Data_Sheet_1_Moringa oleifera leaf ethanolic extract benefits cashmere goat semen quality via improving rumen microbiota and metabolome.docx

BackgroundArtificial insemination (AI) is an effective reproductive technique to improve the performance of cashmere goats and prevent the spread of diseases, and the quality of the semen determines the success of AI. The potential of Moringa oleifera leaf powder (MOLP) and Moringa oleifera leaf ethanolic extract (MOLE) to improve semen quality has been reported, but the underlying mechanisms remain unclear. For the purpose, 18 mature male cashmere goats were randomly assigned into three groups: the control (CON), MOLP, and MOLE groups. The CON group received distilled water orally; the MOLP group was orally treated with 200 mg/kg body weight (BW) MOLP; and the MOLE group was orally treated with 40 mg/kg BW MOLE.
ResultsResults showed that MOLE contained long-chain fatty acids and flavonoids. Treatment with MOLP and MOLE increased the activities of the serum catalase, superoxide dismutase, and glutathione peroxidase (P < 0.05), enhanced the total antioxidant capacity (P < 0.05), and reduced the serum malondialdehyde level (P < 0.05). At the same time, MOLE increased the contents of serum gonadotropin releasing hormone and testosterone (P < 0.05). Moreover, MOLE significantly increased sperm concentration, motility, and viability (P < 0.05). Meanwhile, MOLE raised the Chao1 index (P < 0.05) and altered the composition of the rumen microbiota; it also raised the relative abundance of Treponema (P < 0.05) and Fibrobacter (P < 0.05) and reduced the relative abundance of Prevotella (P < 0.1). Correlation analysis revealed the genus Prevotella was significantly negatively correlated with sperm concentration, as well as sperm motility and viability. Furthermore, MOLE significantly increased the rumen levels of the steroid hormones testosterone and dehydroepiandrosterone (P < 0.05), as well as the polyunsaturated fatty acids (PUFAs) alpha-Linolenic acid, gamma-Linolenic acid, docosapentaenoic acid, and 9-S-Hydroperoxylinoleicacid (P < 0.05).
ConclusionsOral MOLE supplementation can improve semen quality by increasing the antioxidant capacity and altering the rumen microbiota and metabolites of cashmere goats. Moreover, the MOLP supplementation could enhance the antioxidant capacity of cashmere goats.

背景 人工授精（Artificial insemination, AI）是提升绒山羊生产性能、防控疾病传播的有效生殖技术，而精液品质是人工授精成功与否的核心决定因素。已有研究表明，辣木叶粉（Moringa oleifera leaf powder, MOLP）与辣木叶乙醇提取物（Moringa oleifera leaf ethanolic extract, MOLE）具备改善精液品质的潜力，但具体作用机制仍未明确。为阐明该机制，本研究将18只成年雄性绒山羊随机分为3组：对照组（CON）、MOLP组与MOLE组。对照组经口灌服蒸馏水；MOLP组按200 mg/kg体重（body weight, BW）灌服辣木叶粉；MOLE组按40 mg/kg体重灌服辣木叶乙醇提取物。
结果 检测显示，MOLE中含有长链脂肪酸与黄酮类物质。与对照组相比，MOLP与MOLE处理均可显著提高血清过氧化氢酶（catalase）、超氧化物歧化酶（superoxide dismutase）及谷胱甘肽过氧化物酶（glutathione peroxidase）的活性（P < 0.05），增强总抗氧化能力（total antioxidant capacity）（P < 0.05），并降低血清丙二醛（malondialdehyde）水平（P < 0.05）。同时，MOLE可显著提升血清促性腺激素释放激素（gonadotropin releasing hormone）与睾酮（testosterone）含量（P < 0.05）。此外，MOLE显著提高了精子浓度、活力与活率（P < 0.05）。进一步分析发现，MOLE可提升Chao1指数（P < 0.05）并改变瘤胃微生物群组成；其还显著提高了密螺旋体属（Treponema）与纤维杆菌属（Fibrobacter）的相对丰度（P < 0.05），降低了普雷沃氏菌属（Prevotella）的相对丰度（P < 0.1）。相关性分析表明，普雷沃氏菌属与精子浓度、精子活力及活率均呈显著负相关。此外，MOLE显著提高了瘤胃中睾酮（testosterone）与脱氢表雄酮（dehydroepiandrosterone）等类固醇激素的水平（P < 0.05），以及多不饱和脂肪酸（polyunsaturated fatty acids, PUFAs）α-亚麻酸、γ-亚麻酸、二十二碳五烯酸与9-S-过氧羟基亚油酸的含量（P < 0.05）。
结论 经口补充MOLE可通过提升绒山羊的抗氧化能力、调节瘤胃微生物群及其代谢产物，进而改善精液品质。此外，补充MOLP可增强绒山羊的抗氧化能力。