Avaliação dos efeitos probióticos de cepas clínicas de Lactobacillus spp. sobre diferentes espécies de Candida
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2018-02-05
Autores
Santos, Rafaella Braga [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
Devido ao aumento das infecções causadas por Candida albicans e espécies não-albicans, surge a necessidade de novas estratégias terapêuticas, como a identificação de cepas bacterianas com características probióticas. Assim, o objetivo foi avaliar a ação antimicrobiana de cepas clínicas de Lactobacillus paracasei, Lactobacillus fermentum e Lactobacillus rhamnosus sobre diferentes espécies de Candida (C. albicans, C. glabrata, C. krusei e C. tropicalis). Foram utilizadas cepas de Lactobacillus isoladas da cavidade bucal de indivíduos livres de cárie e cepas de Candida isoladas de lesões de candidose orofaríngea obtidas em estudos anteriores. Para estudo in vitro, foram formados biofilmes monoespécies de Candida (Grupo controle) e biofilmes mistos de Candida e Lactobacillus, nos quais os lactobacilos foram acrescentados antes da cepa de Candida (Grupo Profilático), após a cepa de Candida (Grupo Terapêutico) ou ao mesmo tempo da cepa de Candida (Grupo Simultâneo). Após 48 h de incubação, foi realizada a contagem do número de células viáveis de Candida (UFC/mL). Para estudo in vivo, os micro-organismos foram inoculados em larvas de G. mellonella e o desenvolvimento da candidose foi monitorado pela curva de sobrevivência, estudando-se os grupos experimentais descritos acima. Nos resultados in vitro, foi verificado que os efeitos inibitórios mais significativos de Lactobacillus sobre Candida ocorreram nos grupos profiláticos. As cepas de C. tropicalis e C. krusei foram mais sensíveis a ação de Lactobacillus do que C. albicans e C. glabrata. Além disso, foi verificado que as três espécies de Lactobacillus estudadas (L. paracasei, L. rhamnosus e L. fermentum) tiveram ação inibitória sobre Candida. Em relação ao estudo in vivo, também foi verificado que os grupos profiláticos tiveram maiores efeitos inibitórios sobre a candidose experimental em relação aos grupos terapêuticos, levando a um aumento significativo nas taxas de sobrevivência das larvas. Em relação as cepas estudadas, os efeitos profiláticos de Lactobacillus sobre a candidose, foram encontrados, de forma semelhante, para todas as cepas de Candida e Lactobacillus estudadas. Assim, concluiu-se que as cepas clínicas de L. paracasei 28.4, L. rhamnosus 5.2 e L. fermentum 20.4 foram capazes de inibir a formação de biofilme e desenvolvimento de candidose causados por C. albicans, C. krusei, C. tropicalis e C. glabrata, principalmente quando Lactobacillus foi administrado de forma profilática.
Due to the increase of infections caused by Candida albicans and non-albicans species, there is a need for new therapeutic strategies, such as the identification of bacterial strains with probiotic characteristics. The objective of this study was to evaluate the antimicrobial action of Lactobacillus paracasei, Lactobacillus fermentum and Lactobacillus rhamnosus strains on different Candida species (C. albicans, C. glabrata, C. krusei and C. tropicalis). We used Lactobacillus strains isolated from the oral cavity of caries-free and Candida strains isolated from lesions of oropharyngeal candidiasis. For in vitro study, monospecies of Candida (Control Group) and mixed biofilms of Candida and Lactobacillus were prepared, in which the lactobacillus were attached before the Candida strain (Profile Group), after a Candida (Therapeutic Group) strain or same time as the Candida strain (Simultaneous Group). After 48 h of incubation, we counting the number of viable Candida cells (CFU/mL). For in vivo study, we inoculate the microorganisms in larvae of G. mellonella and the development of the application for monitoring by survival curve, studying the experimental groups described above. In the in vitro results, we find that the most significant inhibitory effects of Lactobacillus on Candida occurred in the prophylactic groups. The strains of C. tropicalis and C. krusei were more sensitive to the action of Lactobacillus than C. albicans and C. glabrata. In addition, it we verified that the three species of Lactobacillus studied (L. paracasei, L. rhamnosus and L. fermentum) had an inhibitory action on Candida. In relation to the in vivo study, we verified that the prophylactic groups had more inhibitory effects on an experimental candidose in relation to the therapeutic groups, leading to a significant increase in the survival rates of the larvae. In relation to the studied strains, the prophylactic effects of Lactobacillus on candidiasis were similarly for all Candida and Lactobacillus strains studied. Thus, it concluded that the clinical strains of L. paracasei 28.4, L. rhamnosus 5.2 and L. fermentum 20.4 were able to inhibit a biofilm formation and development of Candida caused by C. albicans, C. krusei, C. tropicalis and C. glabrata, especially when Lactobacillus was given prophylactically. Keywords:
Due to the increase of infections caused by Candida albicans and non-albicans species, there is a need for new therapeutic strategies, such as the identification of bacterial strains with probiotic characteristics. The objective of this study was to evaluate the antimicrobial action of Lactobacillus paracasei, Lactobacillus fermentum and Lactobacillus rhamnosus strains on different Candida species (C. albicans, C. glabrata, C. krusei and C. tropicalis). We used Lactobacillus strains isolated from the oral cavity of caries-free and Candida strains isolated from lesions of oropharyngeal candidiasis. For in vitro study, monospecies of Candida (Control Group) and mixed biofilms of Candida and Lactobacillus were prepared, in which the lactobacillus were attached before the Candida strain (Profile Group), after a Candida (Therapeutic Group) strain or same time as the Candida strain (Simultaneous Group). After 48 h of incubation, we counting the number of viable Candida cells (CFU/mL). For in vivo study, we inoculate the microorganisms in larvae of G. mellonella and the development of the application for monitoring by survival curve, studying the experimental groups described above. In the in vitro results, we find that the most significant inhibitory effects of Lactobacillus on Candida occurred in the prophylactic groups. The strains of C. tropicalis and C. krusei were more sensitive to the action of Lactobacillus than C. albicans and C. glabrata. In addition, it we verified that the three species of Lactobacillus studied (L. paracasei, L. rhamnosus and L. fermentum) had an inhibitory action on Candida. In relation to the in vivo study, we verified that the prophylactic groups had more inhibitory effects on an experimental candidose in relation to the therapeutic groups, leading to a significant increase in the survival rates of the larvae. In relation to the studied strains, the prophylactic effects of Lactobacillus on candidiasis were similarly for all Candida and Lactobacillus strains studied. Thus, it concluded that the clinical strains of L. paracasei 28.4, L. rhamnosus 5.2 and L. fermentum 20.4 were able to inhibit a biofilm formation and development of Candida caused by C. albicans, C. krusei, C. tropicalis and C. glabrata, especially when Lactobacillus was given prophylactically. Keywords:
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Palavras-chave
Lactobacillus, Candida. Probiótico, Galleria mellonella, Biofilme, Probiotic, Biofilm