Diagnosis and management of Cryptococcus neoformans var. grubii detected in an oral mass in a pink-tongued skink (Cyclodomorphus gerarrdii).

Cryptococcus is an encapsulated yeast that is found in both yeast and hyphal forms. Cryptococcus neoformans and C. gattii are the most medically important species, causing disease in both immunocompromised and immunocompetent individuals. A large, friable, sublingual mass was surgically resected from the oral cavity of a pink-tongued skink (Cyclodomorphus gerarrdii). Histopathology, fungal culture and PCR testing with Sanger sequencing confirmed granulomatous inflammation containing large numbers of yeasts identified as Cryptococcus neoformans var. grubii. Surgical excision and treatment with oral amphotericin B and terbinafine was unsuccessful in managing the infection. On Day 67 after surgery, Cryptococcus DNA was detected by PCR in the blood but not oral, cloacal or skin swabs. The skink was euthanised 72 days after surgery due to anorexia, weight loss and progressive neurological signs of disease. Necropsy results showed disseminated cryptococcosis, including meningoencephalitis and ventriculitis. Two in-contact pink-tongued skinks remained asymptomatic and PCR-negative during 198 days of observation. This case suggests Cryptococcus infections should be considered for oral masses presenting with or without neurological signs in skinks. Further investigation is required to determine the best treatment options for disseminated cryptococcosis in reptiles. This report describes the third reported case of Cryptococcus in a reptile and the first case of cryptococcosis in a pink-tongued skink. It is also the first report of Cryptococcus in a reptile identified to the variety level using PCR, including in whole blood samples.

Isolation of Cryptococcus neoformans var. grubii (serotype A) and C. magnus from the nasal lining of free-ranging quokkas (Setonix brachyurus).

Cryptococcus species are environmental yeasts, with a worldwide distribution and remarkable environmental adaptation. Although many species do not cause disease, C. neoformans and C. gattii are causative agents of cryptococcosis, a life threatening infection and a significant public health problem worldwide. Infection especially affects immunocompromised animals and humans. In wildlife, cryptococcosis appears to be more prevalent in captive populations. The objective of this study was to assess whether apparently healthy quokkas (Setonix brachyurus) harbor Cryptococcus spp. Using cultural and molecular methods, we studied yeasts isolated from nasal swabs collected from 130 free-ranging quokkas on Rottnest Island (RI, n = 97) and the mainland (n = 33) of Western Australia. Unspeciated Cryptococcus spp. (from four quokkas), C. neoformans var. grubii (serotype A) (two quokkas) and C. magnus (one quokka) were isolated from the nasal lining of apparently healthy quokkas from RI. Cryptococcus neoformans var. grubii was isolated from animals captured in a human-populated area on RI. There was no significant effect of the presence of Cryptococcus on the results of haematology, blood chemistry, peripheral blood cell morphology or clinical examination. To the best of our knowledge, this is the first documented isolation of C. neoformans var. grubii (serotype A) and C. magnus in a free-ranging macropod in Western Australia. The public health implications of this finding should be further explored.

Reptarenaviruses in apparently healthy snakes in an Australian zoological collection.

BACKGROUND: Inclusion body disease (IBD) is a disease of snakes with a global distribution and has recently been shown to be caused by reptarenaviruses. Testing for this group of viruses in asymptomatic snakes allows the association between infection and disease to be further elucidated. METHODS: A reptarenavirus was detected by RT-PCR in a reticulated python (Malayopython reticulatus) from an Australian zoological collection that was open-mouth breathing and had erythematous oral mucosa. Another 27 pythons, 4 elapids, 2 colubrids and 2 boas from this collection were then screened. From these animals, swabs, whole blood and/or tissue were tested for reptarenaviruses by RT-PCR. Additionally, blood films from 10 snakes were examined by light microscopy for the presence of inclusion bodies. The majority of samples were collected over a 484-day period. RESULTS: A total of 8 animals were RT-PCR-positive (8/36 = 22.2%): 6 were pythons, 1 was a corn snake (Pantherophis guttatus) and 1 was a Madagascar tree boa (Sanzinia madagascariensis). From them, 57 samples were collected, but only one from each animal was RT-PCR-positive (8/57 = 14.0%). From all 36 animals in this study, 8/182 samples were RT-PCR-positive (4.4%). Inclusion bodies were not recognised in any of the blood films. Only the reticulated python showed signs of illness, which improved without any further intervention. All other RT-PCR-positive snakes were apparently healthy throughout the duration of the study. CONCLUSION: This study showed a weak association between the presence of reptarenaviruses and disease. Testing serially collected swab and whole-blood samples increased the number of animals in which reptarenaviruses were detected.

A Case-control Study of Haemorrhagic Septicaemia in Buffaloes and Cattle in Karachi, Pakistan, in 2012.

A retrospective epidemiological case-control study was performed in Karachi, Pakistan, from January to April 2013. The owners of 217 dairy cattle and buffalo farms from six different locations in Karachi were interviewed. The aim of the study was to identify risk factors associated with the presence of haemorrhagic septicaemia (HS). Farms with a history of at least one instance of sudden death in a dairy animal during 2012 and a positive clinical HS diagnosis (made by local veterinarians) were defined as cases. Farms having no history of sudden deaths in 2012 were defined as controls. Univariable analyses were initially conducted, and factors with P ≤ 0.25 were offered to a multivariable logistic regression model to identify putative risk factors. The final multivariable logistic model contained five factors. Vaccination was found to be a protective factor (OR = 0.22) along with the length of time cattle were kept on farm (months). For every extra month cattle were kept, the odds of HS disease were reduced by a factor of 0.9. In contrast, for every extra animal in a herd, the risk of infection increased by a factor of 1.01. Supplying underground water and the presence of foot and mouth disease on the farm increased the risk by 2.90 and 2.37, respectively. To understand the epidemiology of HS in Karachi dairy herds, more in-depth research is required to study the risk and protective factors identified in this survey and to evaluate risk mitigation strategies, where possible.