CHAPTER 8

Dry-Mount Fecal Cytology

Amy L. Weeden, Heather L. Wamsley

There are several diagnostic tests of feces that may be necessary

during a complete evaluation of patients with gastrointestinal
signs. Optimal fecal assessment, including potential tests (e.g.,
wet-mount fecal cytology, dry-mount fecal cytology, bacterial
culture, fecal antigen detection methods, fecal flotation, fecal
sedimentation, Baermann technique) and their required sample
handling, diagnostic indications, and interpretations, have been
compiled elsewhere (Broussard, 2003). Dry-mount fecal cytol-
ogy (air-dried smear) is one component of a thorough diagnos-
tic evaluation of patients with gastrointestinal signs. Since some
fecal pathogens can be morphologically indistinguishable from
incidental nonpathogens, results of dry-mount fecal cytology
should be interpreted in the context of the patient’s clinical pre-
sentation and results of other diagnostic tests, such as wet-mount
fecal cytology, bacterial culture, and fecal antigen detection.
When evaluating dry-mount fecal cytology, it may be use-
ful to apply a systematic approach aimed at evaluation of back-
ground cells and detection of abnormal eukaryotic cells and A
pathogenic microorganisms (Box 8-1).

BOX 8-1  Guidelines for Systematic Method

of Dry-Mount Fecal Cytology Evaluation
1. Evaluate the background bacterial and yeast flora using 100× objective
2. Determine the number of spore-forming bacteria per 100× objective field
3. Examine the smear for other potential pathogens using 50× or 100×
­objectives
a. Algal (e.g., Prototheca)
b. Bacterial (e.g., gull wing–shaped and spiral-shaped bacteria)
c. Fungal (e.g., Histoplasma, Aspergillus, Blastomyces, Candida, Cryptococcus)
d. Oomycetal (e.g., Pythium)
e. Protozoal (e.g., Cryptosporidium, Giardia, Entamoeba, Trichomonas,
Ballintidium)
f. Rare findings (e.g., nematode and trematode eggs [Figs. 8-1A&B] and B
larvae are rarely diagnosed by this method) n FIGURE 8-1  A, Nematode ova. Feces. Dog. Several common types
4. Scan the smear for the presence of host cells of ova are shown. Large dark round - Toxocara canis. Arrows indicate
a. Inflammatory cells—observe types and relative quantities Trichuris vulpis. Arrowheads point to Ancylostoma caninum. (Unstained;
b. Epithelial cells—assess number and morphology (i.e., normal, hyper- IP.) B, Trematode ovum. Platynosumum fastosum (concinnum).
plastic, or neoplastic) Feces. Cat. Ova measure 34 to 50 μm by 20 to 35 μm with a thick wall.
c. Other atypical or neoplastic host cells Arrowhead indicates operculated end. (Unstained; LP.) (A from Purdue
University; B courtesy of Rose Raskin, University of Florida.)

