Metabolic and phenotypic plasticity may contribute for the higher virulence of Trichosporon asahii over other Trichosporonaceae members.

BACKGROUND: The Trichosporonaceae family comprises a large number of basidiomycetes widely distributed in nature. Some of its members, especially Trichosporon asahii, have the ability to cause human infections. This ability is related to a series of virulence factors, which include lytic enzymes production, biofilm formation, resistance to oxidising agents, melanin and glucuronoxylomannan in the cell wall, metabolic plasticity and phenotypic switching. The last two are poorly addressed within human pathogenic Trichosporonaceae. OBJECTIVE: These factors were herein studied to contribute with the knowledge of these emerging pathogens and to uncover mechanisms that would explain the higher frequency of T. asahii in human infections. METHODS: We included 79 clinical isolates phenotypically identified as Trichosporon spp. and performed their molecular identification. Lactate and N-acetyl glucosamine were the carbon sources of metabolic plasticity studies. Morphologically altered colonies after subcultures and incubation at 37°C indicated phenotypic switching. RESULTS AND CONCLUSION: The predominant species was T. asahii (n = 65), followed by Trichosporon inkin (n = 4), Apiotrichum montevideense (n = 3), Trichosporon japonicum (n = 2), Trichosporon faecale (n = 2), Cutaneotrichosporon debeurmannianum (n = 1), Trichosporon ovoides (n = 1) and Cutaneotrichosporon arboriforme (n = 1). T. asahii isolates had statistically higher growth on lactate and N-acetylglucosamine and on glucose during the first 72 h of culture. T. asahii, T. inkin and T. japonicum isolates were able to perform phenotypic switching. These results expand the virulence knowledge of Trichosporonaceae members and point for a role for metabolic plasticity and phenotypic switching on the trichosporonosis pathogenesis.

Production of Secreted Carbohydrates that Present Immunologic Similarities with the Cryptococcus Glucuronoxylomannan by Members of the Trichosporonaceae Family: A Comparative Study Among Species of Clinical Interest.

Glucuronoxylomannan (GXM) participates in several immunoregulatory mechanisms, which makes it an important Cryptococcus virulence factor that is essential for the disease. Trichosporon asahii and Trichosporon mucoides share with Cryptococcus species the ability to produce GXM. To check whether other opportunistic species in the Trichosporonaceae family produce GXM-like polysaccharides, extracts from 28 strains were produced from solid cultures and their carbohydrate content evaluated by the sulfuric acid / phenol method. Moreover, extracts were assessed for cryptococcal GXM cross-reactivity through latex agglutination and lateral flow assay methods. Cryptococcus neoformans and Saccharomyces cerevisiae were used as positive and negative controls, respectively. In addition to T. asahii, the species Trichosporon inkin, Apiotrichum montevideense, Trichosporon japonicum, Trichosporon faecale, Trichosporon ovoides, Cutaneotrichosporon debeurmannianum, and Cutaneotrichosporon arboriformis are also producers of a polysaccharide immunologically similar to the GXM produced by human pathogenic Cryptococcus species. The carbohydrate concentration of the extracts presented a positive correlation with the GXM contents determined by titration of both methodologies. These results add several species to the list of fungal pathogens that produce glycans of the GXM type and bring information about the origin of potential false-positive results on immunological tests for diagnosis of cryptococcosis based on GXM detection.

First description of Candida nivariensis in Brazil: antifungal susceptibility profile and potential virulence attributes.

This study evaluated the antifungal susceptibility profile and the production of potential virulence attributes in a clinical strain of Candida nivariensis for the first time in Brazil, as identified by sequencing the internal transcribed spacer (ITS)1-5.8S-ITS2 region and D1/D2 domains of the 28S of the rDNA. For comparative purposes, tests were also performed with reference strains. All strains presented low planktonic minimal inhibitory concentrations (PMICs) to amphotericin B (AMB), caspofungin (CAS), and voriconazole. However, our strain showed elevated planktonic MICs to posaconazole (POS) and itraconazole, in addition to fluconazole resistance. Adherence to inert surfaces was conducted onto glass and polystyrene. The biofilm formation and antifungal susceptibility on biofilm-growing cells were evaluated by crystal violet staining and a XTT reduction assay. All fungal strains were able to bind both tested surfaces and form biofilm, with a binding preference to polystyrene (p < 0.001). AMB promoted significant reductions (≈50%) in biofilm production by our C. nivariensis strain using both methodologies. This reduction was also observed for CAS and POS, but only in the XTT assay. All strains were excellent protease producers and moderate phytase producers, but lipases were not detected. This study reinforces the pathogenic potential of C. nivariensis and its possible resistance profile to the azolic drugs generally used for candidiasis management.

