To evaluate the effect of pFSH dose on the in vivo embryo production of Dorper ewes in the semi-arid northeast of Brazil, 40 sheep females were distributed into two groups of 20 animals that received intravaginal CIDR for 14 days, and two days before device removal, they received one of the following treatments: in the FSH200 group, the ewes received 200 mg of pFSH; and in the FSH128 group, the ewes received a total of 128 mg in decreasing doses every 12 h. Beginning 12 h after the conclusion of the treatments, estrus detection was performed every four hours using two Dorper rams of proven fertility. The ewes were mated at estrus onset and 24 hours later. Seven days after intravaginal device removal, the superovulatory response was evaluated, and embryo collection was performed using the laparotomy method. The recovered flushings were subjected to embryo searches under a stereomicroscope and classified according to their qualities. Analyses of variance (ANOVAs) and LSD tests were used to compare the different parameters. The data expressed as percentages were analysed by chi-square test. The ovulation rate was higher in the FSH200 group, which had 16.3 ± 0.3 corpora lutea (CL), than in the FSH128 group, which had 11.3 ± 0.3 CL (P<0.05). However, higher fertilization rate (83.6% vs. 62.4%) and higher transferable (86.0% vs. 71.6%) and freezable (67.9% vs. 40.8%) embryo rates were observed in the FSH 128 group compared with the group that received 200 mg. Furthermore, no significant differences in the remaining parameters were observed between the experimental groups (P>0.05), demonstrating that pFSH dose reduction promoted a greater production of freezable and transferable embryos in Dorper ewes subjected to MOET. ResumoCom o objetivo de avaliar o efeito da redução de pFSH na produção in vivo de embriões de ovelhas da raça Dorper exploradas no semiárido do Nordeste do Brasil, 40 fêmeas ovinas foram distribuídas em dois grupos com 20 animais, os quais receberam um CIDR intravaginal por 14 dias e dois dias antes da remoção do dispositivo receberam um tratamento superovulatório proposto: grupo FSH200 onde as ovelhas receberam 200 mg de pFSH, enquanto o grupo FSH128 totalizou 128 mg em doses decrescentes intervaladas por 12 horas. Doze horas após o final do tratamento, foi realizada a detecção de estro, a cada quatro horas, utilizando dois carneiros Dorper, de fertilidade comprovada. As ovelhas foram cobertas no início do estro e 24 horas depois. Sete dias após a retirada do dispositivo intravaginal, foi realizada a avaliação da resposta superovulatória e a colheita de embriões pelo método de laparotomia. O lavado recuperado foi submetido à procura de embriões em estereomicroscópio e classificados de acordo com sua qualidade. Para comparação dos diversos parâmetros, foi utilizada a Análise de Variância (ANOVA), seguida da realização do teste LSD. Os dados expressos em porcentagem foram submetidos ao Teste de Qui-quadrado. A taxa de ovulação foi superior no grupo FSH200 com 16,3 ± 0,3 corpos lúteos (CL) quand...
Knowledge about reproduction of white-lipped peccary is of great importance to assist with the conservation of this species and enable its rational use in captivity. This study aimed to evaluate the effect of ACP-103®, ACP-116® and BTS semen extenders on sperm viability during cooling of Tayassu pecari semen. Five ejaculates from four adult males were chilled. The animals were submitted to the protocols of sedation and anesthesia for semen collection by the electroejaculation method. After collection, the semen was macro- and microscopically assessed and diluted to reach 35x106 spermatozoa/mL in each of the three different extenders tested. The fresh-extended semen was packed in a BotuFLEX® thermal box to keep samples at 15°C for 24 hours. After cooling, the following semen parameters were analyzed: sperm motility, functional and structural integrity of sperm membranes, mitochondrial activity, chromatin condensation, and the thermoresistance test was performed. The parameters sperm motility, structural and functional integrity of sperm membranes, mitochondrial activity, and chromatin condensation were preserved after use of the extenders tested, and were similar to those of in natura semen (p>0.05). Curvilinear velocity (VCL) (p<0.05) was the only parameter with reduced values after cooling regardless of the extender used. The percentage of sperm with normal morphology was greater in samples cooled using the BTS extender (p<0.05). The ACP-103®, ACP-116® and BTS extenders can be used for the cooling and preservation of white-lipped peccary semen at 15°C for 24 hours.
Samples of semen and cervical mucus were provided by 18 couples. Cervical mucus was obtained for each day possible and stored at 4 degrees C until all the samples were collected. Flat capillary tubes were loaded with the mucous samples and spermatozoa from the husband's semen sample were allowed to migrate through the cervical mucus (3 cm column) into culture medium. The spermatozoa recovered after migration through cervical mucus were assayed in vitro with zona-free hamster oocytes. Control experiments were carried out using spermatozoa from the same semen sample but prepared by the swimming-up technique. Altogether, 557 eggs in the control group and 1236 eggs in the experimental group were analysed, and the results demonstrated that the % of sperm penetration, the mean number of sperm decondensations per penetrated egg and the mean number of spermatozoa adhering per egg all had higher values (P less than 0.05) for the control samples than for the experimental samples. We suggest that cervical mucus modifies human spermatozoa, as measured by their interaction with zona-free hamster oocytes.
