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Contents lists available at ScienceDirect

Veterinary Microbiology
journal homepage: www.elsevier.com/locate/vetmic

The clinical significance of Cyniclomyces guttulatus in dogs

with chronic diarrhoea, a survey and a prospective treatment
study
Paul J.J. Mandigers a,*, Mirjam B.H.M. Duijvestijn b, Nynke Ankringa c,
Sofie Maes d, Elise van Essen b, Anky H.W. Schoormans b, Alexander J. German e,
Dirk J. Houwers b
a
Department of Clinical Sciences of Companion Animals, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 108,
3584 CM Utrecht, The Netherlands
b
Department of Infectious Diseases and Immunology, Clinical Infectiology Division, Faculty of Veterinary Medicine, Utrecht University,
Yalelaan 1, 3584 CL Utrecht, The Netherlands
c
Department of Pathobiology, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, 3584 CL Utrecht, The Netherlands
d
Valuepath, Laboratory Veterinary Pathology, PO BOX 144, 6300 AC Valkenburg a/d Geul, The Netherlands
e
Department of Obesity and Endocrinology, University of Liverpool, Leahurst Campus, Chester High Road, NESTON Wirral, CH64 7TE,
United Kingdom

A R T I C L E I N F O A B S T R A C T

Article history: This study surveyed the prevalence of massive numbers of Cyniclomyces guttulatus in
Received 18 February 2014 faecal samples from healthy dogs (18%) and dogs with chronic diarrhoea (14%) suggesting
Received in revised form 8 May 2014 that this yeast has no clinical significance. Subsequently, a total of 57 referred dogs with
Accepted 10 May 2014 chronic diarrhoea were selected because they excreted massive numbers of C. guttulatus
and their initial diagnostic work-up yielded no other direct clues explaining their
Keywords: diarrhoea.
Dogs Treatment with nystatin did not result in any clinical response in 36 out of these 57
Cyniclomyces guttulatus
dogs (63%), although they no longer shed the yeast. However, a response was noted in
Diarrhoea
the remaining 21 (37%) dogs: 13 were ‘responders’, in that their diarrhoea subsided for
Yeast
more than two weeks and the faeces were cleared of the yeast. However, three of these
dogs relapsed repeatedly, with signs of diarrhoea and massive shedding of the yeast.
The other eight dogs were ‘incomplete responders’, whereby faecal quality initially
normalised, but diarrhoea relapsed within two weeks, whilst still not shedding the
yeast. In these cases, further diagnostic work up revealed other co-causes of diarrhoea.
It was concluded that there was no direct evidence that C. guttulatus is a primary
pathogen. However, the results of the prospective treatment study suggest that a
possible role in a minority of cases, perhaps as an opportunist, cannot be ruled out.
ß 2014 Elsevier B.V. All rights reserved.

* Corresponding author. Tel.: +31 302534165.

E-mail address: [email protected] (Paul J.J. Mandigers).

http://dx.doi.org/10.1016/j.vetmic.2014.05.018
0378-1135/ß 2014 Elsevier B.V. All rights reserved.

Please cite this article in press as: Mandigers, P.J.J., et al., The clinical significance of Cyniclomyces guttulatus in dogs with
chronic diarrhoea, a survey and a prospective treatment study. Vet. Microbiol. (2014), http://dx.doi.org/10.1016/
j.vetmic.2014.05.018
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2 P.J.J. Mandigers et al. / Veterinary Microbiology xxx (2014) xxx–xxx

