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。     

维生素E对小鼠体内精子品质及精子DNA的保护作用

旨在研究维生素E对小鼠精子DNA的保护作用及对精液中抗氧化物酶活性的影响。以小鼠为动物模型,选日龄、大小相同的健康雄性小白鼠50只,随机分成5组,在试验组小鼠的每千克日粮中分别添加4、8、12、16g的维生素E,对照组添加量为0,饲喂30d后,通过颈椎脱臼处死小鼠,解剖后通过附睾和曲精细管获得小鼠精液。通过单细胞凝胶电泳及相关检测试剂盒检测分析各组间精液DNA的完整性、精子活力、密度、畸形率、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:每千克日粮中维生素E的添加量为12g时,0级精子百分率与对照组相比差异显著(P0.05);添加量为8~16g时,精子活力与对照组相比差异显著(P0.05);GR和SOD酶的活性与对照组相比差异显著(P0.05)。每千克日粮中维生素E添加量为12~16g时,CAT酶的活性与对照组相比差异显著(P0.05)。综上表明,在饲粮中添加维生素E浓度为12g·kg-1时可以明显提高小鼠精液中CAT、GR、GSH和SOD酶的活性及精子DNA的完整性。     

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。     

VE对4℃保存娟珊牛精液品质的影响

为研究维生素E对4℃保存娟姗牛精液品质和抗氧化保护作用的影响,用假阴道法采集体格健硕的娟姗牛精液,在稀释液中分别添加0 mg/mL、0.01 mg/mL、0.03 mg/mL、0.05 mg/mL、0.07 mg/mL、0.09 mg/mL的维生素E,将新鲜精液稀释至1.47×10~6/mL,于4℃环境下保存48 h,以评价精子活力、精子顶体完整率、谷胱甘肽还原酶(GR)和过氧化氢酶(CAT)活性等精液品质指标。结果显示:娟姗牛精液4℃保存至48 h时,维生素E 0.03 mg/mL添加组娟姗牛精子活力显著高于0.05 mg/mL维生素E添加组(P0.05),极显著高于对照组和其他试验组(P0.01);维生素E 0.03 mg/mL添加组牛精子顶体完整率极显著高于对照组和其他试验组(P0.01);GR酶活性维生素E 0.03 mg/mL添加组牛达71.55 U/L,较0.05 mg/mL维生素E添加组和0.07 mg/mL维生素E添加组高,且差异显著(P0.05),较对照组和其他添加组差异极显著(P0.01);维生素E 0.03 mg/mL添加组牛精子CAT值达5.68 U/mL,极显著高于对照组和其他添加组(P0.01)。说明在精液稀释液中添加0.03 mg/mL的维生素E可明显提高4℃环境保存娟姗牛精液的品质。     

不同抗氧化剂对猪精液冷冻保存效果的影响

在LEY冷冻稀释液基础上分别添加不同浓度维生素C(0、5、10、20、40、60 mmol/mL)、维生素E(0、0.2、0.5、1.0、2.5、5.0 mg/mL)、SOD(0、100、200、400、600 IU/mL)、CAT(0、50、100、200、300 IU/mL)、GSH(0、1、5、10 mmol/mL)等抗氧化剂,检测冷冻-解冻后精子活力、活率、质膜完整率、顶体完整率、平衡后和解冻后精子MDA含量,以观察5种抗氧化剂对猪精液冷冻保存效果的影响。结果表明:在冷冻稀释液中联合添加100 IU/mLSOD和200 IU/mL CAT明显提高冷冻-解冻后精子活力、活率、质膜完整率和顶体完整率(P<0.01)。     

