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SKIN MICROBIOLOGY OF VIETNAMESE MOSSY FROGS (Theloderma corticale)

Amphibians are one of the most endangered vertebrates in the world due to their habitat loss, environmental pollution, climate changes and emerging diseases. By secretion of peptides from skin glands and by composition of skin microbiota, their skin plays an important role in the protection from various diseases. To get more insight into the skin microbiota, skin swabs of eight Vietnamese mossy frogs (Theloderma corticale) situated backstage of the Zagreb Zoo were taken. Samples were taken with sterile swabs without transport media and immediately transported to the laboratory of the Department of Poultry Diseases with Clinic. Swabs were plated on non-selective culture media for detection of aerobic bacteria and incubated 24 hours at room temperature. Isolated bacteria were further identified microscopically, by catalase and oxidase reactions, and by Gram staining. Further identification to genus level was done using MALDI-TOF MS. For fungal detection, swabs were plated on Sabouraud dextrose agar and incubated for five days at room temperature. The identification was based on morphological appearance and microscopic characteristics using lactophenol staining, and by MALDI-TOF MS. Skin swabs were also tested for the presence of Batrachochytrium dendrobatidis, B. salamandrivorans, Ranavirus and Chlamydia spp. by qPCR after DNA extraction using Qiagen Dneasy Blood and Tissue Kit. In total, 13 bacterial and 4 fungal species were isolated, with predominance of Bacillus cereus and Candida catenuata. Although B. cereus is a common isolate of amphibian skin, there is no data about its pathogenic or protective role in amphibians. Detection of C. catenuata could indicate immunodeficiency, which should be corrected by proper husbandry and water quality in captive enclosures. Chytrid fungi, Ranavirus and Chlamydia spp. were not detected. Microbiological screening of amphibian microflora seems to be useful tool for distinguishing between protective and pathogenic microbiota in amphibian conservation efforts.

amphibians, emerging diseases, vietmanese mossy frog, qPCR

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