Resposta superovulatória e produção in vivo de embriões em ovelhas Santa Inês em função da dosagem de FSH
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Data
2016-05-16
Autores
Maciel, Giovanna Serpa [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
O objetivo do presente estudo foi avaliar o efeito da dose de FSH exógeno no tratamento gonadotrófico sobre a resposta superovulatória e produção in vivo de embriões em ovelhas. Vinte e quatro ovelhas Santa Inês foram submetidas à sincronização do estro com dispositivo intravaginal de progesterona (CIDR®) inserido no D0 e permaneceu até D8. Ao início do protocolo (D0) e na retirada do dispositivo intravaginal foram administrados 0,125 mg de análogo sintético da PGF2α (Sincrocio®). O tratamento gonadotrófico teve início 48 horas antes da retirada da progesterona (D6), quando as fêmeas foram aleatoriamente divididas em três grupos experimentais de acordo com a dose total de FSH suíno exógeno: G200 (n=8) - 200 mg; G133 (n=8) - 133 mg; e G100 (n=8) - 100 mg. As doses totais foram administradas em 8 aplicações à intervalos de 12 horas. No D6, as fêmeas receberam 300 UI de eCG. Foram realizadas ultrassonografias modo B e Doppler em cores, com o propósito de acompanhar o crescimento folicular da onda ovulatória, área de vascularização da parede folicular e o momento das ovulações. Seis dias após o início do estro, os embriões foram colhidos e avaliados morfologicamente e quanto ao percentual de células apoptóticas. Realizou-se teste de Kruskal Wallis e pós-teste de Dunns, sendo que as taxas foram comparadas pelo teste do Chi-quadrado. A variável área da vascularização folicular foi comparada entre os tratamentos e momentos pela ANOVA com medidas repetidas no tempo e pós-teste de Tukey, com análises de correlação (Pearson) para o número de folículos e área da vascularização dos mesmos. O nível de significância utilizado para os testes estatísticos foi de 5% (P >0,05). A duração do estro nas ovelhas foi menor no G200 em relação ao G133 e semelhante ao G100 (P = 0,0218). O momento ovulatório ocorreu tardiamente no G200 (P = 0,0037); redução do tamanho do folículo pré-ovulatório e taxa de redução do tamanho do folículo pré-ovulatório foram maiores (P = 0,0027 e 0,0024, respectivamente) para ovelhas do G200 em relação ao G100, entretanto semelhantes ao G133. A área da vascularização folicular variou com relação aos momentos, aumentando gradativamente com pico entre as 216 e 232 horas e posteriormente decaiu (P < 0,001). A taxa de ovulação e falhas anovulatórias apresentaram diferença estatística entre os grupos (P = 0,016), o G100 apresentou maior valor da taxa de ovulação em relação ao G1333 e semelhante ao G200 e o G100 apresentou o menor valor das falhas anovulatórias em relação ao G133 e semelhante ao G200. O número de estruturas não fecundadas e a taxa de não fecundados foram maiores nos animais pertencentes ao G100 (P = 0,008 e 0,0477, respectivamente) em relação ao G200 e semelhante ao G133; o número de embriões viáveis e taxa de viabilidade foram maiores (P = 0,038 e 0,0493, respectivamente) para ovelhas do G200 em relação ao G133, entretanto semelhante ao G100; a porcentagem de células apoptóticas foi maior no grupo G100. Pôde–se concluir que a dose de 200 mg apresentou os melhores resultados das variáveis estudadas, não sendo assim indicado o uso de menores doses de FSH (100 mg e 133 mg) em tratamentos de superestimulação ovulatória em ovelhas Santa Inês.
