METODOLOGIAS DE DESNUDAMENTO PARCIAL DE OÓCITOS BOVINOS MATURADOS E SUBMETIDOS À VITRIFICAÇÃO

Different methods of partial denudation of maturated bovine oocytes were evaluated for in vitro embryo production and vitrification procedures, in three experiments. First the effect of partial denudation by successive pipetting was evaluated. In the second experiment, the denudation by pipetting was compared with the use of hyaluronidase, and finally, the effect of both methods was evaluated in regard to the vitrification of oocytes. Oocytes were submitted to treatments after 22 hours of maturation in TCM 199 medium. For the vitrification, they were firstly exposed for 30 seconds to an equilibrium solution and for 20 seconds to a vitrification solution (20% EG + 20% DMSO + 0.3M SUC), loaded in open pulled straws (OPS) and plunged into liquid nitrogen. The rewarming of the oocytes suspensions was carried out by plunged them in decreasing sucrose concentrations. Oocytes of all treatments were submitted to an additional 2 hours maturation period, followed by fertilization. Presumptive zygotes were cultured in SOFaaci medium. In the first experiment the denudation resulted in a decreased blastocyst rate (28.7% to 20.5% PDiferentes metodologias de desnudamento parcial de oócitos bovinos maturados foram avaliadas na produção in vitro de embriões e na vitrificação, em três experimentos. No primeiro avaliou-se o efeito do desnudamento parcial realizado por sucessivas pipetagens. No segundo, comparou-se o desnudamento através de sucessivas pipetagens, com o realizado pela enzima hialuronidase e no terceiro, avaliou-se a influência das duas metodologias na vitrificação de oócitos. Os oócitos foram submetidos aos tratamentos após 22 horas de maturação em meio TCM 199. Para a vitrificação, foram expostos por 30 segundos a uma solução de equilíbrio e 20 segundos a uma solução de vitrificação (20%EG + 20% Dimitilsufoxido (DMSO) + 0,3MSAC), envasados em palhetas abertas e estiradas (OPS) e mergulhados em N2 líquido. O reaquecimento foi realizado com exposição a concentrações decrescentes de sacarose. Os oócitos de todos os tratamentos foram submetidos a duas horas adicionais de maturação, seguida da fecundação e os possíveis zigotos cultivados em meio SOFaaci. No experimento I foi observado um decréscimo de 28,7% para 20,5% na taxa de blastocistos com o emprego do desnudamento (

The Advertisement Calls and Distribution of Two Sympatric Species of \u3cem\u3eChiasmocleis\u3c/em\u3e (Méhely 1904) (Anura, Microhylidae, Gastrophryninae) from the Atlantic Forest

The advertisement calls of Chiasmocleis cordeiroi and C. crucis are described for populations from the municipalities of Igrapiúna and Camacan, respectively, state of Bahia, Brazil. Both calls consist of multipulsed notes produced in series. Differences between the two calls are: dominant frequency, higher in C. cordeiroi (range 4500-4898 Hz; C. crucis range 4069-4435 Hz); note rate, higher in C. cordeiroi (range 6.20--7.46 s/note; C. crucis range 5.17-5.59 s/note); pulse rate, higher in C. cordeiroi (151.82-194.83 s/note; C. crucis range 125.30- 142.12 s/note); and the structure of the modulation patterns of the notes. Moreover, the advertisement calls of C. crucis and C. cordeiroi are more similar than the calls of all syntopic congeners. Furthermore, the current distribution of both species was extended