247
248 Canine and Feline Cytology
SAMPLE COLLECTION AND PROCESSING
The method of fecal sample collection can affect the following:
the part of the rectum that is represented (luminal vs. mucosal)
and the cellular (eukaryotic and prokaryotic) content of the sam-
ple. The ideal sampling method, desired sample amount, and
subsequent sample handling depends upon the intended fecal
testing (Broussard, 2003). Important goals of sampling for fecal
cytology include examination of a fresh sample (less than 5 min-
utes old) that is representative of the mucosal surface and prepa-
ration of a fecal thin film preparation that is not excessively dense.
Excessively dense areas of fecal thin film preparations are prone
to detachment during staining or they obscure cytologic details.
Because the cellular content of fecal material is dynamic and
will likely continue to change after sample collection, immediate
examination following collection is recommended.
KEY POINT  Preparation of thin fecal film of fresh material is preferred
over a densely packed smear. Results of dry-mount fecal cytology should
be interpreted in view of the patient’s clinical presentation and results of
other diagnostic tests.
KEY POINT  As fecal material readily falls off during staining, it is im-
portant to stain fecal slides at the end of a batch as well as change the
stain solution if a dip technique is used. These precautions will help avoid
bacterial contamination to subsequent specimens being stained.
For fecal cytology, there are a few possible sampling meth-
ods, including collection of unadulterated fecal material, rectal
saline lavage, and rectal scraping. A small amount of fresh fecal
material may be obtained during digital rectal examination or
using a moistened, cotton-tipped applicator or fecal loop if dig-
ital rectal exam is not possible. Immediately collected, voided
feces may also be used; however, voided samples may be more
representative of the luminal portion of the rectum, which is not
ideal for fecal cytology. Voided fecal material is a preferred sam-
ple for other diagnostic tests, such as fecal flotation, fecal sedi-
mentation, or the Baermann technique, since it is more copious
and may be rich in parasitic eggs and cysts (Broussard, 2003).
Usually, if a small amount of fecal material is obtained using a
moistened, cotton-tipped applicator, the cotton tip of the appli-
cator can be gently rolled on the slide to prepare a direct thin
film. Otherwise, slight dilution of the feces may be used to pre-
pare a thin film preparation by placing a drop of sterile, normal
saline on a clean glass microscope slide, adding a very small
amount of fecal material (no larger than a match-head), mixing
with a sterile wooden applicator, and spreading as for other fluid
thin film preparations. Prior to and after spreading, newsprint
should be legible though the preparation.
Rectal saline lavage enriches collection of mucosal material,
including mucus, motile protozoa, and bacteria. Lavage fluid
contains relatively less luminal fecal material and is ideal for
fecal cytology (Broussard, 2003). Properly collected lavage fluid
should have a mud-like consistency, and a single drop can be
used to prepare a thin film preparation.
Material obtained by rectal scraping is often submitted as a
sample for dry-mount fecal cytology, though feces and rectal
scrape material are not synonymous. Rectal scraping with a
swab or blunt spatula is a somewhat more invasive method of
sampling the rectal mucosa—by direct scraping of its surface.
Rectal scraping is typically required to identify some infections
that localize to the deeper portion of the mucosa (e.g., histoplas-
mosis, protothecosis) or to characterize deep mucosal cellular
infiltrates.
Dry-mount fecal cytology can be useful to examine the
microorganism flora and any host cells that may be present (e.g.,
epithelial, inflammatory) and to detect pathogens that may be
present (e.g., bacteria, fungi, algae, oomycetes, or protozoa).
Occasionally, evaluation of dry-mount fecal cytology is diag-
nostic, but more often it aids in ruling out of selected causes of
diarrhea.
KEY POINT  Many abnormalities, particularly those involving the back-
ground flora, are nonspecific, representing incidental findings associated
with other underlying diseases, physiologic processes, or previous antimi-
crobial treatments.
NORMAL OR INCIDENTAL MICROSCOPIC FINDINGS
The background microbial flora should be predominantly
composed of an extremely polymorphic population of sev-
eral different bacilli (Fig. 8-2). There should be fewer than five
spore-forming bacilli per 100× objective field (Broussard, 2003).
Cocci should be absent or only rarely observed. A low number
of extracellular, circular or ovoid, 5 to 10 μm in ­diameter, stip-
pled, variably basophilic yeast with a thin, colorless capsule may
occasionally be seen (Figs 8-3, 8-4). While these yeast structures
are frequently found in diarrheal stools, it is not certain whether
there is a direct causal relationship.
Cyniclomyces guttulatus (formerly Saccharomycopsis gut-
tulata) is part of the normal flora of some rodents and lago-
morphs (rabbits, guinea pigs, and chinchillas) (Zierdt et  al.,
1988). Occasionally, a low number of individual or doublet
Cyniclomyces may be observed as an incidental finding in
canine feces and are thought to represent a nonpathogenic
result of coprophagia (Fig. 8-5). However, there is uncertainty
about whether this yeast is entirely nonpathogenic in dogs. A
few preliminary research or anecdotal reports have associated
n FIGURE 8-2  Normal bacterial flora. A highly polymorphic, mixed
bacterial flora that is composed predominantly of several different bacilli
is expected to be observed in feces and is typically found on an amor-
phous, pale basophilic background that contains either no or few host
cells. (Wright-Giemsa; HP oil.)

