Characterization of Clinical and Environmental Isolates of <i>Cryptococcus neoformans</i>/<i>Cryptococcus gattii</i>Complex Maintained in Yeast Culture Collection in S&#227;o Paulo, Brazil  

作　　者：Pedro Henrique Magalhã es Cardoso Bosco Christiano Maciel da Silva Jairo Ivo dos Santos Rennan Luiz Oliveira dos Santos Diniz Pereira Leite Jú nior Marcos Ereno Auler Luciana da Silva Ruiz Eriques Gonç alves da Silva Dé bora Moreira Carina Domaneschi Francisco de Assis Baroni Marcia de Souza Carvalho Melhem Marilena dos Anjos Martins Claudete Rodrigues Paula 

机构地区：[1]Faculty of Veterinary Medicine and Zootechny-FMVZ-USP, University of Sã o Paulo (USP), Sã o Paulo, Brazil [2]Faculty of Medicine/FMUSP, University of Sã o Paulo (USP), Sã o Paulo, Brazil [3]Federal University of Santa Catarina (UFSC), Florianó polis, Brazil [4]Faculty of Dentistry, University of Sã o Paulo (FOUSP), Sã o Paulo, Brazil [5]Faculty of Medicine, Federal University of Mato Grosso (UFMT), Cuiabá , Brazil [6]Faculty of Pharmacy, University of the Western Center of Paraná (UNICENTRO), Guarapuava, Brazil [7]Institute Adolfo Lutz (IAL), Bauru, Brazil [8]Federal Rural University of Rio de Janeiro (UFRRJ), Seropé dica, Brazil [9]Institute Adolfo Lutz (IAL), Sã o Paulo, Brazil

出　　处：《Open Journal of Epidemiology》2018年第2期76-92,共17页流行病学期刊（英文）

摘　　要：Objective: As isolates of Cryptococcus are frequently kept collection stocks in institutions, sometimes without proper characterization, we sought to determine the genotype profiles, protease and phospholipase activities “in vitro” and the susceptibility testing for azoles and amphotericin B. Methodology: 84 isolates from several regions of Brazil (40 samples from clinical origin and 44 isolates from environmental origin) were maintained at the microorganism’s bank of the Biomedical Science Institute (ICB-USP) of the S&#227;o Paulo University, in S&#227;o Paulo, Brazil. This isolates was submitted fungal strains determination, DNA extraction and purification, determination of genotype by URA5 gene RFLP of CGB-positive isolates, protease and phospholipase activity and susceptibility to antifungals. Results: Of six CGB positive isolates tested by RFLP-PCR, only four presented a genomic profile consistent C. gattii species (VGII), while two other were C. neoformans (VNI and VNIII), indicating the existence of canavanine-resistante C. neoformans isolates in the culture collections. The clinical isolates secreted higher levels of phospholipase and environmental isolates but no differences were observed for the protease levels. Almost all isolates were sensible to azoles and amphotericin B. Conclusion: We point out in this research the existence of C. neoformans strains resistant to canavanine and intrinsic characteristic of C. gatti. These results demonstrate the importance to perform a detailed characterization of isolates kept in culture collections.Objective: As isolates of Cryptococcus are frequently kept collection stocks in institutions, sometimes without proper characterization, we sought to determine the genotype profiles, protease and phospholipase activities “in vitro” and the susceptibility testing for azoles and amphotericin B. Methodology: 84 isolates from several regions of Brazil (40 samples from clinical origin and 44 isolates from environmental origin) were maintained at the microorganism’s bank of the Biomedical Science Institute (ICB-USP) of the S&#227;o Paulo University, in S&#227;o Paulo, Brazil. This isolates was submitted fungal strains determination, DNA extraction and purification, determination of genotype by URA5 gene RFLP of CGB-positive isolates, protease and phospholipase activity and susceptibility to antifungals. Results: Of six CGB positive isolates tested by RFLP-PCR, only four presented a genomic profile consistent C. gattii species (VGII), while two other were C. neoformans (VNI and VNIII), indicating the existence of canavanine-resistante C. neoformans isolates in the culture collections. The clinical isolates secreted higher levels of phospholipase and environmental isolates but no differences were observed for the protease levels. Almost all isolates were sensible to azoles and amphotericin B. Conclusion: We point out in this research the existence of C. neoformans strains resistant to canavanine and intrinsic characteristic of C. gatti. These results demonstrate the importance to perform a detailed characterization of isolates kept in culture collections.

关 键 词：CRYPTOCOCCUS COMPLEX Genotyping Protease Phospholipase ETEST Culture COLLECTION 

分 类 号：R73[医药卫生—肿瘤]