Isolation of filamentous basidiomycetes from respiratory samples in a tertiary care Spanish hospital / Aislamiento de basidiomicetos filamentosos en muestras respiratorias en un hospital español de tercer nivel

Background The clinical significance of the filamentous basidiomycetes isolated from clinical samples is not always clear. Thus, these fungi have been considered environmental contaminants traditionally. Aims To review those clinical cases in which filamentous basidiomycetes from respiratory samples had been isolated. Methods The retrospective study was carried out in a single tertiary care hospital. We recovered all culture-confirmed isolations of filamentous basidiomycetes from respiratory samples (bronchial aspirate [BAS], bronchoalveolar lavage [BAL] and sputum) analyzed between the years 2020 and 2023. Isolates were identified by ITS region sequencing. Results In six patients a filamentous basidiomycete had been isolated from a respiratory sample. The species identified were all different: Fomitopsis sp. (BAS), Trametes ljubarskyi (BAL), Stereum gausapatum (BAS), Porostereum spadiaceum (BAS), Phlebia subserialis (sputum) and Inonotus levis (BAL). All the patients were immunosuppressed or had an underlying disease with pulmonary involvement. None of them received any specific antifungal treatment (in relation with the fungus isolated) and all six improved clinically and were discharged. Conclusions The isolation of filamentous basidiomycetes in these patients had uncertain clinical significance. However, the isolation of any filamentous basidiomycete in respiratory samples from immunosuppressed patients or patients with chronic pulmonary disease is an emerging situation that should be carefully assessed in the context of chronic allergic episodes or suspicion of invasive fungal infections. (AU)

Antecedentes La importancia clínica de los basidiomicetos filamentosos procedentes de muestras clínicas no siempre es clara. Por ello, estos hongos se han considerado tradicionalmente como contaminantes ambientales. Objetivos Describir las características de los pacientes con aislamientos de basidiomicetos filamentosos en muestras respiratorias para intentar aclarar el significado clínico de estos hallazgos. Métodos Se realizó un estudio retrospectivo en nuestro hospital de tercer nivel; fueron incluidos pacientes con el aislamiento de un basidiomiceto filamentoso confirmado en cultivo a partir de una muestra respiratoria (aspirados bronquiales [BAS], lavados broncoalveolares [BAL] y esputos) procesadas entre los años 2020 y 2023. Realizamos la identificación molecular del aislamiento mediante la secuenciación de la región ITS. Resultados En seis pacientes se obtuvo un aislamiento de basidiomiceto filamentoso de una muestra respiratoria, y los seis aislamientos pertenecían a diferentes géneros: Fomitopsis sp. (BAS), Trametes ljubarskyi (BAL), Stereum gausapatum (BAS), Porostereum spadiaceum (BAS), Phlebia subserialis (esputo) e Inonotus levis (BAL). Todos los pacientes sufrían inmunosupresión o alguna enfermedad de base con afectación pulmonar. Ninguno de ellos recibió tratamiento antifúngico específico (en relación con el aislamiento recuperado de la muestra) y finalmente fueron dados de alta tras la mejoría clínica. Conclusiones Los aislamientos de basidiomicetos filamentosos en nuestros pacientes tuvieron un significado clínico incierto. Sin embargo, en los pacientes inmunodeprimidos o con enfermedades pulmonares crónicas, el aislamiento de un basidiomiceto filamentoso en el tracto respiratorio inferior representa una situación emergente que debe evaluarse cuidadosamente en el contexto de un episodio alérgico crónico o ante la sospecha de una infección fúngica invasiva. (AU)

Isolation of filamentous basidiomycetes from respiratory samples in a tertiary care Spanish hospital.

BACKGROUND: The clinical significance of the filamentous basidiomycetes isolated from clinical samples is not always clear. Thus, these fungi have been considered environmental contaminants traditionally. AIMS: To review those clinical cases in which filamentous basidiomycetes from respiratory samples had been isolated. METHODS: The retrospective study was carried out in a single tertiary care hospital. We recovered all culture-confirmed isolations of filamentous basidiomycetes from respiratory samples (bronchial aspirate [BAS], bronchoalveolar lavage [BAL] and sputum) analyzed between the years 2020 and 2023. Isolates were identified by ITS region sequencing. RESULTS: In six patients a filamentous basidiomycete had been isolated from a respiratory sample. The species identified were all different: Fomitopsis sp. (BAS), Trametes ljubarskyi (BAL), Stereum gausapatum (BAS), Porostereum spadiaceum (BAS), Phlebia subserialis (sputum) and Inonotus levis (BAL). All the patients were immunosuppressed or had an underlying disease with pulmonary involvement. None of them received any specific antifungal treatment (in relation with the fungus isolated) and all six improved clinically and were discharged. CONCLUSIONS: The isolation of filamentous basidiomycetes in these patients had uncertain clinical significance. However, the isolation of any filamentous basidiomycete in respiratory samples from immunosuppressed patients or patients with chronic pulmonary disease is an emerging situation that should be carefully assessed in the context of chronic allergic episodes or suspicion of invasive fungal infections.

Filamentous fungi in the airway of patients with cystic fibrosis: Just spectators?

BACKGROUND: There are important advances in the management of bacterial infection in patients with cystic fibrosis (CF), but there are many gaps in the field of fungal infections. AIMS: The aim of this study was to analyse whether chronic respiratory filamentous fungal colonization had clinical impact and whether antifungal treatment can change the disease. METHODS: The prospective, bicentric and descriptive study was carried out within a 3-year follow-up period, with four-month periodicity medical controls. Adult patients from two CF units of tertiary hospitals were included. Clinical, microbiological, analytical and spirometric variables were collected. Quality of life was evaluated in a subgroup, using the Spanish version of the Revised Cystic Fibrosis Quality of Life Questionnaire (CFQ-R). To statistically analyze the evolution of forced expiratory along time (volume of air blown out in 1 second -FEV1-) and the forced vital capacity (FVC), mixed linear models were carried out. RESULTS: From the ninety-eight patients under study, 40 suffered chronic filamentous fungal colonization. The presence of filamentous fungi in airway was associated to an annual fall of FEV1 and FVC of 0.029 and 0.017 litres, respectively (p<0.001). In addition, worse quality of life based on CFQ-R, significant when concerning physical condition and emotional state, was also linked with the fungal colonization. Protocolized antifungal therapy, nebulized or oral, improved FEV1 in 0.023 and 0.024 litres per year, respectively (p<0.001). CONCLUSIONS: Chronic filamentous fungal colonization in patients with CF is associated with a significant annual decline of lung function that persists over time. Chronic antifungal therapy slows down this progression, mainly in the patient with more advanced disease.