A B
n FIGURE 8-3  Incidental yeast. A, A few circular or ovoid, 5 to 10 μm in diameter, stippled, basophilic yeast with a thin, colorless capsule may
occasionally be incidentally observed extracellularly within fecal smears. (Wright-Giemsa; HP oil.) B, Closer magnification of extracellular fecal yeast
that can be observed incidentally in fecal smears. (Wright-Giemsa; HP oil.)
10 m
n FIGURE 8-4  Incidental yeast. Cat. These yeast were present in an
adult cat with diarrhea. They appear both as single and budding forms
with both clear and focally dense staining areas within the structure.
The significance of the yeast is unknown in causing the diarrhea; how-
ever, they occur more commonly in watery stools possibly by producing
a wash effect that releases the organisms more readily. (Wright; HP oil.)
(Courtesy of Rose Raskin, Purdue University.)
this yeast with clinical cases of chronic diarrhea (Houwers
and Blankenstein, 2001; Mandigers, 2007). During evaluation
of some patients with chronic diarrhea, the only abnormality
detected in dry-mount fecal cytology may be the presence of an
extremely large number of Cyniclomyces both individually and
in large mats of numerous budding organisms (Figs. 8-6 and
8-7). The observation of many apparently replicating organ-
isms in a fresh fecal sample may represent a factor contribut-
ing to the ongoing diarrhea or may simply represent abnormal
flora due to underlying disease, physiologic processes, or prior
antimicrobial treatments. At this time, the pathogenicity of this
yeast is unproven, though it is occasionally observed in large
number in the feces of dogs with chronic diarrhea; in such cases
this observation should be reported as an abnormal finding.
It is common to see a variable amount of irregularly shaped,
colorless or amber material and green-blue material with
CHAPTER 8   Dry-Mount Fecal Cytology 249
n FIGURE 8-5  Yeast. An abnormal bacterial flora, containing numer-
ous diplococci, is present on a pale basophilic background that contains
a small amount of irregularly shaped, refractile, amber material and a
single incidental, extracellular fecal yeast (left) and two Cyniclomyces
guttulatus organisms. (Wright-Giemsa; HP oil.)
parallel cell walls, representing digesta and ingested plant mat-
ter, respectively (Fig. 8-8). The background also commonly
contains a variable amount of amorphous basophilic material,
consistent with mucus. The amount of mucus observed depends
on whether the underlying disease is associated with rectal over-
secretion of mucus and whether the sampling method is more
representative of the rectal mucosa or rectal lumen. Being more
representative of the rectal mucosa, samples obtained by rectal
saline lavage or rectal scraping may contain more mucus than
samples collected by other means.
Well-differentiated epithelial cells, including squamous or
low columnar epithelial cells, may be present in variable num-
bers depending on the sample collection method and underlying
disease (Figs. 8-9 and 8-10). With atraumatic sample collection
methods, low numbers of well-differentiated epithelial cells may
be present individually or in small sheets. With more invasive
250 Canine and Feline Cytology

A B
n FIGURE 8-6  Yeast. Cyniclomyces guttulatus. Dog. A, Several budding organisms observed in the feces of a dog with chronic diarrhea.
(Wright-Giemsa; IP.) B, Higher magnification of budding yeast. (Modified Wright; HP oil.) (B, Courtesy of Kristin Fisher, Purdue University.)

n FIGURE 8-7  Yeast. Large mat of budding Cyniclomyces guttulatus n FIGURE 8-8  Plant material. Green-blue ingested plant matter with
associated with amorphous grey-brown ingesta and squamous epithe- parallel cell walls is commonly seen in canine and feline fecal smears.
lial cells. (Wright-Giemsa; IP.) (Wright-Giemsa; HP oil.)

n FIGURE 8-9  Rectal epithelium. A single large, tightly cohesive,
multicellular sheet of low columnar epithelial cells is present in a rectal
scrape direct smear, surrounded by a few individualized anucleate, kera-
tinized squamous epithelial cells and scant streaming nuclear material
from lysed cells. (Wright-Giemsa; IP.)
n FIGURE 8-10  Columnar epithelium. A single small lymphocyte
(left) is present in the fecal smear of a dog with diarrhea along with
three well-differentiated low columnar epithelial cells on a pale baso-
philic background that contains a pleomorphic bacterial population com-
posed predominantly of small bacilli. (Wright-Giemsa; HP oil.)