1. Introduction (n = 2), from various regions of the Netherlands. At the time

of sampling, breed or type of dog was recorded as well as
The yeast Cyniclomyces guttulatus (formerly called age, gender and faecal consistency.
Saccharomycopsis guttulata) is commonly found in faeces
of rodents and rabbits (Boundy-Mills and Miller, 2011; 2.1.2. Diarrhoeic dogs
Richle and Scholer, 1961; Zierdt et al., 1988). It is a normal These data were obtained by searching the database of
inhabitant of the gastrointestinal (GI) tract of these the Veterinary Microbiological Diagnostic Center at
animals, residing on the mucous layer in the gastric Utrecht University (VMDC) for submissions of canine
fundus and pylorus despite the acidic conditions (pH 2– faeces samples for parasitological examination of which
2.5) (Shifrine and Phaff, 1958). Large numbers of C. the history mentioned ‘diarrhoea’ and ‘massive C. guttu-
guttulatus are passed with the faeces into the environment latus’ as the only result. This search covered the period 1-1-
(Boundy-Mills and Miller, 2011) where they may persist 2005 to 20-9-2006.
for a prolonged periods of time due to their ability to form
ascospores (Shifrine and Phaff, 1958). 2.1.3. Analytical methods
Houwers and Blankenstein (2001) were the first to All faecal samples were analysed within 24–48 h at the
suggest that the yeast might be of clinical significance in VMDC. The analysis was performed with the Centrifuge-
dogs, when they observed large numbers of Cyniclomyces- analysis Sedimentation-Flotation Method (CSF method)
like organisms in approximately 15% of canine faecal using Zinc Sulphate (ZnSO4) as a flotation fluid with a
samples submitted to their diagnostic laboratory for specific gravity of 1.34. Briefly, 1 g of faeces was suspended
parasitological examination, mainly for the investigation in 14 mL of tap water, vortexed and centrifuged for 1 min at
of chronic diarrhoea. As some of these patients responded 2950  g. The supernatant was removed; the sediment
remarkably to oral nystatin, they suggested that the yeast suspended in the ZnSO4 solution, the tube was completely
could be of clinical significance, most probably as a direct filled (to produce a convex meniscus). A cover slide was
result of their massive presence in the gut. They also then placed on top of the tube, which was then centrifuged
suggested that the yeast probably colonised a niche in the for 2 min at 1660  g. The cover slide was carefully
patient’s altered intestinal flora. However, no clinical removed and placed on a microscope slide and examined.
efficacy studies were performed and there was limited
follow up of the treated dogs. In a more recent study, large 2.1.4. Microscopic identification
numbers of C. guttulatus were found in the faeces and Slides were studied using a magnification of 400. Five
stomach washings of three dogs suffering from recurrent different microscopic fields were observed to identify C.
vomiting and diarrhoea, and small numbers in a quarter guttulatus, morphologically identified by its characteristic
of the faecal samples from healthy dogs (Flausino et al., ‘spectacle case’ shape (Fig. 1), and a length of approxi-
2012). To date, two case reports have documented the mately 1/5th of the diameter of a Toxocara canis egg.
successful treatment of chronic diarrhoea associated Numbers of C. guttulatus were semi-quantitatively
and massive C. guttulatus shedding with nystatin, one in a assessed as follows:
cat (Peters and Houwers, 2009) and one in a rabbit
(Hersey-Benner, 2008). - No organisms or <50 organisms in total over five
Therefore, currently, the clinical significance of identi- microscopic fields (negative).
fying large numbers C. guttulatus in the faeces of dogs with - 50 to <200 organisms in total over five microscopic fields
chronic diarrhoea is not known. Hence, a survey on the (intermediate).
prevalence of the yeast in healthy and diarrhoeic dogs was - Clear presence, usually in massive – innumerable –
performed, followed by a prospective study on the effects numbers (positive); >200 organisms in total over five
of nystatin treatment in dogs with chronic diarrhoea microscopic fields (positive).
excreting large numbers of C. guttulatus. Given that the it
was also not clear whether this yeast has the potential to Low numbers (<200) were not regarded as positive
induce inflammation, a hallmark of primary pathogens, scores in view of the possibility that they were just
histopathological changes in gastrointestinal biopsies contaminants from oral intake and, thus, not originating
taken during investigations were also reviewed. The null from an actual infection.
hypothesis was that excretion of large numbers of C.
guttulatus is an insignificant finding, does not any 2.1.5. Culture and polymerase chain reaction
histological changes in the stomach or intestine, and that In order to substantiate the morphological identifica-
treatment with nystatin would be ineffective. tion, an in-house conventional PCR for detection of C.
guttulatus was developed as described by others (Kurtz-
man and Robnett, 1998). Rabbit faeces containing C.
2. Material and techniques
guttulatus was cultured on Sabouraud’s dextrose agar,
2.1. Survey under microaerophilic conditions, at 37 8C for a minimum
of 5 days. Light brown colonies were microscopically
2.1.1. Healthy dog population identified as C. guttulatus. DNA from these colonies and
Over a period of 5 months, a total of 140 faecal samples DNA extracted from the rabbit faeces was subjected to the
were obtained from healthy dogs at walking services PCR; the product had a similar size as the product
(n = 4), veterinary practices (n = 13), and animal shelters described by Kurtzman and Robnett (1998). Subsequently,

