Author(s)

Pedro Henrique Magalhães Cardoso¹, Bosco Christiano Maciel da Silva², Jairo Ivo dos Santos³, Rennan Luiz Oliveira dos Santos⁴, Diniz Pereira Leite Júnior⁵, Marcos Ereno Auler⁶, Luciana da Silva Ruiz⁷, Eriques Gonçalves da Silva⁴, Débora Moreira⁴, Carina Domaneschi⁴, Francisco de Assis Baroni⁸, Marcia de Souza Carvalho Melhem⁹, Marilena dos Anjos Martins⁹, Claudete Rodrigues Paula⁴

¹Faculty of Veterinary Medicine and Zootechny-FMVZ-USP, University of São Paulo (USP), São Paulo, Brazil.
²Faculty of Medicine/FMUSP, University of São Paulo (USP), São Paulo, Brazil.
³Federal University of Santa Catarina (UFSC), Florianópolis, Brazil.
⁴Faculty of Dentistry, University of São Paulo (FOUSP), São Paulo, Brazil.
⁵Faculty of Medicine, Federal University of Mato Grosso (UFMT), Cuiabá, Brazil.
⁶Faculty of Pharmacy, University of the Western Center of Paraná (UNICENTRO), Guarapuava, Brazil.
⁷Institute Adolfo Lutz (IAL), Bauru, Brazil.
⁸Federal Rural University of Rio de Janeiro (UFRRJ), Seropédica, Brazil.
⁹Institute Adolfo Lutz (IAL), São Paulo, Brazil.

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 São Paulo University, in São 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.

Cryptococcus Complex, Genotyping, Protease, Phospholipase, Etest, Culture Collection

Henrique Magalhães Cardoso, P. , Christiano Maciel da Silva, B. , Ivo dos Santos, J. , Luiz Oliveira dos Santos, R. , Pereira Leite Júnior, D. , Ereno Auler, M. , da Silva Ruiz, L. , Gonçalves da Silva, E. , Moreira, D. , Domaneschi, C. , de Assis Baroni, F. , de Souza Carvalho Melhem, M. , dos Anjos Martins, M. and Rodrigues Paula, C. (2018) Characterization of Clinical and Environmental Isolates of Cryptococcus neoformans/Cryptococcus gattii Complex Maintained in Yeast Culture Collection in São Paulo, Brazil. Open Journal of Epidemiology, 8, 76-92. doi: 10.4236/ojepi.2018.82007.