n FIGURE 8-11  Abnormal flora. An abnormal bacterial flora, contain-
ing numerous diplococci and a microcolony (top center) of diplococci, is
present on a pale basophilic background that contains a small amount
of irregularly shaped, refractile, amber material and a single poorly pre-
served, karyolytic neutrophil. (Wright-Giemsa; HP oil.)
n FIGURE 8-12  Abnormal flora. An abnormal bacterial flora, exhib-
iting overgrowth of a single large, plump bacillus and an increased
number of diplococci, is present on a pale basophilic background.
(Wright-Giemsa; HP oil.)
collection methods that may abrade the mucosa (i.e., rectal
scrape or catheterization for saline lavage), the numbers of epi-
thelial cells are expected to be increased and the sheets of cells
will likely be larger. In a sample that has been collected atrau-
matically, observation of a large number of epithelial cells in
large, multicellular sheets should not be considered normal and
should raise concern for underlying mucosal pathology involv-
ing sloughing of apical epithelial cells.
ABNORMAL MICROSCOPIC FINDINGS
Abnormal Flora
Overgrowth of microorganisms is typically a secondary, non-
specific finding associated with other underlying diseases, recent
enteric surgical procedures, abnormal physiologic processes, or
recent antimicrobial administration; however, microbial over-
growth may exacerbate underlying pathology. Primary over-
growth can uncommonly be the primary cause of gastrointestinal
CHAPTER 8   Dry-Mount Fecal Cytology 251
20 m
n FIGURE 8-13  Candidiasis. Dog. Candida pseudohyphae are pres-
ent in fecal material collected from a dog with chronic gastrointestinal
signs after recent exploratory laparotomy. The pale basophilic back-
ground, devoid of the usual bacterial flora, contains a moderate amount
of incidental irregularly shaped, refractile, amber material and deeply
basophilic mucus. (Wright-Giemsa; HP oil.)
20 m
n FIGURE 8-14  Candidiasis. Dog. Same case as in Figure 8-12. Can-
dida pseudohyphae and blastospore (dark round structure at lower right)
in a fecal smear that is devoid of normal background bacterial flora.
(Wright-Giemsa; HP oil.)
disease. There is no way to distinguish between secondary or
primary microbial overgrowth using dry-mount fecal cytology,
which may reveal the presence of a bacterial population that
consists of monomorphic or oligomorphic bacilli, an increased
number of cocci, or an increased number of yeast (e.g., Candida,
Cyniclomyces) (Figs. 8-5, 8-11, 8-12, 8-13, 8-14, 8-15).
Fecal Leukocytes
The presence of fecal neutrophils should be considered an abnor-
mal finding that suggests distal colitis or proctitis (Figs. 8-11,
8-16). Fecal neutrophils should prompt diagnostic consideration
of causes of bacterial enteritis, such as salmonellosis, clostridial
colitis, entericcolibacillosis, campylobacteriosis, and infection by
other invasive or enterotoxigenic bacteria (Broussard, 2003). Cur-
rent recommendations for additional diagnostic testing to confirm
gastrointestinal infection, such as culture or molecular techniques
for detecting bacterial agents or toxins, vary depending on the spe-
cific organism and are described elsewhere (Marks et  al., 2011).
252 Canine and Feline Cytology

n FIGURE 8-15  Abnormal flora. An abnormal microbial flora, exhibit- n FIGURE 8-16  Neutrophilic inflammation. Dog. A large aggregate
ing reduced bacterial polymorphism, an increased number of diplococci, of several neutrophils is present among the microbial flora in a dog with
and numerous budding Cyniclomyces guttulatus is present on a pale diarrhea. (Wright-Giemsa; HP oil.)
basophilic background. (Wright-Giemsa; HP oil.)