Please cite this article in press as: Mandigers, P.J.J., et al., The clinical significance of Cyniclomyces guttulatus in dogs with
chronic diarrhoea, a survey and a prospective treatment study. Vet. Microbiol. (2014), http://dx.doi.org/10.1016/
j.vetmic.2014.05.018
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Fig. 1. C. guttulatus characteristic spectacle case-shape (photo: Barend Blankenstein, VMDC).

six microscopically positive canine faecal samples were albumin, globulins, and protein electrophoresis), serum
subjected to culture and PCR, with the amplicons from the trypsin-like immunoreactivity (TLI), faecal parasitology as
suspect colonies and the faeces having a size similar to that described above, and faecal bacteriology (for Salmonella
of the rabbit isolate. spp. and Campylobacter spp.). All faecal examinations were
performed at the VMDC. If deemed necessary, survey
2.2. Prospective study of referral cases with chronic diarrhoea abdominal radiography and abdominal ultrasonography
with massive excretion of C. guttulatus were performed. Finally, upper gastrointestinal endoscopy
was performed in all patients, as well as ileum- and
2.2.1. Patients colonoscopy in patients where it was deemed to be
From a cohort of 300 dogs referred between January diagnostically relevant. During this procedure, multiple
2005 and September 2006 for the investigation and mucosal biopsies were collected from the stomach (corpus
management of chronic gastrointestinal disease (Mandi- and fundus), upper small intestine (duodenum and, where
gers et al., 2010), a total of 57 patients were selected that feasible, jejunum), ileum and colon for standard histo-
appeared to shed massive numbers of C. guttulatus in their pathological analysis. The results were used in the case
faeces, based on a standard parasitological examination management by excluding cases not consistent with
(CSF) at VMDC. Dogs were included, in this part of the chronic enteropathy. All slides of the included dogs were
study, if clinical signs had been present for at least three subsequently assessed by a European-board-certified
weeks and a complete diagnostic work-up (including veterinary pathologist (SM) and a resident in veterinary
endoscopy) had been performed to exclude other causes. pathology (NA) and graded using recent internationally
Dogs that had received antimicrobials and/or corticoster- accepted criteria (Jergens et al., 2012; Washabau et al.,
oid therapy in the three weeks prior to their referral were 2010). Special attention was given to the presence of the
excluded. All owners were informed of the purpose of the yeast and potentially associated histological changes.
study and consented to their dog participating. All
examinations were performed by the same clinician 2.2.3. Treatment study
(PJJM). All dogs received 150,000 IE nystatin1 q8h PO for five
consecutive days. This dosing regimen was based on the
2.2.2. Diagnostic work-up dose suggested for Candida spp. infection in dogs (Jenkins
A complete history was taken and a physical examina- and Boothe, 1987). No concurrent therapy was adminis-
tion performed. Subsequently, detailed laboratory inves- tered during the treatment with nystatin.
tigations were performed including a complete blood
count, a serum chemistry profile (e.g. urea, creatinine,
alkaline phosphatase, alanine aminotransferase, total 1
Nystatine Labaz1, oral suspension, 100,000 IU/mL, Sanofi-Aventis the
bilirubin, bile acids, calcium, phosphate, total protein, Netherlands BV.

