis. Cache
River Press, Clearwater, FL.
Schulman M.L., Bolton L.A., 1997. Uterine horn
aplasia with complications in two mixed-breed
bitches. J. S. Afr. Vet. Assoc. 68:150–153.
Suwa T., Nyska A., Peckham J.C., et al., 2001. A
retrospective analysis of background lesions and
tissue accountability for male accessory sex organs in
Fischer-344 rats. Toxicol. Pathol. 29:467–478.
Van Esch, E., Buse, E., Weinbauer, G.F., Cline J.M.,
2008. The macaque endometrium with special
reference to the cynomolgus monkey (Macaca
fascicularis). Toxicol. Pathol. 36 (suppl), 67S-100S.
Veit, A.C., Painter, J.T., Miller, R.A., Hardisty, J.F.,
Dixon D., 2008. Characterisation of granular cell
tumors in B6C3F1 mice: a histomorphologic,
immunohistochemical and ultrastructural study. Vet.
Pathol. 45, 654–662.
Weinbauer, G.F., Niehoff, M., Niehaus, M., et al.,
2008. Physiology and endocrinology of the ovarian
cycle in macaques. Toxicol. Pathol. 36 (suppl),
7S-23S.
Westwood, F.R., 2008. The female rat reproductive
cycle: a practical histological guide to staging.
Toxicol. Pathol. 36, 375–384.
Yuan Y., Foley G.L., 2002. In: Handbook of Toxicologic
Pathology, Female reproductive system, eds Haschek
WM, Rousseaux CG, Wallig MS. Academic Press,
London, 2, 2, pp. 847–884.
 SUBJECT INDEX

Page numbers followed by “f ” indicate figures. Alveolar macrophages Auricular chondritis, rats, 35, 36f
minipig, 82, 82f Autolysis artifact
mouse, 50, 50f fixative, 94, 94f
A rats, 21, 21f
Alveolar osseous metaplasia, mouse, 50, 50f
kidney, rats, 29, 30f
lens, artifact, 98, 99f
Abscesses Alveolitis, dogs, 39, 39f
mandibular lymph node, minipig, 82, 82f Amyloidosis
prepuce, hamster, 76, 77f
Accessory sex organs, rat male reproductive system,
lamina propria, mouse, 66, 66f
liver, mouse, 66, 67f
B
104 ovary, rodents, 114, 114f Background changes, non-human primate female
Accessory spleen see Pancreas severe diffuse, hamster, 75, 75f reproductive system, 120–121, 120f
Acid haematin, fixative artifact, 95, 95f testes, rodents, 103, 104f Balantidium coli infection, non-human primates, 6,
Acinar atrophy Angiectasis 6f
lacrimal gland, rats, 35, 35f adrenal glands, rats, 31, 31f Barbiturate intracardiac injection, cardiomyocyte
pancreas see Pancreas liver, rats, 26, 26f artifact, 93, 93f
salivary gland, rats, 24, 24f pituitary gland, rats, 30, 30f Basophilic hypertrophic focus, rat parotid salivary
Acinar hypertrophy, rat lacrimal gland, 35, 35f Anestrus, dogs, 116–117, 117f gland, 24, 24f
Acinar vacuolation, rat pancreas, 25, 25f Ante mortem artifacts, 93 Basophilic tinctorial change, rat pancreas, 25,
Adenohypophysis, rat pituitary gland, 30, 30f Anthracosis, non-human primates, 4 25f
Adenomatous hyperplasia, mouse stomach, 52, 52f Aorta Basophilic tubules see Kidney
Adenomyosis, rodents, 115, 115f cartilage, rats, 17, 17f Basophilic vacuolation, rodent epididymis, 104,
Adipocytes inflammation, mouse, 46, 46f 104f
dog atrial infiltration, 37, 38f intimal degeneration, cynomolgus macaque, 3, Beagle dogs, 37–44
dog salivary glands, 39, 40f 3f brain, 43–44
Adipose tissue mineralization cardiovascular system, 37
degeneration, minipig, 84, 84f dogs, 37, 37f endocrine glands, 41–42
right ventricle, rabbit, 87, 87f rabbit, 87, 87f female reproductive system, 116–118
Adrenal gland(s) Aortic valve thrombosis, hamster, 73, 73f anestrus, 116–117, 117f
angiectasis, rats, 31, 31f Apoptosis, rat thymus, 20, 20f diestrus, 118, 118f
ectopic see Ectopic adrenal gland Arterial mineralization, rats, 20, 20f immaturity, 116, 116f–117f
extramedullary hemopoiesis Arterial plaque, mouse lungs, 50, 50f estrus, 117–118, 117f–118f
marmoset, 11, 11f Atrial thrombosis estrus cyclicity, 116–118
rats, 31, 31f mouse, 45–46, 46f peripuberty, 116
lipogenic pigmentation, mouse, 60, 60f rats, 18 pro-estrus, 117, 117f
medullary hyperplasia, rats, 31, 31f Arteritis young adults, 118, 118f
subcapsular cell hyperplasia, mouse, 60, 60f dogs, 37, 37f gastrointestinal system, 39–40
vacuolation, rabbit, 90 lung, rabbit, 87, 88f hepatobiliary system, 40
X zone, mouse, 59, 59f minipig, 81, 81f hemolymphoreticular system, 38
zona glomerulosa focal hypertrophy, rats, 31, mouse, 46, 46f male reproductive system, 105–108, 105f–106f
32f pancreatic vessel, mouse, 56, 56f immaturity, 105–107
zona glomerulosa vacuolation, dogs, 42, 42f Artifacts, 93–99 lesions in young animals, 107–108
zona reticularis vacuolation, dogs, 42, 42f ante mortem, 93 maturity, 105–107
Adrenal gland cortex CNS, 98–99 peripuberty, 105–107
accessory tissue, mouse, 59, 60f cover slipping, 98, 98f prostate gland, 108
cystic degeneration, rats, 30, 30f cutting, 96–97 testes, 107–108
cysts, rats, 30, 30f environmental factors, 93 musculoskeletal system, 43
ectopic tissue, mouse, 59, 60f fixation, 94–95 nervous system, 43–44
extracapsular tissue, rats, 31, 31f mounting, 97 respiratory system, 38–39
focal hypertrophy, mouse, 60, 60f postmortem/necropsy, 93–94 skin and appendages, 43
hyperplasia processing, 95–96 urinary system, 41
hamster, 75, 75f stains, 97–98, 98f Bedding-associated dermatitis, hamster, 76
rats, 31f Atherosclerosis, non-human primates, Benign stromal polyp, rodent uterus, 115, 115f
lymphoid infiltrate, minipig, 81, 81f 2, 3f Bile ducts
mineralization, non-human primates, 10–11, 11f Atria ectopic, non-human primates, 11, 11f
ossification, rats, 31, 31f adipocyte infiltration, dogs, 37, 38f hyperplasia
pigments, rats, 31, 31f mucinous degeneration, non-human primates, 3, hamster, 73, 75f
vacuolation, rats, 30, 30f–31f 3f rats, 26, 26f
Adreno-hepatic fusion (AHF), non-human primates, thrombosis, hamster, 73, 73f Biliary cysts, hamster, 74, 74f
11, 11f villous mesothelial projections, dogs, 37, 38f Bladder, urinary see Urinary bladder
Age-related lesions Atrophy Blind-ending efferent ducts, dogs, 108, 108f
rat female reproductive system, 112–114, 114f acinar see Acinar atrophy Blood collection trauma, rat tongue, 24, 25f
rat male reproductive system, 103–104 lymph nodes, rats, 18 Bone marrow
Agonal alveolar edema, rabbit, 87, 88f myofibers, rats, 33, 33f serous atrophy, minipig, 84, 84f
