【摘要】：隨著規(guī)�；B(yǎng)豬業(yè)發(fā)展,豬人工授精技術(shù)的普及面不斷擴(kuò)大,而精液保存質(zhì)量的優(yōu)劣決定生產(chǎn)效率高低,其中稀釋劑發(fā)揮著至關(guān)重要的作用。精液常溫保存過程中需要進(jìn)行稀釋,會減弱精液原有抗氧化作用,使過多活性氧無法及時清除,質(zhì)膜易受被損傷,導(dǎo)致精子質(zhì)量下降。同時,微生物的存在也會影響精液保存質(zhì)量。為同時解決抗菌和抗氧化的問題,本研究在稀釋液中添加原花青素(Oligomeric Proantho Cyanidins,OPC)和乳酸鏈球菌素(Nisin),檢測精液保存質(zhì)量、滲透壓、總抗氧化能力、丙二醛含量、酸性磷酸酶活性和抑菌效果等,以期為OPC和Nisin在精液稀釋液中的聯(lián)合添加提供一些理論參考。本研究主要獲得以下結(jié)果:1.豬精液稀釋液中添加不同濃度(10、20、30、40、50、60、70μg/mL)OPC溶液,可優(yōu)化精液保存效果。其中,稀釋液中添加50μg/mL OPC效果最好,常溫保存第5d的精子活率61.63%、質(zhì)膜完整率41.88%、頂體完整率75.88%,顯著高于對照組(P0.05)。常溫保存第5d時精子T-AOC為1.43 U/mL,高于對照組,二者差異不顯著(P0.05);MDA含量為2.37 nmol/mL,顯著低于對照組(P0.05)。另外,稀釋液中添加OPC對稀釋液滲透壓無顯著影響(P0.05)。2.稀釋液中添加不同濃度(10、20、30、40、50μg/mL)Nisin到稀釋液中,能夠在提高精液保存質(zhì)量同時抑制細(xì)菌繁殖。添加40μg/mL Nisin組的常溫保存效果最好,保存到第5d精子活率能達(dá)到60.12%,精子質(zhì)膜完整率為44.98%,頂體完整率為71.78%,顯著高于對照組(P0.05)。稀釋液中添加30μg/mL Nisin抑制金黃色葡萄球菌效果顯著優(yōu)于慶大霉素(P0.05),抑制魯菲不動桿菌效果與乳酸鏈球菌素相當(dāng)(P0.05);稀釋液中添加40μg/mL Nisin抑制大腸桿菌效果顯著低于慶大霉素(P0.05)。3.豬精液稀釋液中聯(lián)合添加50μg/mL OPC和20μg/mL Nisin的常溫保存效果優(yōu)于單獨(dú)添加兩種物質(zhì)組和對照組(P0.05),常溫保存到第5 d精子活率為62.07%,精子質(zhì)膜、頂體完整率分別為56.00%和78.64%,精子線粒體活性為60.46%。第5 dOPC:Nisin=50:20組精子T-AOC為1.32 U/mL,酸性磷酸酶活性為573.34 U/L,均顯著高于對照組和單獨(dú)添加藥物組(P0.05);MDA含量最少,為2.36 nmol/mL,顯著低于其余各組(P0.05)。4.稀釋液中添加OPC-Nisin可顯著抑制金黃色葡萄球菌生長(P0.05),且作用效果優(yōu)于慶大霉素,但對大腸桿菌和魯菲不動桿菌的抑制效果欠佳(P0.05)。綜上所述,添加原花青素和乳酸鏈球菌素能提高精液保存質(zhì)量并延長保存時間。原花青素最適添加濃度為50μg/mL,乳酸鏈球菌素最適添加濃度為40μg/mL,聯(lián)合添加最適濃度為50μg/mL原花青素和20μg/mL乳酸鏈球菌素。
[Abstract]:With the development of large-scale pig industry, the popularization of artificial insemination technology is expanding, and the quality of semen preservation determines the production efficiency, among which diluents play a crucial role. In the process of semen preservation at room temperature, it is necessary to dilute, which will weaken the original antioxidant effect of semen, make too much reactive oxygen species can not be cleared in time, and the plasma membrane will be easily damaged, which will lead to the decline of sperm quality. At the same time, the existence of microbes will also affect the quality of semen preservation. In order to solve the problem of anti-bacterial and anti-oxidation simultaneously, the semen quality, osmotic pressure, total antioxidant capacity and malondialdehyde (MDA) content were determined by adding proanthocyanidin (Oligomeric Proantho Cyanidins (OPC) and Lactic streptococcin (Nisin),) to the diluent. The activity of acid phosphatase and bacteriostatic effect are expected to provide some theoretical reference for the combined addition of OPC and Nisin in semen diluent. In this study, the following results were obtained: 1: 1. The preservation effect of porcine semen was optimized by adding different concentrations of OPC solution (10 ~ (20) C ~ (30) C ~ (40) C ~ (50) C ~ (60) ~ (70) 渭 g/mL) to porcine semen diluent. The sperm motility, plasma membrane integrity and acrosome integrity were 61.63, 41.88 and 75.88, respectively, which were significantly higher than those in the control group (P0.05). At the 5th day of normal temperature preservation, the T-AOC of sperm was 1.43 U / mL, which was higher than that of the control group (P0.05), and the content of MDA was 2.37 nmol / mL, which was significantly lower than that of the control group (P0.05). In addition, the addition of OPC to the diluent had no significant effect on the osmotic pressure of the diluent (P0.05). Adding different concentrations of Nisin (10 ~ 20 ~ 30 ~ 40 ~ (50 渭 g/mL) to the diluent could improve the quality of semen preservation and inhibit the propagation of bacteria. The sperm motility was 60.12%, the sperm plasma membrane integrity rate was 44.98 and the acrosome integrity rate was 71.78, which was significantly higher than that of the control group (P0.05). The inhibitory effect of 30 渭 g/mL Nisin on Staphylococcus aureus was significantly better than that of gentamicin (P0.05), and the inhibitory effect on Acinetobacter rufii was similar to that on Streptococcus lactis (P0.05), and the inhibitory effect of 40 渭 g/mL Nisin on Escherichia coli was significantly lower than that on gentamicin (P0.05). The effect of adding 50 渭 g/mL OPC and 20 渭 g/mL Nisin to porcine semen diluent at room temperature was better than that of the control group (P0.05). The sperm motility was 62.07 and the plasma membrane of sperm was 62.07 on the 5th day after preservation at room temperature. The acrosome integrity rates were 56.00% and 78.64%, respectively. The sperm mitochondrial activity was 60.46%. The sperm T-AOC and acid phosphatase activity in the fifth dOPC:Nisin=50:20 group were 1.32 U / mL and 573.34 U / L, respectively, which were significantly higher than those in the control group and the control group (P0.05), the lowest (2.36nmol / mL), and significantly lower than those in the other groups (P0.05) .4. The addition of OPC-Nisin in diluent significantly inhibited the growth of Staphylococcus aureus (P0.05), and the effect was better than gentamicin, but the inhibitory effect on Escherichia coli and Acinetobacter rufii was not good (P0.05). In conclusion, the addition of proanthocyanidin and lactic acid streptococcin can improve the quality of semen preservation and prolong the storage time. The optimal concentration of proanthocyanidins was 50 渭 g / mL, the optimal concentration of nisin was 40 渭 g / mL, the optimal concentration of proanthocyanidins and 20 渭 g/mL nisin was 50 渭 g/mL and 20 渭 g/mL respectively.
【學(xué)位授予單位】：西北農(nóng)林科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S828

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