Please cite this article in press as: Mandigers, P.J.J., et al., The clinical significance of Cyniclomyces guttulatus in dogs with
chronic diarrhoea, a survey and a prospective treatment study. Vet. Microbiol. (2014), http://dx.doi.org/10.1016/
j.vetmic.2014.05.018
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4 P.J.J. Mandigers et al. / Veterinary Microbiology xxx (2014) xxx–xxx

2.2.4. Follow-up Table 1

Faecal samples analysed for the presence of C. guttulatus and categorised
During the study, patient owners recorded their
into three groups. The total number of dogs found to shed the yeast
observations with respect to gastrointestinal signs and massively was 18%. Although the number of dogs, housed in the animal
any changes to the normal routine. The study performer shelters, found positive was low, the difference between the three groups
(PJJM) viewed this record, at the evaluation visit, is statistically not significant (P = 0.18).
approximately 7 days after the start of the nystatin Negative Intermediate Positive
treatment. Dogs that had responded completely to the
Walking services 21 3 6
nystatin therapy received no additional therapies. All
Animal shelters 18 0 1
owners were asked to submit a faecal sample 14 days after Practitioners 70 3 18
the start of the treatment. If these samples were positive
109 6 25
for C. guttulatus, the nystatin therapy was repeated, if a
negative result was obtained but diarrhoea still occurred,
further diagnostic work-up was performed and other
treatments were initiated. 3.2. Prospective treatment study

2.2.5. Classification of cases Of 300 patients referred during the timeframe of the
Classification of cases was based upon the findings of study, 57 dogs fitted the inclusion criteria. Dog breeds
the follow-up evaluations with reference to the informa- represented included a large variety of breeds such as Akita
tion provided by the owner. If the owner observed no Inu (n = 1), American Bulldog (n = 1), Basset (n = 1), Bernese
change in the frequency and severity of clinical signs after mountain dog (n = 6), Boerboel (n = 1), Bordeaux dog
two courses of nystatin, the dog was regarded a ‘non- (n = 1), Border collie (n = 1), Bouvier (n = 2), Boxer (n = 5),
responder’. If a dog responded, i.e. returned for more than Cane corso (n = 2), Dobermann (n = 2), Shepherd (n = 4),
two weeks to what the owner classified as normal prior to English bulldog (n = 2), Fox (n = 1), German Shepherd
onset of signs, to the first or second nystatin treatment and (n = 4), Golden retriever (n = 7), Great Dane (n = 1), Great
the faecal examination was negative for C. guttulatus, it was Swiss (n = 1), Greyhound (n = 1), Jack Russel Terrier (n = 1),
classified as a ‘responder’. Dogs that only responded Labrador retriever (n = 2), Mongrel (n = 1), Ploski Owczarek
temporarily, i.e. less than two weeks, to the treatment(s) Nizziny (n = 1), Rottweiler (n = 5), Stabij (n = 1) and
were classified as ‘incomplete responders’. Weimeraner (n = 2). Twenty-seven dogs were female of
which 16 were neutered and 30 dogs were males of which
2.2.6. Data handling and statistics six were castrated and their mean age was 5.2  3.43 years
Statistical analyses were performed using a Chi-square (range 3.7 months to 11.1 years; median 5.1 years). Twenty-
test with a computer software program,2 and descriptive seven dogs lived in a larger city, 13 semi-rural and 17 rural.
statistics were used to report baseline data and reported as Twenty-six of the owners had seen their dog eating rabbit
median and range. droppings, whilst the others had either not observed this
behaviour or were uncertain as to whether such behaviour
had occurred. The dogs were fed a variety of diet food (50 out
3. Results 57) as well as home-cooked food (7 out 57).