Other considerations for fecal neutrophils include whipworm

infestation, which is usually associated with hemorrhagic, mucoid
diarrhea; primary inflammatory bowel disease, which is associ-
ated with other infiltrating leukocytes (i.e., lymphoplasmacytic
or eosinophilic inflammation); or inflammation associated with
primary structural disease, which is associated with inflammation
and/or necrosis (e.g., neoplasia).
Eosinophilic inflammation (see Chapter 7) may be observed
with primary inflammatory bowel disease (e.g., eosinophilic gas-
troenterocolitis or eosinophilic colitis), a condition in which the
number of mast cells are also expected to be increased relative to
normal animals (Kleinschmidt et al., 2007). Eosinophilic inflam-
matory bowel disease may occur in dogs or cats of any breed or
age. However, it is more common in young adult animals; and the
Boxer, Doberman Pinscher, and German Shepherd appear to be
predisposed. The diagnosis of eosinophilic inflammatory bowel
disease is possible after other causes of eosinophilic inflamma-
tion have been excluded (Hall and German, 2005). Eosinophilic n FIGURE 8-17  Rectal epithelium via traumatic scraping. Dog. A
inflammation is also likely to be observed as a component of the very large, tightly cohesive, multicellular sheet of low columnar epithe-
mixed inflammatory response to certain infections (e.g., fungal, lial cells is present in the rectal scrape of a dog with chronic diarrhea.
oomycetal, algal, or nematode parasites) or a foreign body. Eosin- A few Cyniclomyces guttulatus organisms that stain pale basophilic are
also visible to the left of the image. (Wright-Giemsa; IP.)
ophils may also infiltrate certain neoplasms (e.g., lymphoma).
Small and intermediate lymphocytes (see Chapter 7) with
or without plasma cells may be observed with any cause of which intracellular microorganisms may be identified. Another
chronic inflammation (e.g., infectious) or with primary inflam- specific consideration, given the anatomic location, is histio-
matory bowel disease (e.g., lymphocytic-plasmacytic enteroco- cytic ulcerative colitis. Histiocytic ulcerative colitis is an inflam-
litis or lymphocytic-plasmacytic colitis). Lymphoplasmacytic matory disease that primarily affects young Boxers and other
inflammatory bowel disease typically occurs in older animals brachycephalic breeds (e.g., French Bulldogs and English Bull-
with increased incidence in German Shepherds, Chinese Shar dogs) and requires histology for diagnosis. This disease has also
Peis, and purebred cats. Unique forms of inflammatory bowel rarely been reported in other dog breeds and cats. The presence
disease also affect Basenjis and Soft-Coated Wheaten Terriers of large macrophages that are strongly periodic acid-Schiff–
(Hall and German, 2005). The morphology of neoplastic lym- positive is recognized in cases of histiocytic ulcerative colitis
phocytes is similar to those observed in other tissues. However, (German et al., 2000; Hostutler et al., 2004).
in cats, lymphocyte morphology may not be useful to distin-
guish lymphocytic inflammation from feline small cell lym- Other Nucleated Mammalian Cells
phoma. A full-thickness enteric biopsy is needed to confirm the The number of epithelial cells observed in a sample depends
diagnosis (Evans et al., 2006; Kleinschmidt et al., 2006). on the collection method. In a sample collected atraumati-
Macrophagic inflammation (see Chapter 7) may be observed cally, observation of a large number of epithelial cells in mul-
with various causes of chronic inflammation or with infectious ticellular sheets should raise concern for underlying mucosal
causes of inflammation (e.g., fungal, oomycetal, or algal) with pathology with sloughing of apical epithelial cells (Fig. 8-17).

