RESUMO
The goal of this investigation was to determine the percentage of male and female bovine embryos produced after oocyte fertilization with Percoll density gradient centrifugation or with self-migration (swim-up) selected semen. In Experiment 1, sperm selection was performed by 90-45% discontinuous Percoll density gradient centrifugation (T1) and swim-up (T2). InExperiment 2, in addition to the discontinuous gradient, a 67.5% continuous gradient, and 5 or 10 min centrifugation were used. Four treatment groups were defined (TI = continuous, 5 min, TII = discontinuous, 5 min, TIII = continuous, 10 min and TIV = discontinuous,10 min). Embryos were sexed using PCR. In Experiment 1,Percoll density gradient centrifugation (n = 185) resulted in 48.6% (n = 90) male and 51.4% (n = 95) female embryos, and swim-up (n = 142) in 58.4% (n = 83) male and 41.6% (n = 59) female embryos. In Experiment 2, the percentages of male and female embryos obtained in TI (n = 93), TII (n = 70), TIII (n = 82) and TIV (n = 82) were 49.5% (n = 46) and 50.5% (n = 47), 57.1% (n = 40) and 42.9% (n = 30), 36.6% (n = 30) and 63.4% (n = 52) and 48.8% (n = 40) and 51.2% (n = 42), respectively. Swim-up (T2) and continuous Percoll gradient centrifugation for 10 min (TIII) showed a deviation toward males (P = 0.044) and females (P = 0.015), respectively, when compared to the 50% expected percentage of each gender. There was no difference in male and female percentages in the other treatment groups from Experiments 1 and 2, when compared to the 50% expected percentage of each gender neither between treatment.(AU)
Assuntos
Animais , Embrião de Mamíferos/embriologia , Capacitação Espermática , DNA/análise , Bovinos/classificação , Cromossomos/análiseRESUMO
The goal of this investigation was to determine the percentage of male and female bovine embryos produced after oocyte fertilization with Percoll density gradient centrifugation or with self-migration (swim-up) selected semen. In Experiment 1, sperm selection was performed by 90-45% discontinuous Percoll density gradient centrifugation (T1) and swim-up (T2). InExperiment 2, in addition to the discontinuous gradient, a 67.5% continuous gradient, and 5 or 10 min centrifugation were used. Four treatment groups were defined (TI = continuous, 5 min, TII = discontinuous, 5 min, TIII = continuous, 10 min and TIV = discontinuous,10 min). Embryos were sexed using PCR. In Experiment 1,Percoll density gradient centrifugation (n = 185) resulted in 48.6% (n = 90) male and 51.4% (n = 95) female embryos, and swim-up (n = 142) in 58.4% (n = 83) male and 41.6% (n = 59) female embryos. In Experiment 2, the percentages of male and female embryos obtained in TI (n = 93), TII (n = 70), TIII (n = 82) and TIV (n = 82) were 49.5% (n = 46) and 50.5% (n = 47), 57.1% (n = 40) and 42.9% (n = 30), 36.6% (n = 30) and 63.4% (n = 52) and 48.8% (n = 40) and 51.2% (n = 42), respectively. Swim-up (T2) and continuous Percoll gradient centrifugation for 10 min (TIII) showed a deviation toward males (P = 0.044) and females (P = 0.015), respectively, when compared to the 50% expected percentage of each gender. There was no difference in male and female percentages in the other treatment groups from Experiments 1 and 2, when compared to the 50% expected percentage of each gender neither between treatment.
Assuntos
Animais , DNA , Capacitação Espermática , Embrião de Mamíferos/embriologia , Bovinos/classificação , Cromossomos/análiseRESUMO
The aim of this study was to increase the bovine embryonic development rate, adding recombinant human growth hormone (rhGH) to maturation medium of bovine oocytes. Oocytes were matured for 24 h in TCM 199 Earle's salts and five treatments were developed: T1, 0.01 IU/ml of recombinant human follicle stimulating hormone (rhFSH); T2, 0.01 IU/ml of rhFSH + 100 ng/ml of rhGH; T3, 0.01 IU/ml of rhFSH + 1000 ng/ml of rhGH; T4, 100 ng/ml of rhGH; and T5, 1000 ng/ml of rhGH at 39 degrees C and 5% of CO(2) in air and saturated humidity. In vitro fertilization from cumulus-oocyte complexes was conducted in TALP-Fert medium (18-22 h) and spermatozoa were selected by Percoll gradient. Zygotes were incubated in SOFaaci medium in 5% of CO(2) in air, 5% of O(2) at 39 degrees C and saturated humidity for 11 days. There was no statistical difference in cleavage rate and embryo production on day 7 and day 9 among treatments. However, the hatching rate increased significantly in the T4 and T5 treatments (11.0 and 12.8%, respectively), compared with the T1 treatment (4.6%) (p < 0.05). Therefore, the rhGH addition to the oocyte maturation medium showed beneficial effects on the hatching rate of in vitro-produced bovine embryos.