Alveolar hemorrhage, rat erythrophagocytosis, ovary, rodents, 112–113, 113f fixative artifact, 96, 96f
20–21, 21f retina germinal centres, non-human primates, 3–4,
Alveolar histiocytes, rats, 21, 21f mouse, 65, 65f 4f
Alveolar histiocytosis, hamster, 73, 73f rats, 35, 35f granulopoiesis, mouse, 48–49, 49f
Alveolar hyperplasia, rodent mammary gland, 115, seminiferous tubules, hamster, 76, 77f thrombus, artifact, 93, 93f
115f thymus see Thymus Bones see Musculoskeletal system
124 Subject Index

Bowman’s capsule
cuboidal metaplasia, non-human primates, 9, 9f
Choroid plexus, rat extramedullary hemopoiesis,
34, 34f
metaplasia, rats, 28 Chronic gastritis, non-human primates, 5, 5f
sexual dimorphism, mouse, 57, 57f Chronic suppurative inflammation, hamster ear, 77,
Brain 77f
beagle dogs, 43–44 Clara cell, rat eosinophilic inclusion, 21, 21f
guinea pigs, 78 Clear-cell focus, hamster liver, 73, 75f
lipomatous hamartoma, mouse, 64, 64f Clitoral glands
mineralization rat female reproductive system, 116, 116f
cynomolgus macaque, 12–13, 13f rodents, 116f
hamster, 77, 77f Coagulative necrosis, mouse liver, 55, 55f
minipig, 84, 84f Coccidiosis, rabbit gall bladder, 89, 89f
mouse, 64, 64f Colitis
rats, 34, 34f marmoset, 6–7, 7f
minipigs, 84 non-human primates, 6, 6f
mouse, 64 Shigella infection, non-human primates, 6, 6f
New Zealand White rabbit, 91 Collagenofibrotic glomerulonephropathy (CFGN),
non-human primates, 12–13 9–10
rats, 33–34 Colloidal plugs, rat urinary bladder, 29, 29f
Branchial cysts, dog thymus, 38, 38f Colonic glands
Bronchioles diverticulum, mouse, 52
mucus mineralization, rats, 22, 22f glandular herniation, marmoset, 6, 7f
neuroendocrine cell hyperplasia, rats, 20–21, herniation, minipig, 83, 83f
21f Congenital/developmental lesions, rat male
Brown pigment, non-human primate lungs, 4, 4f reproductive system, 102
Brunner’s gland, marmoset duodenum, 7, 7f Congenital glaucoma, rabbit, 91
Bulbourethral gland, rodent cystic dilatation, 105f Cornea
Buphthalmia, rabbit, 91 degeneration, mouse, 65, 65f
mineralization, mouse, 65, 65f
Cornificiation, non-human primate vagina, 121, 121f
C Corpora lutea, hamster ovary, 76, 76f
Cortex
Campylobacter infection, non-human primates, 6 adrenal glands see Adrenal gland cortex
Capsular cyst, rat spleen, 19, 19f kidney see Kidney
Caput epididymis, dog intranuclear eosinophilic Cover slipping, artifacts, 98, 98f
inclusions, 108, 108f Cracks/fractures
Cardiomyocytes, barbiturate intracardiac injection cutting artifact, 97, 97f
artifacts, 93, 93f fixative artifact, 94, 95f
Cardiomyopathy freezing artifact, 94, 94f
mouse, 45, 45f Cryptorchidism, rodents, 102
rats, 17, 17f
Cardiovascular system
Cuboidal metaplasia, non-human primate Bowman’s
capsule, 9, 9f
beagle dogs, 37 Cupping, minipig optic disk, 84, 84f
guinea pigs, 78 Cutting artifact, 96–97, 97f
hamsters, 73 Cyclicity-related changes, non-human primate
minipigs, 81 female reproductive system, 119–120,
mouse see Mouse 119f–120f
New Zealand White rabbit, 87 Cyst(s)
non-human primates, 1–3 biliary, hamster, 74, 74f
rats, 17–18 brain
Cartilage mouse, 64, 64f
aorta, rats, 17, 17f rats, 34, 34f
sternum degeneration, hamster, 77, 77f branchial, dog thymus, 38, 38f
tongue, minipig, 82, 82f capsular, rat spleen, 19f
trachea mineralization, rats, 21, 21f follicular, ovary see Ovaries
C-cells hard palate, hamster, 74, 74f
hyperplasia heart, non-human primates, 2, 2f
marmoset thyroid gland, 11, 11f heart valve, rats, 17, 17f
rat thyroid, 32, 32f non-glandular stomach, rats, 23, 23f
infiltrates, dog thyroid, 42, 42f parathyroid gland, dogs, 41–42, 42f
Cell ploidy, rat liver, 27, 28f pars distalis, dogs, 41–42, 42f
Cell rest of Hortega, dogs, 43, 44f pars nervosa, dogs, 41–42, 42f
Central nervous system (CNS), artifacts, 98–99 pituitary gland, rats, 30, 30f
Cerebellum, rat hypoplasia, 34, 34f thymus, mouse, 48, 48f
Cerebrum thyroid, rats, 32
lymphoid infiltrate, minipig, 81, 81f vagina, rodents, 112f
nucleus circularis, rats, 34, 34f Cystic atrophy, rodent preputial gland, 104, 105f
Cervix Cystic degeneration, rat liver, 26, 26f
keratin cysts, rodents, 112f Cystic dilatation
lymphocytic infiltration, non-human primates, bulbourethral gland, rodents, 105f
121, 121f gastric glands, rats, 23, 23f
rat female reproductive system, 114–115 mucosal glands, mouse, 52, 52f
Cholangiofibrosis, rats, 26–27, 27f preputial gland, rodents, 104, 105f
Cholecystitis Cystic follicles, rodents, 112–113, 113f
minipig, 83, 83f Cystic hyperplasia
necrotic, rabbit, 89, 89f endometrium, rodents, 114–115, 114f–115f
Cholelithiasis, hamster, 75, 75f uterus, rodents, 114, 114f
Cholesterol clefts, rat lung, 21, 21f endometrium, dog, 118
Chondromucinous degeneration, sternum see Cystic replacement, rodent ovary, 114, 114f
Sternum Cystitis, dogs, 41, 41f
D
Decalcification, fixative artifact, 95, 95f
Decidual reaction, hamster uterus, 76,
76f
Deciduoma, rodent uterus, 115, 115f
Degeneration
cornea, mouse, 65, 65f
Degenerative joint disease, mouse, 63, 63f
Dehydration artifacts, 96, 96f
Demodex mite infestation, dogs, 43, 43f
Demyelination, rat sciatic nerve, 33, 33f
Dendritic reticular cells, mouse hyperplasia, 47,
48f
Dental dysplasia, rats, 23, 23f
Dermatitis
bedding-associated, hamster, 76
ear margin, dogs, 43, 43f
moist, rabbit, 90
rats, 32f, 33
Developmental cysts, mouse pars distalis, 59,
59f
Diffuse glomerulonephritis, marmoset, 10, 10f
Dilated submucosal glands, rat larynx, 22, 22f
Diestrus, beagle dogs, 118, 118f
Diverticulum
colonic glands, mouse, 52
duodenum, rats, 24, 24f
Ductal hyperplasia, rodent mammary gland, 115,
115f
Ductal mucus epithelial hyperplasia, hamster
pancreas, 75, 76f
Duodenum
Brunner’s gland, marmoset, 7, 7f
diverticulum, rats, 24, 24f
ectopic pancreatic tissue, mouse, 52, 52f
hyperplasia, mouse, 53, 53f
Dysplasia, hamster sternum, 77, 77f
E
Ear
hamsters, 77
minipigs, 84
New Zealand White rabbit, 91
rats, 35–36
Ear margin dermatitis, dogs, 43, 43f
Ectopic adrenal gland
epididymis, non-human primates, 8, 9f
kidney, non-human primates, 8, 9f
