精液稀释液中添加维生素C和E对陆川猪精液冷冻效果的影响

为了研究维生素C和E作为精液冷冻保护剂对陆川猪细管冷冻精液品质和精浆中抗氧化物酶活性影响的影响,在精液稀释液中分别添加0、300、600以及900μg/m L的维生素C或E,对陆川猪稀释精液进行冷冻-解冻处理后,检测精液冻后的精子活力、运动速率、线粒体活性、顶体和质膜完整性、存活时间等常规参数,采用试剂盒测定精浆中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)以及谷胱胺肽过氧化物酶(GSHPx)的活性。结果表明:精液稀释液中添加300μg/m L的维生素C可显著提高冻后精子的体外存活时间以及CAT酶活性(P0.05);同浓度的维生素E仅可提高精子的存活时间(P0.05);添加600μg/m L的维生素C或E可以显著提高解冻精子的活力、线粒体活性、质膜和顶体完整性、存活时间以及抗氧化物酶活性(P0.05),同时明显降低精子畸形率(P0.05);当维生素C或E添加量为900μg/m L时,虽可提高线粒体活性、质膜和顶体完整性、CAT酶活等部分指标参数(P0.05),但精子活力与对照组相比没有差异(P0.05)。提示:冷冻稀释液中添加600μg/m L的维生素C或E可以显著提高陆川猪精液冷冻保存效果。     

稀释液中添加维生素E对宠物犬精液冷冻效果的影响

在宠物犬冷冻精液稀释液中添加质量浓度为0,200,400,600μg/mL的维生素E,以观察对精液冷冻效果的影响。结果表明:冷冻精液稀释液中添加维生素E 400μg/mL时,解冻后精子活力显著高于对照组(P0.05);畸形率极显著低于对照组(P0.01),各浓度维生素E组精子解冻后体外存活时间均极显著高于对照组(P0.01),解冻后精子顶体完整率和冻后精子运动速率(VAP、VSL、VCLL)较对照组都有所提高,但差异不显著(P0.05)。综合各项指标后表明以质量浓度为400μg/mL维生素E组的效果最好。     

亚硒酸钠维生素E对牛冷冻精液品质的影响

本试验研究了亚硒酸钠维生素E对牛冷冻精液品质的影响。以常规Tris卵黄冷冻稀释液中分别添加2.5μL、5μL、7.5μL、10μL稀释后的亚硒酸钠维生素E(注射液)为试验组,以常规Tris卵黄冷冻稀释液为对照组,对解冻后的精液品质进行测定。结果显示添加5μL、7.5μL、10μL亚硒酸钠维生素E能够显著降低冷冻精液的精子畸形率,且10μL组顶体脱落的精子数量显著降低,精子顶体完整率上升。小剂量(<10μL)组改善了精子存活时间,使精子在体外37℃的存活时间延长至8.5h。     

维生素E对秦川牛冷冻精液的抗氧化保护作用

本试验旨在研究维生素E对秦川牛细管冷冻精液品质和精浆中抗氧化酶活性的影响.在稀释液中分别添加0.00、0.02、0.04、0.06、0.08、0.10 mg·mL~(-1)的维生素E,将细管冻精解冻(37.5℃,30 s)后用伟力精子分析系统分析精子活力和精子顶体完整率等标准参数,并用相关试剂盒测定精浆中的超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH)、谷胱甘肽还原酶(GR)和过氧化氢酶(CAT)的活性.结果表明:当维生素E添加量为0.04mg·mL~(-1)时,精子活力与对照组相比差异显著(P<0.05),SOD、GSH、CAT 3种酶活性与对照组相比差异显著(P<0.05).当维生素E的添加量为0.06 mg·mL~(-1)时,冷冻-解冻后精子活力显著高于对照组(P<0.05),顶体完整率极显著高于对照组(P<0.01);GR和CAT酶活性与对照组相比均差异显著(P<0.05).当维生素E的添加量为0.08 mg·mL~(-1)时,精子活力、活率及顶体完整率与对照组相比差异显著(P<0.05),GSH与对照相组比差异极显著(P<0.01),GR及CAT与对照组相比差异显著(P<0.05).在冷冻精液稀释液中添加维生素E可以有效提高冷冻-解冻后秦川牛精液品质及精浆中SOD、CAT、GSH、GR酶的活性,稀释液中维生素E的适宜添加量为0.04～0.08 mg·mL~(-1).     