First description of Candida nivariensis in Brazil: antifungal susceptibility profile and potential virulence attributes

This study evaluated the antifungal susceptibility profile and the production of potential virulence attributes in a clinical strain of Candida nivariensis for the first time in Brazil, as identified by sequencing the internal transcribed spacer (ITS)1-5.8S-ITS2 region and D1/D2 domains of the 28S of the rDNA. For comparative purposes, tests were also performed with reference strains. All strains presented low planktonic minimal inhibitory concentrations (PMICs) to amphotericin B (AMB), caspofungin (CAS), and voriconazole. However, our strain showed elevated planktonic MICs to posaconazole (POS) and itraconazole, in addition to fluconazole resistance. Adherence to inert surfaces was conducted onto glass and polystyrene. The biofilm formation and antifungal susceptibility on biofilm-growing cells were evaluated by crystal violet staining and a XTT reduction assay. All fungal strains were able to bind both tested surfaces and form biofilm, with a binding preference to polystyrene (p < 0.001). AMB promoted significant reductions (≈50%) in biofilm production by our C. nivariensis strain using both methodologies. This reduction was also observed for CAS and POS, but only in the XTT assay. All strains were excellent protease producers and moderate phytase producers, but lipases were not detected. This study reinforces the pathogenic potential of C. nivariensis and its possible resistance profile to the azolic drugs generally used for candidiasis management.

Análise microbiológica de isolados de Fusarium spp. obtidos em um hospital de oncologia do Rio de janeiro, Brasil

Fungos do gênero Fusarium spp. são ubíquos e geralmente não são patogênicos. Algumas espécies termotolerantes podem causar infecção localizada em pele traumatizada e unhas de hospedeiros imunocompetentes. Grave infecção disseminada é comum em pacientes imunossuprimidos, principalmente neutropênicos portadores de neoplasias. Fusarium spp. tem sido relatado como patógeno emergente em centros oncológicos, sendo considerado atualmente a terceira causa de micose invasiva no Instituto Nacional do Câncer \2013 Inca, Rio de Janeiro. É fundamental o diagnóstico precoce do fungo, assim como a correta identificação laboratorial da espécie isolada, para se estabelecer o correto tratamento e o sucesso terapêutico. O objetivo deste estudo foi identificar isolados de Fusarium spp., de origem clínica e ambiental, por técnicas fenotípicas e genotípicas, e o perfil de sensibilidade aos antifúngicos em um hospital de oncologia do Rio de Janeiro, no período de janeiro a setembro de 2014. Foram avaliados 42 isolados de Fusarium spp., sendo 35 isolados clínicos provenientes de 19 pacientes com cultura positiva para Fusarium spp., obtidos principalmente de hemoculturas, e sete isolados ambientais provenientes de diferentes locais do hospital, obtidos durante investigação epidemiológica nas enfermarias onde os pacientes estavam internados. Dezessete pacientes apresentaram fusariose, um paciente teve infecção localizada de pele e outro paciente apenas colonização em lavado brônquico alveolar. A maioria é representada por pacientes pediátricos (58,8%), do gênero masculino (58,8%), neutropênicos (52,9%), com doença de base hematológica (70,6%), internados no hospital e submetidos à quimioterapia (88,2%) Todos os pacientes com fusariose utilizaram cateter de longa duração. Através da amplificação e sequenciamento da região Internally Transcribed Spacer (ITS) e do gene Elongator Factor 1\03B1 (EF), foi possível indentificar três espécies diferentes. O isolado mais frequente foi FOSC - complexo de espécies Fusarium oxysporum (32; 76,2%), seguido de FSSC - complexo de espécies Fusarium solani (9; 21,4%) e FIESC - complexo de espécies Fusarium incarnatum equiseti (1; 2,4%). Foi possível identificar 100% de similaridade entre 31 isolados clínicos e uma amostra ambiental obtida na sala de manutenção de cateter do hospital, identificados como FOSC. Houve boa correlação entre os resultados das identificações morfológica e molecular dos isolados. Anfotericina B foi a única droga com atividade in vitro, com CIM 2-8 \03BCg/ml para os isolados clínicos, sugerindo resistência. Posaconazol não demonstrou inibição do crescimento. Enquanto voriconazol demonstrou valores mais elevados de CIM para FOSC, com MG de 10,7 \03BCg/ml, quando comparado a FSSC, com MG de 8 \03BCg/ml. Não houve diferença significativa entre a sensibilidade dos antifúngicos testados e as espécies identificadas. Os resultados encontrados confirmam a resistência intrínseca natural do gênero