Disseminated Candidiasis in a Young, Previously Healthy, Dog and Review of Literature.

BACKGROUND: The reports on disseminated candidiasis in dogs so far describe at least one predisposing factor. This case report, however, highlights candidiasis in a dog without any known predisposition. PATIENT: A 1.5-year-old intact female Hovawart dog was presented with subcutaneous nodules and polyuria/polydipsia. An excisional biopsy revealed a chronic pyogranulomatous and necrotizing inflammation with mycotic structures. The patient became febrile and lethargic, and developed lameness. METHODS: A physical examination, blood tests, urinalysis, thoracic radiographs, abdominal ultrasonography of the abdomen, fine-needle aspiration biopsies, and a culture of a subcutaneous nodule aspirate were obtained. Selected sections of multiple organs were collected for routine histology postmortem. The isolate and a subcutaneous mass were subjected to molecular identification and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) analysis. RESULTS: Clinical, laboratory, and radiological findings were consistent with a granulomatous chronic systemic inflammation. Cytology and histology showed a pyogranulomatous and necrotizing inflammation with myriads of intra- and extra-cellular yeasts and extracellular hyphae. Culture yielded numerous yeast colonies, which appeared Candida albicans-like, but showed a negative serum test and a low identification in API 20 C AUX. Nucleic acid sequences showed homology with the C. albicans-type strain CBS 562. Multilocus sequence typing (MLST) resulted in a new type with designation DST121. The identification of the isolates was confirmed by MALDI-TOF-MS analysis. CONCLUSION AND CLINICAL IMPORTANCE: Future MLST typing and investigation of virulence can provide further evidence whether this MLST-type is associated with clinical cases of disseminated candidiasis without an apparent predisposing condition.

The clinical significance of Cyniclomyces guttulatus in dogs with chronic diarrhoea, a survey and a prospective treatment study.

This study surveyed the prevalence of massive numbers of Cyniclomyces guttulatus in faecal samples from healthy dogs (18%) and dogs with chronic diarrhoea (14%) suggesting that this yeast has no clinical significance. Subsequently, a total of 57 referred dogs with 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 diarrhoea. Treatment with nystatin did not result in any clinical response in 36 out of these 57 dogs (63%), although they no longer shed the yeast. However, a response was noted in the remaining 21 (37%) dogs: 13 were 'responders', in that their diarrhoea subsided for 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.

Prevalence of the causative agents of equine piroplasmosis in the South West of The Netherlands and the identification of two autochthonous clinical Theileria equi infections.

Equine piroplasmosis (EP) has not been considered indigenous in The Netherlands. However, following the detection of an apparently indigenous subclinical Babesia caballi infection in a horse on Schouwen-Duiveland (an island in the Zeeland Province), a survey was undertaken between May and September 2010 to assess the prevalence of the causative agents of EP in the South-West of The Netherlands. Blood samples from 300 randomly selected horses were tested for specific antibodies against Theileria equi and B. caballi using an indirect fluorescence antibody test (IFAT), and for parasite DNA using a specific polymerase chain reaction combined with reverse line blotting (PCR-RLB). Twelve of the horses (4%) were seropositive for EP. Of these, nine (75%) were positive (titre⩾1:160) for B. caballi alone and three (25%) were also positive for T. equi. PCR-RLB detected T. equi DNA in five horses (1.6%), two of which were seronegative. Four (1.3%) of the positive horses (three positive for T. equi and one for both B. caballi and T. equi) were considered truly indigenous. During the study, two indigenous ponies from a farm situated outside the sampling area were diagnosed with acute clinical piroplasmosis characterized by severe anaemia and pyrexia. Blood smears showed T. equi - like inclusions in red blood cells, and T. equi infection was confirmed in both ponies by PCR-RLB. The initial subclinical B. caballi infection, the survey results and the two acute clinical EP cases confirmed the autochthonous transmission of B. caballi and T. equi infections in The Netherlands.

Antibodies to a novel leptospiral protein, LruC, in the eye fluids and sera of horses with Leptospira-associated uveitis.

Screening of an expression library of Leptospira interrogans with eye fluids from uveitic horses resulted in identification of a novel protein, LruC. LruC is located in the inner leaflet of the leptospiral outer membrane, and an lruC gene was detected in all tested pathogenic L. interrogans strains. LruC-specific antibody levels were significantly higher in eye fluids and sera of uveitic horses than healthy horses. These findings suggest that LruC may play a role in equine leptospiral uveitis.

[A cat with diarrhoea associated with the massive presence of Cyniclomyces guttulatus in the faeces]. / Een geval van diarree geassocieerd met Cyniclomyces guttulatus (brillendoosjesgist) bij de kat.

