不同冷冻保护剂在猪精液冷冻中的作用分析

本实验采用一定浓度的甘油、EG、DMA、DMSO及其两两组合物作为冷冻保护剂。用含冷冻保护制的稀释液将精液稀释后,常温保存,观察精子活率,比较精子生存指数。结果表明：在常温下对精子毒害作用最大的是DMSO,其次是甘油,EG和DMA对精子毒害作用最小。以一定浓度的10种冷冻保护剂将对精液冷冻后观察解冻活率,发现EG和DMA混合保护剂解冻后精子活率最高。     

水牛冷冻精液稀释液中添加SOD、GSH对精子品质的影响

目的分析水牛冷冻精液稀释液中添加SOD、GSH对精子品质的影响,方法于广西畜禽品种改良站中选取摩拉水牛的新鲜精液进行分析研究,并且随机将其分为A组、B组、C组、D组,A组为正常水牛精液,B组添加300IU/mL SOD,C组添加1.0mM GSH,D组添加300IU/mL SOD与1.0mM GSH,对比各组影响结果。结果在不同氧化剂对精子影响方面上,4组的顶体完整率无显著差异,在精子解冻后不同时间段内,B组C组D组精子活率显著优于A组,D组精子活率显著优于B组C组,组间对比差异显著(P0.05)。在抗氧化剂组合对体外受精结果上,D组的精子分裂率、囊胚孵化率与囊胚率显著优于A组,组间对比差异显著(P0.05)。结论在水牛精子中添加SOD与GSH可以良好的延缓精子死亡时间,效果显著。     

绵羊细管冷冻精液及改进输精器的应用效果

前言 人工授精技术的推广,特别是冷冻精液的应用,大大提高了种公畜的繁殖率,提高了畜群的生产水平。黑龙江省针对优良肉用种公羊不足,省家畜繁育指导站进行德国肉用美利奴种公羊和法国夏洛莱种公羊精液冷冻进行试验推广,并采用改进后的弯嘴输精器来解决以往由于绵羊输精部位过浅,造成绵羊冷冻精液受胎率低的矛盾,现将试验效果报告如下： 1 材料与方法 1.1 试验场及试验动物 本试验在绥棱县种羊场饲养的小尾寒羊×乌珠穆泌F1羊群中,随机选择发情正常,体重相近,健康无病,1999年8月13日至8月28日之间发情的21只母羊为试验羊只。     

云岭黑山羊的精液冷冻效果观察

为加速品种的育成和改良步伐,通过云岭黑山羊的精液冷冻效果观察试验,结果表明:(1)两次稀释法比一次稀释法好;(2)解冻0h精子活率与解冻1h后精子活率下降不大,差异不显著。解冻3h后,精子活率活下降且差异显著;(3)解冻后精子活率较差,顶体受损率较高,而精子的畸形率变化不明显;(4)云岭黑山羊与努比山羊除射精量有差别外,其他精液的质量指标差别不大。     

不同冷冻温度对藏獒冷冻精液品质的影响

为探讨藏獒细管冻精最佳冷冻曲线,利用全自动细管精液程控冷冻仪控制冷冻温度变化,测定藏獒精液在不同冷冻曲线下冻精品质。结果表明:冷冻曲线4~-10℃(降温速率3℃/min)、-10~-100℃(降温速率40℃/min)、-100~-140℃(降温速率20℃/min),冻精解冻后精子活力最高,冻精精子顶体完整率最高,冻精品质最好。     

畜禽冷冻精液稀释液的研究进展

畜禽精液冷冻保存是人工授精技术的一个重大发展。在精液冷冻保存中稀释液起关键作用。稀释液通过粘附到精子膜上,保护膜结构,增加膜流动性,调低精子对冷休克的易感性,增加了精子冻存能力及解冻后精子活力。     

鱼类精子冷冻保存研究进展

鱼类精子的长期保存对于遗传育种、生物多样性保护、濒危物种保护、渔业可持续发展具有重要意义,引起了人们越来越广泛的关注和重视.低温生物学的不断研究和发展为精子的长期稳定保存提供了理论和技术方面的支持.近年来鱼类精子的冷冻保存在保存原理和技术方面都取得了极大的成就,实现了多种鱼类精子冷冻保存的技术突破,并对冷冻保存的原理、冷冻损伤机制等方面有了更深刻的认识,在冷冻精子的生产应用方面也越来越趋于成熟稳定.本文仅针对鱼类精子冷冻保存的研究现状及发展前景作以阐述,以期对更加深入广泛的研究工作有所帮助.     