liver, non-human primates, 9, 9f
Ectopic granule cells, rat brain, 34, 34f
Ectopic intestinal tissue, rat stomach, 24, 24f
Ectopic liver, non-human primates, 11, 11f
Ectopic pancreatic tissue
duodenum, mouse, 52, 52f
small intestine, rats, 25, 25f
Ectopic parathyroid glands, rats, 20
Ectopic renal tissue, mouse liver, 53, 53f
Ectopic sebaceous glands, rats, 23
Ectopic thymic tissue
parathyroid gland see Parathyroid glands
rats, 19
Ectopic tissue, dog thyroid, 41–42
Efferent ducts, blind-ending, 108, 108f
Endocardium, mesenchymal proliferation, 17,
17f
Endocrine system
beagle dogs, 41–42
hamsters, 75–76
minipigs, 84
mouse see Mouse
New Zealand White rabbit, 90
non-human primates, 10–12
rats, 30–32
see also specific endocrine glands
Endometrial glands
estrus cycle, non-human primate, 120f
squamous metaplasia, rodents, 112–113, 113f
 Subject Index 125

Endometrium, cystic hyperplasia, 114–115, Fibrous hypoplasia, non-human primate testes, Gastrointestinal system
114f–115f 110f beagle dogs, 39–40
Environmental factors, artifacts, 93 Fibrous osteodystrophy (FOD), marmoset, 12, guinea pigs, 78
Eosinophilic crystals 12f hamsters, 74
rat lung, 22, 22f Fighting, rabbit skin damage, 90 minipigs, 82–83
rat thymus, 20, 20f Filaroides infection, dogs, 39, 39f mouse see Mouse
Eosinophilic granules, mouse, 51, 51f Fixation artifacts, 94–95 New Zealand White rabbit, 88
Eosinophilic inclusions Flank glands, ulceration, 76, 76f non-human primates, 5–7
Clara cell, rats, 21, 21f Focal alveolar epithelial hyperplasia, mouse, 50, rats, 23–25
gall bladder, mouse, 56, 56f 50f Germinal centres
intracytoplasmic, liver, 26, 26f Focal angiectasis, mouse liver, 46, 46f bone marrow, non-human primates, 3–4, 4f
myocardium, non-human primates, 2, 2f Focal atrophy, mouse submandibular salivary glands, lymphofollicular structure, mouse, 48, 48f
nasal turbinate, rats, 20, 20f 51, 51f Glandular herniation, marmoset colonic glands, 6, 7f
tracheal glands, mouse, 49–50, 49f Focal biliary cysts, rats, 26–27, 27f Glaucoma, congenital, 91
urothelium, non-human primates, 10, 10f Focal cortical hypertrophy, mouse adrenal glands, Glial scars, non-human primate brain, 13, 13f
Eosinophilic perivascular infiltration, rat lung, 21, 60, 60f Gliosis
21f Focal cystic dilatation, dog prostate gland, 108, non-human primates, 13, 13f
Eosinophilic Purkinje cells, artifact, 98, 99f 108f–109f optic nerve artifacts, 93, 93f
Eosinophils Focal degeneration, mouse spinal cord, 64, 65f rats, 34
jejunum lamina propria, minipig, 83, 83f Focal epithelial hyperplasia, mouse gall bladder, 56, Glomerular lipid accumulation, dog kidney, 41, 41f
subcapsular sinus, minipig, 82, 82f 56f Glomerulonephritis, mouse, 58
Epicardium, mouse mineralization, 46, 46f Focal erosion Glomerulonephropathy, hamster, 75, 75f
Epidermis forestomach, mouse, 51–52, 52f Glomerulosclerosis
cysts, rats, 33, 33f stomach, hamster, 74, 74f cynomolgus macaque, 9–10, 10f
focal ulceration, mouse, 62, 62f Focal hyperplasia, mouse mammary gland, 62, 62f mouse, 58, 58f
inclusion cyst, mouse, 62, 62f Focal hypertrophy, rat parathyroid gland, 32, 32f rats, 28, 28f
Epidermoid cyst, mouse spinal cord, 64, 64f Focal myocarditis, cynomolgus macaque, 1, 1f Glycogen accumulation, marmoset liver, 8, 8f
Epididymis Focal necrosis Granular cell hyperplasia
basophilic vacuolation, rodents, 104, 104f liver see Liver kidney, hamster, 76, 76f
ectopic adrenal gland, non-human primates, 8, mouse Harderian glands, 66, 66f large intestine, hamster, 76, 76f
9f Focal squamous metaplasia, rabbit prostate gland, uterus, hamster, 76, 76f
epithelial cystic vacuolation, rodents, 103f 90, 90f Granule cells, ectopic, 34, 34f
hyperplasia, hamster, 76, 77f Focal tubular basophilia, dog kidney, 41, 41f Granulomas
lymphoid aggregates, rodents, 103, 104f Focal tubular degeneration, non-human primate lungs, dogs, 39, 39f
pseudoglandular cysts, dogs, 108, 108f testes, 110f non-human primates, 7, 7f
rats, 103–104 Focal tubular dilatation, non-human primate testes, Granulopoiesis, mouse bone marrow, 48–49, 49f
sperm granulomas, rodents, 103, 103f 109f Growth plate dysplasia, marmoset, 12f, 13
Epiphyses, mouse hyperostosis, 63, 63f Focal ulceration Guinea pigs, 77–78
Epithelial atrophy, hamster prostate gland, 76, 77f epidermis, mouse, 62, 62f brain, 78
Epithelial cell hyperplasia non-glandular stomach, rats, 23, 23f cardiovascular system, 78
gall bladder, dogs, 40, 41f Focal villous intimal proliferation, dog myocardial gastrointestinal system, 78
prostate gland, rodents, 104f artery, 37, 37f hepatobiliary system, 78
Epithelial cell vacuolation, dog gall bladder, 40, 40f Follicle dilatation, mouse thyroid, 61, 61f hemolymphoreticular system, 78
Epithelial cysts Follicular cysts musculoskeletal system, 78
heart, cynomolgus macaque, 2, 2f ovary see Ovaries nervous system, 78
vacuolation, epididymis, rodents, 103f skin, rats, 33, 33f reproductive system, 78
Erythrophagocytosis, rat alveolar hemorrhage, Follicular epithelial hypertrophy, rat thyroid, 32, respiratory system, 78
20–21, 21f 32f urinary system, 78
Extracapsular cortical tissue, rat adrenal glands, 31, Folliculitis, dogs, 43, 43f Gut-associated lymphoid tissue (GALT)
31f Food accumulation, rat molars, 23, 23f macrophage aggregates, rabbit, 87, 87f
Extramedullary hemopoiesis Fordyce’s granules, rats, 23 non-human primates, 6
adrenal glands Foreign body, mouse larynx, 49, 49f Warthin–Finkeldey bodies, non-human primates,
marmoset, 11, 11f Foreign body pneumonia, non-human primates, 4, 3, 3f
rats, 31, 31f 4f
choroid plexus, rats, 34, 34f Forestomach, focal erosion, 51–52, 52f
liver, mouse, 53–54, 54f Formalin, perivascular expansion, 94, 94f H
spleen Fungal contamination, 97, 97f
mouse, 48, 48f Hematocysts, dog heart valve, 37, 37f
rats, 19, 19f Hematoxylin and eosin (H&E) artifacts, 97
Eye G Hemopoiesis
extramedullary see Extramedullary hemopoiesis
hamsters, 77
minipigs, 84 Gall bladder liver see Liver
New Zealand White rabbit, 91 coccidiosis, rabbit, 89, 89f Hemorrhage
