ABSTRACT Semen cryopreservation damages sperm due to oxidative stress. This study evaluated the antioxidant capacity of Moringa oleifera seed extract in cryopreserved ram semen and the impact of the extract on sperm characteristics. Semen from eight hair rams (four rams per sampling) was allocated into four groups, according to their treatment prior to cryopreservation: Control (no extract), 0.5 (M0.5), 5.0 (M5.0), and 10.0 (M10) mg/mL of M. oleifera extract. The antioxidant activity (ferric reducing antioxidant power, FRAP) and the spermatic characteristics (sperm viability; progressive motility; fast motility; slow motility; acrosome damage; membrane damage; and mitochondrial activity) were assessed post-thawing. Variables were evaluated with analysis of variance followed by Tukey test. While no significant differences were detected in acrosomal damage, mitochondrial activity, fast-, or slow motility, the antioxidant activity was higher (P < 0.05) in M0.5 and M5 treatments. Viability and progressive motility increased in the M0.5 group (P < 0.05), whereas sperm membrane damage was lower (P < 0.05) in the same treatment. In conclusion, supplementation of ram semen with M. oleifera seed extract enhances antioxidant activity, sperm membrane integrity, viability, and progressive motility after thawing. This suggests that M. oleifera extract could be used as an antioxidant to improve the outcome of semen cryopreservation. Highlights It is widely known that semen cryopreservation induces sublethal damage to sperm, deteriorating spermatic characteristics, which is largely attributed to oxidative stress. There are scarce studies regarding the use of plant extracts as a replacement for conventional antioxidants to conserve sperm viability in cryopreserved ram semen. Addition of Moringa oleifera seed extract to a concentration of 0.5 and 5.0 mg/mL prior to ram semen freezing increased antioxidant activity after cryopreservation. M. oleifera seed extract at 0.5 mg/mL decreased post-thawing damage to the sperm membrane, increasing both viability and progressive motility. M. oleifera seed extract could be potentially used as a replacement for conventional antioxidants added to maintain sperm viability in cryopreserved ram semen.

中文翻译：

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辣木种子提取物对冻存公羊精液抗氧化活性及精子特性的影响

摘要 精液冷冻保存会因氧化应激而损害精子。本研究评估了辣木种子提取物在冷冻保存的公羊精液中的抗氧化能力以及提取物对精子特征的影响。根据冷冻保存前的处理，将八只毛公羊的精液（每次采样四只公羊）分为四组：对照（无提取物）、0.5 (M0.5)、5.0 (M5.0) 和 10.0 (M10) mg/mL 的 M. oleifera 提取物。解冻后评估抗氧化活性（铁降低抗氧化能力，FRAP）和精子特征（精子活力；进行性运动；快速运动；缓慢运动；顶体损伤；膜损伤；和线粒体活性）。通过方差分析和随后的 Tukey 检验来评估变量。虽然在顶体损伤、线粒体活性、快或慢运动方面没有检测到显着差异，但 M0.5 和 M5 处理的抗氧化活性更高（P < 0.05）。M0.5 组的活力和进行性运动增加（P < 0.05），而相同处理的精子膜损伤较低（P < 0.05）。总之，在公羊精液中添加油橄榄种子提取物可增强抗氧化活性、精子膜完整性、活力和解冻后的渐进性运动。这表明 M. oleifera 提取物可用作抗氧化剂以改善精液冷冻保存的结果。亮点 众所周知，精液冷冻保存会对精子造成亚致死损伤，使精子特征恶化，这在很大程度上归因于氧化应激。很少有关于使用植物提取物替代传统抗氧化剂来保存冷冻公羊精液中精子活力的研究。在冷冻公羊精液之前，将辣木种子提取物添加到 0.5 和 5.0 毫克/毫升的浓度会增加冷冻保存后的抗氧化活性。M. oleifera 种子提取物 0.5 mg/mL 减少了对精子膜的解冻后损伤，增加了活力和渐进性运动。M. oleifera 种子提取物可能被用作替代常规抗氧化剂，以维持冷冻保存的公羊精液中的精子活力。冷冻公羊精液前 0 mg/mL 增加冷冻保存后的抗氧化活性。M. oleifera 种子提取物 0.5 mg/mL 减少了对精子膜的解冻后损伤，增加了活力和渐进性运动。M. oleifera 种子提取物可能被用作替代常规抗氧化剂，以维持冷冻保存的公羊精液中的精子活力。冷冻公羊精液前 0 mg/mL 可增加冷冻保存后的抗氧化活性。M. oleifera 种子提取物 0.5 mg/mL 减少了对精子膜的解冻后损伤，增加了活力和渐进性运动。M. oleifera 种子提取物可能被用作替代常规抗氧化剂，以维持冷冻保存的公羊精液中的精子活力。