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。     

不同温度解冻对冻后精子活力及生存时间的影响

不同温度解冻对冻后精子活力及生存时间的影响郝爱玲阿淑艳李淑坤（黑龙江省家畜繁育指导站哈尔滨１５００６０）牛冷冻精液质量的好坏，直接影响受胎率，影响着养牛户的经济效益。而冻后精子活力及生存时间（解冻后３７℃保存４小时精子活力）又是冷冻精液质量中最重要的...     

控制牛冷冻精液中细菌数的综合措施

在牛冷冻精液生产过程中，被细菌污染的精液精子活力下降，生存时间缩短，而且输精后极易造成早期胚胎的死亡，降低受胎率。自2009年01月01日开始实施的《牛冷冻精液国家标准》（GB4143-2008）明确规定牛冷冻精液解冻后，每剂量中细菌数应≤800个。     

解冻温度对精子活力的影响

[目的]为了获得较为理想的解冻效果.[方法]采用不同温度,不同解冻时间,通过镜检解冻后活力的方法.[结果]发现:不同解冻温度对牛冷冻精子复苏的影响,用39.5℃温水解冻30 s精子的活力最高,冷冻精液解冻后精子活力达O.53.用40.O℃温水解冻10s、39.0℃温水解冻30 s精子的活力也较高,冷冻精液解冻后精子活力...     

牛精液新型冷冻保护剂的研究

通过对不同浓度的甘油和乙二醇对牛精液冷冻保存效果的测定,探讨冷冻保护剂乙二醇与传统冷冻保护剂甘油的冷冻效果,并通过对葡萄糖和卵黄及Tris等成分的调整进一步分析了主要营养成分和缓冲体系对牛精液冷冻保存的影响程度。试验表明:1GL和EG对牛精子均有良好的冷冻保护能力,解冻后6h的活力GL组中以GL1.2和GL1.4的活力最高,均为0.50;EG组中以EG0.7的活力最高,达到0.53,高于0.35以上的国标。2其他营养成分的变化对GL和EG保护精子的效果有一定影响,特别是卵黄的作用比较明显;3在同等条件下,GL对精子的冷冻保护效果优于EG,EG组的活力衰竭快于GL组。     

比利时蓝白花牛XY精子分选及冷冻精液制备

本研究利用流式细胞仪分选比利时蓝白花牛X、Y精子并制备冷冻精液。选择平均年龄4岁的健康比利时蓝白花种公牛12头，假阴道法采集精液送至实验室分选，经流式细胞仪分离、冻存、解冻后精液重分析和品质鉴定。X、Y冻精（93.4%和91.1%）性别比例显著高于常规冻精（50%）；分选后精液冻后存活率、活力和顶体完整率与常规冻精差异不显著。本研究结果显示，分选前控制公牛的精液品质（活力≥70%，畸形率≤18%）可以明显改善分选效果；分选后精子纯度和冻后活力满足低剂量人工授精要求（纯度>90%，活力>30%），精子分选对比利时蓝白花牛产业的发展具有重要意义。     

影响猪冻精质量因素的研究进展

精液冷冻保存作为精液长期保存的最佳方法,具有保存时间长、生产成本低等优点,但由于猪精子较其他物种更容易受到低温的损伤,导致猪精液冷冻后精子活力低、受精率低,从而限制了冻精在养猪生产中的广泛使用。为优化猪精液冷冻-解冻方法,提高猪冻精质量,本文基于猪精液冷冻保存的损伤机理,从冷冻前处理、精液稀释液、冷冻和解冻程序等方面对影响猪精液冷冻质量的因素进行了详细阐述和总结。     

海藻糖对猪精液冷冻保存效果的影响

在传统的Tris-柠檬酸-葡萄糖稀释液基础上，分别添加25％、50％、75％、100％的海藻糖，研究不同浓度海藻糖对猪精液冷冻后精子质量的影响。结果表明，海藻糖相对于对照TCG稀释液能够显著改善和提高猪精液的冷冻效果，其最佳添加浓度为25％，冷冻-解冻后猪精子活力、活率、线粒体活性、质膜完整性以及顶体完整率均显著提高（P〈0．05），分别达到41．38％、46．34％、44．56％、43．51％和64．09％。海藻糖可以明显抑制精子获能，获能处理前精子获能率仅为3．68％，而获能处理后达到41．82％，有利于促进精子获能。精液稀释液中甘油的适宜添加浓度为2％，海藻糖只有与甘油共同作用，才能在冷冻-解冻过程更加有效地保护精子。猪精子活力、活率、线粒体活性、质膜完整率、顶体完整率等之间存在极显著的正相关关系（P〈0．01），而与获能处理前精子的获能率存在显著的负相关关系（P〈0．05）。     