奶牛冷冻精液新型稀释液的开发与利用

本试验旨在探究牛冷冻精液稀释液中添加不同物质对精液品质及人工授精能力的影响。方法为在牛冷冻精液稀释液中添加不同成分的精子质膜保护剂、顶体完整保护剂、线粒体保护剂,然后检测精子存活时间、动力学参数、ATP含量和人工授精的情期受胎率。结果表明,白藜芦醇、褪黑激素、适宜浓度的半胱胺可显著增加冷冻-解冻后的精子活力;适宜浓度的脱盐乳清粉、透明质酸、牛血清白蛋白均能有效保护精子顶体;适宜浓度的红景天多糖、谷氨酰胺、海藻糖、可显著提高精子线粒体能量水平,牛磺酸可显著提高冷冻精液活力。利用上述物质按一定比例配制的新型稀释液可以保护精子顶体膜和线粒体膜完整性,较常规冷冻精液解冻后精子体外存活时间延长6～8h;经人工授精试验,青年牛情期受胎率、经产牛情期受胎率比对照高1.9个百分点、3.1个百分点,具备推广应用价值。     

添加维生素B_（12）对西门塔尔牛冷冻精液品质影响的初步观察

为观察稀释液中添加VB12对西门塔尔牛精液品质的影响,进行了对比试验。结果表明：添加VB12的试验组冻精解冻后精子活力、顶体完整率比对照组有极显著提高（P〈0.01）,精子畸形率与GOT活性均明显下降,精子存活时间延长。说明VB12在牛精液冷冻中对牛精子形态结构具有一定保护作用。     

维生素E对湖羊精液低温保存效果研究

为研究维生素E对湖羊精液4℃保存效果的影响,在前期筛选的湖羊精液稀释液中添加不同质量浓度(0.0、0.3、0.6、0.9、1.2、1.5 mg/mL)的维生素E用于湖羊精液冷藏试验,检测湖羊精子活率和顶体完整率,以及精液中总抗氧化能力(Total antioxidant capacity,T-AOC)和丙二醛(Malondialdehyde,MDA)含量。结果表明:维生素E的添加显著减少MDA的生成,提高了总抗氧化能力(T-AOC);但维生素E的添加对精子活率和顶体完整率无显著提高,本试验中0.6 mg/mL的添加质量浓度保存效果与对照组无显著差异,但质量浓度过高或过低保存效果欠佳。因此,在4℃保存条件下,在湖羊精液稀释液中添加维生素E不能提高精液保存质量。     

稀释液中添加维生素E对和田羊冷冻精液品质的影响

在和田羊精液冷冻稀释液中添加不同浓度的维生素E,比较其对和田羊冷冻精液品质的影响。结果表明：维生素E添加量为1．2mg／mL时精子解冻后活率（43．56％）显著高于对照组（34．14％）和其他各添加组（P〈0．05）;顶体完整率（51．39％）极显著高于对照组（40．12％）和其他各添加组（P〈0．01）。说明在冷冻精液稀释液中添加适当浓度维生素E可以减缓冷冻对精子的伤害,有效提高和田羊冷冻一解冻后精子活率,改善冷冻精液的品质。     