[Systemic antifungal drugs]. / Antifúngicos de uso sistémico.

Invasive fungal infections have increased over the last decades and the therapeutic choices to treat them are limited. The antifungal agents currently available are useful and have optimal in vitro activity; however, their activity can be lowered due to the development of fungal resistance. The increase in primary or secondary resistance to some antifungal drugs has led to the search of alternatives such as the combination of drugs or the development of new antifungals. In this paper, the activity of the main families of antifungal drugs, polyenes, azoles, echinocandins, 5-fluorocytosine and other new antifungal drugs, are reviewed. The main resistance mechanisms developed by fungi are also described.

Dermatophytoses in patients attended from a private clinical analysis laboratory in João Pessoa-PB, between 2015 to 2019 / Dermatofitoses em pacientes atendidos em um laboratório privado de João Pessoa-PB, entre 2015 a 2019

Introduction: dermatophytoses or "tineas" are characterized by being mycoses caused by fungi of the genera Epidermophyton, Trichophyton and Microsporum. These mycotic infections can present themselves as a form of lesions that affect the skin, hair and nails of individuals of both genders and all ages. Objective: to elucidate the epidemiological profile of dermatophytoses in patients examined by a private clinical analysis laboratory in João Pessoa-PB, between 2015 and 2019. Methodology: this is an epidemiological, analytical, retrospective and documentary study, in which data collection took place at the Clinical Pathology Laboratory ­ "HEMATO", located in João Pessoa ­ PB. Results: the profile of those affected was predominantly female (58.5%), 18 to 59 years old (38.4%), white (53.6%) and with lesions, mainly in skin glabrous (38.5%), feet (33.3%) and nails (12.8%). When relating the age group to the injury site, it was noticed that injuries on glabrous skin, feet and nails, were more frequent in individuals aged 18 to 59 years, while injuries to the scalp were mostly found in individuals younger than 18 years old. The most prevalent species were M. canis (31.9%) and T.rubrum (31.9%). When correlating the fungal species with the lesion site, it was noted that M. canis was the main agent responsible for lesions in glabrous skin, scalp and hands, while T. rubrum was predominantly observed in nails and T. mentagrophytes in feet. Conclusion: it is concluded that the data present in this research can promote the development of indicators and public policies for the population most susceptible to dermatophytosis.

Introdução: dermatofitoses ou tineas se caracterizam por serem micoses causadas por fungos dos gêneros Epidermophyton, Trichophyton e Microsporum. Essas infecções micóticas podem se apresentar na forma de lesões que acometem pele, pelo e unhas de indivíduos de ambos os gêneros e todas as idades. Objetivo: elucidar o perfil epidemiológico de dermatofitoses de pacientes atendidos por um laboratório privado de análises clínicas em João Pessoa-PB, entre 2015 a 2019. Metodologia: trata-se de um estudo epidemiológico, analítico, retrospectivo e documental, em que a coleta de dados ocorreu no Laboratório de Patologia Clínica ­ HEMATO, localizado em João Pessoa ­ PB. Resultados: o perfil de acometidos foi predominantemente de indivíduos do sexo feminino (58,5%), com 18 a 59 anos de idade (38,4%), brancos (53,6%) e com lesões, principalmente, em pele glabra (38,5%), pés (33,3%) e unhas (12,8%). Ao relacionar a faixa etária com o local da lesão, percebeu-se que lesões em pele glabra, pés e unhas, foram mais frequentes em indivíduos de 18 a 59 anos, enquanto que lesões no couro cabeludo foram majoritariamente encontradas em indivíduos menos de 18 anos. As espécies mais prevalentes foram M. canis (31,9%) e T. rubrum (31,9%). Ao correlacionar a espécie fúngica com o local da lesão, notou-se que M. canis foi o principal agente responsável por lesões em pele glabra, couro cabeludo e mãos, enquanto T. rubrum foi predominantemente observado em unhas e T. mentagrophytes em pés. Conclusão: os dados obtidos nesta pesquisa podem fomentar o desenvolvimento de indicadores e políticas públicas para a população mais susceptível às dermatofitoses.

Monitoramento de fungos anemófilos no ambiente de uma biblioteca no Município de São José do Rio Preto - Sp, Brasil / Anemophilic fungi monitoring in a library environment in the city of São Jose do Rio Preto, Brazil

Os fungos desempenham vários papéis que impactam a humanidade de diversas maneiras. Suas características metabólicas são importantes na biotecnologia, porém, tais microrganismos podem desencadear alguns problemas de saúde pública e até mesmo serem letais. Objetivo: detectar a presença de fungos no acervo de uma biblioteca no município de São José do Rio Preto. Metodologia: foram coletadas quarenta amostras nas superfícies inanimadas (livros, estantes, documentos, mapas, artigos e revistas) das principais salas da biblioteca com o auxílio de swabs umedecidos em solução salina estéril, posteriormente encaminhados ao laboratório de Biomedicina da Universidade Paulista ­ UNIP. As amostras foram semeadas em meio de cultura ágar Sabouraud Dextrose (SDA), tendo adicionado cloranfenicol e incubadas a 30 °C. Foi realizada a colônia gigante em todas as cepas crescidas em SDA para a realização da técnica de microcultivo para a identificação dos fungos, de acordo com o Manual de Detecção e Identificação dos Fungos de Importância Médica da Agência Nacional de Vigilância Sanitária. Resultados: Houve positividade em trinta e uma amostras (78%) e em quatro delas foi observado mais de um tipo de colônia (13%). Das vinte e duas superfícies de livros analisadas, foram isolados e identificados: Aspergillus flavus, Aspergillus niger, Cunninghamella sp., Cladosporium sp., Curvularia sp., Mucor sp. e Nigrospora sp. Nas oito superfícies de estantes: Aspergillus flavus, Aspergillus niger, Aspergillus versicolor, Penicillium sp. e Scopulariopsis sp. e, nos dez documentos: Aspergillus nidulans, Aspergillus sp., Cladosporium sp., Cunninghamella sp. e Trichoderma sp. Conclusão: Os fungos encontrados estão amplamente distribuídos no ambiente como solo e ar e, por diversos fatores, instalam-se em locais como bibliotecas. Em condições favoráveis, podem infectar o homem e causar perdas patrimoniais para os acervos.