The aim of this study was to evaluate the effect of exogenous FSH dose in gonadotropin treatment on superovulatory response and in vivo production of embryos in sheep. Twenty four Santa Inês ewes were submitted to synchronization of estrus with intravaginal progesterone device (CIDR®) inserted into D0 and remained until D8. At the start of the protocol (D0) and in the removal of the intravaginal device was administered 0.125 mg of PGF2a synthetic analog (Sincrocio®). The gonadotrophin treatment was initiated 48 hours prior to removal of progesterone (D6) when the females were randomly divided into three groups according to the total dose of exogenous porcine FSH: G200 (n = 8) - 200 mg; G133 (n = 8) - 133 mg; and G100 (n = 8) - 100 mg. The total doses been administered in 8 applications at intervals of 12 hours. On D6, the females received 300 IU eCG. Were performed B mode ultrasonography and color Doppler, in order to verify follicular growth of ovulatory wave, area of vascularization of the follicular wall and the moment of ovulation. Six days after the the onset of estrus, the embryos were collected and assessed morphologically and regarding the percentage of apoptotic cells. Was performed Kruskal Wallis test and Dunns post-test, and the rates were compared using Chi-square test. The variable area of the follicular vascularization was compared between treatments and moments by ANOVA with repeated measures in time and Tukey post-test, with correlation analysis (Pearson) for the number of follicles and area of the vascularization. The significance level used for the statistical tests was 5% (P> 0.05). The duration of estrus of sheep was lower in G200 compared to G133 and similar to G100 (P = 0.0218). The ovulatory moment occurred later in G200 (P = 0.0037); reduction of the size of the preovulatory follicle and rate of reduction of the size of preovulatory follicles were higher (P = 0.0027 and 0.0024, respectively) for sheep of the G200 compared to G100, though similar to G133. The area of the follicle vascularization varied with relation to moments, gradually increasing with a peak between 216 and 232 hours and then declined (P <0.001). The ovulation rate and anovulatory failures showed statistical differences between groups (P = 0.016), the G100 showed higher ovulation rate in relation to the G1333 and similar to G200 and the G100 showed the lowest value of anovulatory failures in relation to G133 and similar to G200. The number of non-fecundated structures and unfertilized rate were higher in animals pertaining to G100 (P = 0.008 and 0.0477, respectively) compared to the G200 and similar to G133; the number of viable embryos and viability rate was higher (P = 0.038 and 0.0493, respectively) for sheep of the G200 compared to G133, though similar to G100; the percentage of apoptotic cells was higher in the group G100. It was concluded that the dose of 200 mg showed the best results of the studied variables and is therefore not indicated the use of smaller doses of FSH (100 mg and 133 mg) in superstimulation ovulatory treatments in Santa Inês ewes.
The aim of this study was to evaluate the effect of exogenous FSH dose in gonadotropin treatment on superovulatory response and in vivo production of embryos in sheep. Twenty four Santa Inês ewes were submitted to synchronization of estrus with intravaginal progesterone device (CIDR®) inserted into D0 and remained until D8. At the start of the protocol (D0) and in the removal of the intravaginal device was administered 0.125 mg of PGF2a synthetic analog (Sincrocio®). The gonadotrophin treatment was initiated 48 hours prior to removal of progesterone (D6) when the females were randomly divided into three groups according to the total dose of exogenous porcine FSH: G200 (n = 8) - 200 mg; G133 (n = 8) - 133 mg; and G100 (n = 8) - 100 mg. The total doses been administered in 8 applications at intervals of 12 hours. On D6, the females received 300 IU eCG. Were performed B mode ultrasonography and color Doppler, in order to verify follicular growth of ovulatory wave, area of vascularization of the follicular wall and the moment of ovulation. Six days after the the onset of estrus, the embryos were collected and assessed morphologically and regarding the percentage of apoptotic cells. Was performed Kruskal Wallis test and Dunns post-test, and the rates were compared using Chi-square test. The variable area of the follicular vascularization was compared between treatments and moments by ANOVA with repeated measures in time and Tukey post-test, with correlation analysis (Pearson) for the number of follicles and area of the vascularization. The significance level used for the statistical tests was 5% (P> 0.05). The duration of estrus of sheep was lower in G200 compared to G133 and similar to G100 (P = 0.0218). The ovulatory moment occurred later in G200 (P = 0.0037); reduction of the size of the preovulatory follicle and rate of reduction of the size of preovulatory follicles were higher (P = 0.0027 and 0.0024, respectively) for sheep of the G200 compared to G100, though similar to G133. The area of the follicle vascularization varied with relation to moments, gradually increasing with a peak between 216 and 232 hours and then declined (P <0.001). The ovulation rate and anovulatory failures showed statistical differences between groups (P = 0.016), the G100 showed higher ovulation rate in relation to the G1333 and similar to G200 and the G100 showed the lowest value of anovulatory failures in relation to G133 and similar to G200. The number of non-fecundated structures and unfertilized rate were higher in animals pertaining to G100 (P = 0.008 and 0.0477, respectively) compared to the G200 and similar to G133; the number of viable embryos and viability rate was higher (P = 0.038 and 0.0493, respectively) for sheep of the G200 compared to G133, though similar to G100; the percentage of apoptotic cells was higher in the group G100. It was concluded that the dose of 200 mg showed the best results of the studied variables and is therefore not indicated the use of smaller doses of FSH (100 mg and 133 mg) in superstimulation ovulatory treatments in Santa Inês ewes.
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Palavras-chave
Dose FSH exógeno, Superestimulação, Ovino, Dose FSH exogenous, Overstimulation, Ovine