Componentes estruturais da matriz do biofilme de Paracoccidioides spp. e sua interação com diferentes microrganismos
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Data
2020-08-24
Autores
Oliveira, Lariane Teodoro
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Universidade Estadual Paulista (Unesp)
Resumo
A habilidade de formar biofilme por Paracoccidioides brasiliensis ocorre tanto in vitro como in vivo. A paracoccidioidomicose é uma doença sistêmica que envolve, primariamente, os pulmões e, posteriormente, dissemina-se para outros órgãos e sistemas, incluindo a cavidade bucal. As lesões bucais podem estar associadas a várias bactérias e fungos e a co-agregação de Paracoccidioides spp com essas bactérias e fungos ainda não foi demonstrada. Além disso, os princípios básicos da formação de biofilme deste fungo e a sua compreensão molecular e organização estrutural precisam ser mais bem conhecidos, visando melhores abordagens terapêuticas. Assim, o estudo teve como objetivo caracterizar os elementos estruturais da matriz do biofilme monoespécie de Paracoccidioides spp., bem como avaliar a interação desse fungo com as bactérias Streptococcus mutans e Staphylococcus aureus e com o fungo Candida albicans na formação de biofilme. Foram utilizados os isolados P. brasiliensis (Pb18), P. lutzii (Pb01 e 8334) e P. restrepiensis (339 e 192), na fase leveduriforme, avaliando-se a atividade metabólica, a biomassa total e a estrutura tridimensional dos biofilmes. A matriz extracelular dos biofilmes de Paracoccidioides spp. foi extraída por sonicação e foram quantificados os polissacarídeos, proteínas e DNA extracelular (eDNA). Todas as cepas estudadas de Paracoccidioides foram capazes de formar biofilme, apresentando valores maiores de atividade metabólica e de biomassa em 144 horas. A matriz do biofilme de P. brasiliensis foi similar ao de P.lutzii, destacando-se a presença maior de polissacarídeos, seguido de proteínas e de eDNA. Os biofilmes dupla-espécies de Paracoccidioides spp. com S. mutans e S. aureus, de modo geral, mostraram antagonismo evidenciado pelos resultados de biomassa total, atividade metabólica, contagem de colônias e eletromicrografias, os quais mostraram inibição no crescimento dos fungos. Em contraste, a interação de P. brasiliensis com C. albicans mostrou que os dois microrganismos podem coexistir em um mesmo ambiente, conforme análise in vitro e in vivo no modelo Galleria mellonella. Esses dados ampliam o conhecimento associado à dinâmica de crescimento de biofilme fúngico e pode contribuir para a descoberta de novas estratégias terapêuticas para essas infecções.
The ability to form biofilm by Paracoccidioides brasiliensis occurs both in vitro and in vivo. Paracoccidioidomycosis is a systemic disease that primarily involves the lungs and subsequently spreads to other organs and systems, including the oral cavity. Oral lesions can be associated with various bacteria and fungi. The co-aggregation of Paracoccidioides spp with these bacteria and fungi has not yet been demonstrated. In addition, the basic principles of biofilm formation of this fungus and its molecular understanding and structural organization need to be better known, aiming at better therapeutic approaches. Thus, the study aimed to characterize the structural elements of the monospecies biofilm matrix of Paracoccidioides spp., and evaluate the interaction of this fungus with the bacteria Streptococcus mutans and Staphylococcus aureus and with the fungus Candida albicans in the formation of biofilm. The isolates P. brasiliensis (Pb18), P. lutzii (Pb01 and 8334), and P. restrepiensis (339 and 192) were used in the yeast phase, evaluating the metabolic activity, the total biomass, and the three-dimensional structure of the biofilms. The extracellular matrix of the biofilms of Paracoccidioides spp. was extracted by sonication and polysaccharides, proteins, and extracellular DNA (eDNA) were quantified. All studied strains of Paracoccidioides were able to form a biofilm, with higher values of metabolic activity and biomass in 144 hours. The biofilm matrix of P. brasiliensis was similar to P. lutzii, made up of polysaccharides, following proteins, and eDNA. The mixed biofilms of Paracoccidioides spp. with the bacteria S. mutans and S. aureus, in general, showed antagonism evidenced by the results of total biomass, metabolic activity, colony counts, and electromicrographs, which showed inhibition in the growth of fungi. In contrast, the interaction of P. brasiliensis with C. albicans showed that the two microorganisms could coexist in the same environment, according to in vitro and in vivo analysis in Galleria mellonella model. These data expand the knowledge associated with the growth dynamics of fungal biofilm and can contribute to the discovery of new therapeutic strategies for these infections.
The ability to form biofilm by Paracoccidioides brasiliensis occurs both in vitro and in vivo. Paracoccidioidomycosis is a systemic disease that primarily involves the lungs and subsequently spreads to other organs and systems, including the oral cavity. Oral lesions can be associated with various bacteria and fungi. The co-aggregation of Paracoccidioides spp with these bacteria and fungi has not yet been demonstrated. In addition, the basic principles of biofilm formation of this fungus and its molecular understanding and structural organization need to be better known, aiming at better therapeutic approaches. Thus, the study aimed to characterize the structural elements of the monospecies biofilm matrix of Paracoccidioides spp., and evaluate the interaction of this fungus with the bacteria Streptococcus mutans and Staphylococcus aureus and with the fungus Candida albicans in the formation of biofilm. The isolates P. brasiliensis (Pb18), P. lutzii (Pb01 and 8334), and P. restrepiensis (339 and 192) were used in the yeast phase, evaluating the metabolic activity, the total biomass, and the three-dimensional structure of the biofilms. The extracellular matrix of the biofilms of Paracoccidioides spp. was extracted by sonication and polysaccharides, proteins, and extracellular DNA (eDNA) were quantified. All studied strains of Paracoccidioides were able to form a biofilm, with higher values of metabolic activity and biomass in 144 hours. The biofilm matrix of P. brasiliensis was similar to P. lutzii, made up of polysaccharides, following proteins, and eDNA. The mixed biofilms of Paracoccidioides spp. with the bacteria S. mutans and S. aureus, in general, showed antagonism evidenced by the results of total biomass, metabolic activity, colony counts, and electromicrographs, which showed inhibition in the growth of fungi. In contrast, the interaction of P. brasiliensis with C. albicans showed that the two microorganisms could coexist in the same environment, according to in vitro and in vivo analysis in Galleria mellonella model. These data expand the knowledge associated with the growth dynamics of fungal biofilm and can contribute to the discovery of new therapeutic strategies for these infections.
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Palavras-chave
Biofilme, matriz extracelular, Paracoccidioides spp., Biofilme dupla- espécie, extracelular matrix