抗氧化剂在公猪精液冷冻保存中的应用

近几年来,在公猪精液冷冻稀释液中添加抗氧化剂以提高冷冻精液质量的研究受到广泛的关注。添加谷胱甘肽、超氧化物歧化酶和过氧化物酶、维生素E、L-半胱氨酸和L-谷氨酰胺、中草药成分等抗氧化剂可以有效地防止氧化损伤,提高解冻后精子活率、精子成活力、质膜与顶体完整性等,进而提高冷冻保存精液的受精能力。作者综述了抗氧化剂的抗精子冷冻损伤的作用机制及不同抗氧化剂的应用效果,并对其在公猪精液冷冻保存中的应用前景进行了展望。     

牛冷冻精液人工授精若干技术问题认知

本文就牛冷冻精液解冻温度及解冻后精液温度、牛冷冻精液细管的取用、精液细管安装于输精枪的要领、母牛的爬跨行为及生殖道排血现象、母牛受胎率统计、牛冷冻精液解冻温度及评定精子活力温度相关标准的表达等技术操作问题进行了探讨。     

Comparative study of different methods for dog semen cryopreservation and testing under clinical conditions

The extenders and freezing rates from three different freezing protocols were combined and compared to each other in order to study the post-thawing acrosome integrity and fertility of frozen dog sperm. A commercial bovine TRIS-base extender (TRILADYL) and two self-made canine semen extenders (Norwegian and Dutch) were combined with a conventional bovine and two canine freezing regimes, and acrosome integrity of frozen/thawed spermatozoa was assessed by fluorescein isothiocyanate conjugated peanut agglutinin staining (FITC-PNA). Differences between freezing/thawing protocols were reflected in the proportion of cells with acrosomal damage and not based on motility results. It was concluded that during dog semen cryopreservation extenders had less influence on the post-thawing sperm quality than did the freezing rates. The optimal extender/freezing rate combination (TRILADYL/Norwegian) was used in the clinical practice to evaluate the fertility of frozen sperm administered by intrauterine insemination using a surgical approach. The pregnancy rate was 57% (4/7), but the average litter size was low (2.8). This may have been due to the insufficient sperm numbers contained in an insemination dose and/or to the incorrect timing of artificial insemination (AI). The final conclusion is that the commercial bovine extender is useful for freezing dog semen, and the TRILADYL/Norwegian freezing protocol is recommended as the most advantageous combination for the freezing of canine semen in the clinical practice.     

提高奶牛细管冻精精子活力的试验研究

为了提高奶牛细管冻精精子的活力,试验探索了稀释液种类、最佳熏蒸距离、最佳冷冻温度、最佳熏蒸时间、不同解冻温度及时间对精子活力的影响。结果表明:精子在由柠檬酸钠和果糖组成的稀释液中存活时间长,在三羟甲基氨基甲烷稀释液中冷冻后精子活力高于其他稀释液;牛细管冻精最佳熏蒸距离为2.5 cm,时间影响不显著(5～10 min均可);用50℃温水解冻15 s的精子活力比其他解冻温度和时间时的精子活力要好。     

畜禽精液冷冻损伤保护研究进展

精液冷冻过程中液体冰晶化造成的机械损伤和氧化自由基造成的氧化损伤是畜禽冷冻精液产业化的主要限制因素。在冻精中添加外源抗氧化剂维持氧化还原平衡状态以此减轻氧化损伤、通过添加抗冻剂和控制冷冻速率来减少冰晶产生和减轻机械损伤成为畜禽冻精技术研究的重点。本文从不同种类抗氧化剂的抗氧化机理及其在冻精中的应用现状、添加抗冻剂和控制冷冻速率等方面减少机械损伤进行综述,以期为畜禽精液冷冻技术提供理论和技术参考。     

Cryopreservation of stallion semen: Effect of adding antioxidants to the freezing medium on sperm physiology

Cryopreservation of stallion semen has not reached the level of efficiency and positive results described in other species. This is mainly due to the greater sensitivity of stallion sperm to the freezing process, showing higher rates of oxidative stress and plasma membrane damage, which trigger the activation of several cell damage pathways that ultimately culminate in DNA fragmentation and cell death. Therefore, finding molecules that improve the efficiency of this technique in stallion by preventing oxidative stress and cell damage is required. Thus, the aim of the present study was to evaluate the effect of adding three antioxidants (MnTBAP, NAC and FeTPPS) to the freezing medium on the quality and functional parameters of stallion sperm. Semen samples from three stallions frozen with the antioxidants were evaluated in two conditions: (a) adding the antioxidants before freezing, and (b) before and after freezing. Plasma membrane integrity, mitochondrial membrane potential, lipid peroxidation, intracellular ROS levels, membrane lipid disorder, DNA damage, sperm motility and binding to the zona pellucida were assessed. The results showed that MnTBAP was the antioxidant treatment that best controlled the oxidative stress process and post-thaw cell damage, showing higher plasma membrane integrity, mitochondrial membrane potential, sperm motility, number of spermatozoa bound to the zona pellucida of bovine oocytes and lower lipid disorder. Additionally, it was determined that a second post-thaw application of antioxidants is detrimental since induced higher cell damage and lower sperm motility, without showing any beneficial effect on the spermatozoa.     