To evaluate the effect of pFSH dose on the in vivo embryo production of Dorper ewes in the semi-arid northeast of Brazil, 40 sheep females were distributed into two groups of 20 animals that received intravaginal CIDR for 14 days, and two days before device removal, they received one of the following treatments: in the FSH200 group, the ewes received 200 mg of pFSH; and in the FSH128 group, the ewes received a total of 128 mg in decreasing doses every 12 h. Beginning 12 h after the conclusion of the treatments, estrus detection was performed every four hours using two Dorper rams of proven fertility. The ewes were mated at estrus onset and 24 hours later. Seven days after intravaginal device removal, the superovulatory response was evaluated, and embryo collection was performed using the laparotomy method. The recovered flushings were subjected to embryo searches under a stereomicroscope and classified according to their qualities. Analyses of variance (ANOVAs) and LSD tests were used to compare the different parameters. The data expressed as percentages were analysed by chi-square test. The ovulation rate was higher in the FSH200 group, which had 16.3 ± 0.3 corpora lutea (CL), than in the FSH128 group, which had 11.3 ± 0.3 CL (P<0.05). However, higher fertilization rate (83.6% vs. 62.4%) and higher transferable (86.0% vs. 71.6%) and freezable (67.9% vs. 40.8%) embryo rates were observed in the FSH 128 group compared with the group that received 200 mg. Furthermore, no significant differences in the remaining parameters were observed between the experimental groups (P>0.05), demonstrating that pFSH dose reduction promoted a greater production of freezable and transferable embryos in Dorper ewes subjected to MOET. ResumoCom o objetivo de avaliar o efeito da redução de pFSH na produção in vivo de embriões de ovelhas da raça Dorper exploradas no semiárido do Nordeste do Brasil, 40 fêmeas ovinas foram distribuídas em dois grupos com 20 animais, os quais receberam um CIDR intravaginal por 14 dias e dois dias antes da remoção do dispositivo receberam um tratamento superovulatório proposto: grupo FSH200 onde as ovelhas receberam 200 mg de pFSH, enquanto o grupo FSH128 totalizou 128 mg em doses decrescentes intervaladas por 12 horas. Doze horas após o final do tratamento, foi realizada a detecção de estro, a cada quatro horas, utilizando dois carneiros Dorper, de fertilidade comprovada. As ovelhas foram cobertas no início do estro e 24 horas depois. Sete dias após a retirada do dispositivo intravaginal, foi realizada a avaliação da resposta superovulatória e a colheita de embriões pelo método de laparotomia. O lavado recuperado foi submetido à procura de embriões em estereomicroscópio e classificados de acordo com sua qualidade. Para comparação dos diversos parâmetros, foi utilizada a Análise de Variância (ANOVA), seguida da realização do teste LSD. Os dados expressos em porcentagem foram submetidos ao Teste de Qui-quadrado. A taxa de ovulação foi superior no grupo FSH200 com 16,3 ± 0,3 corpos lúteos (CL) quand...
Zona-free hamster oocytes inseminated in vitro with acrosome-reacted guinea-pig spermatozoa were examined with the electron microscope. Guinea-pig spermatozoa, in the vicinity of the oocytes, consistently lacked the whole acrosome including the equatorial segment region. In cross fertilization sperm-egg membrane fusion does not differ significantly from that of normal fertilization. However, it was sometimes possible to observe protrusions of oocyte cytoplasm containing sperm chromatin in the process of dispersion.
RESUMO Objetivou-se avaliar o efeito do ácido docosa-hexaenoico (DHA), associado ou não ao Trolox®, na refrigeração de sêmen de garanhões da raça Mangalarga Marchador. Foram refrigerados 10 ejaculados nos diluidores: D1) BotuSêmen® (BS; controle); D2) BS + 30ngmL-1 de DHA (BS30DHA); D3) BS30DHA + 40µM de Trolox® (BS30DHA40T); D4) BS + 50ngmL-1 de DHA (BS50DHA); D5) BS50DHA + 40µM de Trolox® (BS50DHA40T). Após 48 horas de refrigeração, foram avaliados os parâmetros de movimento espermático, a integridade estrutural e funcional da membrana plasmática e a longevidade espermática. Todos os diluidores testados preservaram, de forma semelhante, a motilidade, a linearidade, a retilinearidade, a amplitude do deslocamento lateral da cabeça, a frequência do batimento flagelar cruzado, o percentual de hiperativos e a integridade estrutural e funcional da membrana espermática (P>0,05). O diluidor BS50DHA foi superior ao BS30DHA40T em preservar a VCL e a VSL e foi superior ao BS30DHA40T e ao BS50DHA40T em preservar a VAP e o índice de oscilação (P<0,05). Conclui-se que o uso do Trolox® em diluidores utilizados para refrigeração de sêmen de garanhões contendo ácido docosa-hexaenoico, nas concentrações propostas, não é indicado por alterar parâmetros de movimento espermático considerados importantes para a fertilidade.
RESUMO Queixada (Tayassu pecari) é um mamífero neotropical, classificado como vulnerável devido à caça e à destruição de seu habitat. Em razão das dificuldades na reprodução em cativeiro, técnicas de reprodução assistida podem ser aplicadas em programas de repovoamento da espécie. Dessa forma, objetivou-se identificar o melhor protocolo de teste hiposmótico (HOST) para avaliar a integridade funcional dos espermatozoides de queixada. O sêmen de quatro machos adultos foi coletado com auxílio de eletroejaculador após contenção física e protocolo de sedação e anestesia. O sêmen foi avaliado quanto às características macro e microscópicas e diluído nas seguintes soluções hiposmóticas: água destilada (0mOsmol/L), sacarose (50, 100, 150mOsm/L) e frutose (50, 100, 150mOsm/L). Cada amostra foi incubada em duplicata, e uma sofreu fixação em solução de citrato de sódio formolizado a 4%. Duzentos espermatozoides foram avaliados por amostra e classificados em reativos ou não ao HOST. Todas as soluções testadas foram semelhantes em identificar o percentual de espermatozoides reativos, independentemente de a amostra ser ou não fixada (P>0,05). Dessa forma, pode-se usar água destilada como HOST por este apresentar resultados similares e por ser um teste mais barato.
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