3.1. Prevalence study 3.3. Clinical signs and baseline diagnostic findings
A total of 140 faecal samples of clinically healthy dogs
The preceding median duration of the clinical signs was
were analysed: 115 (79%) samples were negative or
142 days (range 23–2588 days). Clinical signs of small
intermediate and 25 (18%) were C. guttulatus positive.
bowel diarrhoea were seen in 47 out 57 dogs (82%), large
Dogs housed in animal shelters tended to have a lower
bowel diarrhoea in 5 out 57 (9%), and signs of both small
number of positive faecal samples (1 out 19; 5%) compared
and large bowel diarrhoea were seen in 5 out 57 (9%). No
to the dogs sampled from walking services (6 out 30; 20%)
significant abnormalities were seen on routine haemato-
and the veterinary practices (18 out 91; 20%) (Table 1).
logical and serum biochemical analysis, except for
There was no statistical significant difference noted
hypoalbuminaemia, which was identified in 14 out of 57
(P = 0.18). Between 1-1-2005 and 20-9-2006, VMDC
dogs (25%).
received 1564 canine faecal samples for parasitological
analysis. The vast majority originated from dogs with
3.4. Histopathology
diarrhoea lasting for more than a week. A total of 215 (14%)
samples were reported as massive shedders.
Gastric biopsies were available of 23 of the 47 dogs that
There was no statistical significant difference in the
presented with chronic small bowel diarrhoea. Lympho-
prevalence of C. guttulatus in samples from healthy dogs
plasmacytic inflammation was identified in six dogs, with
and those with diarrhoea (P = 0.18). Neither was there a
eosinophilic gastritis being identified in a further six.
correlation for C. guttulatus excretion with age, gender or
However, C. guttulatus was seen on the inner surface of the
breed in the 140 healthy dogs.
stomach in only three cases, and there was no evidence of
mucosal invasion or intracellular organisms. Duodenal/
jejunal biopsies were available from all 47 dogs with
2
SPSS 22, IBM company. chronic small bowel diarrhoea and from an additional two

Please cite this article in press as: Mandigers, P.J.J., et al., The clinical significance of Cyniclomyces guttulatus in dogs with
chronic diarrhoea, a survey and a prospective treatment study. Vet. Microbiol. (2014), http://dx.doi.org/10.1016/
j.vetmic.2014.05.018
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dogs with mixed diarrhoea. The biopsies of six dogs 4. Discussion

