蛋氨酸碘对猪精液常温保存效果的研究
发布时间：2018-09-03 12:44
【摘要】：猪精液常温保存技术在猪人工授精过程中起着非常关键的作用,而精液稀释剂的质量决定精液的常温保存效果。在采精和稀释过程中,常常会因外界环境或操作疏忽使猪精液受到某些微生物的污染,而精液稀释液中恰恰含有丰富的利于微生物生长繁殖的营养物质,大量的微生物会对精液品质造成影响。为了抑制微生物的生长,常规方法是在稀释剂中添加抗生素。然而,在生产实践中缺乏科学的抗生素添加方案,使得抗生素不断被滥用,导致病原菌耐药性问题日益突出。为了解决这一问题,本研究探究了一种新型的营养素型杀菌消毒药物——蛋氨酸碘在精液中的抑菌效果。首先进行蛋氨酸碘的体外杀菌试验,然后鉴定猪鲜精中的细菌种类,接着探究蛋氨酸碘对猪精液中细菌及精液品质的影响,最后比较蛋氨酸碘与常规抗生素对猪精液常温保存效果的影响。以期为蛋氨酸碘可替代抗生素作为精液稀释剂中的一种新型抗菌剂提供有力依据,同时为我国精液稀释剂自主研发提供一定的理论支持。本研究主要获得以下结果:1.蛋氨酸碘对大肠杆菌作用时,高浓度组(1:5000)作用1 min杀菌率可达100%;中浓度组(1:10000)作用10 min杀菌率为100%;低浓度组(1:20000)作用10 min杀菌率为81.71%。对沙门氏菌作用时,高浓度组(1:5000)作用1 min杀菌率可达100%;中浓度组(1:10000)作用5 min杀菌率为100%;低浓度组(1:20000)作用10 min杀菌率为89.32%。对金黄色葡萄球菌作用时,高浓度组(1:5000)作用5 min杀菌率可达100%;中浓度组(1:10000)作用10 min杀菌率为100%;低浓度组(1:20000)作用10 min杀菌率为78.39%。因此,高浓度组(1:5000)和中浓度组(1:10000)可以达到杀菌目的,而低浓度组(1:20000)达不到杀菌目的。2.猪鲜精中主要含有7类细菌,分别是盐单胞菌属(Halomonas)占62.21%,不动杆菌属(Acinetobacter)占11.71%,弯曲菌属(Campylobacter)占8.64%,普氏菌属(Prevotella)占8.55%,链球菌属(Streptococcus)占4.45%,拟杆菌属(Bacteroides)占2.38%,葡萄球菌属(Staphylococcus)占1.83%。3.在猪精液稀释液中添加0~400?L/L蛋氨酸碘时,随着浓度的增加杀菌效果越好。但是,蛋氨酸碘的添加量在200、400?L/L时,精液保存效果受到严重影响,精子活率、质膜完整率、顶体完整率均显著低于对照组(P0.05),而添加量在25、50、100?L/L时随着蛋氨酸碘浓度的增加精液的保存效果越好,并且添加100?L/L蛋氨酸碘时保存效果最佳,精子活率、质膜完整率、顶体完整率均显著高于对照组(P0.05)。可见,添加100μL/L蛋氨酸碘可有效抑制精液中细菌的生长,并且可有效提高精液的常温保存效果。4.精液保存1~6 d时,蛋氨酸碘组精子活率总体上高于磺胺组,在保存5、6 d时差异显著(P0.05);保存1~6 d时,磺胺组精子活率高于青霉素组,在保存3、6 d时差异显著(P0.05)。精液保存1、2 d时,蛋氨酸碘组精子质膜完整率低于磺胺组,但差异不显著(P0.05);保存3~6 d时,蛋氨酸碘组精子质膜完整率高于磺胺组,在保存5、6 d时差异显著(P0.05);保存1~6 d时,磺胺组精子质膜完整率高于青霉素组,在保存4、5、6 d时差异均显著(P0.05)。精液保存2、3 d时,蛋氨酸碘组精子顶体完整率低于磺胺组,但差异不显著(P0.05);保存1、4、5、6 d时,蛋氨酸碘组精子顶体完整率高于磺胺组,在保存5、6 d时差异显著(P0.05);保存2~6 d时,磺胺组精子顶体完整率高于青霉素组,在保存3、5、6 d时差异均显著(P0.05)。可见,蛋氨酸碘对精液保存效果最佳,磺胺次之,青霉素较差。
[Abstract]:At * * the preservation of pig semen at normal temperature plays a key role in the process of artificial insemination, and the quality of semen diluent determines the preservation of semen at normal temperature. In the process of spermatozoa * dilution, the semen of pigs is often contaminated by some microorganisms because of external environment or operation negligence. In order to inhibit the growth of microorganisms, the conventional method is to add antibiotics in diluents. However, the lack of scientific antibiotic addition program in production practice makes antibiotics abused constantly, resulting in the problem of antibiotic resistance of pathogenic bacteria is becoming increasingly prominent. In order to solve this problem, this study explored the bacteriostatic effect of a new nutrient type bactericidal disinfectant methionine iodine in semen. First, methionine iodine in vitro was bactericidal test, then identified the species of * * in pig semen, and then explored the effect of methionine iodine on the bacteria and semen quality in pig semen. * the effect of methionine iodine and conventional antibiotics on the preservation of pig spermatozoa at room temperature was compared. In order to provide a strong basis for methionine iodine to replace antibiotics as a new antibacterial agent in semen diluent, and provide some theoretical support for the independent