Fungi play many roles that impact humankind in different ways. Their metabolic characteristics are important in biotechnology; however, these microorganisms can trigger some public health problems or may even be lethal. Objective: detect the presence of fungi in the collection of a public library in the city of São José do Rio Preto, Brazil. Methods: a total of forty samples were collected from inanimate surfaces (books, shelves, documents, maps, articles and magazines) located in the main rooms of the library with swabs soaked in sterile saline solution and sent to the Universidade Paulista ­ UNIP laboratories. The samples were plated in Sabouraud Dextrose Agar (SDA) supplemented with chloramphenicol and incubated at 30 °C. The colonies that grew in SDA were isolated in Potato Dextrose Agar for performing the slide culture technique for the identification of the fungi, performed according to the Manual of Detection and Identification of Fungi of Medical Importance from the Brazilian Health Surveillance Agency (ANVISA). Results: Thirty-one samples (78%) were positive, and in four of them more than one fungus genus was observed (13%). From the twenty-two book surfaces analyzed, the following fungi were isolated and identified: Aspergillus flavus, Aspergillus niger, Cunninghamella sp., Cladosporium sp., Curvularia sp., Mucor sp. and Nigrospora sp. On the eight shelves: Aspergillus flavus, Aspergillus niger, Aspergillus versicolor, Penicillium sp. and Scopulariopsis sp. The ten documents analyzed presented the following fungi: Aspergillus nidulans, Aspergillus sp., Cladosporium sp., Cunninghamella sp. and Trichoderma sp.. Conclusion: These fungi are widely distributed in the environment such as in the soil and air, and due to several factors, they colonize public places, such as libraries. In favorable conditions, they may infect humans and cause diseases.

Significativo aumento de onicomicosis por hongos filamentosos no dermatofitos en red de salud privada en Santiago de Chile entre los períodos 2008-2009 y 2016-2017 / Significant increase in onychomycosis due to Filamentous Fungi Non Dermatophytes in Network of Private Health in Santiago de Chile among the periods 2008-2009 and 2016-2017

INTRODUCCION: La onicomicosis corresponde a una patología prevalente causada por hongos dermatofitos, levaduras y en menor proporción, hongos filamentosos no dermatofitos (HFND). Se ha reportado un aumento a nivel mundial en la incidencia de onicomicosis por HFND1 , sin embargo, en Chile no hay registros de ello. OBJETIVO: Comparar la epidemiología local de onicomicosis por HFND en una red privada de Santiago de Chile entre dos períodos de tiempo. Material y métodos: Se realizó un estudio transversal de análisis de los registros de laboratorio de la Red de Salud UC-Christus entre los períodos 2008- 2009 y 2016-2017. RESULTADOS: Se obtuvieron un total de 9.579 muestras en ambos períodos, 4.985 entre 2008-2009 y 4.594 entre 2016-2017, con 3.442 (36%) cultivos positivos para hongos, 1.831 (36,7%) en el primer período y 1.611 (35%) en el segundo. Del total 40 muestras (1,2%) fueron de HFND, destacando que la prevalencia aumentó significativamente entre ambos períodos: 8 (0,4%) y 32 (2,0%) cultivos, respectivamente (p<0,0001). Del total, las especies de HFND más prevalentes fueron: 23 Fusarium sp (57,5%), 8 Scopulariopsis sp (20%) y 7 Acremonium sp (17,5%). En pacientes con cultivo positivo para HFND, se buscaron asociaciones probables en los antecedentes a través de la ficha clínica, sin encontrar resultados significativos. CONCLUCION: Los HFND son la tercera causa de onicomicosis y su prevalencia fue 1,2%. Las especies más frecuentes fueron Fusarium sp, Scopulariopsis sp, Acremonium sp. La prevalencia de HFND aumentó significativamente en un período de 6 años.

INTRODUCTION: Onychomycosis is a frequent nail disease caused mainly by dermatophytes, in less proportion yeast and last non-dermatophyte molds (NDM). NDM onychomycosis is an increasing problem worldwide, though in Chile there is no epidemiological registry about it. OBJETIVE: The aim of the study was to determine the local epidemiology of NDM onychomycosis. MATERIAL Y METHODS: We did a transversal cohort study in the Red de Salud UC-Christus between 2008-2009 and 2016-2017. RESULTS: Of the 9,579 clinically suspected cases of onychomycosis, 4,985 in 2008-2009 and 4,594 in 2016-2017, 3,448 (36%) cultures were positive in total, 1,831 (36.7%) in the first period and 1,611 (35%) in the second. Only 40 cultures (1.2%) were NDM in total, but the prevalence significantly increased between both periods with 8 (0.45%) and 32 (2.0%) cultures, respectively (p y (p<0.0001). Among the total of NDM, the most prevalent species were: 23 Fusarium sp (57.5%), 8 Scopulariopsis sp (20%) and 7 Acremonium sp (17.5%). We searched for possible associations of patients with NDM onychomycosis and morbid history, but found no significant result. COMCLUSION: NDM are the third cause of onychomycosis and its prevalence of 1.2% was predominantly from toe nails. The most frequent species were Fusarium sp, Scopulariopsis sp. and Acremonium sp. The prevalence of NDM increased significantly in a period of 6 years.