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C对湖羊精液低温保存效果的影响

试验旨在探讨添加不同浓度维生素C的精液稀释液,对湖羊精液液态保存效果的影响。用假阴道法采集湖羊种公羊精液,并用添加不同浓度(0、1、3 mg/mL)维生素C的稀释液进行稀释,于4℃条件下冷藏,在0、24、48、72、96 h后对精子活力、精液的精浆中谷胱甘肽过氧化物酶(GSH-Px)活力、过氧化氢酶(CAT)活力、总抗氧化能力(T-AOC)和丙二醛(MDA)含量进行检测。结果表明,与对照组相比,添加维生素C的试验1组和试验2组精浆中MDA含量显著降低(P0.01或P0.05),T-AOC水平、GSH-Px活力、CAT活力极显著提高(P0.01);试验1组精子活力极显著提高(P0.01),而试验2组精子活力却在24 h内极显著降低(P0.01)。说明稀释液中添加1 mg/mL维生素C有助于提高湖羊精液低温保存的质量。     

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)。     

维生素E提高绵羊颗粒冻精品质的机理研究

以无角多赛特公羊为试验对象,采用2步稀释法,在冷冻精液稀释液中添加维生素E,对维生素E提高绵羊颗粒冻精品质的机理进行初探.试验1证明,试验组的活率(0.435)极显著高于对照组(0.365)(P＜0.01),其精子顶体总异常率(33.72%)极显著低于对照组(44.35%)(P＜0.01),且顶体完整率与冻精活率相关极显著(r＝0.662,P＝0.037).添加维生素E可降低冷冻对精子顶体的损害程度,冷冻精液品质得到显著改善.试验2结果表明添加维生素E后AST、ALP、GGT、CK、LDH与ALT的活性均与对照组无显著差异.但对照组与试验组比较,GOT及CK释放量与解冻精子的活率负相关下降(分别为r＝-0.536,r＝-0.209及r＝-0.419,r＝-0.051),LDH的释放量与解冻精子的活率之间正相关(r＝0.123与r＝0.507).试验3结果证明在绵羊冷冻精液稀释液中添加维生素E可以显著提高精子顶体酶的活力,分别为82.91 mu/ml和99.82 mu/ml(P＜0.05).绵羊冷冻精液精子顶体酶活力与精子活率呈正相关(r=0.490),与精子畸形率呈负相关(r=-0.122),冷冻精子活率随着精子顶体酶活力的提高而提高.     

精浆对猪精子冷冻效果影响的研究

为优化猪精子冷冻技术,提高解冻后精子的活力和受精能力,本试验分别以含精浆浓度为10%、20%和30%的冷冻保护剂处理精子,以冷冻前、解冻后精子活力和质膜完整性,解冻后精子进行体外受精(IVF)的卵裂率和囊胚率等作为检测指标,同时以含有卵黄的Tris-柠檬酸－葡萄糖(Tris-citric acid-glucose,TCG)冷冻基础液作为对照研究精浆对猪冷冻精子的保护作用。结果显示,在含有10%精浆浓度的稀释液中,冷冻前质膜完整性,解冻后精子活率、质膜完整性、IVF囊胚率相对于对照组均显著提高(P<0.05);当含有10%精浆的冷冻精液解冻后用于人工授精时,与配母猪妊娠率、窝产仔数、窝产活仔数等仍显著低于鲜精授精组(P<0.05)。上述结果表明,含10%精浆的冷冻保护剂能提高精子的冷冻后活力和IVF胚胎发育率,但用于人工授精配种与鲜精相比还有一定差距。     

