稀释液中添加精氨酸和牛血清白蛋白对驴冻精品质的影响研究

Effect of arginine and bovine serum albumin added to the diluent on the quality of donkey frozen semen

为了探究在驴精液冷冻稀释液中添加不同浓度的精氨酸和牛血清白蛋白(BSA)对德州驴冷冻精液品质的影响,并找出两种试剂混合最佳添加浓度,试验用分别添加不同浓度的精氨酸(0.5,1.0,1.5,2.0,2.5 mg/mL)和BSA(1,2,3,4,5 mg/mL)的常规驴精液冷冻稀释液对德州驴精液进行稀释,稀释后的精液液氮冷冻保存30 d后解冻,检测精液的精子活率、顶体完整率及质膜完整率,找出最佳浓度;然后选择精氨酸和BSA最佳浓度及其左右相邻浓度,两两组合成9个浓度组合对德州驴精液进行稀释,然后根据上述步骤找出最佳浓度组合。结果表明:单独添加精氨酸时,浓度为1.5 mg/mL时精液的精子活率、顶体完整率及质膜完整率较优于其他浓度,其中顶体完整率与未添加精氨酸相比显著提高(P<0.05)。单独添加BSA时,浓度为3 mg/mL时精液的精子活率、质膜完整率较优于其他浓度,但对顶体完整率无显著改善作用,其中精子活率、顶体完整率与未添加BSA相比极显著提高(P<0.01);精氨酸和BSA联合添加时(除1.5 mg/mL×3 mg/mL、1.5 mg/mL×4 mg/mL、2.0 mg/mL×4 mg/mL组合外)对驴精液的精子活率、顶体完整率、质膜完整率都有一定程度的改善作用,其中1.0 mg/mL×2 mg/mL组合的精子活率和顶体完整率效果最好,质膜完整率较高;1.0 mg/mL×3 mg/mL组合的精子质膜完整率最高。说明联合添加精氨酸与BSA在一定程度上可以提高德州驴精液冷冻后精子品质,二者联合有一定的协同作用,最佳组合浓度为精氨酸1.0 mg/mL、BSA 2 mg/mL或3 mg/mL。

In order to explore the effect of various concentrations of arginine and bovine serum albumin(BSA)added to the diluent on the quality of Dezhou donkey frozen semen, and to find out the optimal mixed concentration of the two reagents, in the experiment, Dezhou donkey semen was diluted with conventional frozen diluent of donkey semen with different concentrations of arginine(0.5, 1.0, 1.5, 2.0, 2.5 mg/mL) and BSA(1, 2, 3, 4, 5 mg/mL).The diluted semen was cryopreserved with liquid nitrogen for 30 days and then thawed to determine the sperm viability, acrosome integrity and plasma membrane integrity of semen, so as to find the optimal concentration.Then the optimal concentrations of Arg and BSA and their adjacent concentrations were selected.Dezhou donkey semen was diluted 9 concentrations in pairs, and the optimal combination concentration was found according to the above steps. The results showed that when Arg was added alone, the sperm viability, acrosome integrity and plasma membrane integrity of semen at the concentration of 1.5 mg/mL were better than those at other concentrations, and compared with no arginine, the acrosome integrity rate was significantly improved(P<0.05).When BSA was added alone, the sperm viability and plasma membrane integrity of semen at the concentration of 3 mg/mL were better than those at other concentrations, but there was no significant effect on the acrosome integrityrate;compared with no BSA, the sperm viability and acrosome integrity were significantly improved(P<0.01).The combined supplementation of Arg and BSA(except 1.5 mg/mL×3 mg/mL, 1.5 mg/mL×4 mg/mL and 2.0 mg/mL×4 mg/mL) could improve the sperm viability, acrosome integrity and plasma membrane integrity of donkey semen to a certain extent, among which the combination of 1.0 mg/mL×2 mg/mL had the best effect on sperm viability and acrosome integrity, and plasma membrane integrity was good.The plasma membrane integrity rate in the combination of 1.0 mg/mL×3 mg/mL was the highest.The results indicated that the combined addition of Arg and BSA could improve the quality of frozen Dezhou donkey semen to some extent.The combination of the two reagents had a certain synergistic effect, and the best combination concentration was arginine 1.0 mg/mL and BSA 2 mg/mL or 3 mg/mL.