20 m
A n FIGURE 8-19  Creatorrhea. Dog. Two fragments of undigested
20 m
B Creatorrhea is the presence of undigested muscle fibers in
C tion (Broussard, 2003; Marks et al., 2011).
n FIGURE 8-18  Lymphoma. Dog. Same case A-C. This is a rectal
scraping from a dog with multicentric lymphoma. In addition to finding
a monomorphic population of lymphoblasts in the rectal scraping, the
diagnosis of lymphoma was confirmed by cytology of two peripheral
lymph nodes. A, Two lymphoblasts are shown with a neutrophil for
size comparison. B, A mitotic lymphoblast is seen at the left next to
an intermediate lymphocyte. The purple granular material in the back-
ground is consistent with lubricant. C, Several lymphoblasts are shown.
An anucleate squamous cell is seen at the top center. There is abundant
purple granular lubricant and streaming lysed nuclear material in the
background. (Wright-Giemsa; HP oil.) (Bar = 10 μm.)
CHAPTER 8   Dry-Mount Fecal Cytology 253
20 m
skeletal muscle are pictured on a background of mixed bacteria in a
rectal scrape from a dog with chronic diarrhea and hematochezia. The
fragment at the right shows striations typical of skeletal muscle. This
finding may be seen in patients with maldigestion or hypermotility dis-
orders. (Aqueous Romanowsky; HP oil.)
If inflammation is absent, epithelial cells should be cytologi­
cally well differentiated. Criteria for assessment of atypia in
epithelial cells are similar to those in other tissues; inflam-
mation can induce a hyperplastic response associated with
cytoplasmic and/or nuclear changes that can be cytologically
indistinguishable from those observed with neoplasia. In such
cases histopathology may provide definitive diagnosis. In sam-
ples obtained by rectal scrape, other neoplastic cell types may
also be observed, such as those from lymphoma (Fig. 8-18),
mast cell tumor, or gastrointestinal mesenchymal tumor (see
Chapter 7).
feces; they are recognized microscopically and may be seen with
diseases that causes maldigestion or hypermotility (Mundt and
Shanahan 2011). This may be observed on either wet-mount or
dry-mount fecal cytology preparations (Fig. 8-19).
Potential Microbial Pathogens
In addition to the microbial flora that is expected in feces, poten-
tial pathogens may also be present. In some instances definitive
cytologic diagnosis of infectious disease underlying a patient’s
gastrointestinal signs is possible (e.g., fungal, algal, oomycetal,
or protozoal infection). However, by dry-mount fecal cytology
alone, pathogenic bacteria are morphologically indistinguish-
able from nonpathogens that may be incidentally observed. For
bacterial culture of feces, unique sample collection, handling,
and culture conditions are essential for enteric pathogen detec-
Spore-forming bacteria can be an incidental finding in feces
(Figs. 8-20, 8-21, 8-22). There are typically no more than five
spore-forming bacteria per 100× objective field. An increased
number of spores can be observed in dogs with diarrhea. How-
ever, a direct cause-effect relationship remains arguable. There
is a poor correlation between the number of fecal spores per
100× objective field and the detection of Clostridium perfrin-
gens enterotoxin (Broussard, 2003). There are a few potential
explanations for this poor correlation. Clostridia spp. are not
the only bacteria that sporulate; common soil Bacillus spp. are
also large sporulating bacilli. Not all sporulated Clostridia spp.
254 Canine and Feline Cytology
n FIGURE 8-20  Sporulating bacilli. Dog. An increased number
of spore-forming bacilli (more than five per 100× objective field) are
present in the feces of a dog with diarrhea. These bacilli may either be
Bacillus spp. or Clostridium spp. Individualized spores are visible along
with the sporulated bacilli, which, in this field, often contain a terminally
located spore that renders a “tennis racket” appearance. When the
spores are centrally located, sporulated bacilli may have a “safety pin”
appearance. (Wright-Giemsa; HP oil.)
n FIGURE 8-21  Sporulating bacilli. Dog. Same case as in Fig. 8-20.
Malachite green is a microbiologic stain used to identify the presence
of bacterial spores, such as in Bacillus spp. or Clostridium spp. Steam is
included in the staining process to permeabilize the hard, dehydrated,
multilamellar spore walls. A pink safranin counterstain is used to distin-
guish the spores. (Malachite green/Safranin; HP oil.)
produce Clostridium perfringens enterotoxin (Broussard, 2003).
Sporulating bacteria may form spores during a delay in sam-
ple processing. Also, quantitative culture has revealed that up
to 75% of normal dogs harbor C. perfringens without detectable
fecal Clostridium perfringens enterotoxin (Broussard, 2003).
Some recommend that greater than five spore-forming bacteria
per 100× objective field is considered an abnormal finding, and
concurrent fecal neutrophils further support bacteria-induced
diarrhea (Broussard, 2003). However, a positive enzyme-linked
immunosorbent assay (ELISA) for C. perfringens or C. difficile
n FIGURE 8-22  Clostridial colitis. Fecal smear. Numerous neu-
trophils were present upon scanning of this direct fecal smear. An
increased number of large, rod-shaped bacterial endospores with a clear
center and an increased density predominantly on one end (“safety pin”
appearance) are the notable feature (short arrows). The bacterial mor-
phology is consistent with Clostridium perfringens. Occasional organ-
isms can normally be seen, but greater than five organisms per 100×
objective field is considered abnormal (Twedt, 1992). Confirmation was
made by measurement of the enterotoxin in the feces (Twedt, 1992;
Marks et al., 1999). A degenerate neutrophil (long arrow) and epithelial
cell (asterisk) are present. (Wright; HP oil.) (Courtesy of Denny Meyer
and Dave Twedt, Colorado State University.)
20 m
n FIGURE 8-23  Campylobacteriosis. Dog. A neutrophil is seen along
with two miniscule, pleomorphic, gull wing–shaped bacteria (arrow-
heads) in a fecal smear from a dog with diarrhea and Campylobacter spp.
isolated by bacterial culture. The bacteria observed in this fecal smear
from a dog with confirmed Campylobacter infection are much smaller
than those observed in Figs. 8-26 and 8-27. (Wright-Giemsa; HP oil.)
enterotoxins with positive culture or PCR is more conclusive for
clostridial diarrhea (Marks et al., 2011).
Pleomorphic fecal bacteria that exhibit gull-wing and
spiral morphology on dry-mount fecal cytology include
treponeme-like bacteria, Serpulina spp., Helicobacter spp.,
Anaerobiospirillum spp., and Campylobacter spp. (Figs. 8-23,
8-24, 8-25, 8-26, 8-27). It is unusual to observe pleomorphic
gull-wing and spiral-shaped bacteria in routine dry-mount fecal
cytology, and this should be reported as an abnormal finding
when observed in large numbers. These organisms are quite