research and development of semen diluent in China. The following results were obtained in this study: 1. The bactericidal rate of high concentration group (1:5000) for 1 minute was 100%; that of medium concentration group (1:10000) for 10 minutes was 100%; that of low concentration group (1:20000) for 10 minutes was 81.71%; that of high concentration group (1:5000) for 1 minute was 100%; that of medium concentration group (1:10000) for 5 minutes was 100%. The bactericidal rate of high concentration group (1:5000) for 5 minutes was 100%; medium concentration group (1:10000) for 10 minutes was 100%; low concentration group (1:20000) for 10 minutes was 78.39%. 0) it can achieve bactericidal purpose, while low concentration group (1:20000) can not achieve bactericidal purpose *.2. pig fresh essence contains 7 kinds of bacteria, namely, Halomonas, 62.21%, Acinetobacter (Acinetobacter), 11.71%, Campylobacter (Campylobacter), 8.64%, Prevotella (8.55%), and Streptococcus (Streptococcus) 4.45%. Bacteroides (Bacteroides) accounted for 2.38%. Staphylococci (Staphylococcus) accounted for 1.83%.3.. When adding 0~400 * L/L methionine iodine in the pig semen dilution, the germicidal efficacy increased with increasing concentration. However, when methionine iodine was added at 200400 L/L, semen preservation effect was seriously affected, sperm viability, plasma membrane integrity and acrosome integrity. The sperm survival rate, plasma membrane integrity rate, acrosome integrity rate were significantly higher than those of the control group (P Sperm viability of methionine-iodine group was higher than that of sulfonamide group on the whole, and the difference was significant (P 0.05) on the 5th and 6th day, and the sperm viability of sulfonamide group was higher than that of penicillin group on the 1st and 6th day, and the difference was significant (P 0.05) on the 3rd and 6th day. Sperm plasma membrane integrity rate of methionine iodine group was lower than that of sulfonamide group at day 1, but the difference was not significant (P 0.05); Sperm plasma membrane integrity rate of methionine iodine group was higher than that of sulfonamide group at day 3 to 6, and the difference was significant at day 5 and 6 (P 0.05); Sperm plasma membrane integrity rate of sulfonamide group was higher than that of penicillin group at day 1 to 6, and the difference was significant at day 4, 5 and 6 (P 0.05). Sperm acrosome intact rate of methionine iodine group was lower than that of sulfonamide group at 2,3 days of preservation, but the difference was not significant (P 0.05); Sperm acrosome intact rate of methionine iodine group was higher than that of sulfonamide group at 1,4,5,6 days of preservation, and the difference was significant at 5,6 days of preservation (P 0.05); Sperm acrosome intact rate of sulfonamide group was higher than that of penicillin group at 2 to 6 days of preservation, and the difference was significant at 3,5,6 days (P0.05). Methionine iodine has the best effect on semen preservation, followed by sulfamethoxazole and penicillin.
