Andrology Laboratory (ANDROLAB), Departament of Veterinary Medicine, Universidade Federal Rural de Pernambuco, Recife, Pernambuco, Brazil.
Andrology Laboratory (ANDROLAB), Departament of Veterinary Medicine, Universidade Federal Rural de Pernambuco, Recife, Pernambuco, Brazil.
J Equine Vet Sci. 2023 Jul;126:104489. doi: 10.1016/j.jevs.2023.104489. Epub 2023 Mar 30.
Semen cryopreservation ensures the storage of stallion genetics for an unlimited time. The improvement of extenders with new antioxidant substances can optimize the properties of post-thawed semen. The study aimed to investigate the addition effect of medium-molecular-weight carboxymethylchitosan (CQm) derivates to freezing diluent of stallion sperm after freezinf/thawing. Twice a week, five ejaculates of four stallions were obtained, totalizing 20 ejaculates. Semen was diluted in commercial freezing extender (Botucrio) supplemented with CQm: control (0), 0.75, 1.5, and 3 mg/mL. Samples were filled in straws (0.5 mL) and submitted to freezing and storage at -196°C. Thawing was performed at 37°C/30 s, and the samples of each group were analyzed for kinetics, plasma membrane integrity, acrosome membrane integrity, and mitochondrial membrane potential . The addition of 1.5 and 3 mg/mL CQm showed lower values (P < .05) of total motility (TM), progressive motility (PM), curvilinear velocity (VCL), straight line velocity (VSL), average path velocity (VAP) and wobble (WOB), comparing to control group. Besides, it was observed lower (P < .05) percentages of sperm with intact acrosomes in the group treated with 3 mg/mL of CQm than control group. In conclusion, high concentration of medium-molecular-weight carboxymethylchitosan to freezing diluent damages kinematic and acrosome of stallion sperm after freezing/thawing.
精子冷冻保存可确保种马遗传物质的无限期储存。通过添加新型抗氧化物质的 extender 可以优化解冻后精液的性能。本研究旨在探讨中分子量羧甲基壳聚糖(CQm)衍生物在冷冻稀释剂中的添加对冻融后种马精子的影响。每周两次,从 4 匹种马中获得 5 次射精,共 20 次射精。精液用商业冷冻稀释液(Botucrio)稀释,添加 CQm:对照组(0)、0.75、1.5 和 3mg/mL。将样品装入 straw(0.5mL)中,进行冷冻并在-196°C下储存。解冻在 37°C/30 秒进行,分析每组样品的动力学、质膜完整性、顶体膜完整性和线粒体膜电位。添加 1.5 和 3mg/mL 的 CQm 后,总活力(TM)、前向运动活力(PM)、曲线速度(VCL)、直线速度(VSL)、平均路径速度(VAP)和摆动性(WOB)的总活力(TM)、前向运动活力(PM)、曲线速度(VCL)、直线速度(VSL)、平均路径速度(VAP)和摆动性(WOB)均低于对照组(P<0.05)。此外,与对照组相比,添加 3mg/mL CQm 的组中具有完整顶体的精子百分比较低(P<0.05)。综上所述,高浓度的中分子量羧甲基壳聚糖对冷冻稀释剂会损害冻融后种马精子的运动能力和顶体。