rats, 35–36 duplication, rabbit, 89, 89f optic nerve, artifacts, 93, 93f
eosinophilic inclusions, mouse, 56, 56f thymus, rats, 20, 20f
epithelial cell hyperplasia, dogs, 40, 41f Hemosiderin
F epithelial cell vacuolation, dogs, 40, 40f
focal epithelial hyperplasia, mouse, 56, 56f
liver, marmoset, 8, 8f
spleen, rats, 19, 19f
Fatty vacuolation, mouse liver, 54, 54f Gastric erosion see Stomach Hair contamination, 94, 94f
Female reproductive system, 110–111 Gastric glands Hair embolus, rats, 22, 22f
beagle dogs see Beagle dogs cystic dilatation, rats, 23, 23f Hamsters, 73–79
non-human primates see Non-human primates Helicobacter, dogs, 40, 40f cardiovascular system, 73
rats see Rats herniation, hamster, 74, 74f ear, 77
Fibroadenoma, rodent mammary gland, 116f Gastric heteropia, dogs, 40, 40f endocrine glands, 75–76
Fibrosing alveolitis, dogs, 39, 39f Gastric infarction, non-human primates, 5–6 eye, 77
Fibrosis Gastric mucosa gastrointestinal system, 74
herniated liver lobe, rats, 27, 27f hypertrophy, mouse, 52, 52f hepatobiliary system, 74–75
liver, marmoset, 8, 8f mineralization hemolymphoreticular system, 73
middle ear, hamster, 77, 77f dogs, 40, 40f musculoskeletal system, 77
myocardium, marmoset, 2, 2f hamster, 74, 74f reproductive system, 76
126 Subject Index

Hamsters (continued)
respiratory system, 73
I glomerular lipid accumulation, dogs, 41, 41f
granular cell hyperplasia, hamster, 76, 76f
skin and appendages, 76 Immaturity histiocytic sarcoma, rats, 28, 28f
urinary system, 75 dog female reproductive system, 116, 116f–117f hyaline casts, hamster, 75, 75f
Harderian glands dog male reproductive system, 105–107 hyaline droplets, rats, 28, 28f
focal necrosis, mouse, 66, 66f minipig male reproductive system, 110 inflammation, mouse, 57–58
squamous hyperplasia, rats, 35, 35f non-human primate female reproductive system, interstitial lymphocytes, minipig, 83, 83f
Harderianization, lacrimal gland 119 juvenile glomerulus, dogs, 41, 41f
mouse, 66, 66f rat female reproductive system, 111 lymphocytic infiltration, mouse, 57, 57f
rats, 35, 35f rat spermatogenesis, 101–102 mineralization
Hard palate Inclusion cyst, mouse epidermis, 62, 62f dogs, 41, 41f
cyst formation, hamster, 74, 74f Inflammation marmoset, 4–5, 5f
mineralization, hamster, 74, 74f aorta, mouse, 46, 46f minipig, 83, 83f
Hassall’s corpuscles, dog thymus, 38, 38f chronic suppurative, hamster ear, 77, 77f mouse, 57
Heart artery, mural hypertrophy, 17–18, 18f kidney, mouse, 57–58 rabbit, 89, 89f
Heart valve lacrimal gland, rats, 35, 35f rats, 28, 29f
cyst, rats, 17, 17f liver, non-human primates, 8, 9f multinucleate cells, cynomolgus macaque, 10, 10f
hematocysts, dogs, 37, 37f multifocal subpleural, rats, 22, 22f tubular dilatation, hamster, 75, 75f
Helicobacter infections muscle tubular epithelial cells, nuclear brick inclusions,
gastric glands, dogs, 40, 40f minipig, 84, 84f dogs, 40, 40f
non-human primates, 5 rats, 33, 33f tubular hypertrophy, rats, 29, 29f
Hemosiderotic plaque, dog spleen, 38, 38f myocardium, cynomolgus macaque, 1, 1f Kidney cortex
Hepatobiliary system preputial gland, rodents, 104, 104f cysts, mouse, 57, 57f
beagle dogs, 40 prostate gland, rodents, 104, 104f interstitial lymphoplasmacytic infiltration,
guinea pigs, 78 Information sources, mouse, 45 non-human primates, 9, 9f
hamsters, 74–75 Inherited buphthalmia, rabbit, 91 lipomatous transformation, rats, 29, 29f
minipigs, 83 Interstitial glands, rabbit ovary, 90, 90f lymphoid infiltrate, minipig, 81, 81f
New Zealand White rabbit, 88–89 Interstitial lymphocytes, minipig kidney, 83, 83f pigment accumulation, mouse, 57, 57f
non-human primates, 8–9 Interstitial lymphoplasmacytic infiltration, tubular epithelium, lipofuscin pigments, 28, 28f
rats, 25–27 non-human primate kidney cortex, 9, 9f tubular epithelium vacuolation, dogs, 41, 41f
Hepatocytes Interstitial nephritis, non-human primates, 9 Kidney pelvis, rat transitional epithelium
intracytoplasmic eosinophilic inclusions, mouse, Interstitial pneumonia, non-human primates, 4, 4f hyperplasia, 29, 29f
54, 54f Intestinal tissue, rat stomach, 24, 24f Knee joints, synovial hyperplasia, 63, 63f
intracytoplasmic erythrocytes, mouse, 54, 54f Intimal degeneration, cynomolgus macaque aorta, 3,
intranuclear eosinophilic inclusions, mouse, 54, 3f
54f
intranuclear inclusions, hamster, 74, 74f
Intracytoplasmic colloid, rat thyroid, 32, 32f
Intracytoplasmic eosinophilic inclusions, mouse
L
lipofuscin pigment, rats, 27, 27f hepatocytes, 54, 54f Lacrimal glands
multinucleate Intracytoplasmic erythrocytes, mouse hepatocytes, acinar atrophy, rats, 35, 35f
mouse, 54f 54, 54f acinar hypertrophy, rats, 35, 35f
rats, 27, 27f Intracytoplasmic hyaline inclusions, mouse olfactory Harderianization
necrosis epithelium, 49, 49f mouse, 66, 66f
minipig, 83, 83f Intrahepatic bile duct, hypertrophy, 55, 56f rats, 35, 35f
rats, 26–27, 27f Intranuclear eosinophilic inclusions inflammation, rats, 35, 35f
non-glandular stomach, rats, 26, 26f caput epididymis, dogs, 108, 108f mesenchymal proliferation, mouse, 66, 66f
nuclear brick inclusions, dogs, 40, 40f hepatocytes, mouse, 54, 54f Lamina propria
rarefaction, barbiturate injection artifact, 93, 93f Intranuclear inclusions, hamster hepatocytes, 74, amyloidosis, mouse, 66, 66f
vacuolation 74f macrophages, non-human primates, 6, 6f
mouse, 54, 55f Iris, mineralization, 65, 65f Large intestine, granular cell hyperplasia, 76, 76f
rabbit, 88, 89f Ischemic adipose tissue, mineralization, 23, 24f Laryngeal cartilage, ossification, 22, 22f
Herniated liver lobe, rat fibrosis, 27, 27f Islet cell hyperplasia see Pancreas Larynx
Herniation dilated submucosal glands, rats, 22, 22f
colonic glands, minipig, 83, 83f foreign body, mouse, 49, 49f
gastric glands, hamster, 74, 74f
Histiocytic sarcoma, rat kidney, 28, 28f
J Left ventricle, dilatation, 73, 73f
Lens
Histiocytosis, alveolar, 73, 73f Jejunum lamina propria, minipig eosinophils, 83, autolysis
Hyaline casts, hamster kidney, 75, 75f 83f artifacts, 98, 99f
Hyaline deposits Joint mice, rats, 33, 33f rats, 35, 35f