Epidemiología de la enfermedad fúngica invasora por hongos filamentosos en el período 2005 a 2015, en un hospital universitario en Santiago, Chile / Epidemiology of invasive fungal disease by filamentous fungi in the period 2005 to 2015, in a university hospital in Santiago, Chile

Resumen Introducción: La enfermedad fúngica invasora (EFI) por hongos filamentosos es cada vez más frecuente. Objetivo: Estudiar la epidemiología de la EFI en adultos hospitalizados en nuestro centro. Metodología: Estudio retrospectivo de pacientes adultos de un hospital universitario en Santiago, Chile, con EFI por hongos filamentosos entre enero de 2005 y diciembre de 2015. Resultados: Se identificaron 125 episodios, siendo 48% categoria probada, 40% probable y 11% posible según criterios EORTC/MSG, incidencia global 0,47 x 1.000 egresos, 57% pacientes masculinos y edad de 50 ± 16 años. El 66,4% tenía patología hematológica, 11,2% trasplante de órgano sólido, 11,2% enfermedad reumatológica, 11,2% otra condición. Los factores de riesgo fueron neutropenia 44%, corticoterapia 21%, inmunosupresores 13%. Los hongos más frecuentemente identificados fueron Aspergillus spp (53,6%), Mucorales (16%), Fusarium spp (8,8%), Alternaria spp (5,6%), otros filamentosos (3,2%). Todos recibieron antifúngicos, 82% monoterapia, 18% terapia combinada, hubo defocación quirúrgica en 90% de mucormicosis. La mortalidad global fue 42%. Al comparar 2005-2009 vs 2010-2015, hubo un aumento significativo de la incidencia y una tendencia a menor mortalidad en el segundo período. Conclusiones: Durante un período de 10 años, observamos un aumento de la incidencia de EFI por filamentosos, aspergilosis fue la etiología más frecuente y la mortalidad global fue 42%.

Background: Invasive fungal disease (IFD) due to filamentous fungi is increasingly common. Aim: To study the epidemiology of EFI in hospitalized adults in our center. Methods: Retrospective study of adult patients of a university hospital in Santiago, Chile, with EFI due to filamentous fungi between January 2005 and December 2015. Results: 125 episodes were identified, being 48% proven, 40% probable and 11% possible according to EORTC/MSG criteria, overall incidence was 0.47/1,000 admissions, 57% male patients and age 50 ± 16 years. 66.4% had hematological pathology, 11.2% solid organ transplant, 11.2% rheumatology diseases, 11.2% other conditions. The risk factors were neutropenia 44%, corticosteroid therapy 21%, immunosuppressants 13%. The most frequent mould identified were Aspergillus spp (53.6%), Mucorales (16%), Fusarium spp (8.8%), Alternaria spp (5.6%) and other filamentous (3.2%). All received antifungals, 82% monotherapy, 18% combined therapy, there was surgical defocation in 90% of mucormycosis. The overall mortality was 42%. When comparing 2005-2009 vs 2010-2015, there was a significant increase in incidence and a tendency to lower mortality in the second period. Conclusions: Over a period of 10 years, we observed an increase in the incidence of EFI by filamentous, aspergillosis was the most frequent etiology and the overall mortality was 42%.

Prevalência de infecções fúngicas em um laboratório de análises clínicas da cidade de Veranópolis, Rio Grande do Sul / Prevalence of the fungal infecctions in a clinical analysis laboratory in the city of Veranópolis, Rio Grande do Sul

Objetivo: As infecções fúngicas, atualmente, constituem um problema de saúde pública, devido à elevada prevalência e frequência com que acometem a população. Fatores predisponentes como condições climáticas, região geográfica e características culturais e socioeconômicas favorecem a ocorrência destas infecções. O objetivo do presente estudo foi determinar a prevalência das infecções fúngicas em pacientes atendidos em um laboratório de análises clínicas na cidade de Veranópolis, Rio Grande do Sul (RS). Métodos: Foi realizado um estudo retrospectivo, por meio da consulta ao banco de dados, de 403 laudos de exames micológicos, direto e cultural, realizados no laboratório durante o período de abril de 2014 a abril de 2017. Resultados: Dentre os laudos analisados no estudo, houve positividade em 251 (62,28%) exames micológicos diretos (EMD) e em 226 (56,08%) exames culturais. As micoses superficiais e cutâneas constituíram as principais infecções diagnosticadas, sendo os fungos dermatófitos os agentes etiológicos mais isolados (69,47%), representados majoritariamente pela espécie Trichophyton mentagrophytes, seguidos por leveduras (20,35%), especialmente as pertencentes ao gênero Candida. Houve predomínio de infecções em pacientes do sexo feminino (64,54% EMD e 61,5% cultura) e adultos de 31 a 60 anos foram os mais acometidos pelas micoses diagnosticadas (58,96% EMD e 60,62% cultura). Conclusão: O conhecimento do perfil epidemiológico pode servir como guia na correta identificação do patógeno no diagnóstico laboratorial, influenciando diretamente no prognóstico e na escolha da conduta terapêutica adequada.

Objective: The fungal infections are a public health issue, because ofthe high prevalence and frequency that they victimize the population. Predisposal factors as bioclimatic conditions, geographic region and cultural and socioeconomic characteristics favor the occurrence of this infections, generating, this way, variation in the prevalence of species in each region. The goal of the present study was to determinate the prevalence of fungal infections in patients in a clinical analysis laboratory in the city of Veranópolis, Rio Grande do Sul (RS). Methods: A retrospective study was made, using the consult of data basis, of 403 direct and culture mycological examination reports made in the laboratory during the period of April 2014 to April 2017. Results: In the analyzed examination reports in the study, there was positivity in 251 (62,28%) mycological direct exams (EMD) and, in 226 (56,08%) cultural exams. The superficial and cutaneous mycosis constituted the mainly diagnosed infections, being the dermatophytic fungi the most isolated etiological factors (69,47%), represented mainly, by the Trichophyton mentagrophytes species, followed by yeasts (20,35%), especially those who belong to the Candida genre. There was prevalence of infections in women patients (64,54% EMD and 61,5% culture). Adults among 31 and 60 years old were the most victimized by the diagnosed mycosis (58,96% EMD e 60,62% cultural). Conclusion: Knowledge of the epidemiological profile can serve as guide in the correct identification of the pathogen in the laboratory diagnosis, influencing directly in the prognosis and in the choice of the right therapeutic conduct

Cutaneous mycobiota of boid snakes kept in captivity / Microbiota cutânea de serpentes Boidae mantidas em cativeiro

