绵羊精液稀释液中添加PUFA对冷冻精液品质的影响

通过在绵羊精液冷冻稀释液葡3-3稀释液(葡萄糖-柠檬酸盐-卵黄)中添加富含多不饱和脂肪酸(PUFA)的深海鱼油,研究其对绵羊精子冷冻-解冻过程的作用.     

稀释液中添加维生素E对绵羊冷冻精液品质的影响

采用两步稀释法 ,在绵羊冷冻精液中添加维生素E ,精液稀释平衡后精子活率 ,试验组 (0 .6 35 )高于对照组(0 .5 75 ) ,但无显著性差异 (P >0 .0 5 ) ;而解冻后 ,试验组活率 (0 .4 35 )极显著高于对照组 (0 .36 5 ) (P <0 .0 1) ;精子顶体总异常率 ,试验组 (33.72 % )极显著低于对照组 (4 4 .35 % ) (P <0 .0 1) ,其中试验组顶体膨胀率和脱落率与对照组分别为 1.2 8% ,2 3.0 8%与 2 .6 8% ,2 8.96 %。添加维生素E可以降低冷冻对精子顶体的损害程度 ,提高精液品质。试验也证明在解冻后保存 1h内 ,精子活率呈现缓慢下降的趋势 ,但 1h后活率急剧下降 ,因而 ,绵羊冻精颗粒解冻后应在 1h内输精完毕     

稀释液中添加多不饱和脂肪酸(PUFA)对绵羊精液冷冻效果的影响

以葡3-3稀释液(葡萄糖-柠檬酸盐-卵黄)为基础,添加富含多不饱和脂肪酸(PUFA),且含较高比例DHA和EPA的深海鱼油作为冷冻绵羊精液的新型添加剂,进而研究PUFA对绵羊精子冷冻-解冻过程优良的保护作用.研究结果表明,绵羊精液冷冻稀释液中添加3%PUFA的处理组,同时添加1%VE(V/V)作为抗氧化剂,经冷冻-解冻对精子的损害较小,解冻后活率和顶体完整率分别比对照组1(0)显著提高6%和10%(P＜0.05),精清中LDH酶(乳酸脱氢酶)活力和GOT酶(谷-草转氨酶)活力比对照组1(0)显著降低65%和15%(P＜0.05).     

稀释液中添加维生素E对绒山羊冷冻精液品质的影响

在绒山羊精液冷冻稀释液中添加不同浓度的维生素E,比较其对绒山羊冷冻精液品质的影响。结果表明:维生素E添加量为0.8～1.2 mg/mL时精子解冻后活率和顶体完整率显著高于对照组和其他各添加组(P<0.05);维生素E添加量为0.8～2.0 mg/mL时,畸形率显著低于对照组和其他各添加组(P<0.05);维生素E添加量为1.2 mg/mL时,质膜完整率显著高于对照组和其他各添加组(P<0.05)。说明适量的添加维生素E可以提高绒山羊精液冷冻的效果,且稀释液中维生素E的适宜添加量为0.8～1.2 mg/mL。     

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。     

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。     

不同甘油浓度下稀释液中添加棉子糖对绒山羊冷冻精液品质的影响

在甘油浓度为3%和6%的冷冻精液稀释液中添加不同浓度的棉子糖,旨在优化绒山羊冷冻稀释液配方。结果表明:甘油浓度为3%时,山羊精液稀释液的抗冻保护效果较好,在此基础上添加10 mmol/L棉子糖,精子活率、顶体完整率、生存指数、质膜完整率均高于其他实验组(P<0.05),并且畸形率显著降低(P<0.05);在脂质过氧化反应和抗氧化酶活性方面,该实验组CAT活性高于其他实验组(P<0.05),而MDA浓度低于其他实验组(P<0.05)。     

不同类型稀释液对驴冷冻精液品质的影响

[目的]:对比分析各类型稀释液对驴冷冻精子活力、畸形率的影响,根据冻精解冻效果,筛选出适合驴精子冷冻的最佳稀释液,最适宜稀释比例,为驴冷冻精液研发提供参考.[方法]:实验共计14 d,采集6头种公驴精液,每份原精均分为3等份,分别用3种稀释液稀释,从而选取最优稀释液;按1:0.75、1:1.5、1:3、1:6不同比例混...     