维生素E对4℃保存藏猪精液品质和抗氧化保护作用的影响

试验旨在研究维生素E对4℃藏猪精液品质和抗氧化保护作用影响,采用电刺激法采集健康藏猪精液,在改进的猪精液低温稀释液为基础液中,每1 000 m L添加维生素E 0、3.0、4.0、5.0、6.0 g,将藏猪种公猪新鲜精液做4倍稀释后于4℃环境保存至第5天,以藏猪精子活力、精子畸形率、精子顶体完整率和质膜完整率为评价精液品质指标,以超氧化物歧化酶(Superoxidas dismutase,SOD)活力和丙二醛(Maleic dialdehyde,MDA)含量为评价抗氧化保护作用指标,探讨维生素E对4℃藏猪精液品质影响。结果显示:维生素E 5.0 g/L添加组藏猪精子活力、精子畸形率、精子顶体完整率和质膜完整率均极显著好于其他试验组(P0.01)。精浆超氧化物歧化酶活力和丙二醛含量以5.0 g/L维生素E添加组为最高;同时发现,5.0 g/L维生素E添加组精子超氧化物歧化酶活力与其他各组间差异不显著。总之,在改进的猪精液稀释液中添加5.0 g/L的维生素E可明显改善4℃环境保存藏猪精液品质和抗氧化保护作用,此试验研究为今后藏猪精液低温保存的生产应用奠定了基础。     

生物类黄酮和维生素C对猪精液冷冻效果的影响

为了研究冷冻稀释液中添加不同浓度的抗氧化剂生物类黄酮和维生素C对猪精液冷冻保存效果的影响,试验分别在冷冻稀释液中添加0,0.4,0.6,0.8,1.0 mg/mL的生物类黄酮和0,2,4,6,8 mg/mL的维生素C以及二者最佳配伍浓度的混合物,分别检测各试验组精液冷冻解冻后的活率、质膜完整率、顶体完整率、线粒体活率等指标。结果表明:除了添加4,6 mg/mL维生素C顶体完整率与对照组相比差异不显著(P>0.05)外,添加0.4,0.6,0.8,1.0 mg/mL生物类黄酮或2,4,6,8 mg/mL维生素C精子活率、质膜完整率与对照组相比差异均显著(P<0.05),其中添加0.8 mg/mL生物类黄酮和6 mg/mL维生素C效果最好,但随着添加浓度的增加冷冻效果逐渐降低。说明在猪精液冷冻稀释液中联合添加生物类黄酮和维生素C,解冻后精液质量显著高于对照组(P<0.05)。     

辅酶Q10对绒山羊精液冷冻保存效果的影响

旨在探讨辅酶Q10对绒山羊精液冷冻保存效果的影响。利用添加不同浓度辅酶Q10(4、40、400?滋g/mL)的精液冷冻稀释液对绒山羊精液样本进行冷冻保存,待冷冻精液解冻后,采用流式细胞仪和计算机辅助精液分析系统(CASAS)分别检测不同精液样本的精子活率、质膜完整率、顶体完整率、DNA完整率、线粒体膜电位和细胞内ROS水平。结果表明,当冷冻稀释液中添加浓度为40μg/mL辅酶Q10时,经历冷冻—解冻过程的绒山羊精液样本的精子活率、质膜完整率、顶体完整率均显著高于对照组(P<0.05);在冷冻稀释液中添加浓度为40μg/mL或400μg/mL的辅酶Q10均能显著提高线粒体膜电位并降低细胞内ROS水平(P<0.05)。综上所述,在冷冻稀释液中添加40μg/mL的辅酶Q10能够显著提高绒山羊精子抗氧化能力和冷冻保存效果。     

冷冻稀释液中添加大豆卵磷脂对梅花鹿冻融精子质量的影响

【目的】探究在冷冻稀释液中添加大豆卵磷脂代替10%卵黄对梅花鹿精液冷冻保存效果的影响,为梅花鹿人工授精体系的完善提供参考。【方法】采用电刺激法采集梅花鹿精液,以精液冷冻稀释液中分别添加1%、2%、3%、4%和5%大豆卵磷脂代替10%卵黄作为试验组,添加20%卵黄作为对照组,分别进行各组精液冷冻保存。5 d后,进行精液解冻,检测解冻后各组精子的活力、质膜完整率、顶体完整率、线粒体活性、存活时间,筛选合适浓度的大豆卵磷脂。选取4～5岁健康雌性梅花鹿,肌肉注射300 IU孕马血清促性腺激素(PMSG)和0.4 mg氯前列醇钠进行同期发情处理,发情后第20 h用20%卵黄组与筛选出的大豆卵磷脂组冻精进行人工输精,输精后30 d使用B超检测仪检测妊娠情况,统计妊娠率。【结果】与对照组相比,1%大豆卵磷脂组冻融后的精子活力、向前活动力、快速前进活力、活率、质膜完整率、顶体完整率及线粒体活性均显著提高(P<0.05);随着稀释液中大豆卵磷脂浓度的增加,其冻融后精子活力、向前活动力、快速前进活力、活率、质膜完整率、顶体完整率以及线粒体活性呈下降趋势,精子存活时间也随浓度的增加而减少。1%大豆卵磷...     