Boids are large, constrictor snakes that feed mostly on mammals, reptiles, and birds. These animals are commonly raised as pets, and their improper handling can favor the emergence of fungal infections, which can lead to dermatological diseases that are undiagnosed in nature. Here, we isolate and identify the filamentous fungi that compose the mycobiota of the scales of boid snakes kept in captivity at the Biological Museum of the Butantan Institute. Thirty individuals of four species were evaluated: four Eunectes murinus, twelve Boa constrictor constrictor, seven Corallus hortulanus, and seven Epicrates crassus. Microbiological samples were collected by rubbing small square carpets on the snake scales. We isolated five genera of fungi: Penicillium sp. (30%), Aspergillus sp. (25%), Mucor sp. (25%), Acremonium sp. (10%), and Scopulariopsis sp. (10%). Approximately half of the snakes evaluated had filamentous fungi on the scales, but only 12% of the individuals were colonized by more than one fungal genus. We found no dermatophytes in the evaluated species. Our results provide an overview of the fungal mycobiota of the population of boids kept in the Biological Museum, allowing the identification of possible pathogens.(AU)

Boidae são serpentes grandes e constritoras, que comem normalmente mamíferos, répteis e aves. São comumente criadas como animais de estimação, e seu manuseio inadequado pode favorecer infecções fúngicas emergentes, as quais podem causar doenças dermatológicas que não são diagnosticadas na natureza. Foram isolados e identificados os fungos filamentosos da microbiota das escamas das serpentes Boidae mantidas em cativeiro no Museu Biológico do Instituto Butantan. Trinta indivíduos de quatro espécies foram avaliados: quatro Eunectes murinus, 12 Boa constrictor constrictor, sete Corallus hortulanus e sete Epicrates crassus. As amostras microbiológicas foram coletadas por fricção, em pequenos quadrados de carpete das escamas das serpentes. Isolaram-se cinco gêneros de fungos: Penicillium sp. (30%), Aspergillus sp. (25%), Mucor sp. (25%), Acremonium sp. (10%) e Scopulariopsis sp. (10%). Aproximadamente metade das serpentes avaliadas tinham fungos filamentosos nas escamas e apenas 12% dos indivíduos foram colonizados por mais de um gênero. Não foram encontrados dermatófitos nas espécies avaliadas. Os resultados mostraram uma visão geral da microbiota fúngica da população de boídeos mantidos no Museu Biológico, o que permitiu a identificação de possíveis patógenos.(AU)

Caracterização de queijos coloniais - fungos filamentosos e características físico-químicas / Colonial cheese characterization - filamentous fungi and physicochemical characteristics

Este estudo teve como objetivo isolar e identificar fungos filamentosos de queijos coloniais comercializados na região metropolitana de Porto Alegre/RS, assim como avaliar suas características físico-químicas. Para isso, foram adquiridos dez amostras de queijos coloniais, as quais foram porcionadas e encaminhadas para análise microbiológica e físico-quimica. Os resultados apontaram que o fungo filamentoso mais prevalente nos queijos coloniais avaliados foi o Aspergillus fumigatus presente em três das 10 amostras. Já os resultados médios para os teores de gordura e umidade foram respectivamente 27,0g/100g e 56,99%. O pH médio foi de 5,46. Os resultados das análises demostraram que há grande variabilidade entre os queijos avaliados, sendo necessárias mais pesquisas sobre o tema, a fim de permitir a definição de suas características e propor legislação específica.

Avaliação microbiológica e parasitológica de barras de cereais comercializadas em Taubaté, SP / Microbiological and parasitological evaluation of cereal bars marketed in Taubaté, SP

O objetivo do presente estudo foi avaliar a presença de fungos, bactérias e partes de insetos de 30 barras de cerais, sendo 10 tradicionais, 10 lights e 10 diets, de diferentes marcas, comercializadas no município de Taubaté, São Paulo. As amostras foram avaliadas quanto à contagem total de bactérias aeróbias mesófilas e fungos e contagem de larvas, ovos e demais sujidades. Os resultados apontaram que todas as amostras estavam contaminadas com bactérias e fungos, sendo que algumas amostras apresentaram elevadas contagens de bactérias aeróbias mesófilas. Todas as amostras apresentaram fragmentos de insetos.

The aim of the present study was to evaluate the presence of fungi, bacteria and parts of insects of 30 cereal bars, 10 of which were traditional, 10 lights and 10 diets of different brands marketed in the city of Taubaté, São Paulo. Samples were evaluated for total counts of aerobic mesophilic bacteria, fungi and counts of larvae, eggs and other contaminants. The results indicated that all samples were contaminated with bacteria, fungi, and some samples showed high counts of aerobic mesophilic bacteria. All samples showed fragments of insects.

Cellulolytic and lipolytic fungi isolated from soil and leaf litter samples from the Cerrado (Brazilian Savanna) / Hongos celulolíticos y lipolíticos aislados a partir de muestras de suelo y de hojarasca del Cerrado (Sabana brasileña)

Abstract The Brazilian savanna, known as the Cerrado, is a biome with a high degree of endemism, with the potential to house many microorganisms suitable for biotechnological exploitation, especially fungi. The Cerrado soil, which is usually acidic, is a favorable environment for the growth of fungi capable of degrading lignocellulosic materials. The aim of the present study was to isolate cellulolytic filamentous fungi native to the Cerrado. Samples of soil and leaf litter were collected in three points of Cerrado State Park, located in the South of Brazil, during the rainy season in September 2014. Samples were stored in sterile plastic bags, transported at room temperature and kept at 4 ºC for three days. Filamentous fungi were isolated by successive inoculations in PDA (maintained at 30 ºC). Cellulase activity was tested in CMC (carboxymethyl cellulose) medium and lipase activity was assessed in medium containing phenol red and tween 20 (incubated at 37 °C), and in medium supplemented with Rhodamine B (kept at 30 °C). We isolated a total of 28 strains, 25 produced cellulase, detected with lugol in strains grown in CMC medium. The isolates were identified morphologically (color, form of growth) and by sequencing of the 18S rRNA region, with both techniques producing congruent results. One strain of Colletotrichum boninense and one strain of Trichoderma sp., both isolated from soil samples, presented the highest cellulolytic activity. All strains exhibited lipolytic activity, with enzyme production and activity influenced by temperature. The present study revealed new strains of known filamentous fungi that can be applied in biomass degradation. These strains are suitable for optimization of culture conditions, which could lead to the economic feasibility of the process. Rev. Biol. Trop. 66(1): 237-245. Epub 2018 March 01.

