SummaryInformation on donkey sperm bioenergetics, kinetics and oxidative status is scarce even though crucial for development of reproductive technologies and germplasm conservation. For these reasons, it is interesting to monitor sperm kinetics, bioenergetics, and oxidative status during sperm storage at +4°C and with several sperm extenders and concentrations. Donkey semen was collected from three jackasses, three times each. It was diluted with four extenders (Kenney, Equiplus, INRA96 or Hippex), set at three sperm concentrations (30, 50 or 70 × 106 spermatozoa/ml) and evaluated for its functionality after 0, 3, 24, 48 and 72 h storage at +4°C. Sperm kinetics was analyzed by Sperm Computer Analysis; sperm bioenergetics was assessed by mitochondrial membrane potential (MMP); sperm oxidative status was evaluated by lipid peroxidation (LPO), anti-LPO potential and nitroblue tetrazolium (NBT) assays. Incubation produced a progressive (P < 0.01) decline in sperm kinetics and MMP, whereas parameters related to oxidative status either increased (LPO, NBT) or decreased (anti-LPO). The anti-LPO potential was the index better related to sperm motility and kinetics. Extenders proved to be differently (P < 0.01) effective in preserving sperm kinetics, MMP, and oxidative status. The concentration of 30 × 106 spermatozoa/ml provided an optimum preservation of sperm functions. Significant correlations emerged between most parameters examined. This study identified reference criteria for storing donkey spermatozoa at +4°C. A low sperm concentration together with a proper extender are crucial requirements for optimum sperm cryopreservation efficiency. Field trials are, however, required to validate these findings, making them operational in practice.

中文翻译：

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+4°C保存的驴精子的运动学、生物能和氧化评估

摘要关于驴精子生物能量学、动力学和氧化状态的信息很少，尽管对于生殖技术的发展和种质保护至关重要。由于这些原因，在 +4°C 下精子储存过程中监测精子动力学、生物能量学和氧化状态以及使用几种精子扩展剂和浓度是很有趣的。从三头公驴身上采集驴精液，每头 3 次。用四种稀释剂（Kenney、Equiplus、INRA96 或 Hippex）稀释，设置为三种精子浓度（30、50 或 70 × 106精子/ml) 并在 +4°C 下储存 0、3、24、48 和 72 小时后评估其功能。通过精子计算机分析分析精子动力学；通过线粒体膜电位 (MMP) 评估精子生物能量学；通过脂质过氧化 (LPO)、抗 LPO 电位和硝基蓝四唑 (NBT) 测定评估精子氧化状态。孵化产生了渐进式（磷< 0.01) 精子动力学和 MMP 下降，而与氧化状态相关的参数要么增加（LPO，NBT）要么减少（抗 LPO）。抗LPO电位是与精子活力和动力学相关性更好的指标。扩展器被证明是不同的（磷< 0.01) 有效保持精子动力学、MMP 和氧化状态。浓度30×106精子/毫升提供了精子功能的最佳保存。大多数检查的参数之间出现了显着的相关性。该研究确定了在 +4°C 下储存驴精子的参考标准。低精子浓度和适当的稀释剂是最佳精子冷冻保存效率的关键要求。然而，需要实地试验来验证这些发现，使其在实践中具有可操作性。