Professional Documents
Culture Documents
Cyniclomyces Guttulatus
Cyniclomyces Guttulatus
Veterinary Microbiology
journal homepage: www.elsevier.com/locate/vetmic
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.
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
G Model
VETMIC-6624; No. of Pages 7
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
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
G Model
VETMIC-6624; No. of Pages 7
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
G Model
VETMIC-6624; No. of Pages 7
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
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
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