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Introduction: Dermatophytoses are superficial mycoses, and the identification of their etiological agents is routinely performed by culture and microscopic features, which is time-consuming and relies on personnel expertise. Molecular approaches have been developed to provide faster and reliable results; therefore, this study aimed to identify dermatophytes isolated from Alagoas state patients, employing phenotypical and molecular methods. Methodology: Clinical samples for morphological identification were collected from private and public laboratories and cultivated on Sabouraud dextrose agar. DNA extraction was followed by ITS amplicon analysis after restriction enzyme digestion DdeI (ITS-RFLP). Results: Out of fourteen representative strains, ITS-RFLP with DdeI efficiently identified Microsporum canis, Nannizzia gypsea, and Trichophyton rubrum, while species of the complex T. tonsurans/T. mentagrophytes presented the same restriction pattern. After genotyping, 2 T. tonsurans and 1 Microsporum sp. strain were reclassified as T. rubrum. Conclusions: RFLP of ITS-region followed by DdeI digestion produced faster and relatively reliable results than classic methods; however, this method has not been as efficient for closely related dermatophytes cryptic species.

Respiratory diseases are commonly related to grain mills in association with fungi. Pigeons (Columba livia) are a potential source of pathogenic fungi and a public health concern, causing economic damages in urban areas. We aimed to determine the tracheal and pulmonary mycobiota of pigeons captured in a grain mill, identifying potentially pathogenic fungi. Captured pigeons were euthanized for the collection of tracheal secretion and lungs in saline suspensions for culture (100µL) in Sabouraud dextrose agar with chloramphenicol (48h/28°C). Yeasts were evaluated after Gram staining in microscopy and cultured in Hicrome®Candida agar, while lactophenol cotton blue solution was used in filamentous fungi/molds. Mycological identification keys were used for the genus determination. 207 fungal (9 genus) were isolated from the trachea and lungs of 21 pigeons captured: 199 molds (96.14%) and 8 yeast (3.86%). Aspergillus spp., Rhizopus spp., Penicillium spp. and Candida spp. were the most prevalent, mainly in lungs. We confirmed yeasts and molds from pigeons, increasing the potential risks for chronic or acute diseases after contact with several fungi from pigeons. A greater control of the presence of pigeons in the work environment is necessary. Enfermedades respiratorias se relacionan comúnmente con los molinos de granos en asociación con hongos. Las palomas (Columba livia) son una fuente potencial de hongos patógenos y un problema de salud pública, causando daños económicos en zonas urbanas. Nosotros determinamos la micobiota traqueal y pulmonar de palomas capturadas en molino de granos, identificando hongos potencialmente patógenos. Las palomas capturadas fueron sacrificadas para la recolección de secreción traqueal y pulmones en suspensiones salina para cultivo (100 µl) en agar Sabouraud dextrosa con cloranfenicol (48h/28°C). Las levaduras se evaluaron después de la tinción de Gram en microscopía y se cultivaron en agar Hicrome®Candida, mientras que lactofenol azul se usó en hongos filamentosos. Se utilizaron claves de identificación micológicas para la determinación del género. 207 hongos (9 géneros) fueron aislados de la tráchea y pulmones de 21 palomas capturadas: 199 mohos (96.14%) y 8 levaduras (3.86%). Aspergillus spp., Rhizopus spp., Penicillium spp. y Candida spp. fueron los más prevalentes, principalmente en pulmones. Confirmamos levaduras y mohos de palomas, aumentando los riesgos potenciales de enfermedades crónicas o agudas tras el contacto con varios hongos de palomas. Es necesario un mayor control de la presencia de palomas en el entorno laboral. As doenças respiratórias são comumente relacionadas aos moinhos de grãos em associação com fungos. Visitantes frequentes devido ao hábito granívoro, pombos (Columba livia) são vetores em potencial de fungos patogênicos e um problema de saúde pública, causando prejuízos econômicos em áreas urbanas. Nós objetivamos determinar a micobiota traqueal e pulmonar de pombos capturados em moinho de grãos, identificando fungos potencialmente patogênicos. Os pombos capturados foram eutanasiados para coleta de secreção traqueal e pulmões em suspensões salinas para cultivo (100µL) em ágar Sabouraud dextrose com cloranfenicol (48h/28°C). As leveduras foram avaliadas após coloração de Gram em microscopia e cultivadas em ágar Hicrome®Candida, enquanto a solução azul algodão de lactofenol foi utilizada em fungos filamentosos. Chaves de identificação micológica foram utilizadas para a determinação do gênero. Um total de 207 fungos (9 gêneros) foram isolados da traqueia e pulmões de 21 pombos capturados: 199 filamentosos (96,14%) e 8 leveduras (3,86%). Aspergillus spp., Rhizopus spp., Penicillium spp. e Candida spp. foram os mais prevalentes, principalmente nos pulmões. Confirmamos leveduras e fungos filamentosos potencialmente patogênicos no sistema respiratório dos pombos, o que pode aumentar os riscos potenciais para doenças crônicas ou agudas após contato com os diversos gêneros identificados. É necessário um maior controle da presença de pombos no ambiente de trabalho.

Passeriformes and Psittaciformes birds and pigeons (Columba livia) are known to be reservoirs of microorganisms, and their stool allows fungi development. Since accumulated avian excreta can interfere with public health, this study aimed to perform a molecular screening of medically important Candida species in pigeon droppings in public places and birds raised in captivity. Excreta collected from captive birds (3 residences) and pigeons (4 districts) were inoculated on Sabouraud dextrose agar with chloramphenicol for Gram staining and subculture on Hicrome® Candida. Three DNA extraction methods were performed for comparison (commercial kit, in-house and by boiling) and PCR to screen 6 clinically important Candida species among the isolates. The correlation between phenotypic and molecular methods was calculated by kappa/K. Only 6 C. parapsilosis (20%) were identified from captive birds’ feces among 30 isolates (80% not identified), while pigeons’ feces harbored a greater diversity, with the 6 pathogenic species confirmed among 41 isolates: C. albicans (31.70%/13), C. krusei (14.63%/6), C. tropicalis (14.63%/6), C. parapsilosis (17.10%/7), C. glabrata (14.63%/6) and C. guilliermondii (7.31%/3); 100% correlation between tested methods (K = 1) for the first 3 species. Boiling DNA extraction method was fast and efficient to obtain viable DNA from Candida spp. for PCR. Our results indicate that pigeon droppings harbor more potentially pathogenic species than birds in residential captivity, which probably have non-albicans Candida less frequently isolated in infectious processes. The greater availability of nutrients may have contributed to a diversity of Candida spp. in feces from public environments.