RESUMO
Trichophyton (T.) verrucosum is a highly pathogenic dermatophyte causing zoonotic bovine ringworm that is transmissible to humans. The virulence factors subtilisin (Sub)3 and Sub6 are discussed to contribute to disease manifestation but no protein expression study is available for T. verrucosum. We used customized antibodies (against Trichophyton-species, Sub3 and Sub6) to examine skin biopsies of infected cattle via immunofluorescence stainings. Both virulence factors Sub3 and 6 were solely expressed by conidia and not only found in epidermal but also in dermal and hair structures. The anti-T-antibody reliably detected the fungus and proved more sensitive compared to histological stains. LAY SUMMARY: We examined the zoonotic dermatophyte Trichophyton (T.) verrucosum in bovine skin and studied two important virulence factors called subtilisin (Sub)3 and Sub6 that T. verrucosum produces and secretes using immunolabeling.
Assuntos
Doenças dos Bovinos/microbiologia , Pele/microbiologia , Subtilisina/genética , Tinha/veterinária , Trichophyton/genética , Trichophyton/patogenicidade , Animais , Biópsia/veterinária , Bovinos , Doenças dos Bovinos/diagnóstico , Imunofluorescência , Pele/patologia , Esporos Fúngicos/genética , Esporos Fúngicos/patogenicidade , Subtilisina/classificação , Tinha/microbiologia , Fatores de Virulência/genética , Zoonoses/microbiologiaRESUMO
The zoophilic dermatophyte Trichophyton verrucosum is the most important causative agent of bovine dermatophytosis. Additionally, it causes profound and poorly healing skin infections in humans indicating the high zoonotic potential. The objective of this study was to establish differentiation of T. verrucosum from other dermatophytes by mass spectrometry and to identify distinct features of the mass spectra. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was successful for identification of this pathogen only after extension of the database of the manufacturer with spectra from T. verrucosum strains, which were identified as such by sequencing of the internal transcribed spacer (ITS) region. MALDI-TOF MS analysis was conducted with 46 field isolates from cattle, two live vaccine strains, and 10 isolates from humans identified as T. verrucosum by sequence analysis of the ITS region. The results suggest a very good agreement of both methods. Comparison with the mass spectra of 68 strains of other keratinophilic fungi revealed that most T. verrucosum wild-type isolates showed a characteristic peak at 7950-7954 m/z, which was missing in the spectra of other keratinophilic fungi and the live vaccine strains. The spectra of T. verrucosum were most similar to the spectra of T. benhamiae, an emerging zoophilic dermatophyte. In summary, MALDI-TOF MS is a powerful and reliable tool to identify T. verrucosum.
Assuntos
Doenças dos Bovinos/microbiologia , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Tinha/veterinária , Trichophyton/classificação , Trichophyton/fisiologia , Zoonoses/microbiologia , Animais , Bovinos , Análise por Conglomerados , DNA Fúngico/genética , DNA Espaçador Ribossômico/genética , Bases de Dados Genéticas , Humanos , Técnicas de Tipagem Micológica/veterinária , Reprodutibilidade dos Testes , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/veterinária , Tinha/microbiologia , Trichophyton/química , Trichophyton/genéticaRESUMO
Trichophyton benhamiae is an emerging zoonotic dermatophyte. We present a case of a small animal stock infected with two Trichophyton species. T. benhamiae was isolated from 15 out of 26 (58%) guinea pigs including two morphologically different phenotypes. Eight guinea pigs were infected with T. benhamiae and T. mentagrophytes simultaneously. The animals showed alopecia and crusts or no clinical signs at all. T. benhamiae was not detected in rats, rabbits and mice kept in the same stock.