This is the first report documenting the presence of a high number of Cyniclomyces guttulatus yeasts in the faeces of a cat. The animal was initially presented with acute complaints of vomiting and diarrhoea. The patient responded well to oral salazosul-fapyridine but the stools remained soft and C. guttulatus yeasts were still present. After a course of nystatin (15,000 IU/kg bw q24 PO for 4 days) the stools were normal and no yeasts were found anymore (centrifugation/flotation/zinc sulphate). C. guttulatus occurs naturally in the digestive tract of rabbits, guinea pigs, chinchillas, rats and mice. It is occasionally found in massive numbers in the faeces of dogs with diarrhoea; part of these patients respond well to nystatin treatment. Recent experience indicates that the most effective dosage of nystatin for dogs and cats is 50.000 IU/kg q24 PO for 4 days.

Strain variation within Campylobacter species in fecal samples from dogs and cats.

To investigate the incidence of co-colonization of different strains of Campylobacter species present in canine and feline stool samples, isolates were recovered by culture from 40 samples from dogs (n=34) and cats (n=6). Animals were of different ages, with diarrhoea or without clinical signs. Three isolation procedures were used: two selective agars and a filtration method. In each stool sample, multiple colonies were identified to the species level by PCR, subsequently genotyped by Amplified Fragment Length Polymorphism (AFLP) and pattern similarities (451 isolates) were calculated to investigate their phylogenetic relationships. Genetic heterogeneity of strains in individual stool samples was detected within the species Campylobacter jejuni, C. upsaliensis and C. helveticus, though to a different degree in dogs and cats. In 3 of the 34 (9%) canine samples, more than one genotype of the same Campylobacter species was present, while strain variation was detected in four of the six feline samples. The results show that preferably, multiple colonies should be analyzed in molecular epidemiological and aetiological studies.

A new PCR followed by MboI digestion for the detection of all variants of the Clostridium perfringens cpb2 gene.

Clostridium perfringens which is a causative agent of several diseases in animals and humans is capable of producing a variety of toxins. Isolates are typed into five types on the basis of the presence of one or more of the four major toxins genes, i.e. cpa, cpb, etx, and iap. A decade ago another toxin termed beta2 (beta2) and its gene (cpb2) were identified. Two alleles of cpb2 are known and a possible link between differences in gene expression and allelic variation has been reported. A correlation between the level of expression and the origin of the isolates has also been suggested. The demonstration and typing of the cpb2 gene in the genome of isolates can be seen as a vital part of research on the role of the beta2 toxin in the pathogenesis of disease. This study describes a PCR with a single primer set which in contrast to published primer sets recognizes both alleles. Subsequent restriction enzyme analysis of the PCR product enables typing of the alleles. Applying this protocol on a total of 102 isolates, a sub-variant was found which occurred only in C. perfringens isolates from pigs and appeared to be the predominant variant found in C. perfringens isolates from this species.

Class 1 integrons in Enterobacteriaceae isolated from clinical infections of horses and dogs in the Netherlands.

Integrons in gentamicin- and cotrimoxazole-resistant Enterobacteriaceae from dogs and horses with clinical infections were analyzed by conserved segment PCR-RFLP. Five distinct integron types were found, most of which have previously been reported in Enterobacteriaceae isolated from humans and farm animals, indicating that resistance genes are exchanged between the reservoirs in humans, farm animals, and companion animals.

Alpha4-integrin (CD49d) expression on bovine peripheral blood neutrophils is related to inflammation of the respiratory system.

Neutrophil emigration from the pulmonary vasculature, is mediated by cellular adhesion molecules (CAM) expressed on the outer membranes of endothelial cells and neutrophils. Although beta(2)-integrin-dependent migration is a major mechanism of neutrophil migration, which was demonstrated by extensive invasion of neutrophils in pulmonary tissue of calves suffering from a genetic deficit in expression of beta(2)-integrins, termed bovine leukocyte adhesion deficiency (LAD), the role of alternative CAM is still unclear. We investigated whether an alternate CAM for beta(2)-integrin function, i.e. the alpha(4)-integrin, was expressed on peripheral blood neutrophils of calves. As we detected basal but significant expression, the effect of naturally acquired pulmonary infection on the expression of either integrin was determined, as an indication for its function in the migration process. In our experiments, basal expression of alpha(4)-integrins on peripheral blood neutrophils from clinically healthy calves was detected. On neutrophils of calves, experiencing field outbreaks of enzootic bronchopneumonia, higher expression of the alpha(4)-integrin was detected, which returned to normal after successful treatment of the disease. In addition, its level of expression was linearly related to plasma acute phase protein (haptoglobin) concentrations, which is a sensitive parameter for severity of respiratory inflammation. Increased expression of the alpha(4)-integrin on peripheral blood neutrophils during pulmonary inflammation indicates a role for this CAM in neutrophil migration in the lung.