抗氧化剂对马精子低温及冷冻保存效果的影响

试验旨在研究不同抗氧化剂对马精子低温保存及冷冻效果的影响。选取6匹英纯血种公马作为试验动物,以INRA82液作为基础稀释液（对照组）,分别添加谷氨酰胺（0.015 g/mL）、甘氨酸（0.019 g/mL）、半胱氨酸（0.024 g/mL）、甲硫氨酸（0.015 g/mL）、牛磺酸（0.063 g/mL）、维生素C（0.4 mg/mL）、维生素E（0.5 mg/mL）、褪黑素（0.001 mg/mL）制备成含有不同抗氧化剂的低温保存和冷冻保存稀释液,将浓缩处理后的马精子分别置于上述稀释液中保存或冷冻,检测低温保存48 h后精子运动参数,评价抗氧化剂对精子低温保存的影响;检测精子冻融后运动参数、精子质膜完整性、线粒体膜电势及顶体完整性,评价抗氧化剂对精子冷冻效果的影响。结果表明,精子低温保存48 h后,添加牛磺酸组活精子比例和前向运动精子比例显著高于对照组（P<0.05）,添加维生素C组前向运动精子比例显著高于对照组（P<0.05）。冷冻解冻后结果表明,添加不同抗氧化剂没有改善精子冻融后活精子比例和前向运动精子比例,但添加甲硫氨酸显著延长了精子体外存活能力（P<0.05）;添加不同抗氧化剂精子质膜完整性与对照组间无显著差异（P>0.05）,但甲硫氨酸和甘氨酸组有高于对照组的趋势;添加维生素E和甲硫氨酸组精子线粒体膜电势显著高于对照组（P<0.05）,不同抗氧化剂组精子顶体完整性与对照组间均无显著差异（P>0.05）。结果提示,低温保存时添加牛磺酸和维生素C可以提高精液低温保存效果;冷冻时添加甲硫氨酸能提高精子质膜完整性和线粒体膜电势,且能够延长精子冻融后的存活时间。     

Effects of Antioxidants on the Quality of Hypothermic Preservation and Frozen-thawed Equine Semen

This study was aimed to evaluate the effects of various antioxidants, namely glutamine (0.015 g/mL), glycine (0.019 g/mL), cysteine (0.024 g/mL), methionine (0.015 g/mL), taurine (0.063 g/mL), vitamin C (0.4 mg/mL), vitamin E (0.5 mg/mL) and melatonin (0.001 mg/mL) on equine sperm quality after chill or freeze-thaw.Semen were collected from 6 adult thoroughbred stallions, INRA82 was used as the base extender (control group), adding INRA82 with different antioxidants was used in experimental group. Assess the effect of antioxidants on semen by detecting motion parameters after storage at 5℃ for 48 h. Motion parameters, plasma membrane integrity (PMI) and mitochondrial membrane potential were used to evaluate semen quality after thawing. The extender supplemented with 25 mmol/L taurine led to higher TM and PM, and supplemented with 0.4 mg/mL vitamin C obtained significant higher PM compared with control group (P<0.05) after storage at 5℃ for 48 h. The freeze extender supplemented with 0.5 mg/mL vitamin E or 0.015 g/mL methionine significantly increased the mitochondrial membrane potential compare with control group (P<0.05). No significant differences were observed for PMI and acrosomes integrity rate after frozen-thawed (P>0.05), but there was a trend that PMI of adding methionine and glycine group was higher than control group. The results suggested that extender supplemented with taurine and vitamin C could improve the semen preservation effect, and the extender supplemented with methionine could improve plasma membrane integrity, mitochondrial membrane potential of thawing sperm, and also could prolong the survival time of frozen thawed sperm.     