kidney, rats, 28, 28f Joints see Musculoskeletal system degeneration see Lenticular degeneration
lymphoid follicles, dogs, 38, 38f Juvenile glomerulus, dog kidney, 41, 41f Lenticular degeneration
nasal septum, mouse, 49, 49f hamster, 77, 77f
Hydronephrosis, rats, 28, 28f mouse, 65, 65f
Hyperostosis, epiphyses, 63, 63f
Hyperplasia
K rats, 35, 35f
Leydig cell hyperplasia, rodents, 103, 104f
adrenal cortex, hamster, 75, 75f Karyomegaly Lipid vacuolation, liver see Liver
bile duct see Bile ducts cynomolgus macaque, 1, 2f Lipofuscin pigments
dendritic reticular cells, mouse, 47, 48f mouse liver, 54, 54f hepatocytes, rats, 27, 27f
ductal mammary gland, rodents, 115f Keratin cysts kidney cortical tubular epithelium, rats, 28, 28f
duodenum, mouse, 53, 53f cervix, rodents, 112f Lipogenic pigmentation, mouse adrenal glands, 60,
lymphoid, rabbit bladder, 90, 90f muscle, rats, 33, 33f 60f
neuroendocrine cell, hamster, 73, 74f spinal cord, rats, 34, 34f Lipomatosis, mouse, 47, 47f
pelvic urothelium, rats, 29, 29f Kidney Lipomatous hamartomas, mouse brain, 64, 64f
rete epithelium, rodents, 103, 104f autolysis artefact, rats, 29, 30f Lipomatous transformation, rat renal cortex, 29,
Hypertrophy basophilic tubules 29f
gastric mucosa, mouse, 52, 52f minipig, 83, 83f Liver
intrahepatic bile duct, mouse, 55, 56f rabbit, 89–90, 90f amyloidosis, mouse, 66, 67f
Hypoplasia corticomedullary mineralization, rats, 28, 29f angiectasis, rats, 26, 26f
cerebellum, rats, 34, 34f ectopic adrenal gland, non-human primates, 8, 9f cell ploidy, rats, 27, 28f
testes, rodents, 102 ectopic tissue, mouse liver, 53, 53f clear-cell focus, hamster, 73, 75f
Hypospermatogenesis, dogs, 107, 107f focal tubular basophilia, dogs, 41, 41f coagulative necrosis, mouse, 55, 55f
 Subject Index 127

cystic degeneration, rats, 26, 26f

ectopic, non-human primates, 11, 11f
M ear, 84
endocrine system, 84
ectopic adrenal gland, non-human primates, 9, 9f Macrophages eye, 84
ectopic renal tissue, mouse, 53, 53f aggregates, gut-associated lymphoid tissue gastrointestinal system, 82–83
extramedullary hemopoiesis, mouse, 53–54, 54f (GALT), 87, 87f hepatobiliary system, 83
fatty vacuolation, mouse, 54, 54f alveolar see Alveolar macrophages hemolymphoreticular system, 81–82
fibrosis, marmoset, 8, 8f lamina propria, non-human primates, 6, 6f male reproductive system, 110
focal angiectasis, mouse, 46, 46f mesenteric lymph node, rats, 18, 18f immaturity, 110
focal necrosis Male reproductive system, 101–110 maturity, 110
mouse, 55, 55f dogs see Beagle dogs peripuberty, 110
non-human primates, 8, 9f minipigs see Minipigs musculoskeletal system, 84
glycogen accumulation, marmoset, 8, 8f non-human primates see Non-human primates nervous system, 84
hemopoiesis rats see Rats reproductive system, 84
marmoset, 8, 8f Mammary glands female reproductive system, 121
rats, 26–27, 27f alveolar hyperplasia, rodents, 115, 115f respiratory system, 82
hemosiderin, marmoset, 8, 8f ductal hyperplasia, rodents, 115, 115f skin and appendages, 84
inflammation, non-human primates, 8, 9f fibroadenoma, rodents, 116f urinary system, 83
intracytoplasmic eosinophilic inclusions, rats, 26, focal hyperplasia, mouse, 62, 62f Moist dermatitis, rabbit, 90
26f New Zealand White rabbit, 90 Molars, food accumulation, 23, 23f
karyomegaly, mouse, 54, 54f estrus cycle, non-human primate, 119f Mounting, artifacts, 97
lipid vacuolation rats, 115 Mouse, 45–72
marmoset, 8, 8f Vater Pacinian corpuscle, cynomolgus macaque, brain, 64
non-human primates, 8, 8f 13, 13f cardiovascular system, 45–46
lobe torsion, rats, 27, 27f Mandibular lymph node endocrine glands, 59–61
marmoset, 8 abscess, minipig, 82, 82f gastrointestinal system, 50–53
microgranuloma, mouse, 55, 55f mast cells, rats, 18, 18f information sources, 45
necrosis, marmoset, 8f plasmacytosis, rats, 18, 19f hemolymphoreticular system, 46–49
periportal fat vacuolation, rats, 26, 26f Mast cells, rat mandibular lymph node, 18, 18f musculoskeletal system, 62–63
tension lipidosis Maturity nervous systems, 64
non-human primates, 8, 8f dog male reproductive system, 105–107 pancreas, 56
rats, 25, 26f minipig male reproductive system, 110 reproductive system see Rats
Luminal crystals, mouse thyroid, 61, 61f rat spermatogenesis, 101–102 respiratory system, 49–50
Lungs Medullary hyperplasia, rat adrenal glands, 31, 31f stock/strain differences, 45
mineralization Mercuric chloride–formaldehyde fixation, artifacts, systemic disease, 66
marmoset, 4–5, 5f 95 urinary system, 57–59
minipig, 82, 82f Mesenchymal proliferation Mucinous degeneration, non-human primate atria,
smooth muscle mineralization, dogs, 38, 38f endocardium, rats, 17, 17f 3, 3f
Lymph follicles, non-human primates, 3–4, 4f lacrimal gland, mouse, 66, 66f Mucosal glands, cystic dilatation, 52,
Lymph nodes Mesenteric adipose tissue necrosis, rats, 23, 24f 52f
atrophy, rats, 18 Mesenteric lymph node Mucus cell hyperplasia, dogs, 40, 40f
hyalinisation, non-human primates, 3–4, 4f macrophages, rats, 18, 18f Multifocal subpleural inflammation, rats, 22,
Russell bodies, rats, 18, 19f sinus ectasia, mouse, 47, 47f 22f
sinus dilatation, rats, 18, 18f sinus erythrocytosis, mouse, 47, 47f Multilocular biliary cyst, mouse, 55, 55f
tattoo pigment, rats, 19, 19f Metabolic disease, New Zealand White rabbit, 91 Multinucleate cells, cynomolgus macaque kidney,
Lymphocytic epididymitis, dogs, 107, 108f Metaplasia 10, 10f
Lymphocytic infiltration Bowman’s capsule, rats, 28 Multinucleate giant cells, dog testes, 107,
cervix, non-human primates, 121, 121f cuboidal, Bowman’s capsule, 9, 9f 107f
eye, non-human primates, 13, 13f osseous see Osseous metaplasia Multinucleate hepatocytes see Hepatocytes
kidney, mouse, 57, 57f Microgranuloma, mouse liver, 55, 55f Mural hypertrophy, rat heart artery, 17–18,
thyroid gland, rabbit, 90 Middle ear fibrosis, hamster, 77, 77f 18f
urinary bladder, rats, 29, 29f Mineralization Muscles
Lymphocytic orchitis, dogs, 107, 108f adrenal cortex, non-human primates, 10–11, inflammation, rats, 33, 33f
Lymphocytic thyroiditis, dog thyroid, 42, 42f 11f keratin cysts, rats, 33, 33f
