【摘要】：豬精液常溫保存技術在豬人工授精過程中起著非常關鍵的作用,而精液稀釋劑的質量決定精液的常溫保存效果。在采精和稀釋過程中,常常會因外界環(huán)境或操作疏忽使豬精液受到某些微生物的污染,而精液稀釋液中恰恰含有豐富的利于微生物生長繁殖的營養(yǎng)物質,大量的微生物會對精液品質造成影響。為了抑制微生物的生長,常規(guī)方法是在稀釋劑中添加抗生素。然而,在生產實踐中缺乏科學的抗生素添加方案,使得抗生素不斷被濫用,導致病原菌耐藥性問題日益突出。為了解決這一問題,本研究探究了一種新型的營養(yǎng)素型殺菌消毒藥物——蛋氨酸碘在精液中的抑菌效果。首先進行蛋氨酸碘的體外殺菌試驗,然后鑒定豬鮮精中的細菌種類,接著探究蛋氨酸碘對豬精液中細菌及精液品質的影響,最后比較蛋氨酸碘與常規(guī)抗生素對豬精液常溫保存效果的影響。以期為蛋氨酸碘可替代抗生素作為精液稀釋劑中的一種新型抗菌劑提供有力依據,同時為我國精液稀釋劑自主研發(fā)提供一定的理論支持。本研究主要獲得以下結果:1.蛋氨酸碘對大腸桿菌作用時,高濃度組(1:5000)作用1 min殺菌率可達100%;中濃度組(1:10000)作用10 min殺菌率為100%;低濃度組(1:20000)作用10 min殺菌率為81.71%。對沙門氏菌作用時,高濃度組(1:5000)作用1 min殺菌率可達100%;中濃度組(1:10000)作用5 min殺菌率為100%;低濃度組(1:20000)作用10 min殺菌率為89.32%。對金黃色葡萄球菌作用時,高濃度組(1:5000)作用5 min殺菌率可達100%;中濃度組(1:10000)作用10 min殺菌率為100%;低濃度組(1:20000)作用10 min殺菌率為78.39%。因此,高濃度組(1:5000)和中濃度組(1:10000)可以達到殺菌目的,而低濃度組(1:20000)達不到殺菌目的。2.豬鮮精中主要含有7類細菌,分別是鹽單胞菌屬(Halomonas)占62.21%,不動桿菌屬(Acinetobacter)占11.71%,彎曲菌屬(Campylobacter)占8.64%,普氏菌屬(Prevotella)占8.55%,鏈球菌屬(Streptococcus)占4.45%,擬桿菌屬(Bacteroides)占2.38%,葡萄球菌屬(Staphylococcus)占1.83%。3.在豬精液稀釋液中添加0~400?L/L蛋氨酸碘時,隨著濃度的增加殺菌效果越好。但是,蛋氨酸碘的添加量在200、400?L/L時,精液保存效果受到嚴重影響,精子活率、質膜完整率、頂體完整率均顯著低于對照組(P0.05),而添加量在25、50、100?L/L時隨著蛋氨酸碘濃度的增加精液的保存效果越好,并且添加100?L/L蛋氨酸碘時保存效果最佳,精子活率、質膜完整率、頂體完整率均顯著高于對照組(P0.05)。可見,添加100μL/L蛋氨酸碘可有效抑制精液中細菌的生長,并且可有效提高精液的常溫保存效果。4.精液保存1~6 d時,蛋氨酸碘組精子活率總體上高于磺胺組,在保存5、6 d時差異顯著(P0.05);保存1~6 d時,磺胺組精子活率高于青霉素組,在保存3、6 d時差異顯著(P0.05)。精液保存1、2 d時,蛋氨酸碘組精子質膜完整率低于磺胺組,但差異不顯著(P0.05);保存3~6 d時,蛋氨酸碘組精子質膜完整率高于磺胺組,在保存5、6 d時差異顯著(P0.05);保存1~6 d時,磺胺組精子質膜完整率高于青霉素組,在保存4、5、6 d時差異均顯著(P0.05)。精液保存2、3 d時,蛋氨酸碘組精子頂體完整率低于磺胺組,但差異不顯著(P0.05);保存1、4、5、6 d時,蛋氨酸碘組精子頂體完整率高于磺胺組,在保存5、6 d時差異顯著(P0.05);保存2~6 d時,磺胺組精子頂體完整率高于青霉素組,在保存3、5、6 d時差異均顯著(P0.05)�？梢�,蛋氨酸碘對精液保存效果最佳,磺胺次之,青霉素較差。
[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.
【學位授予單位】：西北農林科技大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：S828

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本文編號：2220015