番茄红素对秦川牛精液冷冻保存及鲜精品质影响

旨在探究番茄红素对秦川牛精液冷冻保存和新鲜精液品质的影响,为番茄红素作为新型精液稀释液添加剂和种公畜日粮添加剂提供依据。本研究采集3头年龄3~5岁健康状况良好秦川种公牛的新鲜精液混合,用0、1、2、3、4 μmol·L^-1浓度的番茄红素稀释液冷冻保存;利用计算机辅助分析仪、低渗肿胀检测法、花生凝集素荧光标记检测法、罗丹明荧光检测法等检测精子的运动性能、顶体完整率、质膜完整性、线粒体活性和抗氧化性能;在秦川种公牛日粮中添加15 mg·kg^-1 BW番茄红素,检测饲喂前后秦川种公牛的新鲜精液品质和血清抗氧化指标。结果表明,在秦川牛精液冷冻保存过程中添加番茄红素可以显著提高冷冻-解冻后精子的活力、顶体完整率、质膜完整性和线粒体活性以及抗氧化酶CAT和GSH-Px酶活性,显著降低精子中MDA含量,且番茄红素的最适添加浓度为2 μmol·L^-1(P<0.05);饲喂15 mg·kg^-1 BW番茄红素后,与饲喂之前相比,秦川种公牛的精子活力显著提高(P<0.05),精子畸形率显著降低(P<0.05),血清中抗氧化酶SOD、CAT、GSH-Px酶活性显著增加(P<0.05),MDA含量显著降低(P<0.05)。综上所述,一定浓度的番茄红素对精子在冷冻过程中造成的精子损伤和秦川牛机体应激造成的繁殖性能降低具有良好的保护作用,这可能与番茄红素通过抗氧化作用减少精子在冷冻-解冻过程中的氧化损伤和提高动物机体的抗氧化能力有关。     

奶牛性控精液与常规精液抗氧化物酶活性比较分析

通过比较常规精液和性控精液在活力、顶体完整率及精浆中四种抗氧化酶类活力的差异,对性控精液的品质进行综合评定。对用流式细胞仪分离后的性控精液和常规精液细管解冻(37.5℃,30s),分别分析精子活力和精子顶体完整率,并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。结果表明性控精液精子在活力显著低于常规精液,且精浆中四种抗氧化酶类的活力也都显著低于常规精液。     

维生素B12、E和C在家畜精液保存中的应用

家畜精液保存时,向稀释液中添加一定量的维生素B12、E或C,在提高精液品质方面具有重要的作用。精液稀释液中添加维生素B12,具有维持精子顶体完整率,延长精子体外存活时间和提高解冻后的精子活力的作用。维生素E是重要的抗氧化剂,能通过抑制自由基的形成,阻止诱发的膜脂质发生过氧化,从而维持精子顶体膜的完整,提高精子顶体完整率和降低精子畸形率;维生素E还能防止精子发生冷休克,提高冻精解冻后的精子活力,从而提高受精率。维生素C作为抗氧化物质,能有效地防止冷冻精子磷脂发生过氧化,提高解冻后的精子活力,延长精子在体外存活的时间;维生素C的抗氧化作用还表现在能够保护精子细胞中的遗传基因DNA免受氧化破坏。     

Effects of Exogenous Glutathione on the Quality of Chilled Bull Semen

Glutathione (GSH) at concentrations of 0.0 (control), 0.5, 1.0, 2.0 and 3.0 mM was added to chilled bull semen to determine its effects on the keeping quality of semen used for artificial insemination (AI). The semen was preserved with egg yolk citrate extender. All samples were stored at 4-8 degrees C for 5 days. Sperm motility and proportion of abnormal acrosome were assessed daily. Sperm motility was significantly (p < 0.01) higher in the semen treated with 0.5 mM glutathione than in untreated semen on each day. The optimum sperm motility (>or=50%) for AI was retained significantly (p < 0.01) for 3 days in 0.0, 0.5, 1.0 and 2.0 mM glutathione treated semen, whereas in 0.3 mM glutathione-treated semen, sperm motility was 46.8% for 3 days. Acrosomal damage was significantly (p < 0.01) reduced after addition of 0.5 mM GSH in the preserved semen. Bull semen can be preserved in chilled condition for 5 days with 0.5 mM GSH with sperm motility above 40% and 12% acrosome abnormality.     