Lymphofollicular structure, mouse germinal centres, aorta Muscle spindle, nuclear bag fiber, 63, 64f
48, 48f dogs, 37, 37f Musculoskeletal system
Lymphoid aggregates, rodent epididymis, 103, rabbit, 87, 87f beagle dogs, 43
104f artery, rats, 20, 20f guinea pigs, 78
Lymphoid follicles brain see Brain hamsters, 77
bladder, dogs, 41, 41f bronchiole mucus, rats, 22, 22f minipigs, 84
hyaline deposits, dogs, 38, 38f cornea, mouse, 65, 65f mouse, 62–63
vagina, non-human primates, 121, 121f epicardium, mouse, 46, 46f New Zealand White rabbit, 91
Lymphoid hyperplasia gastric mucosa, hamster, 74, 74f non-human primates, 12
bladder, rabbit, 90, 90f hard palate, hamster, 74, 74f rats, 33
Peyer’s patch, rats, 25, 25f iris, mouse, 65, 65f see also specific features
thymus, mouse, 48, 48f kidney see Kidney Mycotic oesophagitis, marmoset, 5, 5f
Lymphoid infiltrates lungs see Lungs Myocardial artery, focal villous intimal proliferation,
adrenal cortex, minipig, 81, 81f nasal turbinate, rats, 20, 20f 37, 37f
cerebrum, minipig, 81, 81f nasal turbinate mucus, rats, 22, 22f Myocardial degeneration, cynomolgus macaque, 1,
renal cortex, minipig, 81, 81f pelvic transitional epithelium, rats, 29, 29f 2f
urinary bladder, mouse, 58, 58f Peyer’s patches, rats, 19, 19f Myocardial lymphoid cells, rats, 17, 17f
Lymphoma tissue, cutting artifact, 96, 96f prostate gland see Prostate gland Myocarditis, focal, 1, 1f
Hemolymphoreticular system salivary gland, minipig, 82, 82f Myocardium
beagle dogs, 38 spinal cord meninges, dogs, 43, 43f degeneration, cynomolgus macaque, 2, 2f
guinea pigs, 78 strangulated/ischemic adipose tissue, rats, 23, eosinophilic inclusions, non-human primates, 2,
hamsters, 73 24f 2f
minipigs, 81–82 tongue see Tongue fibrosis, marmoset, 2, 2f
mouse see Mouse urinary bladder, non-human primates, 10, 10f inflammatory cells, cynomolgus macaque, 1,
New Zealand White rabbit, 87 Minipigs, 81–85 1f
non-human primates, 3–4 brain, 84 Myodegeneration, minipig, 84, 84f
rats, 18–20 cardiovascular system, 81 Myofibers, atrophy, 33, 33f
128 Subject Index

N O focal hypertrophy, rats, 32, 32f

minipig, 84, 84f
Nasal septum, hyaline material, 49, 49f Oesophagostomum infection, non-human primates, ultimobranchial cyst, mouse, 61, 61f
Nasal turbinates 7, 7f Parotid salivary gland, basophilic hypertrophic
eosinophilic inclusions, rats, 20, 20f Estrus, dog female reproductive system, 117–118, focus, 24, 24f
mineralization, rats, 20, 20f 117f–118f Pars distalis
mucus mineralization, rats, 22, 22f Estrus cycle cysts, dogs, 41–42, 42f
Necrosis dog female reproductive system, 116–118 developmental cysts, mouse, 59, 59f
focal, liver, 8, 9f mammary gland, non-human primate, 119f focal hyperplasia, pituitary gland, 30, 30f
hepatocytes ovary, non-human primate, 119f Pars nervosa, cysts, 41–42, 42f
minipig, 83, 83f rats, 111–112, 111f–112f Passalurus ambiguus ova, rabbit, 88, 88f
rats, 26–27, 27f uterus, non-human primate, 119f–120f Pelger–Huet syndrome, rabbit, 87
liver, marmoset, 8, 8f vagina, rats, 111f Pelvic transitional epithelium, mineralization, 29,
submucosal, stomach, 5–6, 6f Olfactory epithelium, intracytoplasmic hyaline 29f
Necrotic cholecystitis, rabbit, 89, 89f inclusions, 49, 49f Pelvic urothelium, hyperplasia, 29, 29f
Neonates, rat brain, 34, 34f Optic disk, cupping, 84, 84f Penile lesions, rats, 104
Nephropathy, rats, 28, 28f Optic nerve Periarteritis
Nervous system atrophy, rats, 35, 35f dogs, 37, 37f
beagle dogs, 43–44 gliosis, artifacts, 93, 93f minipig, 81, 81f
guinea pigs, 78 hemorrhage, artifacts, 93, 93f mouse, 46, 46f
minipigs, 84 neuritis, cynomolgus macaque, 13, 13f Perimesenteric plexus, minipig, 82, 82f
mouse, 64 Osseous metaplasia Periodontal gingivitis, mouse, 50, 51f
New Zealand White rabbit, 91 lung parenchyma Periportal fat vacuolation, rat liver, 26, 26f
non-human primates, 12–13 dogs, 38, 38f Peripuberty
rats, 33–34 rats, 22, 22f dog female reproductive system, 116
Nesidioblastosis, dog pancreas, 42, 42f rats, 28 dog male reproductive system, 105–107
Neuroblasts, rats, 34, 34f Ossification minipig male reproductive system, 110
Neuroendocrine cell hyperplasia adrenal cortex, rats, 31, 31f rat female reproductive system, 111
bronchiole, rats, 20–21, 21f laryngeal cartilage, rats, 22, 22f rat spermatogenesis, 101–102
hamster, 73, 74f Osteomyelitis, marmoset, 12, 12f Perivascular cuffing
New Zealand White rabbit, 87–91 Osteophytes, dog lung parenchyma, 38, 38f brain, non-human primates, 13, 13f
brain, 91 Ovaries CNS, dogs, 43, 43f
cardiovascular system, 87 amyloid replacement, rodents, 114, 114f Perivascular melanin, non-human primate brain, 13,
ear, 91 atrophy, rodents, 112–113, 113f 13f
endocrine system, 90 corpora lutea, hamster, 76, 76f Perivasculitis, non-human primates, 3, 3f
eye, 91 cystic replacement, rodents, 114, 114f Peyer’s patches
gastrointestinal system, 88 follicular cysts lymphoid hyperplasia, rats, 25, 25f
hepatobiliary system, 88–89 non-human primates, 121, 121f mineralization, rats, 19, 19f
hemolymphoreticular system, 87 rodents, 112f Pigments
mammary glands, 90 interstitial gland, rabbit, 90, 90f adrenal cortex, rats, 31, 31f
metabolic disease, 91 estrus cycle, non-human primate, 119f renal cortical tubules, mouse, 57, 57f
musculoskeletal system, 91 pseudopregnancy, rodents, 112–113, 113f Pinch artifact, 94, 94f
nervous system, 91 rats, 114 Pituitary gland
pancreas, 89 tubulostromal hyperplasia, rodents, 114, adenohypophysis, rats, 30, 30f
reproductive system, 90 114f angiectasis, rats, 30, 30f
respiratory system, 87 Ovotestis, rodents, 112–113, 113f cyst, rats, 30, 30f
skin and appendages, 90 pars distalis focal hyperplasia, rats, 30, 30f
urinary system, 89–90 Rathke’s pouch remnant, rats, 30, 30f
Non-glandular stomach
focal ulceration, 23, 23f
P Plant material contamination, 93, 94f
Plasmacytic infiltrates, rat salivary gland, 24, 24f
hepatocytes, 26, 26f Pancreas Plasmacytosis
Non-glandular stomach, rats, cyst, 23, 23f accessory spleen mandibular lymph node, rats, 18, 19f