Studies on the Effect of Supplementing Boar Semen Cryopreservation Media with Different Avian Egg Yolk Types on in Vitro Post-thaw Sperm Quality

Fertility after insemination of cryopreserved boar semen is currently below that of fresh semen. In an attempt to improve the post-thaw motility and acrosome integrity of boar sperm, semen was frozen using an adapted Westendorf method in which the chicken egg yolk was replaced by either duck or quail egg yolk. The different composition of the yolk types, particularly the amount of cholesterol, fatty acids and phospholipids, were thought to potentially afford a greater level of protection to sperm against damage during freezing and thawing. Sperm frozen in medium containing chicken egg yolk displayed higher motility immediately after thawing, but there was no difference in the motility of sperm frozen with different types of egg yolk 3 or 6 h after thawing and maintenance at 37 degrees C. Sperm frozen in media containing chicken or duck egg yolk had a higher proportion of intact acrosomes immediately after thawing than sperm frozen in medium containing quail egg yolk, but 6 h after thawing and maintenance at 37 degrees C the sperm that had been frozen in medium containing chicken egg yolk had a higher proportion of intact acrosomes than the sperm frozen in media containing duck or quail egg yolk. Analysis of the composition of the different yolk types showed that the basic components of the yolks were similar, but the ratios of fatty acids and phospholipid classes differed. Duck egg yolk had more monounsaturated fatty acids (MUFA) than chicken egg yolk, which had more MUFA than quail egg yolk. Duck egg yolk contained more phosphotidylinositol (PI) than chicken or quail egg yolks and quail egg yolk contained more phosphotidylserine than either chicken or duck egg yolks. The differences in post-thaw motility and acrosome integrity of boar sperm when frozen in media containing the different types of egg yolk may be due to the variation in composition.     

Study on the Role of AMPK Activator in Cryopreservation of Sheep Semen

This study aimed to investigate the effects of AMPK activators metformin (Met) and acadesine (AICAR) on sheep semen cryopreservation. Firstly, Met and AICAR with different concentrations (0, 100, 200, 300, 400, 500 μmol·L^-1) were added into the frozen diluent. After freezing and thawing, the optimal concentration (400 μmol·L^-1 Met, 200 μmol·L^-1 AICAR) were selected based on sperm motility, motion performance and membrane integrity; Secondly, the collected semen was froze using different frozen diluents (control group:diluent; Met group:diluent + 400 μmol·L^-1 Met; AICAR group:diluent + 200 μmol·L^-1 AICAR). After thawing, the sperm AMPK protein expression, acrosomal enzyme activity, metabolic index, mitochondrial function and antioxidant enzyme activity were examined. The results showed that the addition of 400 μmol·L^-1 Met and 200 μmol·L^-1 AICAR could significantly improve sperm motility, motion performance and membrane integrity(P<0.05) after thawing. In 400 μmol·L^-1 Met group, the total sperm motility, acrosome integrity rate and plasma membrane integrity rate were 43.20%, 91% and 46%, respectively. Compared with the control group, the level of AMPK phosphorylation in the sperm of the Met and AICAR groups was significantly increased after thawing (P<0.05), the acrosome enzyme activity of sperm was significantly increased(P<0.05); the pyruvate level was significantly decreased (P<0.05), the lactate dehydrogenase activity, lactic acid content, and ATP content were significantly increased(P<0.05). Compared with the control group, the dilution of Met group and AICAR group was more conducive to maintain mitochondrial membrane potential (P<0.05), increase ATPase and antioxidant enzyme activity (P<0.05). The addition of appropriate concentrations of Met and AICAR could improve semen quality of cryopreservation in sheep.     

