GSH和SOD对猪精液常温保存效果的研究

发布时间：2018-06-17 03:52

本文选题：猪精液 + 稀释剂　；参考：《西北农林科技大学》2015年硕士论文

【摘要】：精液保存技术在猪人工授精过程中起着非常关键的作用,而猪精液常温保存稀释剂的质量决定着精液的保存效果。国外关于猪精液常温保存稀释剂配方成分及其比例的研究比较深入,而我国在猪精液稀释剂研发方面起步较晚,目前我国生猪养殖企业和养殖户使用的猪精液常温保存稀释剂主要是进口稀释剂,不利于我国生猪产业的健康发展。因此,开发研制出国内优良猪精液稀释剂配方,对于降低养猪成本、增加养猪收益、促进我国养猪业的健康发展有着重大的意义。本试验对猪精液常温保存的机理进行深入研究,探索分析不同物质对猪精子的保护机制,研究抗氧化物质GSH、SOD对精液保存效果的影响;同时研制出三种猪精液稀释剂配方,筛选出保存效果最好的自配稀释剂;最后对保存效果最好的自配稀释剂进行实际应用效果评价试验,同时与商品稀释剂进行比较。本研究主要获得以下结果:1.猪精液稀释剂BTS中加入1 mmol/L的GSH能够减少精子的氧化损伤,明显降低精液中MDA、H2O2含量(P0.05),显著提高精液SOD活性(P0.05);Modena稀释剂Modena中加入1 mmol/L和5 mmol/L GSH精子有效保存时间和精子活率无明显差异(P0.05),精子有效保存时间为5 d,精子活力分别为0.60和0.63。GSH的最适添加量为1 mmol/L。2.Modena稀释液中添加200 IU/mL SOD可显著提高猪精液常温保存效果,降低精液中MDA和H2O2含量(P0.05),增强精液中SOD活性(P0.05),保存4 d后精子活率为0.657;BTS稀释液中添加300 IU/mL的SOD,可显著提高猪精液常温保存效果,保存4 d后精子活力为0.653。添加适量的SOD能提高精液的保存效果,SOD过量反而会对精子产生毒害作用。3.在3种自配稀释剂中,自配稀释剂III号保存效果最好,保存5 d后精子活力为0.60;自配稀释剂III号与Modena保存效果不显著(P0.05);自配稀释剂II号与BTS保存效果不显著,有效保存时间均为4 d(P0.05);自配稀释剂I号,保存时间最短,但在1~2 d内保存效果较好。在精液刚稀释完成时,Modena、BTS、自配稀释剂I、II、III处理组其质膜完整率、顶体完整率没有差异(P0.05)。保存1 d后,Modena、自配稀释剂I、III号处理组其质膜完整率、顶体完整率显著高于BTS、自配稀释液II号,且自配稀释剂I号处理组其质膜完整率、顶体完整率最高。保存2 d后,自配稀释液I号处理组其质膜完整率、顶体完整率最低,下降最快,显著低于其他4个稀释剂处理组(P0.05)。保存3~5 d后,Modena、自配稀释剂III号处理组其质膜完整率、顶体完整率显著高于其他三种稀释剂处理组(P0.05)。4.本试验应用自配稀释剂III号在大约克、长白、杜洛克猪上进行受胎试验,自配稀释剂III号对于大白猪、长白猪实际应用效果较好,受胎率在90%以上,受胎效果与市场上应用效果优良的商品稀释剂相比差异不显著,可以用于实践生产。
[Abstract]:Sperm preservation technology plays a very important role in the process of artificial insemination of pigs, and the quality of the porcine semen preserving diluent determines the preservation effect of the semen. The research on the composition and proportion of the porcine semen preserving diluent in foreign countries is comparatively thorough, but in our country it started late in the research and development of the porcine semen thinner. The normal temperature preservation diluents used by pig breeding enterprises and farmers are mainly imported diluents, which are not conducive to the healthy development of the pig industry in China. Therefore, the development and development of the domestic fine porcine semen thinner formula is important to reduce the cost of pig raising, increase the income of pig raising, and promote the healthy development of the pig industry in China. This experiment studied the mechanism of pig semen preservation at normal temperature, explored the protection mechanism of different substances on pig sperm, studied the effect of antioxidant GSH, SOD on the preservation of semen, and developed three kinds of porcine semen thinner formula, and screened out the best preservative with the best preservation effect. Finally, the best preservation effect was made. The results were compared with the commercial diluents. The main results of this study were as follows: 1. the addition of 1 mmol/L GSH in the porcine semen BTS could reduce the oxidative damage of sperm, reduce the MDA and H2O2 content (P0.05) in semen, significantly improve the sperm SOD activity (P0.05), Modena thinner Mod. The effective preservation time and sperm viability of 1 mmol/L and 5 mmol/L GSH sperm were not significantly different (P0.05), the effective preservation time of sperm was 5 d, and the optimum addition of sperm 0.60 and 0.63.GSH to 1 mmol/L.2.Modena diluents with 200 IU/mL SOD could significantly increase the preservation effect of the pig semen at normal temperature, and reduce the MDA and H2 in the semen. O2 content (P0.05) increased the activity of SOD in semen (P0.05), the survival rate of sperm was 0.657 after 4 D, and SOD of 300 IU/mL in BTS diluted solution could significantly improve the preservation effect of the pig semen at room temperature. After 4 D, the sperm vitality was added to 0.653. and SOD could improve the preservation effect of semen, while SOD excess would poison sperm in 3 species. The self matched diluent III was best preserved and the sperm motility was 0.60 after preservation of 5 d; the preservation effect of self matched diluent III and Modena was not significant (P0.05); the preservation effect of self matched diluent II and BTS was not significant, the effective preservation time was 4 d (P0.05), and the saving time was the shortest, but the preservation effect was in 1~2 D. Better. When the semen was just diluted, Modena, BTS, I, II, III treatment group had no difference in the integrity of the plasma membrane, and the integrity of the acrosome was not different (P0.05). After 1 D, Modena, the self diluent I, the III No. treatment group, the integrity of the plasma membrane, the acrosome integrity rate was significantly higher than that of BTS, and the self matching diluent number treatment group was finished with the membrane. The integrity rate of the acrosome was the highest. After the preservation of 2 D, the integrity rate of the plasmalemma, the acrosome integrity rate was the lowest, and the decrease was the fastest, significantly lower than the other 4 diluent treatment groups (P0.05). After the preservation of 3~5 D, Modena, the complete rate of the plasmalemma, the acrosome integrity rate was significantly higher than the other three kinds of diluents treatment. Group (P0.05).4. in this experiment, the self matched diluent III was used to carry out the pregnancy test on the great York, Changbai and Duroc pigs. The self matched diluent III has good practical application effect for the white pig and the long white pig, and the pregnancy rate is above 90%. The difference of the pregnancy effect is not significant compared with the commercial thinner with good application effect on the market. Produce.
【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S828

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