Resumen La sabana brasileña conocida como Cerrado, es un bioma con alto grado de endemismo con el potencial de albergar muchos microorganismos de alto interés biotecnológico. Los hongos producen muchas enzimas que se aplican ampliamente en procesos industriales. El suelo del Cerrado, que suele ser ácido, es un ambiente favorable para el crecimiento de hongos capaces de degradar materiales lignocelulósicos. El objetivo del presente estudio fue aislar los hongos celulolíticos filamentosos nativos del Cerrado. Muestras de suelo y de hojarasca fueron recolectadas en el Parque Estatal del Cerrado, ubicado en el sur de Brasil. De 28 cepas aisladas, 25 produjeron celulasa que fue detectada con lugol. Las cepas fueron cultivadas en medio CMC (carboximetilcelulosa). Los aislados fueron identificados morfológicamente (color, forma de crecimiento) y por secuenciación de la región 18S rRNA, produciendo resultados congruentes. Una cepa de Colletotrichum boninense y una cepa de Trichoderma sp., ambas aisladas de muestras de suelo, presentaron la mayor actividad celulolítica. Todas las cepas mostraron actividad lipolítica, la producción y la actividad se vieron influidos por la temperatura. El presente estudio revela nuevas cepas de hongos filamentosos conocidos con potencial de aplicación en la degradación de la biomasa, sin embargo, la optimización de las condiciones de cultivo es necesaria para lograr la viabilidad económica.

Qualidade microbiológica de açaí industrializado / Microbiological quality of industrialized açai

Amostras de açaí (Euterpe oleracea Mart.) tipo fino congeladas e sem adição de quaisquer outros ingredientes foram adquiridas no comércio varejista da cidade do Rio de Janeiro. Foram analisadas microbiologicamente 54 amostras, pertencentes a 6 diferentes marcas (A, B, C, D, E e F). Todas as amostras analisadas encontravam-se dentro do prazo de validade estabelecido pelo fabricante e as empresas eram registradas no MAPA. Os micro-organismos analisados foram: Salmonella, Escherichia coli, fungos filamentosos e leveduras, de acordo com o preconizado pela legislação brasileira vigente. Considerando-se o limite máximo possível estabelecido pela lei, 2 lotes de cada marca B, E e F e 1 lote de cada marca C e D apresentaram contagens acima do permitido para fungos filamentosos e leveduras. Apenas 2 amostras apresentaram contagens de Escherichia coli, ambas estando fora do máximo estabelecido pela legislação vigente. Constatou- -se ausência de Salmonella em todas as amostras analisadas. A presença de fungos filamentosos, leveduras e Escherichia coli pode ser um indicativo de práticas inadequadas durante a fabricação do açaí. Assim, é importante ressaltar a necessidade de um controle rigoroso da cadeia do frio, uma vez que esses micro-organismos podem ser responsáveis pela deterioração do produto.

[Role of G-protein alpha sub-units in the morphogenic processes of filamentous Ascomycota fungi]. / Papel de las subunidades alfa de proteínas G en los procesos morfogénicos de hongos filamentosos de la división Ascomycota.

The phylum Ascomycota comprises about 75% of all the fungal species described, and includes species of medical, phytosanitary, agricultural, and biotechnological importance. The ability to spread, explore, and colonise new substrates is a feature of critical importance for this group of organisms. In this regard, basic processes such as conidial germination, the extension of hyphae and sporulation, make up the backbone of development in most filamentous fungi. These processes require specialised morphogenic machinery, coordinated and regulated by mechanisms that are still being elucidated. In recent years, substantial progress has been made in understanding the role of the signalling pathway mediated by heterotrimericG proteins in basic biological processes of many filamentous fungi. This review focuses on the role of the alpha subunits of heterotrimericG proteins in the morphogenic processes of filamentous Ascomycota.

Bioprospecting Aspergillus section Nigri in Atlantic Forest soil and plant litter / Bioprospecção de Aspergillus seção Nigri em solo e serrapilheira da Mata Atlântica

The use of fungi as a source of enzymes has become widespread in various industrial and commercial areas, and Aspergillus section Nigri has significant potential for producing enzymes. The aim of this study was to isolate Aspergillus section Nigri from plant litter and soil from the Atlantic Forest biome and evaluate it with regards to hydrolytic enzyme production. The trials for producing the enzymes were carried out in Petri dishes, using different culture mediums adapted for microbial growth and with the respective substrates for inducing enzyme production - cellulase (carboxymethyl cellulose), protease (skimmed milk), amylase (soluble starch), pectinase (citrus pectin), and phytase (Pikovskaya medium). Forty-two fungi were isolated, 16.7% derived from the plant litter layer and 83.3% derived from soil at a depth of 0 to 5 cm and 5 to 10 cm. All of the isolated lineages presented amylase, protease, and phytase production, with 90.4% producing cellulase and no lineage producing pectinase. From the results, the significant potential for Atlantic Forest fungi as hydrolytic enzyme producers could be perceived. The enzymatic activity evaluations presented a satisfactory result when compared with the scientific literature.(AU)

A utilização dos fungos como fonte de enzimas vem adquirindo status de destaque nas mais variadas áreas industriais e comerciais, e os Aspergillus membros da seção Nigri possuem significativo potencial para produção de enzimas. Os objetivos deste estudo foram isolar e avaliar Aspergillus da seção Nigri de serrapilheira e solos do bioma Mata Atlântica quanto à produção de enzimas hidrolíticas. Os ensaios para produção das enzimas foram realizados em placas de Petri, utilizando diferentes meios de cultivo, adequados ao crescimento microbiano e com a presença dos respectivos substratos indutores à produção das enzimas - celulases (carboximetilcelulose), proteases (leite desnatado), amilases (amido solúvel), pectinases (pectina cítrica) e fitase (meio Pikovskaya). Foram isolados 42 fungos, sendo desse total 16,7% provenientes da camada de serrapilheira e 83,3% provenientes do solo na profundidade de 0 a 5 cm e 5 a 10 cm. Todas as linhagens isoladas apresentaram produção de amilases, protease e fitase, 90,4% produziram celulase, e nenhuma linhagem produziu pectinase. Com esses resultados, percebeu-se significativo potencial dos fungos da Mata Atlântica como produtores de enzimas hidrolíticas. As avaliações da atividade enzimática apresentaram resultado satisfatório quando comparados à literatura científica.(AU)

