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The objective of the study was to determine the optimal conditions for obtaining species-specific surface antigens of dermatophytes (the present authors’ methodology), as well as their protein profile analysis. The studies included the clinical isolates of the following strains: Microsporum canis, Trichophyton verrucosum, Trichophyton mentagrophytes and Microsporum gypseum. The analyzed strains were cultured on Sabouraud’s solid medium for 7 and 21 days at a temperature of 25°C (M. canis, M. gypseum) and at 37°C (T. verrucosum, T. mentagrophytes). Surface antigens were obtained from this material according to the present authors’ methodology, the established elution time of antigen fraction was 1.3 and 24 h. The obtained protein fractions were stored as a lyophilize at a temperature of -20°C. The protein profiles of each antigen preparation were determined by SDS PAGE technique after Laemmli. The documents and analysis of the fractions obtained were performed with Gel-Doc (Bio-Rad). The studied preparations exhibited from 8 to 18 components of 190 kDa - 14.8 kDa molecular weight, while their qualitative and quantitative composition depended on the conditions of preparation obtainment and fungus species. The comparative analysis of dermatophyte protein profiles comprised the selected preparations obtained after the 24 hours’ elution and a week of fungus culture. Besides the common components (70. 35 and 25 kDa), the examined surface antigens contained the following species-specific fractions: a band of 27.7 kDa molecular weight was characteristic for M. canis, 107 and 87.3 kDa for M. gypseum and for T. mentagrophytes - 73.6; 59.4 and 45.6 kDa. The isolation and detailed characteristics of these proteins are likely to facilitate a quick and more specific diagnostics of dermatophytoses, as well as a thorough recognition of fungus pathogenicity mechanisms.
Dermatophytosis in dogs is variable and can clinically mimics other skin diseases. Diagnosis based on clinical presentation may be misleading and additional laboratory tests are needed for a final diagnosis. A systematic diagnostic procedure can prevent a wrong diagnosis and allow for the right treatment.
Interactions between selected geophilic fungi (keratinofytic and keratinophylic species) and pathogenic dermatophytes were examined in dual cultures on Sabouraud glucose agar (SGA). It was demonstrated that Trichophyton mentagrophytes var. mentagrophytes together with T. mentagrophytes var., quinceanum are the species most sensitive to the influences of the geophilic fungi. Microsporum persicolor was found to be more resistant to these influences. Only a few geophilic species inhibited the growth of M. canis on SGA. Keratinophilic species, unable to initiate hair decomposition but utilize the products of this process, exerted the larger inhibitory effect on the pathogenic dermatophytes than keratinofytic fungi.
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