XXI ALAM
Resumo:297-1


Poster (Painel)
297-1Freezing methodology for maintenance isolated from enviromental Cryptococcus neoformans.
Autores:Edson Maria Torres (UNESP - Universidade Estadual Paulista Júlio Mesquita Filho) ; Valter Luiz Iost Teodoro (UNESP - Universidade Estadual Paulista Júlio Mesquita Filho) ; Fernanda Patrícia Gullo (UNESP - Universidade Estadual Paulista Júlio Mesquita Filho) ; Janaina de Cássia Orlandi Sardi (UNESP - Universidade Estadual Paulista Júlio Mesquita Filho) ; Rosangela Aparecida Moraes Silva (UNESP - Universidade Estadual Paulista Júlio Mesquita Filho) ; Ana Marisa Fusco-almeida (UNESP - Universidade Estadual Paulista Júlio Mesquita Filho) ; Maria José Soares Mendes-giannini (UNESP - Universidade Estadual Paulista Júlio Mesquita Filho)

Resumo

Introduction: Cryptococcus neoformans is the pathogen most commonly found in fungal infections of the central nervous system (CNS), is distributed globally, isolated from soil, contaminated with mainly broiler chickens, pigeons, in particular, besides wood decomposition, fruits, vegetables and hollow trees, affecting mainly immunocompromised hosts, especially HIV-positive patient, with this important risk factor for cryptococcosis. Given this it is necessary to the preservation of environmental and clinical strains for retrospective and prospective studies of this fungus. Mycology collections are useful for fungal studies focused on biology, etiology, epidemiology, resistance, virulence, molecular typing. The maintenance of Cryptococcus neoformans by successive samplings favor contamination and reduces the virulence of the pathogen. Objective: This study aimed to evaluate the efficacy of preservation of 28 environmental strains of C. neoformans var. grubii (VNI) identified by PCR / RFLP and 02 ATCC strains of C. neoformans (90012 and 24065). Method: These samples after 48 hours of repiques on Sabouraud dextrose agar were grown in sterile vials containing brain heart infusion broth (BHI) at 20% glycerol. The vials were incubated at 300C for 48 hours, then deposited on refrigeration for 07 days and then placed in a freezer at – 20º for 30 days. Next the samples were passed for the defrosting and plated on Sabouraud dextrose agar. Since then the same samples were frozen at – 80ºC for 30 days and also defrosted and plated on Sabouraud dextrose agar to check viability. Results: 100% of the samples frozen at-20ºC and - 80ºC were viable after 30 days of freezing. Discussion and Conclusions: The methodology used for the freezing of the strains was easy to perform and storage, low cost and efficient conservation and can be used for maintenance of mycology collections of Cryptococcus neoformans, ensuring the integrity of the fungus for retrospective and prospective studies.


Palavras-chave:  Cryptococcus neoformans, Freezing methodology, Maintenance of isolated