Interações entre diferentes espécies de Candida na candidose experimental em modelos de invertebrados e vertebrados

• Ano de defesa: 2013
• Autor(a) principal: Rossoni, Rodnei Dennis [UNESP]
• Tipo de documento: Dissertação
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Estadual Paulista (Unesp)
• Palavras-chave em Português: Candida; Candidiase; Patogenicidade; Modelos animais em pesquisa
• Link de acesso: http://hdl.handle.net/11449/108849

The aim of this study was to evaluate in vitro and in vivo interactions between different Candida species through the formation of biofilms in 96-well plates, induction of experimental candidiasis in Galleria mellonella model and immunosuppressed mice. We studied the standard strains of the following species: C. albicans (ATCC 18804), C. krusei (ATCC 6258) and C. glabrata (ATCC 90030). Monotypic biofilms of each species and heterotypic biofilms were performed on the bottom of 96-well plate for 48 h. Then, the amount of biofilm was analyzed by determining the number of colony forming units (CFU/mL). The larvae of G. mellonella were inoculated with homotypic and heterotypic suspensions of Candida (105 cells/mL) and incubated at 37°C. For 5 days, the number of dead larvae was assessed daily for survival curve analysis. In another experiment, after 0, 2, 4, 8, 12 and 24 h of infection with Candida, hemolymph of worms was extracted for counting the fungal cells (CFU/mL). For induction of oral candidiasis in mice, the animals were inoculated with homotypic or heterotypic microbial suspensions containing 108 cells/mL. After 48 h of the last inoculation, samples of the tongue were collected and seed in HiCrome chromogenic agar for counting of CFU/mL recovered from the oral cavity. Then, the animals were euthanized and their tongues removed for macroscopic and microscopic analysis. The analysis of Candida biofilms in vitro CFU/mL, the hemolymph of G. mellonella and recovery of mice were made by ANOVA, Tukey test or Student’s t test. The analysis of the survival curve was performed using GraphPad Prism using the Log-rank test (Mantel - Cox). Kruskal-Wallis or Mann-Whitney were applied for evaluation scores of macroscopic and histological analysis (p≤0,05). The results of biofilms in vitro have demonstrated a lower number of CFU/mL of C. albicans biofilms in heterotypic biofilm compared ...