AMPK激活剂在绵羊精液冷冻保存中的作用研究

旨在研究AMPK激活剂二甲双胍（metformin,Met）和阿卡地新（acadesine,AICAR）对绵羊精液冷冻保存效果的影响。本研究首先在冷冻基础稀释液中分别添加不同浓度（0、100、200、300、400、500 μmol·L^-1）的Met和AICAR,冷冻解冻后根据精子活力、运动性能和结构完整性指标筛选出最佳的添加浓度（400 μmol·L^-1 Met、200 μmol·L^-1 AICAR）;然后分别使用不同的冷冻稀释液（对照组：稀释液;Met组：含400 μmol·L^-1 Met的稀释液;AICAR组：含200 μmol·L^-1 AICAR的稀释液）冷冻精液,解冻后检测精子中AMPK蛋白表达、顶体酶活性、代谢指标、线粒体功能以及抗氧化酶活性。结果表明,稀释液中添加400 μmol·L^-1 Met和200 μmol·L^-1AICAR均可显著提高解冻后精子活力、运动性能及精子结构完整性（P<0.05）,其中400 μmol·L^-1 Met组精子总活力达43.20%,顶体完整率为91%,质膜完整率为46%。与对照组相比,Met组和AICAR组解冻后精子中AMPK磷酸化水平显著升高（P<0.05）;顶体酶活性显著提高（P<0.05）;丙酮酸水平显著下降（P<0.05）,乳酸脱氢酶活性、乳酸以及ATP含量均显著升高（P<0.05）;与对照组相比,Met和AICAR组稀释液更有利于维持线粒体膜电位（P<0.05）,提高ATP酶（P<0.05）以及抗氧化酶的活性（P<0.05）。添加适当浓度的AMPK激活剂可以提高绵羊精液冷冻保存的效果。     

Effects of Bovine Serum Albumin on Boar Sperm Quality During Liquid Storage at 17°C

This study aimed to investigate the effects of bovine serum albumin (BSA) on boar sperm quality during liquid storage at 17°C. Boar semen samples were collected and diluted with Modena containing different concentrations (0, 1, 2, 3, 4, 5 and 6 g/l) of BSA, and sperm motility, plasma membrane integrity, acrosome integrity, total antioxidative capacity (T‐AOC) activity and malondialdehyde (MDA) content were measured and analysed. The results showed that Modena supplemented with 3, 4 and 5 g/l BSA could improve boar sperm motility, effective survival time and plasma membrane integrity (p < 0.05), decrease MDA content (p < 0.05), while no statistical difference was observed for sperm acrosome integrity and T‐AOC activity among these three groups (p > 0.05). The semen sample diluted with Modena containing 4 g/l BSA could achieve optimum effect, and sperm survival time was 7.5 days. After 7 days preservation, sperm motility, plasma membrane integrity and acrosome integrity were 54%, 49% and 78%, respectively. T‐AOC activity and MDA content were 1.03 U/ml and 17.5 nmol/ml, respectively. In conclusion, Modena supplemented with BSA reduced the oxidative stress and improved the sperm quality of boar semen during liquid storage at 17°C, and 4 g/l BSA was the optimum concentration. Further studies are required to obtain more concrete results on the determination of antioxidant capacities of BSA in liquid preserved boar semen.     

不同浓度大豆卵磷脂替代蛋黄对东佛里生奶绵羊精液冷冻保存效果的影响

本试验皆在研究添加不同浓度大豆卵磷脂（SL）冷冻保存东佛里生奶绵羊精液的效果。我们在Tris基础稀释液中,添加18%蛋黄为对照组,添加0.5%、1%、1.5%、2%、2.5%SL设为试验组,检测冷冻精液解冻后的精子活率和顶体完整率。结果显示,添加0.5%、2.5% SL冷冻稀释液稀释的精液,解冻后精子活率和顶体完整率与其他组之间存在显著差异（P<0.05）;添加18%蛋黄和1%～2% SL冷冻稀释液稀释的精液,冷冻解冻后精子活率和顶体完整率之间无显著差异（P>0.05）;添加18%蛋黄和1.0%～1.5% SL冷冻稀释液稀释后的精液,进行人工授精后母羊的妊娠率与对照组无显著差异（P>0.05）。因此,大豆卵磷脂可以作为冷冻保护剂用于东佛里生奶绵羊精液的冷冻保存,其最佳添加浓度为1～2%（g／L）。