engleski

Biochemical and molecular identification of Malassezia spp. isolated from the healthy or inflamed animal external auditory canal

Malassezia yeasts are recognized as opportunistic pathogens causing different skin disorders in warm-blooded vertebrates. Thirteen Malassezia species are now recognized but their distribution in mammals and birds remains largely unknown. In the present study, we collected and identified Malassezia isolates from different animal species in Croatia. Swabs from dogs, cats and bears were examined and strains collected for investigation. In addition, six Malassezia cultures were obtained from healthy ear canals of pigs. Yeasts were cultured on modified Dixon agar and incubated for 4-5 days at 31º ; C. For specific identification we used morphological characteristics and physiological tests based on the ability of Malassezia yeasts to assimilate polyoxyethylene sorbitan esters (Tween 20, 40, 60 and 80) and Cremophor EL (castor oil). In addition, catalase and β -glucosidase activities were evaluated. Molecular analyses were performed using conventional PCR method followed with restriction fragment length analysis (RFLP) based on digestion of PCR products with three restriction endonucleases: CfoI, HaeII and MspI. For biochemical and molecular identification, six different CBS Malassezia strains were used as controls. Results from the samples obtained from dogs, cat and bear yielded clearly distinguishable molecular patterns typical for Malassezia pachydermatis. Cultures obtained from pigs did not yield clear results, showing variable results in Cremophor EL assimilation. However, RFLP analysis for all six samples yielded a pattern similar to Malassezia furfur control strains CBS 1878 and CBS 7019. Acknowledgment: This study was performed during supported by FEMS Research Fellowship grant visit of Suzana Hađina to Service de Parasitologie-Mycologie, Ecole Nationale Vétérinaire d'Alfort, Maisons-Alfort, France.

Malassezia; animals; external auditory canal

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