L-肉碱对猪精液冷冻保存效果的研究

本试验用5%二甲基甲酰胺(DMF)替代甘油作为冷冻保护剂,研究不同浓度(0、0.02、0.04、0.06、0.08 mg/mL)L-肉碱对猪精液冻后常规指标(精子活率、线粒体活性、顶体完整性、质膜完整性)、过氧化氢酶(CAT)和总抗氧化酶(T-AOC)活性以及丙二醛(MDA)含量的影响。结果表明:添加0.04和0.06 mg/mL的L-肉碱可有效改善猪精液冷冻后效果。0.04 mg/mL组可显著提高精子冻后活率和线粒体活性(P<0.05);0.06 mg/mL组可显著提高顶体完整性和质膜完整性(P<0.05)。添加0.06 mg/mL L-肉碱可显著提高冷冻后精子内T-AOC酶活性并且抑制MDA的产生(P<0.05),但CAT活性与0.04 mg/mL组差异不显著。在冷冻稀释液中添加0.06 mg/mL L-肉碱可以提高猪精液冷冻保存效果。     

纳米化红景天多糖对猪精液冷冻效果的影响

在冷冻稀释液中分别添加分数为1％、5％、10％、15％、20％的纳米化红景天多糖（Nonomaterialsrhodiolasa—chalinensispolysaccaride,NRSP）溶液,以添加6mg／L红景天多糖（Rhodiolasachalinensispolysaccaride,RSP）为对照组,不添加RSP为空白组,检测其对猪冷冻精子活力、顶体完整率、质膜完整率等生理结构指标,以及精子抗氧化能力的影响。结果表明,添加体积分数为15％NRSP组的精子活力（0．69）、顶体完整率（51．42％）、质膜完整率（25．95％）均高于其他组,并显著高于空白组（P〈0．05）。此外添加15％NRSP组的解冻后直线运动精子百分比（46．33％）要显著高于其他各组（P〈0．05）;添加6mg／LRSP组丙二醛（MDA）浓度最低（10．83±0．39）／2mol／L,并且显著低于空白组（31．85土1．36）μmol／L,（P〈0．05）,但与15％的NRSP组（11．60±0．42）μmol／L无显著差异（P〉0．05）。添加20％的RNSP组超氧化物歧化酶（SOD）活力最高（343．05-＋-19．64）μ／mgprot,显著高于其他添加组（P〈0．05）。精子活力、顶体完整率、质膜完整率之间存在显著的正相关关系（P〈0．05）,而精子活力和顶体完整性与MDA浓度呈显著负相关（P〈0．05）,与SOD活力呈显著正相关（P〈0．05）,质膜完整率与两者含量相关性不显著（P〉0．05）。     

荷斯坦牛和西门塔尔牛冻精品质及其体外受精后早期胚胎发育的比较研究

为探讨荷斯坦牛和西门塔尔牛冻精的精液品质及体外受精后胚胎发育能力的差异,利用目测法、低渗膨胀法和考马斯亮蓝染色法评估了荷斯坦牛和西门塔尔牛冻精的活力、质膜完整率和顶体完整率,并比较了二者冻精体外受精后胚胎的卵裂率和囊胚率。结果表明,荷斯坦牛和西门塔尔牛冻精的活力(30.4%和27.2%)、质膜完整率(41.96%和36.22%)和顶体完整率(77.02%和73.02%)均无显著差异(P>0.05),但荷斯坦牛冻精体外受精后的卵裂率(57.5%和48.6%)和囊胚率(30.3%和23.2%)显著高于西门塔尔牛冻精(P<0.05)。提示,不同品种公牛精液体外受精后的发育能力有显著差异(P>0.05)。     

Antioxidant supplementation,effect on post‐thaw spermatozoan function in three sturgeon species

High levels of reactive oxygen species (ROS), which may be associated with reduced sperm quality, can be detected during cryopreservation of sperm of some species. Our objective was to investigate whether the addition of antioxidants to cryopreservation extenders influenced post‐thaw sperm characteristics and fertility in Acipenser dabryanus, Acipenser sinensis and Acipenser baerii. Prior to freezing, sperm samples were diluted with a base extender as control or in extender supplemented with catalase (CAT), glutathione (GSH), cysteine (NAC), ascorbic acid (VC) or their paired combinations. Protective concentrations of CAT, GSH and VC in the three species were 25 U/ml, 0.25‐0.5 mg/ml and 0.5 mg/ml, respectively. Cysteine showed no protective effect against ROS. The addition of CAT, GSH and VC positively affected either acrosome or membrane integrity of post‐thawed sperm in the three species, as well as spermatozoan motility in A. sinensis. The combination of antioxidants did not show a positive synergistic effect. This study suggested that the use of antioxidants in the cryopreservation of sturgeon sperm has potential to decrease intracellular ROS, and consequently preserve acrosome and membrane integrity, as well as spermatozoan motility.     