稀释液中添加棉子糖对冻融后绵羊精液品质的影响

本试验目的是研究稀释液中添加不同浓度的棉子糖对冻融后绵羊精液品质的影响。选用10只健康乌珠穆沁种公羊,采集精液经检验合格在37 ℃下混匀,精液样品平均分成5份,用含有0、5、10、15、20 mmol/L的不同浓度棉子糖稀释液稀释。采用0.25 mL细管法冷冻,将冷冻后的细管精液置于37 ℃水浴锅中解冻15 s后进行相关指标评价及酶(SOD、GSH、MDA、GSH-Px和CAT)的测定。结果表明,当棉子糖的添加量为15 mmol/L时精子活率显著高于对照组(P＜0.05),畸形率显著低于对照组(P＜0.05),弯尾率显著高于对照组(P＜0.05),SOD活力显著提高(P＜0.05),MDA也显著低于对照组(P＜0.05)。     

冷冻精液稀释液保护剂成分探究

为了长时间的保存种畜精液并且能够扩大精液的剂量,在鲜精中加入一定的保护溶液,即通常所说的“稀释液”。尽管目前稀释液的种类较多,其目的都是为了保护精子有一个适于的存活环境,它需要维持适合的渗透压和电解质平衡,保证精子细胞膜的完整不受到损害,维持精清的环境,能供给精子营养,提高其耐低温性,抗冻性,精子的活力和顶体完整,降低精子畸形,因此在稀释液中常加入一些保护剂成分。     

绵羊冷冻精液研究进展

随着肉羊产业化、工厂化的发展,冷冻精液人工授精技术亟待应用于生产中.绵羊精液冷冻保存技术虽然得到了快速发展,但目前仍处在试验研究或小范围的应用阶段.因此,绵羊精液冷冻技术还需要深入、系统的研究.文章从绵羊精液冷冻保存稀释液中的添加剂、冷冻速率、冷冻-解冻对绵羊冷冻精液品质的影响,精液输精对受胎率的影响等方面进行了综述,为精液冷冻保存研究提供一定参考.     

TCM199对陇东绒山羊细管精液的冷冻效果

目的研究陇东绒山羊细管精液冷冻时,通过在葡萄糖一柠檬酸盐稀释液中添加TCM199,观察其对陇东绒山羊细管冷冻精冷冻效果的影响。方法选用4个试验组和对照组进行对比试验,试验组是在稀释液基础上分别添加TCM199标准液,含量为0．5％、1％、3％、5％（V／V）。结果表明：添加19／6的TCM199标准液的试验组（B）效果较好,其冷冻解冻后精子活力高于对照组和其它试验组,但差异不显著（P〉0．05）,质膜完整率试验组和对照组相比差异不显著（P〉0．05）,在精子穿卵活力试验中（SPA）,试验组和对照组相比差异不显著（P〉0．05）。但在细管解冻后不同时间段活力测定试验中,解冻后5h、8h、12h,试验组（B）和对照组活力差异显著（P〈0．05）,且随着时间延长呈下降趋势。因此制作陇东绒山羊精液细管冷冻时,添加1％TCM199标准液具有比较好的效果。     

白藜芦醇对绵羊冷冻精液质量的影响

试验旨在研究白藜芦醇对绵羊冷冻精液质量的改善效果。采用假阴道法采集6只云南半细毛羊精液,用含不同浓度（0、0.1、1、10、20 μmol/L）白藜芦醇的Optidyl稀释液稀释后进行细管分装,低温平衡和液氮气相预冻后,在液氮中保存30 d。解冻后测定精子活力、质膜完整性、磷脂酰丝氨酸（PS）分布、顶体完整性和活性氧等指标。结果表明,解冻后10 μmol/L白藜芦醇组精子总活力、直线运动百分率、精子弯尾率分别为76.14%±0.97%、43.56%±0.91%、43.24%±1.68%,均显著高于其他各组（P<0.05）;而20 μmol/L白藜芦醇组精子总活力、直线运动百分率、精子弯尾率分别为21.78%±0.79%、25.23%±1.34%、4.84%±0.68%,均显著低于其他各组（P<0.05）。10 μmol/L白藜芦醇组精子顶体完整性最高,为50.47%±0.91%,显著高于其他各处理组（P<0.05）。PS分布结果表明,10 μmol/L白藜芦醇组正常精子百分率为46.43%±2.95%,显著高于20 μmol/L组（31.14%±3.56%,P<0.05）,与其他各组无显著性差异（P>0.05）。20 μmol/L白藜芦醇组PS标记率（39.82%±3.38%）显著高于其他处理组（P<0.05）。活性氧试验结果表明,10 μmol/L白藜芦醇组正常精子（63.57%±0.71%）显著高于其他各组（P<0.05）;而20 μmol/L白藜芦醇组正常精子（32.45%±1.42%）显著低于其他各组（P<0.05）。综上,在冷冻稀释液中添加白藜芦醇可以改善绵羊冷冻精液品质,这与白藜芦醇的抗氧化特性有关。但是,白藜芦醇的冷冻保护效果具有明显的浓度依赖性,其最佳作用浓度为10 μmol/L,过高浓度的白藜芦醇反而加重精子的冷冻损伤。此外,对于白藜芦醇对绵羊精子的抗冻保护效果仍然需要体外受精或人工授精验证。     

