【摘要】 试验旨在探讨绵羊冻精中乳铁蛋白的最适添加浓度。选取3只健康的杜博种公羊，通过体外采精技术获得精液。分别将0,10,100,1 000 mg/L乳铁蛋白添加到绵羊冻精保存溶液中，研究对精子运动能力、质膜完整性和线粒体功能的影响；进一步对添加最佳剂量乳铁蛋白的冻精进行体外受精试验，以鲜精体外受精作为对照组（鲜精组），1组未添加乳铁蛋白，2组添加乳铁蛋白最佳剂量，统计早期胚胎发育各阶段比例；同时观察冷冻精子形态和卵裂过程。结果表明：(1)在各浓度处理中，冻精中添加100 mg/L乳铁蛋白的精子运动能力、质膜完整性、线粒体膜电势均表现最佳。(2)鲜精组囊胚发育率最高，为30.0%;1组（未添加乳铁蛋白）囊胚发育率最低，为24.6%;2组（添加100 mg/L乳铁蛋白）相比1组囊胚发育率有所改善，为27.7%。(3)2组（添加100 mg/L乳铁蛋白）的冻精保存0 h时，精子呈密集直线运动，头部完整，尾部摆动均匀；保存10 h时呈分散运动，超过80%精子质膜完整。综上所述，在精子冷冻保存液中添加100 mg/L乳铁蛋白能够有效改善绵羊精子活力，提升质膜完整性，维持线粒体高膜电势，有效促进体外受精过程中胚胎发育。更多还原

【Abstract】 This study aimed to investigate the optimal addition concentration of lactoferrin in sheep frozen semen. Three healthy Dober male sheep were selected, and semen was obtained through in vitro insemination techniques. Lactoferrin concentrations of 0, 10,100 and 1,000 mg/L were added to sheep frozen semen preservation solutions to assess their effects on sperm motility, plasma membrane integrity and mitochondrial function. In vitro fertilization(IVF) experiments were conducted using frozen semen with the optimal lactoferrin concentration, with fresh semen IVF serving as the control group(fresh semen group). The group 1 received no lactoferrin, while the group 2 received the optimal additive dose. The proportions of early embryonic development stages were statistically analyzed, and the morphology of frozen sperm and cleavage processes were observed. Results demonstrated:(1) Among all concentration treatments, adding 100 mg/L lactoferrin to frozen semen resulted in optimal sperm motility, plasma membrane integrity, and mitochondrial high membrane potential.(2) The fresh semen group exhibited the highest blastocyst rate(30. 0%), followed by group 1(no lactoferrin added) with the lowest rate(24. 6%). The group 2(100 mg/L lactoferrin added) showed improved blastocyst rates compared to group 1, reaching 27. 7%.(3)At 0 hour post-preservation, sperm in the 100 mg/L lactoferrin group(group 2) exhibited dense linear motility, intact heads and uniform tail oscillation. After 10-hour preservation, sperm displayed dispersed motility, with over 80% maintaining plasma membrane integrity. In conclusion, adding 100 mg/L lactoferrin to sperm cryopreservation solution can effectively improve the motility of sheep semen, enhance plasma membrane integrity, maintain mitochondrial high membrane potential and promote embryo development during in vitro fertilization(IVF).更多还原