Influence of vitamin E and vitamin E-selenium combination on arginase activity,nitric oxide level and some spermatological properties in ram semen

The effects of vitamin E and vitamin E-selenium combination on seminal plasma arginase activity and nitric oxide level and some spermatological properties in rams were investigated in this study. For control group, animals were injected intramuscularly with physiological saline. For vitamin E group, rams were injected intramuscularly with 300 mg/ram vitamin E. For vitamin E + selenium group, animals were injected intramuscularly with 5 ml/ram vitamin E + selenium. The semen was collected by artificial vagina at 1st, 4th, 24th, 48th and 72nd hr after administration in each group. Significant decreases in seminal plasma arginase activity (at 1st, 24th and 48th hr), nitric oxide level (at 72nd hr) and abnormal sperm rate (at 1st, 24th and 72nd hr), and significant increases in semen volume (at 24th hr), semen mass activity (at 24th and 48th hr), sperm motility (at 24th, 48th and 72nd hr) and concentration (at 1st hr) were observed in vitamin E group compared with control group. Similarly, significant increase in semen volume (at 1st, 24th and 48th hr), mass activity, (at 48th hr), motility (at 48th and 72nd hr) and concentration (at 4th, 24th and 48th hr), and significant decrements in abnormal sperm rate (at 1st, 24th, 48th and 72nd hr), seminal plasma nitric oxide level (at 1st, 4th, 24th and 48th hr) and semen pH (at 24th and 48th hr) were detected in vitamin E + selenium group in comparison to the control group. As a result, it is suggested that vitamin E and/or vitamin E + selenium applications may improve reproductive performance.     

Effect of Se and VE Supplement on Semen Quality,Antioxidant Enzyme Activities and Heat Shock Protein Expression of Goat in High Temperature Season

This experiment was conducted to study the effect of selenium and vitamin E supplement on semen quality, antioxidant enzyme activities and heat shock protein expression of goat in Hainan high temperature season.16 adult Hainan Black goat with good health and approximate weight were randomly divided into 4 groups, fed with basal diet(control group), basal diet+0.5 mg/kg Se(Se group), basal diet+100 mg/kg VE(VE group), and basal diet+0.5 mg/kg Se+100 mg/kg VE(Se+VE group), respectively.The experimental period was 93 d.Semen samples were collected in the last week of the experiment on two consecutive days.The semen quality, antioxidant enzyme activities and heat shock protein expression were analyzed.The results showed that compared with control group, the ejaculate volume was not significantly affected by Se or VE supplement(P>0.05).Sperm density and sperm motility were increased significantly by Se and VE supplement(P<0.05), and the abnormal rate was decreased extremely significantly(P<0.01).The goats fed with Se and VE also had higher activities of GSH-Px(P<0.01), SOD(P<0.05), CAT(P<0.05) and T-AOC(P<0.01), and lower MDA concentration in seminal plasma(P<0.05).The relative expression levels of HSP70 and HSP90 mRNA in supplement groups were decreased extremely significantly(P<0.01).However, there were some certain differences between the Se and VE supplement groups on semen quality and heat shock protein expression.In conclusion, the supplementation of Se and VE could help to improve goat semen quality by increasing the sperm density, sperm motility, the antioxidant enzyme activities, and decreasing the abnormal rate in hot season of Hainan.Finally, Se and VE supplement had good effects on relieving the environment heat stress.