Correlation between in vitro sperm kinetic, oxidative stress assessments and field fertility of cryopreserved bull semen

Lis Marques, D. Missio, K. Mattos, F. Cibin, F. G. Leivas, Daniela Dos Santos Brum
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Abstract

This study assessed kinetic parameters and oxidative stress in bull sperm after post-thaw (PT) or after sperm selection by Percoll™ gradient, and thermo resistance test (SS + TRT) to identify useful indicators of field fertility. For the experiment, commercial doses of frozen semen were obtained from six Aberdeen Angus bulls. Three of the bulls were classified as high fertility and three as low fertility according to the IFert™ index provided by the international breeding company CRV Lagoa. Pooled semen samples were distributed between two treatment groups for analysis: post-thaw (PT) or sperm selection (SS) (Percoll™) and thermal resistance test (SS + TRT). The samples were evaluated using sperm kinetics (CASA) (motility %, progressive motility %, VCL µm/s, VSL µm/s, VAP µm/s, LIN %, STR % and WOB%), production of reactive oxygen species (ROS), lipid peroxidation, superoxide dismutase (SOD) enzyme activity and total antioxidant capacity. Data were analyzed using Two-Way ANOVA, considering the fertility index, the treatment used in the samples as effects, and the interaction between these factors. When a significant effect was observed, the values were compared using the Bonferroni test. A Pearson Correlation analysis was performed between the fertility indices and the sperm parameters analyzed in vitro, to evaluate the relationship between sperm quality and the fecundity rate obtained by the bulls. Sperm kinetic parameters, including total motility, progressive motile, and beat cross-frequency, were higher in low fertility compared to high fertility bulls (P < 0.05). However, curvilinear velocity was greater in high fertility bulls followed by SS + TRT. Straight-line velocity, average path velocity, linearity, and beat cross-frequency beat were higher in high fertility bulls after SS + TRT. Reactive oxygen species was correlated with fertility after SS. In addition, there was a decrease in lipid peroxidation was observed only in high fertility bulls. However, lipid peroxidation and high fertility were correlated after PT and SS + TRT. The combination of in vitro sperm kinetic parameters predicted in vivo fertility more accurately than individual kinetic parameters. The lipid peroxidation of sperm is an important indicator of fertility in bulls. High fertility bulls appeared to be more susceptible to lipid peroxidation, which was only reduced in high fertility bulls, suggesting that their sperm can repair the damage induced by oxidative stress.
冷冻保存公牛精液的体外精子动力学、氧化应激评估与现场生育能力之间的相关性
本研究评估了公牛精子在解冻后(PT)或经 Percoll™ 梯度精选后的动力学参数和氧化应激,以及耐热试验(SS + TRT),以确定田间繁殖力的有用指标。在实验中,从六头阿伯丁安格斯公牛身上获得了商业剂量的冷冻精液。根据国际育种公司 CRV Lagoa 提供的 IFert™ 指数,其中三头公牛被归类为高繁殖力,三头为低繁殖力。汇集的精液样本被分配到两个处理组进行分析:解冻后(PT)或精子选择(SS)(Percoll™)和热阻测试(SS + TRT)。使用精子动力学(CASA)(运动率、渐进运动率、VCL µm/s、VSL µm/s、VAP µm/s、LIN %、STR % 和 WOB%)、活性氧(ROS)产生、脂质过氧化、超氧化物歧化酶(SOD)酶活性和总抗氧化能力对样本进行评估。数据采用双向方差分析,将肥力指数、样品中使用的处理方法以及这些因素之间的交互作用视为效应。当观察到明显的影响时,使用 Bonferroni 检验对数值进行比较。对繁殖力指数和体外分析的精子参数进行了皮尔逊相关分析,以评估精子质量和公牛受胎率之间的关系。与高繁殖力公牛相比,低繁殖力公牛的精子动力学参数(包括总运动率、渐进运动率和搏动交叉频率)较高(P < 0.05)。然而,高繁殖力公牛的曲线速度更高,其次是 SS + TRT。使用 SS + TRT 后,高繁殖力公牛的直线速度、平均路径速度、线性度和交叉频率节拍都更高。活性氧与SS后的繁殖力相关。此外,仅在高繁殖力公牛中观察到脂质过氧化减少。然而,PT 和 SS + TRT 后,脂质过氧化与高繁殖力相关。与单个动力学参数相比,体外精子动力学参数组合能更准确地预测体内繁殖力。精子的脂质过氧化是公牛繁殖力的一个重要指标。高繁殖力公牛似乎更容易受到脂质过氧化的影响,只有高繁殖力公牛的脂质过氧化才会减少,这表明它们的精子可以修复氧化应激引起的损伤。
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