The Effects of Resveratrol on the Quality of Frozen Sheep Semen

The effects of resveratrol on the quality of frozen-thawed sheep semen were studied in this study.Semen was collected from six Yunnan semi-fine wool sheep using artificial vagina.The semen was diluted with the Optidyl extender supplemented with resveratrol with a concentration of 0,0.1,1,10,or 20 μmol/L,followed by loading into plastic straws and equilibration at a low temperature.Then,the straws were pre-frozen in liquid nitrogen vapor,followed by storage in liquid nitrogen for 30 days.After thawing for 30 seconds in a 37 ℃ water bath,the parameters including sperm motility,acrosome integrity,membrane integrity,distribution of phosphatidylserine (PS) and ROS were measured.The results showed that the post-thaw sperm total motility,progressive motility and the rate of sperm with curved tail were 76.14%±0.97%,43.56%±0.91% and 43.24%±1.68% in the 10 μmol/L resveratrol group,respectively,which were significantly higher than those in the other groups (P<0.05).However,when the concentration of resveratrol in the freezing extender was 20 μmol/L,the post-thaw total motility,progressive motility and the rate of sperm with curved tail were 21.78%±0.79%,25.23%±1.34% and 4.84%±0.68%,respectively,which were significantly lower than those in the other groups (P<0.05).The acrosome integrity of sperm frozen in the 10 μmol/L resveratrol group was best,which was 50.47%±0.91% and significantly higher than the other treatments (P<0.05).The results of PS distribution showed that the percentage of viable sperm in the 10 μmol/L resveratrol group was 46.43%±2.95%,and significantly higher than that in the 20 μmol/L group (31.14%±3.56%,P<0.05),but there was no significant difference with the other groups (P>0.05).In addition,the PS labeling rate of sperm in the 20 μmol/L resveratrol group (39.82%±3.38%) was significantly higher than those of the other groups (P<0.05).In terms of ROS production,this study demonstrated that the rate of viable sperm (ROS and PI were negative) in the 10 μmol/L resveratrol group (63.57%±0.71%) was significantly higher than that of the other groups (P<0.05),while the rate of viable sperm (32.45%±1.42%) in the 20 μmol/L resveratrol group was significantly lower than those of the other groups (P<0.05).In conclusion,the post-thaw quality of sheep semen could be improved after the addition of resveratrol to the freezing extenders,which might be related to the antioxidant properties of resveratrol.But,the cryoprotective effects of resveratrol dependents on its used doses,and the optimal concentration was 10 μmol/L based on this present study.However,excessively high concentration of resveratrol could aggravate cryodamage on sperm.In addition,the protective effects of resveratrol on sheep sperm still needs to be verified by in vitro fertilization and artificial insemination.     

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。     

Effect of L-lysine Added in Diluent on the Fresh Semen Quality of Dorper Sheep

In order to explore the effect of different L-lysine levels added in semen dilution on the fresh Dorper sheep quality stored at liquid state,3 health Dorper ram were used to collect semen which were equal-packaged and added to solution containing different levels (0,0.1,0.2,0.3,0.4 and 0.5 g) L-lysine in 120 mL diluent and at 0,6,24,30,48,54,72 and 78 h,the sperm motility,membrane integrity and acrosome integrity were detected.The results showed that the sperm motility in groups added 0,0.1 and 0.2 g L-lysine were higher than other groups,and that in 0.4 and 0.5 g L-lysine groups decreased faster than other groups;The sperm motility in 0.5 g L-lysine group reduced to 0 at 30 h,and it reduced to 0 at 48 h in the group added 0.4 g L-lysine;The effective survival time and survival index in the group added 0.1 g L-lysine was higher than other groups (P< 0.05);The membrane integrity of 0.1 g group was the highest,that had no significant difference with control group from 0 to 54 h (P >0.05),while after 72 h the two groups had significant differences (P< 0.05).The membrane integrity in 0.4 and 0.5 g groups were the worst,and that was significant different with other groups (P< 0.05);The acrosome integrity of 0.1 g group was the highest which had no significant difference with control group (P >0.05),0.4 and 0.5 g groups were the worst,the difference between them was not significant (P >0.05),while was extremely significant difference with other groups except for 0 and 6 h (P< 0.01).The results suggested that dilution added a high concentration of L-lysine could inhibit sperm quality,when the count of L-lysine was more than 0.2 g,sperm quality was significantly decreased,adding 0.1 g L-lysine could improve Dorper sheep fresh sperm quality stored at liquid state.     

