目的研究不同稀释剂和冷冻方法对冷冻保存后解冻精子质量的影响,寻找一种廉价实用的用于农场条件下人工授精的Huram精液冷冻方法。从五只胡公羊收集射精。在实验I中,用八种不同的冷冻稀释剂(基本稀释剂A,B,C,D,E,F,G,和H)。稀释和冷却后,将样品装入0.25mL吸管中,用液氮熏蒸法冷冻.在实验二,使用稀释剂C作为基础稀释剂,使用液氮熏蒸和两种程控冷却方法冷冻精液。为了进行分析,根据运动性参数(总运动性(TM),渐进运动(PM)),生物动力学特性(直线速度(VSL),平均路径速度(VAP),曲线速度(VCL),头部侧向位移(ALH)的振幅,摆动运动系数(WOB),平均运动度(MAD)),活性氧(ROS)水平,膜和顶体的完整性。在实验I中,稀释剂C具有较高的TM,PM,和顶体和膜的完整性和较低的ROS相比,其他延长剂(p<0.05),稀释剂A稀释剂C表现出更高(p<0.05)的VCL,VAP,ALH,WOB,和MAD与稀释剂B相比,D,E,和F.在实验II中,TM和所有生物动力学特性在三种冷冻方法中没有显示出显著差异(p>0.05)。液氮熏蒸导致更高的(p<0.05)PM,膜完整性,顶体完整性,与程序相比,ROS水平较低。总之,与其他稀释剂相比,用稀释剂C稀释的解冻精液具有更高的质量。与使用稀释剂C的程控冷却方法相比,液氮熏蒸显示出优异的精液冷冻保存效果。
The purpose of this study was to investigate the effects of different diluents and freezing methods on the quality of thawed sperm after cryopreservation and find an inexpensive and practical method for freezing Hu ram semen for use in inseminations under farm conditions. Ejaculates were collected from five Hu rams. In experiment I, ejaculates were diluted with eight different freezing diluents (basic diluents A, B, C, D, E, F, G, and H). After dilution and cooling, the samples were loaded into 0.25 mL straws and frozen using the liquid nitrogen fumigation method. In experiment II, diluent C was used as the basic diluent and the semen was frozen using liquid nitrogen fumigation and two program-controlled cooling methods. For analysis, frozen samples were evaluated in terms of motility parameters (total motility (TM), progressive motility (PM)), biokinetic characteristics (straight-line velocity (VSL), average path velocity (VAP), curvilinear velocity (VCL), amplitude of lateral head displacement (ALH), wobble movement coefficient (WOB), average motion degree (MAD)), reactive oxygen species (ROS) level, and membrane and acrosome integrity. In experiment I, diluent C had higher TM, PM, and acrosome and membrane integrity and lower ROS compared to other extenders (p < 0.05) except diluent A. Diluent C exhibited higher (p < 0.05) VCL, VAP, ALH, WOB, and MAD compared to diluents B, D, E, and F. In experiment II, TM and all biokinetic characteristics did not show significant differences (p > 0.05) amongst the three freezing methods. Liquid nitrogen fumigation resulted in higher (p < 0.05) PM, membrane integrity, acrosome integrity, and lower ROS level compared to the program. In conclusion, the thawed semen diluted with diluent C had higher quality compared to other diluents. The liquid nitrogen fumigation demonstrated superior semen cryopreservation effects compared to the program-controlled cooling method using diluent C.