Bioprospection and characterization of the amylolytic activity by filamentous fungi from Brazilian Atlantic Forest / Bioprospecção e caracterização da atividade amilolítica de fungos filamentosos da Mata Atlântica Brasileira

Abstract Filamentous fungi are widely diverse and ubiquitous organisms. Such biodiversity is barely known, making room for a great potential still to be discovered, especially in tropical environments - which are favorable to growth and species variety. Filamentous fungi are extensively applied to the production of industrial enzymes, such as the amylases. This class of enzymes acts in the hydrolysis of starch to glucose or maltooligosaccharides. In this work twenty-five filamentous fungi were isolated from samples of decomposing material collected in the Brazilian Atlantic Forest. The two best amylase producers were identified as Aspergillus brasiliensis and Rhizopus oryzae. Both are mesophilic, they grow well in organic nitrogen-rich media produce great amounts of glucoamylases. The enzymes of A. brasiliensis and R. oryzae are different, possibly because of their phylogenetical distance. The best amylase production of A. brasiliensis occurred during 120 hours with initial pH of 7.5; it had a better activity in the pH range of 3.5-5.0 and at 60-75°C. Both fungal glucoamylase had wide pH stability (3-8) and were activated by Mn2+. R. oryzae best production occurred in 96 hours and at pH 6.5. Its amylases had a greater activity in the pH range of 4.0-5.5 and temperature at 50-65ºC. The most significant difference between the enzymes produced by both fungi is the resistance to thermal denaturation: A. brasiliensis glucoamylase had a T50 of 60 minutes at 70ºC. The R. oryzae glucoamylase only had a residual activity when incubated at 50°C with a 12 min T50.

Resumo Fungos filamentosos são organismos amplamente diversificados e ubíquos. Esta biodiversidade ainda é pouco caracterizada, desta forma, há um grande potencial a ser descoberto, sobretudo em biomas tropicais, que favorecem o crescimento e diversificação de espécies. Fungos filamentosos são extensivamente utilizados para a produção industrial de enzimas, como as amilases. Esta classe de enzimas atua na hidrólise do amido em glicose ou maltooligossacarídeos. Neste trabalho 25 cepas de fungos filamentosos foram isoladas a partir de amostras de material em decomposição coletados na Mata Atlântica Brasileira. As duas cepas que produziram mais amilases foram identificadas como Aspergillus brasiliensis e Rhizopus oryzae. Ambos os fungos são mesofílicos, crescem bem em meio de cultivo rico em nitrogênio orgânico, e produziram grande quantidade de glucoamilase. As enzimas de A. brasiliensis e R. oryzae possuem características distintas, possivelmente devido à distância filogenética das espécies. A produção de amilase mais expressiva de A. brasiliensis ocorreu em 120 horas de cultivo e pH inicial de 7,5; possui maior atividade em temperaturas entre 60-75ºC e pH entre 3,5-5,0. Ambas glucoamilases fúngicas obtiveram ampla estabilidade de pH (3-8) e foram ativadas por Mn2+. A melhor produção de R. oryzae ocorreu em 96 horas de cultivo e pH 6,5. Suas amilases são mais ativas na faixa de pH de 4,0-5,5 e temperatura entre 50-60ºC. A diferença mais significativa dentre as enzimas produzidas pelos fungos selecionados é a resistência à desnaturação térmica, tendo a glucoamilase de A. brasiliensis um T50 de 60 minutos a 70ºC, já a glucoamilase de R. oryzae somente obteve atividade residual quando incubada a 50°C, com um T50 de apenas 12 minutos.

Fungal production of the anti-leukemic enzyme L-asparaginase: from screening to medium development / Produção fúngica da enzima antileucêmica L-asparaginase: da triagem ao desenvolvimento de um meio de cultivo

The treatment of acute lymphoblastic leukemia is challenging due to side effects, efficacy of the available drugs, and costs. Utilization of L-asparaginase as a therapeutic agent is essential to increase survival of patients. However, costs are elevated and the bacterial forms of the enzyme cause reactions that result in its inhibition by the immune system. Therapeutics alternatives may be searched among eukaryote producers, like fungi. Twelve strains of filamentous fungi were evaluated regarding expression of L-asparaginase activity. The profile of nitrogen assimilation and radial growth were determined for strains which showed higher production ratios. Three media were formulated after selection of the carbon source and carbon/nitrogen ratio that better induced L-asparaginase expression by Penicillium sp. and Fusarium sp. The enzyme activity produced in liquid media reached 8.3 U min.-1 mL-1 (Penicillium sp. T6.2) and 11.4 U min.-1 mL-1 (Fusarium sp.) after 72 hours of cultivation in Bacelar-1 medium. These data show that good producers can be found among fungi, and adjustment of productive processes may offer an alternative to implement eukaryote L-asparaginase production.

O tratamento da leucemia linfoblástica aguda é desafiador devido aos efeitos adversos, à eficácia dos fármacos disponíveis e aos custos envolvidos. A utilização de L-asparaginase como agente terapêutico é fundamental para aumentar a sobrevida do paciente. Porém, seu custo é elevado e as formas bacterianas da enzima causam reações que resultam na sua inibição pelo sistema imune. Alternativas terapêuticas podem ser buscadas entre produtores eucariontes, como os fungos. Doze linhagens de fungos filamentosos foram avaliadas quanto à expressão de atividade de L-asparaginase. O perfil de assimilação de nitrogênio e o crescimento radial foram determinados para as linhagens com maior razão de produção. Três meios foram formulados após a seleção da fonte de carbono e da razão carbono/nitrogênio que melhor induziram a expressão de L-asparaginase por Penicillium sp. e Fusarium sp. A atividade enzimática produzida em meio líquido alcançou 8,32 U min.-1 mL-1 (Penicillium sp. T6.2) e 11,45 U min.-1 mL-1 (Fusarium sp.) após 72 horas de cultivo no meio Bacelar-1. Esses dados mostram que bons produtores podem ser encontrados entre os fungos e que ajustes nos processos produtivos podem oferecer uma alternativa para a implementação da produção de L-asparaginase eucarionte.

