Department of Basic Science, Faculty of Veterinary Medicine, Urmia Branch, Islamic Azad University, Urmia, Iran.
Vet Med Sci. 2024 Sep;10(5):e31548. doi: 10.1002/vms3.1548.
The vulnerability of buffalo sperm to cryoinjury necessitates the improvement of sperm cryo-resistance as a critical strategy for the widespread use of assisted reproductive technologies in buffalo.
The main aim of the present study was to evaluate the effects of different concentrations of rutin and chlorogenic acid (CGA) on buffalo semen quality, antioxidant activity and fertility during cryopreservation.
The semen was collected and pooled from the 3 buffaloes using an artificial vagina (18 ejaculations). The pooled sperm were divided into nine different groups: control (Tris-based extender); 0.4, 0.6, 0.8 and 1 mM rutin (rutin + Tris-based extender); and 50, 100, 150 and 200 µM CGG (CGA + Tris-based extender). Sperm kinematics, viability, hypo-osmotic swelling test, mitochondrial activity, antioxidant activities and malondialdehyde (MDA) concentration of frozen and thawed buffalo sperm were evaluated. In addition, 48 buffalo were finally inseminated, and pregnancy was rectally determined 1 month after insemination.
Compared to the control group, adding R-0.4, R-0.6, CGA-100 and CGA-150 can improve total and progressive motility, motility characteristics, viability, PMF and DNA damage in buffalo sperm. In addition, the results showed that R-0.4, R-0.6, CGA-50, CGA-100 and CGA-150 increased total antioxidant capacity, catalase, glutathione peroxidase and glutathione activities and decreased MDA levels compared to the control group. Furthermore, it has been shown that adding 150 µM CGA and 0.6 mM rutin to an extender can increase in vivo fertility compared to the control group.
In conclusion, adding rutin and CGA to the extender improves membrane stability and in vivo fertility of buffalo sperm by reducing oxidative stress.
水牛精子对冷冻损伤很敏感，因此提高精子的抗冷冻能力是广泛应用水牛辅助生殖技术的关键策略。
本研究的主要目的是评估不同浓度芦丁和绿原酸（CGA）对水牛精液质量、抗氧化活性和冷冻保存期间生育力的影响。
使用人工授精阴道（18 次射精）从 3 头水牛中收集和混合精液。将混合后的精子分为 9 个不同的组：对照组（基于 Tris 的稀释液）；0.4、0.6、0.8 和 1 mM 芦丁（芦丁+基于 Tris 的稀释液）；和 50、100、150 和 200 µM CGA（CGA+基于 Tris 的稀释液）。评估冷冻和解冻水牛精子的运动学、活力、低渗肿胀试验、线粒体活性、抗氧化活性和丙二醛（MDA）浓度。此外，最终对 48 头水牛进行了授精，并在授精后 1 个月经直肠确定妊娠。
与对照组相比，添加 R-0.4、R-0.6、CGA-100 和 CGA-150 可以提高水牛精子的总运动和向前运动、运动特性、活力、PMF 和 DNA 损伤。此外，结果表明，与对照组相比，R-0.4、R-0.6、CGA-50、CGA-100 和 CGA-150 增加了总抗氧化能力、过氧化氢酶、谷胱甘肽过氧化物酶和谷胱甘肽的活性，降低了 MDA 水平。此外，与对照组相比，添加 150 µM CGA 和 0.6 mM 芦丁到稀释液中可以提高体内生育力。
总之，在稀释液中添加芦丁和 CGA 可以通过降低氧化应激来提高水牛精子的膜稳定性和体内生育力。