牛精液冷冻保护剂研究进展

牛的人工授精技术是目前应用最为广泛的动物繁殖技术之一,对牛的遗传改良做出了巨大贡献。但是,在冷冻一解冻过程中仍然有大约40％～50％的精子失去活性,限制良种公牛种用性能的发挥。论文在分析精子冷冻保存原理的基础上,阐述了渗透性、非渗透性冷冻保护剂以及低密度脂蛋白对牛精子的冷冻保护作用,以期为开发新型冷冻保护剂的研究提供一定参考,进一步提升牛冷冻精液的质量。     

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）。     

A comparison of duck and chicken egg yolk for cryopreservation of stallion sperm

OBJECTIVE: Duck and chicken egg yolk were compared for their protective effects against cold shock during the cryopreservation of stallion sperm in a lactose-EDTA-glycerol cryodiluent. DESIGN: A completely randomised design was used. Procedure Ejaculates from five stallions (n = 14 ejaculates) were split and diluted to either 20 or 200 x 10(6) sperm/mL in a lactose-EDTA extender containing either duck or chicken egg yolk. The extended semen was then frozen in liquid nitrogen. The percentage of sperm total motility and forward progressive motility were assessed before freezing and at 0 and 1 hr after thawing. Morphology data were also collected at 0 and 1 hr post thaw. RESULTS: Total and forward progressive motility were higher when the sperm were frozen in the presence of duck rather than chicken egg yolk. Furthermore, the total and forward progressive motility and percentage of morphologically normal sperm were higher when frozen at a concentration of 200 than 20 x 10(6)/mL. CONCLUSION: The results of this study demonstrate that the motility parameters of stallion sperm are improved when the semen is frozen in lactose EDTA extender supplemented with duck egg yolk rather than chicken egg yolk. Moreover, sperm motility and the percentage of morphologically normal sperm were higher after freezing at a concentration of 200 x 10(6)/ml rather than 20 x 10(6)/ml.     

冻存液添加蜂王浆主蛋白对精子品质的影响

目的 精液冷冻保存作为人工授精不可或缺的一个技术环节,对优质畜禽种群的繁衍和保存有着至关重要的意义。目前,因精液冻存时冻存液成分、冷冻方法和外部氧化应激等因素影响,导致冻存精液品质不一。蜂王浆（RJ）已被证明能提高动物精液品质,蜂王浆主蛋白（MRJPs）作为RJ主要生物活性成分物质,具有多种生物活性和抗氧化能力。方法 为提高冻存精子品质,本研究开展了在公牛精液冻存液中添加不同浓度（0 g/25 mL、0.01 g/25 mL、0.02 g/25 mL、0.03 g/25 mL、0.04 g/25 mL）的MRJPs冻干粉,对冻存48 h后解冻精子的总活力、前进性活力、畸形率等相关参数进行了观察评估。结果 添加MRJPs呈浓度依赖性方式显著降低精子总活力、快速前进性活力、缓慢前进性活力和直行性活力（P<0.05）,而且精子畸形率和精子原地移动活力与对照组相比无显著差异（P> 0.05）。结论 精液冻存液中添加MRJPs会抑制冻存精子活力,因此,对MRJPs能以何种方式提高精液品质的研究还需要进一步开展。     

抗氧化剂对家畜精液冷冻保存的应用研究进展

 在家畜精液冷冻稀释液中加入抗氧化剂以提高冷冻精液质量的研究受到广泛关注,通过添加抗氧化剂降低精子在冷冻保存过程中的氧化损伤,保护精子质膜、精子顶体和DNA的完整性,提高冷冻-解冻后精子的受精能力。论文针对目前主要的一些抗氧化剂在家畜精液上应用研究的现状,对精子氧化损伤机理和常用抗氧化剂研究进行综述,期望对家畜精液冷冻保存的相关研究提供一定的理论依据和参考。     

Is sperm cryopreservation in absence of permeable cryoprotectants suitable for subfertile donkeys?

Sperm from fertile donkeys have been successfully frozen in absence of permeable cryoprotectants. The aim of this study was to determine whether this cryopreservation method is suitable for subfertile donkeys in comparison to conventional sperm freezing with glycerol. Ejaculates were collected from four Andalusian Donkeys: three fertile and one subfertile. Semen was frozen with an extender containing glycerol (GLY), or adding instead sucrose 0.25 molar and 1% bovine serum albumin (SUC) as non‐permeable cryoprotectants. After thawing, samples were assessed for total (TM, %) and progressive (PM, %) sperm motility by CASA, plasma membrane integrity (PMI, %) by epifluorescence microscopy and DNA integrity (DFI, %) by SCSA. Results (mean ± SD) were compared between extenders in fertile and subfertile donkeys using the Student's t test. No differences between GLY and SUC treatments were found in the fertile group for the sperm parameters assessed. In subfertile donkey ejaculates, GLY resulted in significantly higher values than SUC for TM (25.5 ± 3.1 vs. 19.6 ± 1.9) and PM (13.3 ± 5.1 vs. 4.0 ± 1.2), respectively. In conclusion, considering all the sperm parameters assessed, sperm freezing in absence of permeable cryoprotectants may not be still an option for cryopreservation of subfertile donkey sperm.