engleski

Atypical Malassezia yeast isolates from captive tapirs

Objective: Malassezia yeasts are well known opportunistic microorganisms. However, their transition from the opportunistic to pathogenic yeast stage still remains unknown. This genus colonizes the skin of healthy domestic and wild animals. At present, there is no many data about the prevalence of Malassezia species in wild animals. During the epidemiological study of the colonization of these species of the wild animals kept in zoological park, we isolated Malassezia species from tapirs. The aim of this study was to explore morphological, biochemical and molecular characteristics of Malassezia yeasts isolated from the external auditory canal and skin of four tapirs. Methods: Malassezia colonies were grown on modified Dixon agar for 14 days at 32°C and identified based on colony appearance and microscopic features. In order to distinguish their lipid dependency, isolates were subcultured on Sabouraud dextrose agar (SDA). In addition catalase, urease and esculin test were performed. Genomic DNA was isolated using zymolyase enzyme and commercial DNA isolation kit. Molecular characterization was performed amplifying ITS-1 region with 18SF1 and 5.8SR1 primers, while F63 and LR3 primers were used for the amplification of D1/D2 regions of the LSU rRNA gene. Results: Malassezia isolates grew slowly on Dixon agar and did not grow on SDA. All colonies showed similar morphological and biochemical characteristics. They were catalase and urease positive and able to split esculin. By using ITS-1 and LSU primers, the sequences of expected size were amplified ; 282 bp and 640 bp, respectively. NCBI BLAST analysis of LSU sequences had 95% identity to M. pachydermatis. Additionally, ITS-1 marker exhibited 79% identity (89% sequence coverage) with M. pachydermatis. Conclusion: These results indicate that isolates could belong to the lipid dependent M. pachydermatis. However, lipid dependency needs to be thoroughly evaluated. Further molecular and phylogenetic analysis will be employed in order to clarify identification of examined isolates.

Malassezia yeast, external auditory canal, skin, tapirs

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