showed no abnormalities. All remaining dogs showed
various abnormalities, generally mild-to-marked lympho- Although C. guttulatus was initially described more than
plasmacytic infiltrates (19), mild-to-marked eosinophilic 50 years ago (Parle, 1956, 1957; Shifrine and Phaff, 1958) it
infiltrates (10), and mild-to-moderate mixed (i.e. lympho- only became the subject of clinical studies in animals
plasmacytic and eosinophilic) infiltrates (13). One dog had during the last decade. Some authors had suggested on the
a multifocal granulomatous lymphangitis. None in none of basis of a clinical response to nystatin treatment that the
the biopsies C. guttulatus was seen. massive presence of the yeast in GI tract could play a
Colonic biopsies were available from 10 dogs, five of causative role in cases of chronic diarrhoea in dogs
which were normal, four had mild eosinophilic colitis, and (Houwers and Blankenstein, 2001) and cats (Peters and
one had ulcerative colitis. Again, none of these biopsies Houwers, 2009). One study presented circumstantial
showed C. guttulatus. As result, there was no host evidence that it could act as an opportunistic pathogen
inflammatory reaction to the yeast in any of the dogs. in dogs, but it was also found in 21% of wet mounts of the
faeces of 60 healthy dogs (Flausino et al., 2012). However,
3.5. Effect of nystatin treatment others saw the presence of the yeast as a clinically non-
significant finding in dogs (Boundy-Mills and Miller, 2011).
Thirty-six out of fifty-seven dogs (63%) did not clinically This is the first study investigating the prevalence of the
respond to the first treatment with nystatin, yet all but two yeast C. guttulatus in dogs suffering from chronic diarrhoea
faecal samples taken 14 days after the start of the and those without, combined with data regarding medical
treatment were found to be clear of C. guttulatus. The intervention to clear this yeast infection. Microscopic
two positive dogs were treated again and, although they detection of the yeast was confirmed using a species-
did not respond, the repeat faecal samples were clear. specific PCR.
According to the owners, clinical signs did resolve in 21/ In the present study in the Netherlands, the observed
57 (37%) dogs after the first treatment, 13 of which had prevalence in healthy dogs is 18%, which is comparable
negative faecal results and were, thus, classified as with the prevalence in Brazil (Flausino et al., 2012). To
‘responders’. The remaining eight of these responding exclude any regional, sex or breed bias faecal samples were
dogs, continued to shed large numbers of C. guttulatus in obtained from different regions in the Netherlands, and
their faeces, despite resolution of signs, and diarrhoea different breeds and housing situations. It is, therefore,
relapsed, within two weeks. At this stage, a second nystatin concluded that the yeast has a relatively high prevalence in
course was administered, and signs resolved in three of healthy dogs. Since most samples contained massive
these eight dogs, with faecal samples being negative. numbers it is likely that C. guttulatus actually multiplied
However, these three dogs returned to the clinic after in the dogs and thus that the dogs were truly colonised. It
several months, again with diarrhoea and massive excre- should be noted that the CSF method used in this study
tion of the yeast. They were treated once more, and again leads to considerable enrichment and may, therefore, be a
responded with negative faecal samples and resolution of more sensitive method than wet mount detection.
clinical signs. This cycle of events occurred many times The environmental source of this organism is unclear,
thereafter. Hence, these three were also classified as but one possibility is exposure to rabbit faeces. This is in
‘responders’. line with finding that the organism was less commonly
Four dogs out of these eight responding dogs still isolated from dogs in animal shelters that usually only
shedding C. guttulatus, only recovered completely after an have access to secluded outdoor areas. Given that rabbits
additional antibiotic treatment. These dogs were classified are ubiquitous in areas of countryside, it is likely that at
as ‘incomplete responders’. One dog out of these eight least some of the pet dogs not excreting C. guttulatus were
responding dogs still shedding C. guttulatus, was subse- also exposed. So, the negative dogs were almost certainly
quently found to be positive for T. canis and completely exposed but did not become infected, possibly as a result of
recovered after appropriate therapy with fenbendazol.3 absence of particular conditions in the stomach lining or
This dog was also classified as an ‘incomplete responder’. the lack of a niche in the gastrointestinal environment.
Therefore, of the 21 dogs in which owners reported a The prevalence of the yeast in dogs suffering from
clinical response to nystatin, 16 were classified as chronic diarrhoea was 14%, which corroborates the 15% in
‘responders’, and five as ‘incomplete responders’. Six of the earlier study (Houwers and Blankenstein, 2001), and is
the ‘responders’ were followed for more than a year, and statistically not significantly different. This finding, sup-
they remained free of diarrhoea; three of these dogs ports the null hypothesis that massive excretion of C.
returned for review at regular intervals and were each time guttulatus may not have clinical significance. Thus, the
again shedding the yeast. However, the remaining seven presence of chronic diarrhoea, whatever the cause, does
‘responders’, subsequently returned after signs of diar- not increase the likelihood of infection with the yeast,
rhoea had relapsed after several weeks (at least >4); in all which is interesting. As it has been suggested that
cases, their faecal samples were still free of the yeast, and extensive colonisation with C. guttulatus may exacerbate
further examination of these patients revealed various an existing imbalance in gastrointestinal function and
other causes. trigger or exacerbate diarrhoea (Houwers and Blanken-
stein, 2001).
The effect of monotherapy with oral nystatin was
3
Fenbendazol 50 mg/kg for 3 days. Brandname Panacur1 MSD. prospectively studied in a group of 57 dogs with chronic

Please cite this article in press as: Mandigers, P.J.J., et al., The clinical significance of Cyniclomyces guttulatus in dogs with
chronic diarrhoea, a survey and a prospective treatment study. Vet. Microbiol. (2014), http://dx.doi.org/10.1016/
j.vetmic.2014.05.018
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6 P.J.J. Mandigers et al. / Veterinary Microbiology xxx (2014) xxx–xxx