咖啡因对猪精液井水保存效果的试验报告

贺州市八步区是广西的养猪强区，去年存栏母猪85269头，外调肉猪1092095头。我区很重视猪的良种推广工作，首先在农村全面推广猪人工授精，建立猪人工输精网络，实行统一供应优良猪精液。在推广这一技术中应用猪精液井水（14～20℃）保存方法尤为必要和实用。为此，笔者从2002年4月开始至2003年10月，对公猪精液井水保存添加咖啡因的保存效果进行了试验，现将结果报告如下。     

N-乙酰半胱氨酸显著提高猪精液冷冻保存效果

延边大学农学院魏世宝等，以手握法采集的猪精液为试验材料，以解冻后精子的活率、顶体完整率、低渗膨胀率、运动学参数（VAP、LIN、ALH、BCF）和膜脂质过氧化反应为评定标准，旨在探讨N-乙酰半胱氨酸对猪精液冷冻保存效果的影响。结果表明，在冷冻稀释液中添加1mmol／L的N-乙酰半胱氨酸可以显著地提高解冻后精子的活率和顶体完整率，     

海藻糖对施氏鲟精子冷冻保存效果的影响及其冷冻损伤机理的初步探究

为了解抗冻剂对施氏鲟(Acipenser schrenckii)精子冷冻保存效果的影响及其冷冻损伤机理,比较了添加不同浓度海藻糖和蔗糖作为冷冻保护剂的精子稀释液处理后施氏鲟精子冷冻复苏的活力、快速运动时间和寿命。结果表明,添加60 mmol·L^-1海藻糖1.0 mmol·L^-1氯化钾的稀释液处理后,精子冻后快速运动时间和寿命与鲜精相比无显著差异(P>0.05);且活力较其他处理组有所提高,达到(26.67±3.32)%,但仍显著低于鲜精(P<0.05)。利用透射电镜和扫描电镜对施氏鲟精子超低温冷冻保存前后的超微结构进行观察,并对其能量代谢酶和抗氧化酶活进行测定和比较,结果表明,低温冻存造成施氏鲟精子的膜系统和细胞器(主要为线粒体和轴丝)损伤;能量代谢酶[总ATP酶、肌酸激酶(CK)、琥珀酸脱氢酶(SDH)、乳酸脱氢酶(LDH)]和抗氧化酶[过氧化氢酶(CAT)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-PX)]活性均显著下降(P<0.05),而抗氧化酶谷胱甘肽还原酶(GR)活性显著上升(P<0.05),表明...     

离子浓度和一氧化氮供体对池塘养殖鳗鲡精子活力的影响

鳗鲡是世界性的优质养殖鱼类,20世纪80年代以来养鳗业迅速发展,已成为水产产业化程度最高的养殖品种之一,但养殖所需的苗种完全靠天然捕捞。早在1934年Boucher等[1]就用孕妇尿促使雄性欧洲鳗(A.anguilla)精巢成熟,Yamamoto和Yamauch于1974年对日本鳗鲡进行人工催熟获得精子[2],但多年来鳗鲡精子质量和数量一直在波动(数量JOURNALOFFISHERIESOFCHINA　　　　　　　　　　　Vol.27,No.4　为0.1～0.4g,运动精子为20%～50%)[3,4],因此,如何稳定和提高精子的质量和数量的研究是非常必要的。绝大多数鱼类的受精过程是在体外条件下进…     

分光光度法测定俄罗斯鲟精子密度标准的研究

为了标准化精子超低温冷冻保存和人工授精程序,建立了分光光度法测定俄罗斯鲟精子密度的方法,比较了不同波长(380 nm、530 nm、780 nm)下吸光度(A)与精子密度(C)的关系。结果表明,分光光度法的检测下限为3×106cells/mL,且检测上限随波长的增加而上升,当精子密度为3×106~1.5×109cells/mL时,530 nm为最适检测波长,吸光度与精子密度呈对数回归关系,其回归方程为:A530=-8.560+1.323 lgC(R2=0.971)。     

Standardization of photometric measurement of sperm concentration from diploid and tetraploid Pacific oysters, Crassostrea gigas (Thunberg)

To provide necessary standardization of procedures for cryopreservation of sperm, a spectrophotomeric method was developed to determine the sperm concentration of diploid and tetraploid Pacific oysters, Crassostrea gigas. Wavelengths of 380, 550, 581 and 780 nm were compared, and 550 and 581 nm were found to be the most sensitive and reliable. A linear relationship between sperm concentration and photometric absorbance was observed for sperm concentrations between 2 × 10⁷ and 2 × 10⁹ cells mL^?1. The regression equation for the standard curve at 550 nm for sperm of diploid oysters was Y=?8.528+1.165 log X. The equation for sperm of tetraploid oysters was Y=?8.844+1.236 log X. The equation at 581 nm for sperm of diploid oysters was Y=?8.07+1.104 log X. The equation at 581 nm for sperm of tetraploid oysters was Y=?8.331+1.167 log X. Comparisons derived from the standard curves at 581 nm between observed values and the predicted values indicated good agreement for sperm from diploid (coefficient of determination, r²=0.983) and tetraploid (r²=0.980) oysters.     

不同盐度激活液、K+浓度及保存时间对日本鳗鲡精子活力的影响

在6种不同盐度(34、32、30、28、26和22)激活液、3种不同K+浓度(25 mmol/L、30 mmol/L和35 mmol/L)稀释液和不同保存时间(0 h、24 h、48 h、72 h和96 h)条件下,对日本鳗鲡(Anguilla japonica)精子的活力进行观察和测定.结果表明,激活液盐度为30时精...     