稀释液中添加L-赖氨酸对杜泊羊鲜精液品质的影响

为了探讨添加不同水平L-赖氨酸的精液稀释液对杜泊羊鲜精液态保存下品质的影响,试验选用健康的杜泊种公羊3只,采集精液,等量分装,分别加入到含不同水平(0、0.1、0.2、0.3、0.4和0.5 g) L-赖氨酸的120 mL稀释液中,在0、6、24、30、48、54、72和78 h对精子活力、质膜完整率和顶体完整率进行检测。结果表明,添加0、0.1、0.2 g L-赖氨酸组精子活力较其他添加水平组高,添加0.4和0.5 g L-赖氨酸组精子活力下降速度快,添加0.5 g L-赖氨酸组精子活力在30 h时降为0,添加0.4 g L-赖氨酸组精子活力在48 h时降为0;添加0.1 g L-赖氨酸组精子有效存活时间和生存指数最高,与其他组间差异显著(P< 0.05);质膜完整率0.1 g组最高,贮存0~54 h间与对照组间差异不显著(P >0.05),72 h后与对照组间出现显著性差异(P< 0.05),0.4和0.5 g组最差,与其他组间差异显著(P< 0.05);顶体完整率0.1 g组最高,与对照组间差异不显著(P >0.05),0.4和0.5 g组最差,二者差异不显著(P >0.05),除贮存0和6 h外,与其他组间差异极显著(P< 0.01)。结果提示,稀释液中添加高浓度的L-赖氨酸对精液品质有抑制作用,当L-赖氨酸添加水平≥0.2 g,精子品质显著下降,添加0.1 g L-赖氨酸有助于提高杜泊羊鲜精液态保存的品质。     

鸡精液稀释配方对低温保存精子活力和输精效果的影响

为了完善中国南方地区鸡场公鸡精液的保存技术、提高精液的利用率,本试验研究了在低温（4 ℃）保存的条件下,不同的保存时间（0、4、8、24 h）、不同的稀释液配方（原精液、配方Ⅰ和配方Ⅱ）对精液的精子活力变化以及人工输精繁殖效果的影响。选用33周龄黄鸡母鸡192只、公鸡42只,192只母鸡依笼号分为12组（3种处理精液×4个保存时间）,每组16只母鸡,公鸡不分组。统一采精后用两种不同稀释液稀释、低温（4 ℃）保存至0、4、8、24 h后观察精子活力,并对母鸡输精,分组收集鸡蛋,对各组受精率、出雏率、健雏率进行比较分析,以原精液低温保存作为对照。结果显示,两种稀释液组的精子低温（4 ℃）保存4、8、24 h,其精子活力极显著高于原精液组（P<0.01）,配方Ⅱ组精子的活力高于配方Ⅰ组（P>0.05）;两种配方稀释液组的精液低温（4 ℃）保存4 h,输精受精率高于原精液组（P<0.05）;两种配方稀释液组的精液在低温（4 ℃）保存8 h,输精受精率极显著高于原精液组（P<0.01）。表明这两种精液稀释液更有利于精液保存,经稀释后的精液可以显著提高受精率,可为中国南方鸡场种公鸡精液保存技术的完善提供有力的数据支撑。     

绵羊精液冷冻保存研究进展

绵羊精液保存技术目前虽然已进入生产应用阶段,但冷冻精液人工授精受胎率低仍是限制绵羊冷冻精液进行生产推广的重要因素。目前,对于绵羊精液冷冻的研究主要集中在冷冻稀释液组分和冷冻程序等方面。文章针对国内外有关绵羊精液冷冻保存稀释液、冷冻程序、解冻方法和冷冻损伤等研究进展作一综述,旨在对今后精液冷冻保存研究提供一定的参考。