【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】：2016
【分类号】：S828
本文编号：2220015
[Abstract]:At * * the preservation of pig semen at normal temperature plays a key role in the process of artificial insemination, and the quality of semen diluent determines the preservation of semen at normal temperature. In the process of spermatozoa * dilution, the semen of pigs is often contaminated by some microorganisms because of external environment or operation negligence. In order to inhibit the growth of microorganisms, the conventional method is to add antibiotics in diluents. However, the lack of scientific antibiotic addition program in production practice makes antibiotics abused constantly, resulting in the problem of antibiotic resistance of pathogenic bacteria is becoming increasingly prominent. In order to solve this problem, this study explored the bacteriostatic effect of a new nutrient type bactericidal disinfectant methionine iodine in semen. First, methionine iodine in vitro was bactericidal test, then identified the species of * * in pig semen, and then explored the effect of methionine iodine on the bacteria and semen quality in pig semen. * the effect of methionine iodine and conventional antibiotics on the preservation of pig spermatozoa at room temperature was compared. In order to provide a strong basis for methionine iodine to replace antibiotics as a new antibacterial agent in semen diluent, and provide some theoretical support for the independent research and development of semen diluent in China. The following results were obtained in this study: 1. The bactericidal rate of high concentration group (1:5000) for 1 minute was 100%; that of medium concentration group (1:10000) for 10 minutes was 100%; that of low concentration group (1:20000) for 10 minutes was 81.71%; that of high concentration group (1:5000) for 1 minute was 100%; that of medium concentration group (1:10000) for 5 minutes was 100%. The bactericidal rate of high concentration group (1:5000) for 5 minutes was 100%; medium concentration group (1:10000) for 10 minutes was 100%; low concentration group (1:20000) for 10 minutes was 78.39%. 0) it can achieve bactericidal purpose, while low concentration group (1:20000) can not achieve bactericidal purpose *.2. pig fresh essence contains 7 kinds of bacteria, namely, Halomonas, 62.21%, Acinetobacter (Acinetobacter), 11.71%, Campylobacter (Campylobacter), 8.64%, Prevotella (8.55%), and Streptococcus (Streptococcus) 4.45%. Bacteroides (Bacteroides) accounted for 2.38%. Staphylococci (Staphylococcus) accounted for 1.83%.3.. When adding 0~400 * L/L methionine iodine in the pig semen dilution, the germicidal efficacy increased with increasing concentration. However, when methionine iodine was added at 200400 L/L, semen preservation effect was seriously affected, sperm viability, plasma membrane integrity and acrosome integrity. The sperm survival rate, plasma membrane integrity rate, acrosome integrity rate were significantly higher than those of the control group (P Sperm viability of methionine-iodine group was higher than that of sulfonamide group on the whole, and the difference was significant (P 0.05) on the 5th and 6th day, and the sperm viability of sulfonamide group was higher than that of penicillin group on the 1st and 6th day, and the difference was significant (P 0.05) on the 3rd and 6th day. Sperm plasma membrane integrity rate of methionine iodine group was lower than that of sulfonamide group at day 1, but the difference was not significant (P 0.05); Sperm plasma membrane integrity rate of methionine iodine group was higher than that of sulfonamide group at day 3 to 6, and the difference was significant at day 5 and 6 (P 0.05); Sperm plasma membrane integrity rate of sulfonamide group was higher than that of penicillin group at day 1 to 6, and the difference was significant at day 4, 5 and 6 (P 0.05). Sperm acrosome intact rate of methionine iodine group was lower than that of sulfonamide group at 2,3 days of preservation, but the difference was not significant (P 0.05); Sperm acrosome intact rate of methionine iodine group was higher than that of sulfonamide group at 1,4,5,6 days of preservation, and the difference was significant at 5,6 days of preservation (P 0.05); Sperm acrosome intact rate of sulfonamide group was higher than that of penicillin group at 2 to 6 days of preservation, and the difference was significant at 3,5,6 days (P0.05). Methionine iodine has the best effect on semen preservation, followed by sulfamethoxazole and penicillin.
【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】：2016
【分类号】：S828
【相似文献】
相关会议论文 前1条
1 叶志辉;屠波;;蛋氨酸碘的应用研究[A];中国科学技术协会2008防灾减灾论坛论文集[C];2008年
相关硕士学位论文 前1条
1 王健;蛋氨酸碘对猪精液常温保存效果的研究[D];西北农林科技大学;2016年
，本文编号：2220015
本文链接：https://www.wllwen.com/yixuelunwen/dongwuyixue/2220015.html