Spectrophotometric determination of sperm concentration and short‐term cold‐storage of sperm in Atlantic croaker Micropogonias undulatus L. broodstock

The objective of this study was to optimize the methodology for spectrophotometric determination of sperm concentration in Atlantic croaker Micropogonias undulatus L. milt and to estimate its potential for short‐term cold‐storage. The spectrophotometric determination of sperm concentration was evaluated using milt samples from six males serially diluted in Hank's balanced salt solution at 200 mOsm kg^?1 (HBSS). The predictive power of regression models between sperm concentration and absorbance was determined from 200 to 500 nm and found to be highest within the visible spectrum despite a peak of milt absorbance at 288 nm. Absorbance reading at 400 nm was selected for further analysis to maximize the absorbance of the sample hence the sensitivity of the method while minimizing the impact of potential sample contamination with blood. The standard‐curve of correlation between sperm absorbance at 400 nm and concentration was validated and held an accuracy ranging between ?7.40% and +4.56% across males. Total sperm motility duration and the proportion of motile spermatozoa were significantly higher in milt samples diluted 1:3 in HBSS than in the undiluted control during up to 30 h of cold‐storage.The methodologies investigated in this study can be applied to optimize sperm usage and achieve predictable artificial fertilization protocols in Atlantic croaker.     

Effect of methanol concentration and thaw rate on the viability and fertility of cryopreserved Arctic char,Salvelinus alpinus (L.), spermatozoa

In two trials, Arctic char (Salvelinus alpinus) semen was frozen in 0.5 mL straws using extenders consisting of 0.3 M glucose and 10%, 12.5% or 15% methanol. Cryopreserved semen was thawed by immersing straws in 25 °C water for 17 s (11.6 °C s^?1) or in 5 °C water for 60 s (3.3 °C s^?1). The viability of the frozen–thawed semen was measured by determining post‐thaw motility and sperm membrane integrity. Two fertility trials were also conducted. There was no effect of trial or thaw rate on post‐thaw sperm viability or fertility. Use of 15% methanol in the extender resulted in the highest overall percentage of sperm motility and fertility. Use of 12.5% methanol as a cryoprotectant resulted in a higher per cent post‐thaw motility and a lower percentage of dead cells than did 10% methanol. Thus, levels of methanol higher than the commonly used 10% are beneficial for cryopreserving Arctic char sperm.     

Development of a spectrophotometric technique for sperm quantification in the spermcasting Australian flat oyster Ostrea angasi Sowerby

Sperm quantification is vitally important when sperm concentration is required for standardization of different fertilization treatments in a hatchery. Although the haemocytometer method is generally used to determine sperm concentration, the procedure is tedious and the attributes are not suitable for handling a large number of sperm samples within a short period. In this study, the efficiency of sperm concentration determination was improved in the spermcasting oyster Ostrea angasi Sowerby by optimizing the regression model and parameters critical to spectrophotometric reading. Although sperm concentration can be estimated in a wide range of wavelengths, the 350‐nm wavelength produced the best fit to the regression model (y = 1 × 10^?8 x + 0.163; r² = 0.996). In addition, the sperm counts estimated with this model were similar to the haemocytometer counts. The reading repeatability of this technique was further validated with samples from different individuals. Comparisons with literature suggest that when the spectrophotometric technique is applied to a new species for estimating sperm concentration, the regression relationship between sperm concentration and wavelength reading should be reassessed due to species‐specific discrepancy.     

Effect of ascorbic acid supplement in vitro onrainbow trout sperm viability

We evaluated motility and fertilizing ability of rainbow trout Oncorhynchus mykiss semen obtained from fish fed diets without ascorbic acid and a diet supplemented with 870 mg kg^{minus 1} of ascorbyl monophosphate. Semen was stored in vitro on ice (0 °C) during 14 days. The spermatozoa from the supplemented group had the highest motility and lowest decline in fertilizing ability after storage. Lack of a positive effect of exogenous vitamin C on semen in fish deficient in ascorbic acid (milt was supplemented with 50 mg l^–1 of ascorbic acid) suggests that the positive effect of ascorbic acid on semen quality is related to its long-term effects during spermatogenesis.     

圆口铜鱼精浆的理化性质分析

对圆口铜鱼(Coreius guichenoti)精浆离子和氨基酸成分及精液生理特性进行了检测分析。结果显示,圆口铜鱼精液pH值为7.3,呈弱碱性;精液浓度为39.7%,精子密度为5.3×10~9个/mL;精浆离子以Na~+含量88.7 mmol/L最高,其次是K~+,之后为Ca~(2+)、Mg~(2+)、Zn~(2+)、Cu~(2+);精浆水解氨基酸总量为2 872.69μmol/100mL,其中以脯氨酸含量最高,蛋氨酸、酪氨酸和组氨酸含量最低。该结果填补了圆口铜鱼繁殖生物学的相关数据,为圆口铜鱼规模化人工繁育提供了基础资料。