Non-human primates, 1–15 non-human primates, 7, 8f mouse, 47, 47f
brain, 12–13 rabbit, 89, 89f Pleural lung tag, rats, 21, 21f
cardiovascular system, 1–3 acinar atrophy Pneumoconiosis, non-human primates, 4
ectopic liver, 11, 11f mouse, 56, 56f Polyarteritis, rabbit lung, 87, 88f
endocrine system, 10–12 rats, 25, 25f Polycystic kidney, rats, 28, 28f
female reproductive system, 119–121 acinar vacuolation, rats, 25, 25f Postmortem/necropsy, artifacts, 93–94
background changes, 120–121, 120f atrophy, dogs, 42, 42f Pregnancy toxemia, rabbit, 91
cyclicity-related changes, 119–120, 119f–120f basophilic tinctorial change, rats, 25, 25f Prepuce abscess, hamster, 76, 77f
immaturity, 119 ductal mucus epithelial hyperplasia, hamster, 75, Preputial gland
puberty, 119 76f cystic atrophy, rodents, 104, 105f
gastrointestinal system, 5–7 ectopic see Ectopic pancreatic tissue cystic distension, rodents, 104, 105f
hepatobiliary system, 8–9 islet cell hyperplasia inflammation, rodents, 104, 104f
hemolymphoreticular system, 3–4 hamster, 75, 75f Processing artifacts, 95–96
male reproductive system, 109–110, 109f mouse, 61, 61f Pro-estrus, dog female reproductive system, 117,
incidental findings, 109–110, 109f–110f rats, 32, 32f 117f
seasonal changes, 109, 109f mouse, 56 Prolapse, mouse rectum, 53, 53f
musculoskeletal system, 12 nesidioblastosis, dogs, 42, 42f Prostate gland
nervous system, 12–13 New Zealand White rabbit, 89 dogs, 108
respiratory system, 4–5 zymogen degranulation focus, rats, 25, 25f epithelial atrophy/mineralization, hamster, 76,
urinary bladder, 10 zymogen granules, rats, 32, 32f 77f
urinary system, 9–10 Pancreatic vessel, arteritis, 56, 56f epithelial hyperplasia, rodents, 104f
Basophilic hypertrophic focus, mouse salivary Paneth cells, zymogen granules, 52–53, 53f focal cystic dilatation, dogs, 108, 108f–109f
glands, 51, 51f Parathyroid glands focal squamous metaplasia, rabbit, 90, 90f
Nuclear bag fiber, muscle spindle, 63, 64f cysts, dogs, 41–42, 42f inflammation, rodents, 104, 104f
Nuclear brick inclusions ectopic, rats, 20 mineralization
hepatocytes, dogs, 40, 40f ectopic thymic tissue hamster, 76, 77f
renal tubular epithelial cells, dogs, 40, 40f marmoset, 11–12, 12f non-human primates, 110f
Nucleus circularis, rat cerebrum, 34, 34f mouse, 60–61, 61f rodents, 104, 105f
 Subject Index 129

Pseudoglandular cysts, dog epididymis, 108, 108f Rete epithelium, hyperplasia, rodents, 103, 104f chondromucinous degeneration
Pseudopregnancy Rete ovary cyst, rodents, 112f dogs, 43, 43f
dogs, 118, 118f Retina mouse, 63, 63f
ovary, rodents, 112–113, 113f atrophy rats, 33, 33f
uterus, rodents, 112–113, 113f mouse, 65, 65f dysplasia, hamster, 77, 77f
vagina, rodents, 112–113, 113f rats, 35, 35f Stock/strain differences, mouse, 45
Puberty, non-human primate female reproductive rosettes, rats, 35, 35f Stomach
system, 119 Rhabdomyomatosis, guinea pig, 78, 78f adenomatous hyperplasia, mouse, 52, 52f
Pulmonary acinar ectasia, rats, 22, 22f Right ventricle, adipose tissue infiltration, 87, 87f focal erosion, hamster, 74, 74f
Pulmonary thrombi, non-human primates, 4, 4f Russell bodies, rat lymph node, 18, 19f gastric erosion
Purkinje cells, eosinophilic, 98, 99f minipig, 82, 83f
non-human primates, 5, 5f
S squamous cyst, rats, 23, 23f
R Salivary glands
submucosal eosinophils, rats, 23, 23f
submucosal necrosis, non-human primates, 5–6,
Rathke’s pouch remnant, rat pituitary gland, 30, acinar atrophy, rats, 24, 24f 6f
30f adipocytes, dogs, 39, 40f Strangulated adipose tissue, mineralization, 23,
Rats, 17–36 mineralization, minipig, 82, 82f 24f
brain, 33–34 basophilic hypertrophic focus, mouse, 51, 51f Striated (cardiac) muscle, mouse lungs, 50, 50f
cardiovascular system, 17–18 plasmacytic infiltrates, rats, 24, 24f Subarticular bone cyst, rats, 33, 33f
ear, 35–36 serous focus, rats, 24, 24f Subcapsular cell hyperplasia, mouse adrenal glands,
endocrine glands, 30–32 squamous metaplasia, minipig, 82, 82f 60, 60f
eye, 35–36 see also specific salivary glands Subcapsular sinus, eosinophils, 82, 82f
female reproductive system, 111–116 Salmonella infection, non-human primates, 6 Subcutaneous abscess, mouse, 62, 62f
aging animals, 112–114, 114f Sarcocystis cysts, non-human primates, 12, 12f Subepithelial arteriovenous anastomosis, dog
cervix, 114–115 Schweiggel–Seidel sheaths, minipig spleen, 81, 81f tongue, 39, 39f
clitoral gland, 116, 116f Sciatic nerve Subepithelial inflammation, dog tongue, 39, 39f
immaturity, 111 demyelination, rats, 33, 33f Submandibular salivary glands
mammary gland, 115 Renault bodies, dogs, 43, 43f focal atrophy, mouse, 51, 51f
estrus cyclicity, 111–112, 111f–112f Seasonal changes, non-human primate male sexual dimorphism, mouse, 51, 51f
ovaries, 114 reproductive system, 109, 109f Submucosal eosinophils, rat stomach, 23, 23f
peripuberty, 111 Sebaceous glands Submucosal necrosis, non-human primates stomach,
uterus, 114–115 ectopic, rats, 23 5–6, 6f
vagina, 115 Seminal vesicle, distension, 104, 105f Subpleural lymphatics, dogs, 39, 39f
young animals, 112, 112f Serous focus, rat salivary gland, 24, 24f Synovial hyperplasia, mouse knee joint, 63, 63f
gastrointestinal system, 23–25 Sperm granulomas, rodent epididymis, 103, 103f Syphacia obvelata infection, mouse, 53, 53f
hepatobiliary system, 25–27 Severe diffuse amyloidosis, hamster, 75, 75f Systemic disease, mouse, 66
hemolymphoreticular system, 18–20 Sexual dimorphism
male reproductive system, 101–104 Bowman’s capsule, mouse, 57, 57f
accessory sex organs, 104
age-related lesions, 103–104
submandibular salivary glands, mouse, 51, 51f
Shigella infection, colitis, 6, 6f
T
congenital/developmental lesions, 102 Sinus dilatation, rat lymph nodes, 18, 18f Talcum powder, artifact, 93
epididymides, 103–104 Sinus ectasia, mouse mesenteric lymph node, 47, Tattoo pigment, rat lymph node, 19, 19f
penile lesions, 104 47f Tension lipidosis, liver see Liver
residual bodies, 103f Sinus erythrocytosis, mouse mesenteric lymph Testes
tubular degeneration/atrophy, 102–103, node, 47, 47f amyloid deposition, rodents, 103, 104f