Influencia del campo magnético sobre la distribución de los hongos en el aire de un local cerrado / Influence of magnetic field on fungi distribution in the air of a closed local

Los campos eléctricos y magnéticos pueden atraer microorganismos, por lo tanto el objetivo de este estudio fue analizar la influencia del campo magnético sobre la distribución de los hongos en el aire de un local cerrado. Se colocaron placas de Petri con agar Extracto de Malta en un local cerrado aplicándose un campo magnético de 10 mT generado por una bobina de 60 Hz/220 V. Se realizó un diseño multifactorial D-óptimo de tres factores (tiempo de exposición al aire, distancia al centro de la bobina y tiempo de tratamiento con campo magnético) y se determinó el recuento de hongos filamentosos en unidades formadoras de colonias por metro cúbico de aire, según el método de sedimentación propuesto por Omeliansky. Se obtuvo un modelo cuadrático en los rangos evaluados, que indicó que el tratamiento aplicado tuvo influencia significativa sobre la distribución de las esporas debido al aumento del recuento de los hongos, cuyo crecimiento resultó estimulado con respecto a los controles. El mayor efecto estimulante se observó a 1 m de distancia de la bobina (0,4 mT) y a 2 h de exposición, alcanzándose concentraciones fúngicas que pudieran ser peligrosas para la salud humana según la Organización Mundial de la Salud.

Electric and magnetic fields can attract microorganisms; based on those premises the aim of this study was to analyze the influence of a magnetic field on the fungi distribution in the air of a closed environment. Uncovered Petri dishes with Malt Extract Agar were placed in a closed area where a magnetic field of 10 mT generated by a coil of 60 Hz / 220 V was applied. A multifactorial D-optimum design considering three elements (air exposure time, distance to the center of the coil and time of magnetic field application) to determine the amount of filamentous fungi, expressed as the number of colony forming units by cubic meter of air, according to the sedimentation method proposed by Omeliansky. A quadratic model of the evaluated ranges was obtained, which indicated that the treatment applied showed significant influence on fungal spore distribution demonstrated by the increased fungal count which growth was stimulated by the treatment applied when compared to the controls. The largest stimulant effect was observed at 1 m of distance from the coil (0.4 mT) and after 2 h of exposure, conditions that permitted the growth of fungal concentrations that could be dangerous for human health according to the World Health Organization.

Produção de proteases por fungos filamentosos isolados do cerrado do centro-oeste brasileiro / Protease production by filamentous fungi isolated from the midwestern Brazilian Cerrado

Proteases ácidas pertencem a um importante grupo de enzimas industriais produzidas por fungos filamentosos, com aplicações na indústria de alimentos, de couro, farmacêutica e de cosméticos. O objetivo principal deste trabalho foi avaliar a produção de proteases ácidas extracelulares de fungos filamentosos isolados do solo do cerrado do centro-oeste brasileiro. Inicialmente, foi realizada uma triagem para avaliar a capacidade de 17 linhagens de fungos quanto à produção de protease em meio de cultura contendo Agar-leite. O fungo Aspergillus foetidus foi selecionado como melhor produtor de protease ácida extracelular. Visando à otimização da produção de proteases pelo fungo selecionado, avaliou-se a influência de diversos fatores no cultivo (pH, temperatura, agitação e diferentes fontes de nitrogênio e carbono). Após essa etapa, um planejamento experimental estatístico foi realizado com as variáveis independentes temperatura, pH inicial do meio e fonte de carbono e nitrogênio. A produção máxima de protease foi encontrada (63,7 U/mL) nas condições: pH inicial do meio igual a 7,0 a 28 ºC, 150 rpm em peptona 2% (p/v). Os estudos em biorreator demonstraram produção de protease nas condições de agitação e aeração iguais à 300 rpm e 1,0 vvm, após 120 h de cultivo. Os ensaios com diferentes temperaturas para a estimativa dos parâmetros termodinâmicos demonstraram que a protease ácida produzida pelo fungo é altamente estável apresentando máxima atividade em pH 5,0 e temperatura ótima igual a 55ºC. E, finalmente, para a purificação da enzima foi realizada cromatografia de gel-filtração. A enzima apresentou massa molecular de 50,6 kDa, e a análise do zimograma confirmou a atividade proteolítica. Além disso, a protease purificada foi inibida pelo composto pepstatina, indicando uma característica de protease ácida. Esses resultados obtidos demonstram um fungo filamentoso produtor de uma nova protease ácida com potencial aplicação para indústria farmacêutica e de cosméticos

The acid proteases belong to the most important group of industrial enzymes produced by filamentous fungi, with applications in the food, leather, pharmaceutical and cosmetics industries. This study aimed the evaluation of extracellular acid proteases production from filamentous fungi isolated from different samples of the midwestern Brazil cerrado. Initially, a screening was performed to assess the ability of the 17 strains of yeast for production of protease-agar medium containing milk culture. The Aspergillus foetidus was selected as the best producer. Aimed at optimizing the production of proteases by the selected fungus, first evaluated the influence of various factors on the cultivation (pH, temperatura, agitation and different sources of nitrogen and carbon). After this step, a statistical experimental design was carried out with the independent variables temperatura, initial pH of the medium and source of carbon and nitrogen. The best conditions for protease production were (63.7 U / mL): initial pH values greater than 7.0, at 28 °C, 150 rpm peptone 2% (w/v). Aiming future production of this protease in industrial scale, studies have shown better in bioreactor protease production under the conditions of agitation and aeration equal to 300 rpm and 1.0 vvm, after 120 h of cultive. The tests at different temperaturas to estimate the thermodynamic parameters showed that the acid protease produced by the fungus is highly stable with maximum activity at pH 5.0 and optimum temperatura of 55 °C. And finally, for the purification of the enzyme were performed gel-filtration chromatography. The enzyme had a molecular mass of 50.6 kDa, and the analysis of the zymogram showed a proteolytic band. Furthermore, the purified protease was inhibited by pepstatin compound, indicating a feature of acid protease. These results demonstrate a new filamentous fungus producing acid protease with potential application to pharmaceuticals and cosmetics