diarrhoea and massive faecal shedding of C. guttulatus. The guttulatus might lead to diarrhoea through an alternative
overall response rate observed in this study was 21/57 mechanism. However, further studies would be needed to
(37%) of which 16 were categorised as ‘responders’, after examine such a possibility.
one or repeated treatment courses, and five as ‘incomplete
responders’ that became diarrhoeic again within weeks. 5. Conclusion
Notably, a Cyniclomyces-negative faecal sample was seen
in every ‘responder’, whereas ‘incomplete responders’ once In conclusion, the survey results and the histology
again showed massive faecal shedding after several weeks. supported the null hypothesis, and there was no direct
Conversely, the fact that only 2/36 ‘non-responders’ shed evidence that C. guttulatus is a primary pathogen. However,
the yeast after a single nystatin course, suggested that the the results of the prospective treatment study did
therapy was at least effective in eliminating it from the demonstrate that some dogs with chronic diarrhoea and
gastrointestinal tract. Therefore, the five ‘incomplete massive C. guttulatus shedding improve after nystatin
responders’ probably relapsed because either the yeast therapy. Thus, a possible role in a minority of cases,
was not entirely eliminated or re-infection occurred perhaps as an opportunist, cannot be ruled out. Further
rapidly, which is obviously hard to prevent in the average studies, including randomised controlled trials, would now
dog walking situation. be recommended in order better to define the true
In the case 6/16 ‘responders’, the clinical response was significance of this organism and the benefit of therapeutic
long-lasting suggesting that the C. guttulatus might have eradication.
had a causal role (either primary or secondary) of in the
clinical signs. In another three dogs, a clear association Conflict of interest
between signs if diarrhoea and the presence of C. guttulatus
in the faeces, in that massive yeast shedding was noted The authors report no conflict of interest.
only when these dogs had diarrhoea. Such a finding again
suggests a possible role for C. guttulatus in some cases of
Acknowledgements
chronic diarrhoea. However, in the remaining seven
‘responders’, the diarrhoea relapsed after more than two
The authors thank all owners for their participation in
weeks without massive yeast excretion. Further investiga-
this study and Mrs. Marieke van der Kooy with her help in
tion of these patients revealed various other causes, which
preparing this manuscript.
were subsequently successfully leading to clinical resolu-
tion. The significance of C. guttulatus in these cases is less References
clear, be it an incidental finding or a possible exacerbating
factor. Boundy-Mills, K., Miller, M.W., 2011. Cyniclomyces van der Walt & D.B.
It is possible that the nystatin was acting against other Scott (1971). In: Kurtzman, C.P., Fell, J.W., Boekhout, T. (Eds.), The
Yeasts, A Taxonomic Study. Elsevier, San Diego, pp. 357–360.
elements of the gastrointestinal microbiota and not Flausino, G., Leal, P.D., McIntosh, D., Amaral, L.G., Teixeira Filho, W.L.,
against C. guttulatus. However, since nystatin affects Flausino, W., Lopes, C.W., 2012. Isolation and characterization of
ergosterol-binding sites which are only present in fungal Cyniclomyces guttulatus (Robin) Van Der Walt and Scott, 1971 in dogs
in Brazil. Curr. Microbiol. 65, 542–546.
cell wall, it is unlikely that bacterial components of the Foster, M.L., Dowd, S.E., Stephenson, C., Steiner, J.M., Suchodolski, J.S.,
gastrointestinal microbiota were affected. Although a 2013. Characterization of the fungal microbiome (mycobiome) in
recent molecular analysis of the fungal microbiome fecal samples from dogs. Vet. Med. Int. 2013, 658373.
Hersey-Benner, C., 2008. Diarrhea in a rabbit. Cyniclomyces guttulatus
demonstrated that the DNA of many fungal species can
yeast. Lab Anim. 37, 347–349.
be present in canine faeces (Foster et al., 2013), only low Houwers, D.J., Blankenstein, B., 2001. Cyniclomyces guttulatus and diar-
numbers of yeasts can usually be cultured from jejunal rhea in dogs. Tijdschr. Diergeneesk. 126, 502.
chyme and faeces of healthy dogs (Mentula et al., 2005). Jenkins, W.L., Boothe, D.M., 1987. Amphotericin B, nystatin, flucytosine,
imidazoles, griseofulvin. In: Johnston, D.E. (Ed.), The Bristol Handbook
Further, if substantial numbers of other fungal species of Antimicrobial Therapy. , Evansville.
(including yeasts) were a causative factor, they arguably Jergens, E., Evans, R., Suchodolski, J., Allenspach, K., Garden, O.A., Day, M.,
should have been identifiable in either faecal samples or Ackermann, M., Hostetter, J., Steiner, J.M., 2012. Design of a simplified
histopathologic model for gastrointestinal inflammation in dogs and
tissue biopsies. Thus, if the nystatin were having a genuine cats. In: ECVIM-CA Congress, Maastricht, p. 262.
effect, it is most likely to have been due to eradication of Kurtzman, C.P., Robnett, C.J., 1998. Identification and phylogeny of asco-
the C. guttulatus. That said, the trial was uncontrolled, and mycetous yeasts from analysis of nuclear large subunit (26S) ribo-
somal DNA partial sequences. Anton. Leeuw. 73, 331–371.
it is possible that the improvement were coincidental to Mandigers, P.J., Biourge, V., van, d.I., Ankringa, N., German, A.J., 2010. A
this therapy. Arguably, a prospective randomised con- randomized, open-label, positively-controlled field trial of a hydro-
trolled trial would be a better means of verifying that the lyzed protein diet in dogs with chronic small bowel enteropathy. J.
Vet. Intern. Med. 24, 1350–1357.
response was genuine.
Mentula, S., Harmoinen, J., Heikkila, M., Westermarck, E., Rautio, M.,
Although inflammatory changes were identified in the Huovinen, P., Kononen, E., 2005. Comparison between cultured
histological examination of both gastric and intestinal small-intestinal and fecal microbiotas in beagle dogs. Appl. Environ.
Microbiol. 71, 4169–4175.
biopsies, the lack of mucosal invasion suggested that they
Parle, J.N., 1956. The growth of Saccharomycopsis guttulata. Anton. Leeuw.
were not the direct effect of C. guttulatus. Thus, if this yeast 22, 237–242.
does have a pathogenetic role in cases of chronic diarrhoea, Parle, J.N., 1957. Yeasts isolated from the mammalian alimentary tract. J.
it is probably not through invasion into, or adherence to, Gen. Microbiol. 17, 363–367.
Peters, S., Houwers, D.J., 2009. A cat with diarrhoea associated with the
the inner surface layers of the GI tract. It is of course massive presence of Cyniclomyces guttulatus in the faeces. Tijdschr.
possible that the presence of massive numbers of C. Diergeneesk. 134, 198–199.

