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Molecular identification of animal – derived Candida species and their susceptibility to miconazole determined by the broth microdilution method

Miconazole (MCZ) is an antifungal drug very often used in topical treatment of local yeast infections in animals. Azole resistance in Candida isolates in human medicine is a well-known fact for decades. Although Candida infections are not so frequently reported in animals, there is growing evidence of resistance to both C. albicans and some nonalbicans Candida species such as C. tropicalis. In this study, we identified thirteen Candida species isolated from healthy and diseased animals to the species level and evaluated their susceptibility to miconazole using the broth microdilution method. Species identification was performed using phenotypic and molecular methods. Polymerase chain reaction was performed with ITS-1 and ITS-4 primers for the amplification of ITS1, ITS2 and 5.8S rDNA region followed with phylogenetic analysis. Susceptibility testing against MCZ was determined according to the European Committee on Antimicrobial Susceptibility Testing guidelines for yeasts. The range of tested MCZ concentrations were from 0.0625 to 32 µg/mL. Testing was performed in triplicate on three separate microplates for each sample. The most commonly isolated species was C. albicans with six isolates, followed with two isolates of C. tropicalis and C. palmioleophila, and one isolate of C. krusei, C. lusitaniae and C. zeylanoides. Minimum inhibitory concentrations (MIC) values were in the range from < 0.06 to 2 µg/mL. The activity of C. krusei was the highest (2 µg/mL) followed with C. zeylanoides (1 µg/mL) in comparison with the rest of the isolates. In conclusion, C. albicans was dominant species isolated from animal samples. The obtained range of MIC values indicate various susceptibility of tested Candida species. Further investigation of a number of Candida isolates from animal species is needed to identify susceptibility patterns of this yeast in veterinary settings.

Candida species, susceptibility, miconazole, broth microdilution method

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