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The objective of the present study was to evaluate the antifungal activity of Enizol, a new disinfecting preparation with enilkonazole as the active substance. For both the studies in vitro and in vivo the initial concentration of Enizol was constituted by an aqueous solution of the preparation at a ratio of 1:100. The investigations covered 34 strains of the following mould fungi: A. fumigatus (n=5), A. versicolor (n=3), Penicillium spp. (n=5), Cladosporium spp. (n=4), Scopulariopsis spp. (n=3), Fusarium spp. (n=4), Alternaria spp. (n=5), Mucor spp. (n=5), as well as 10 strains of yeast-like fungi: Candida albicans (n=5) and Candida non-albicans (n=5). The minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC) of Enizol were determined in vitro according to NCCLS M27-A and by a cylinder dilution method. The MIC values for these organisms appeared to be differentiated and ranged from 0.07 µgml-1 (A. versicolor) up to 37.5 µgml⁻¹ (Mucor spp.). A group of extremely sensitive fungi comprised Aspergillus spp. (0.07 - 1.2 µgml⁻¹), Penicillium spp. (0.07 - 1.2 µgml⁻¹) and Alternaria spp. (2.4 µgml⁻¹); Cladosporium spp. genus (4.75 µgml-1) indicated the medium susceptibility toward the inhibitory activity of Enizol, whereas Fusarium spp. (9.5 µgml⁻¹), Mucor spp. (19.0 - 37.5 µgml⁻¹) and Scopulariopsis spp. (19.0 µgml⁻¹) had the lowest values. The antifungal efficacy of the studied preparation is confirmed by its lethal characteristics. The minimal fungicidal concentrations (MFC) were differentiated subject to the species of fungus studied. At the same time the sensitivity of the anascogenic yeast Candida genus was analyzed and consequently the fungi were classified among the organisms relatively resistant to (9.5 µgml⁻¹ MIC, 37.5 µgml⁻¹ MFC) Enizol activity. The studies in vivo confirmed the sensitivity of fimbriate fungi to the preparation and, usually, this was consistent with the tests in vitro. At the same time the antifungal efficacy of Enizol in vivo was demonstrated towards the fungi Candida genus, which indicates its usability as a lethal preparation in an environment where animals stay.
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.
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