102f–103f Skin atrophic seminiferous tubules, hamster, 76, 77f
musculoskeletal system, 33 beagle dogs, 43 dogs, 107–108
nervous system, 33–34 hamsters, 76 epididymis hyperplasia, hamster, 76, 77f
respiratory system, 20–22 minipigs, 84 fibrous hypoplasia, non-human primates, 110f
skin and appendages, 33 New Zealand White rabbit, 90 focal tubular degeneration, non-human primates,
spermatogenesis, 101–104, 101f–102f rats, 33 110f
immaturity, 101–102 Small intestine, ectopic pancreas, 25, 25f focal tubular dilatation, non-human primates,
maturity, 101–102 Spermatogenesis, 101, 101f–102f 109f
peripuberty, 101–102 rats see Rats hypoplasia, rodents, 102
urinary system, 28–29 Spinal cord multinucleate giant cells, dogs, 107, 107f
Rectum, prolapse, 53, 53f epidermoid cyst, mouse, 64, 64f tissue distortion, fixative artifact, 95, 95f
Refluxed seminal colloid plug, mouse urinary focal degeneration, mouse, 64, 65f tubular hypoplasia
bladder, 58, 59f keratin cysts, rats, 34, 34f dogs, 107, 107f
Renal cortex see Kidney cortex meninges mineralization, dogs, 43, 43f minipig, 110f
Renault bodies, dog sciatic nerve, 43, 43f Spleen Thrombosis
Reproductive system accessory see Pancreas aortic valve, hamster, 73, 73f
female see Female reproductive system capsular cyst, rats, 19, 19f atrial, hamster, 73, 73f
guinea pigs, 78 extramedullary hemopoiesis see Extramedullary umbilical blood vessel, minipig, 83, 83f
hamsters, 76 hemopoiesis Thrombus
male see Male reproductive system hemosiderin, rats, 19, 19f bone marrow, artifact, 93, 93f
minipigs, 84 hemosiderotic plaque, dogs, 38, 38f lung, rabbit, 87, 88f
New Zealand White rabbit, 90 Schweiggel–Seidel sheaths, minipig, 81, 81f Thymus
Residual bodies, rat male reproductive system, Squamous cyst, rat stomach, 23, 23f apoptosis, rats, 20, 20f
103f Squamous epithelium atrophy
Respiratory system thymus, rats, 19, 20f hamster, 73, 73f
beagle dogs, 38–39 trachea, dogs, 38, 38f rats, 20, 20f
guinea pigs, 78 Squamous hyperplasia, rat Harderian glands, 35, branchial cysts, dogs, 38, 38f
hamsters, 73 35f cyst, mouse, 48, 48f
minipigs, 82 Squamous metaplasia ectopic, parathyroid gland see Parathyroid glands
mouse see Mouse endometrial glands, rodents, 112–113, 113f eosinophilic crystals, rats, 20, 20f
New Zealand White rabbit, 87 salivary gland, minipig, 82, 82f hemorrhage, rats, 20, 20f
non-human primates, 4–5 Stains, artifacts, 97–98, 98f Hassall’s corpuscles, dogs, 38, 38f
rats, 20–22 Sternum lymphoid hyperplasia, mouse, 48, 48f
see also specific components cartilage degeneration, hamster, 77, 77f squamous epithelium, rats, 19, 20f
130 Subject Index
Thyroid gland
C-cell hyperplasia
marmoset, 11, 11f
rats, 32, 32f
C-cell infiltrates, dogs, 42, 42f
cysts, rats, 32
ectopic tissue, dogs, 41–42
follicle dilatation, mouse, 61, 61f
follicular epithelial hypertrophy, rats, 32, 32f
intracytoplasmic colloid, rats, 32, 32f
luminal crystals, mouse, 61, 61f
lymphocytic infiltration, rabbit, 90
lymphocytic thyroiditis, dogs, 42, 42f
venipuncture damage, minipig, 84, 84f
Tissue contamination, 94–97, 94f, 96f–97f
Tissue distortion artifact, testicles, 95, 95f
Tongue
blood collection trauma, rats, 24, 25f
cartilage, minipig, 82, 82f
mineralization
hamster, 74, 74f
rats, 18, 18f
subepithelial arteriovenous anastomosis, dogs, 39,
39f
subepithelial inflammation, dogs, 39, 39f
Trachea
cartilage mineralization, rats, 21, 21f
squamous epithelium, dogs, 38, 38f
Tracheal glands, eosinophilic materials, 49–50,
49f
Transitional epithelium hyperplasia, rat kidney
pelvis, 29, 29f
Tubular degeneration/atrophy, rat male reproductive
system, 102–103, 102f–103f
Tubular dilatation, hamster kidney, 75, 75f
Tubular hypoplasia see Testes
Tubulostromal hyperplasia, rodent ovary, 114,
114f
U 12f
Ulceration
flank glands, hamster, 76, 76f
rats, 32f, 33
skin, minipig, 84, 84f
Ultimobranchial cyst, mouse parathyroid gland, 61,
61f
Umbilical blood vessel, thrombosis, 83, 83f
Urachal artery thrombosis, dog bladder, 41, 41f
Urinary bladder
colloidal plug, rats, 29, 29f
lymphocytic infiltration, rats, 29, 29f
lymphoid infiltrates, mouse, 58, 58f
non-human primates, 10
refluxed seminal colloid plug, mouse, 58, 59f
urothelial hyperplasia, mouse, 58, 58f
Urinary system
beagle dogs, 41
guinea pigs, 78
hamsters, 75
minipigs, 83
mouse see Mouse
New Zealand White rabbit, 89–90
non-human primates, 9–10
rats, 28–29
see also Kidney; Urinary bladder
Urothelial hyperplasia, mouse urinary bladder, 58,
58f
Urothelium
eosinophilic inclusions, non-human primates, 10,
10f
vacuolation, non-human primates, 10, 10f
Uterus
benign stromal polyp, rodents, 115, 115f
cystic hyperplasia, rodents, 114, 114f
decidual reaction, hamster, 76, 76f
deciduoma, rodents, 115, 115f
granular cell hyperplasia, hamster, 76, 76f
estrus cycle, non-human primate,
119f–120f
pseudopregnancy, rodents, 112–113, 113f
rats, 114–115
V Zona glomerulosa
Vaccine granuloma, non-human primate muscle, 12,
Vacuolation
adrenal cortex, rats, 30, 30f–31f
adrenal glands, rabbit, 90
fixation artifact, 98, 98f
hepatocytes see Hepatocytes
processing artifacts, 96, 96f
urothelium, non-human primates, 10, 10f
Vagina
cornificiation, non-human primates, 121,
121f
cysts, rodents, 112f
lymphoid follicles, non-human primates, 121,
121f
estrus cyclicity, rats, 111f
pseudopregnancy, rodents, 112–113, 113f
rats, 115
Vaso vasorum, rats, 18, 18f
Vater Pacinian corpuscle, cynomolgus macaque
mammary gland, 13, 13f
Venipuncture damage, minipig thyroid, 84, 84f
Ventricle, adipose tissue infiltration, 87, 87f
Ventricular dilatation, rabbit brain, 91
Villous mesothelial projections, dog atria, 37,
38f
W
Warthin–Finkeldey bodies, non-human primate
gut-associated lymphoid tissue (GALT), 3,
3f
Wasting marmoset syndrome (WMS), 6–7
X
X zone, mouse adrenal glands, 59, 59f
Y
Yersinia infection, non-human primates, 6
Z
focal hypertrophy, 31, 32f
vacuolation, 42, 42f
Zona reticularis vacuolation, dog adrenal glands, 42,
42f
Zymogen degranulation focus, rat pancreas, 25,
25f
Zymogen granules
pancreas, rats, 32, 32f
Paneth cells, mouse, 52–53, 53f