Please cite this article in press as: Mandigers, P.J.J., et al., The clinical significance of Cyniclomyces guttulatus in dogs with
chronic diarrhoea, a survey and a prospective treatment study. Vet. Microbiol. (2014), http://dx.doi.org/10.1016/
j.vetmic.2014.05.018
G Model
VETMIC-6624; No. of Pages 7

P.J.J. Mandigers et al. / Veterinary Microbiology xxx (2014) xxx–xxx 7

Richle, R., Scholer, H.J., 1961. Saccharomycopsis guttulata in rabbits: cultural guidelines for the evaluation of gastrointestinal inflammation in com-
properties and possible significance. Pathol. Microbiol. 24, 783–793. panion animals. J. Vet. Inter. Med./Am. Coll. Vet. Intern. Med. 24, 10–26.
Shifrine, M., Phaff, H.J., 1958. On the isolation, ecology and taxonomy of Zierdt, C.H., Detlefson, C., Muller, J., Waggie, K.S., 1988. Cyniclomyces
Saccharomycopsis guttula. Anton. Leeuw. 24, 193–209. guttulatus (Saccharomycopsis guttulata)—culture, ultrastructure and
Washabau, R.J., Day, M.J., Willard, M.D., Hall, E.J., Jergens, A.E., Mansell, J., physiology. Anton. Leeuw. 54, 357–366.
Minami, T., Bilzer, T.W., 2010. Endoscopic, biopsy, and histopathologic

Please cite this article in press as: Mandigers, P.J.J., et al., The clinical significance of Cyniclomyces guttulatus in dogs with
chronic diarrhoea, a survey and a prospective treatment study. Vet. Microbiol. (2014), http://dx.doi.org/10.1016/
j.vetmic.2014.05.018