20 m
n FIGURE 8-24  Campylobacteriosis. Dog. Same case as in Fig. 8-23.
A few miniscule, pleomorphic, gull wing–shaped bacteria (arrowheads) are
seen near a pigmented, squamous epithelial cell. (Wright-Giemsa; HP oil.)
20 m
n FIGURE 8-25  Campylobacteriosis. Dog. Same case as in Fig.
8-23. Gram stain of Campylobacter spp. cultured from the feces indi-
cates that the bacteria are gram-negative bacilli. (Gram stain; HP oil.)
small (0.5 to 1.0 μm × 5 to 10 μm) and can be easily overlooked
during microscopic examination, particularly when these organ-
isms are present in low numbers. The likelihood of observing
these microorganisms is enhanced by high-power examination
of areas of the preparation that contain a lesser amount of back-
ground flora, such as very thin areas of the preparation or areas
of the preparation that are mucus-rich because these microor-
ganisms localize to the mucus-rich mucosal surface. Diarrhea
has been associated with fecal isolation of bacteria from all of
these genera except for the treponeme-like bacteria, and bac-
teria from all of these genera have also been isolated from the
feces of asymptomatic dogs and cats (Bender et al., 2005; Brous-
sard, 2003; De Cock et al., 2004; Malnick et al., 1990; Misawa
et al., 2002; Rossi et al., 2008). Concurrent observation of fecal
neutrophils would support a bacterial cause of the diarrhea.
Protozoal, fungal, or pseudofungal (i.e., algal and oomycetal)
infections are occasionally diagnosed (Figs. 8-28A-D, 8-29, 8-30,
8-31, 8-32, 8-33, 8-34) using dry-mount fecal cytology (Graves
CHAPTER 8   Dry-Mount Fecal Cytology 255
20 m
n FIGURE 8-26  Spiriliform bacteria. Dog. Numerous fine, pleomor-
phic, gull wing–shaped, and spiriliform fecal bacteria consistent with
Serpulina spp. are a bit larger than those shown in Figs. 8-23 through
8-25. The dog presented with chronic mucoid diarrhea. The large num-
ber of spiriliform bacteria in this sample may reflect the mucoid nature
of the diarrhea because bacteria with this morphology are found in
large numbers within mucoid gastrointestinal tract secretions. (Wright-­
Giemsa; HP oil.)
20 m
n FIGURE 8-27  Treponeme-like bacteria. Dog. Numerous darkly
stained, spiriliform fecal bacteria that exhibit several complete convolu-
tions are seen in the fecal smear of a dog with diarrhea. These bacteria are
larger than those shown in Figs. 8-23 through 8-25. Examination of a fecal
wet-mount may be useful to more conclusively identify these as a non-
pathogenic treponeme-like bacterium, which exhibits very rapid forward
motility in the fluid medium (Broussard, 2003). (Wright-Giemsa; HP oil.)
et al., 2005) or rectal scrape (Chapman et al., 2009). Tritricho-
monas foetus (Fig. 8-28) is a protozoal cause of feline diarrhea.
Trophozoites may be observed extracellularly in direct smears of
fresh feces from cats (Payne and Artzer, 2009). Nonsporulated
endospores of the algae Prototheca may appear morphologi-
cally similar to incidentally observed, round or oval, extracel-
lular fecal yeast described previously (Figs. 8-2, 8-3, 8-4, 8-5).
Incidental yeast should not be observed intracellularly within
macrophages, whereas Prototheca should be observed both intra-
cellularly within macrophages (Fig. 8-33) and extracellularly.
256 Canine and Feline Cytology

