Isolation and in vitro culture of trypanosomes from Leptodactylus ocellatus from the Atlantic Forest in a new experimental culture medium.
The purpose of this study was to verify the in vitro development of Trypanosoma sp. isolated from Leptodactylus ocellatus frogs under a new protocol using a biphasic medium composed of Novy, McNeal, and Nicolle (NNN) blood agar medium as a solid phase and liver infusion, brain heart infusion, and tryptose (LIBHIT) medium as a liquid phase. Blood forms, collected by cardiac puncture or after the maceration of different organs, were inoculated in culture tubes containing the biphasic medium composed by NNN and LIBHIT. Trypanosomes were observed 4 days postinoculation; most bloodstream trypomastigotes had differentiated into epimastigotes and amastigotes by this time. Trypomastigotes were again observed in older cultures (7 days). Parasites were successfully subcultured for 8 mo in this medium and successfully cryopreserved. The present study provides a new protocol medium for the isolation and culture of anuran trypanosomes.
Laboratório de Biologia Celular e Ultraestrutura, Departamento de Microbiologia Geral, Instituto de Microbiologia Prof. Paulo de Góes, Centro de Ciências da Saúde, bloco I, Universidade Federal do Rio de Janeiro, Ilha do Fundão, Rio de Janeiro, Brazil.
M
M
Lemos
C S F
CS
Souza
S C Gonçalves
SC
da Costa
T
T
Souto-Padrón
M
M
D'Agosto
D000818:Animals; D001001:Anura / Q000469:parasitology*; D003470:Culture Media; D014345:Trypanosoma / Q000254:growth & development* / Q000302:isolation & purification*; D014352:Trypanosomiasis / Q000469:parasitology / Q000662:veterinary*
D003470:Culture Media