B 10 m

20 m
A

10 m 20 m
C D
n FIGURE 8-28  Tritrichomonas foetus. Cat. Same case A-D. A, Tritrichomonas foetus trophozoite is shown with a central axostyle (asterisk),
undulating membrane (thin arrow), and three anterior flagella (arrowhead). Trichomonads may be diagnosed by direct fecal smear or wet-mount with
iodine stain, but flotation solutions will destroy trophozoites. Tritrichomonas foetus may be difficult to distinguish from Giardia spp. and nonpatho-
genic Pentatrichomonas hominis (Payne and Artzer, 2009). Note the mixed bacilli and treponeme-like bacteria in the background. (Wright-Giemsa;
HP oil.) B, Four Tritrichomonas foetus trophozoites are shown from the feces of a diarrheic cat. Two degenerate neutrophils containing many bacilli
are also present. Note the extracellular treponeme-like bacteria in the background. This patient had a negative ELISA test for Giardia spp. antigen
and responded well to ronidazole treatment for Tritrichomonas spp. (Wright-Giemsa; HP oil.) C, Four Tritrichomonas foetus trophozoites are shown.
Many neutrophils are seen, some of which contain phagocytized bacteria. (Wright-Giemsa; HP oil.) D, Several Tritrichomonas foetus trophozoites are
shown with two neutrophils, containing phagocytized bacteria. (Wright-Giemsa; HP oil.)

20 m
n FIGURE 8-29  Giardiasis. Dog. Giardia trophozoites are shown from
the feces of a dog with diarrhea. These flagellate protozoan organisms are
pyriform with two apical nuclei. It is uncommon to diagnose Giardia using
dry-mount fecal cytology. Giardia trophozoites may be more easily iden-
tified in fecal wet-mounts, although care should be taken to distinguish
them from trichomonads, which have a single nucleus, prominent central
axostyle, and an undulating membrane. As opposed to Giardia cysts, tro-
phozoites that are fecally shed are labile and rapidly die after elimination.
Giardia cysts are usually not detected by dry-mount fecal cytology; fecal
flotation is usually required for this purpose. (Wright-Giemsa; HP oil.)
20 m
n FIGURE 8-30  Entamoebiasis. Dog. Two Entamoeba histolytica
protozoal organisms are present along with a single neutrophil (upper
left corner) in the feces of a dog with diarrhea. (Wright-Giemsa; HP oil.)
(Courtesy of Rick Alleman, University of Florida.)
CHAPTER 8   Dry-Mount Fecal Cytology 257
25 m
n FIGURE 8-31  Blastocystosis. Rectal scraping. Dog. Several
organisms and well-differentiated epithelial cells are present in a back-
ground of mixed bacteria in a rectal scrape from a dog with diarrhea.
The intestinal organism is most consistent with Blastocystis spp., an
algal-like protist (stramenopile). The binucleated forms support this
identification, rather than Iodamoeba bütschlii, which can look similar.
(Aqueous Romanowsky; HP oil.) (Courtesy of Craig Thompson, Purdue
University.)
20 m
n FIGURE 8-32  Cryptococcosis. Dog. Three budding (top) and one
nonbudding (lower right) Cryptococcus organisms are present with a
few bacilli in the feces of a dog with diarrhea. Most forms of Cryptococ-
cus develop a thick, polysaccharide capsule that does not stain and is
represented by the wide, colorless area surrounding the narrow-based–
budding, purple yeast. (Wright-Giemsa; HP oil.)
258 Canine and Feline Cytology
n FIGURE 8-33  Oomycosis. Dog. A branching oomycete is shown
surrounded by neutrophils, nucleoproteinaceous material, and erythro-
cytes in this rectal scraping from a dog with a history of bloody, mucoid
diarrhea and inappetence. Other cytologic findings not shown consisted of
marked mixed inflammation, which was primarily neutrophilic with lesser
macrophagic and eosinophilic inflammation. An oomycete, consistent
with Pythium or Lagenidium spp., was cultured. (Wright-Giemsa; HP oil.)
(Bar = 10 μm.)
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