北极狐、乌苏里貉精液细管冷冻技术研究

为了对濒危珍稀动物种质资源的进行保护,完善精液冷冻技术方案,对北极狐37只次、乌苏里貉32只次的精液进行细管冷冻技术研究,结果表明,北极狐精液用3%柠檬酸三钠—卵黄—甘油作冷冻稀释液,乌苏里貉精液用11%蔗糖—卵黄—甘油作冷冻稀释液,在5℃冰箱内平衡1 h后,进行细管冷冻(液氮-196℃),冻溶粒子活力达4~5级。速冻法优于缓冻法。细管冻精在50℃温水中快速溶化,北极狐精液用3%柠檬酸三钠作解冻液,乌苏里貉精液用7%葡萄糖液作解冻液效果较佳。     

冷冻精液的温度控制

冷冻精液温度控制是保证冷冻精液质量和提高母畜受胎率的技术关键。在冷冻精液生产过程中,对精液稀释、平衡温度有着严格的要求;在精液冷冻过程中必须快速通过危险温区和按照规定的精液冷冻曲线操作;在使用冷冻精液时,对冷冻精液的保存温度和解冻温度及解冻后精液温度均有特定的要求。     

BMY牛冷冻精液质量控制及影响因素研究

[目的]为了加大BMY牛的推广应用力度和提高人工授精受胎率,开展了BMY牛冷冻精液质量控制研究,优选出冷冻稀释液及得出冷冻精液质量控制技术程序。[方法]采用不同的初始冷冻温度、不同种类的稀释液、不同的冷冻方法对BMY牛精液冷冻及解冻效果进行试验。[结果]不同的初始冷冻温度、不同种类的稀释液、不同的冷冻方法,BMY牛精液冷冻解冻效果各异。初始冷冻温度为-121--140℃时,TRIS液为基础稀释液添加糖类和氨基酸一步法稀释BMY牛精液冷冻6-10 min BMY牛冷冻精液解冻效果较好,用程控冷冻仪冷冻BMY牛精液,同样获得良好的解冻效果,解冻活力达到0.372±0.026。BMY牛冷冻精液平均解冻活力可达0.357±0.029以上,精子解冻复苏率达50%以上。精子畸形率平均为16.8%±4.26%,BMY牛采精成功率为75.3%。[结论]全放牧条件下BMY牛电刺激采精法获得很好效果,精液质量好,更有利于种质资源的保存和利用。     

马冷冻精液受胎率效果调查

<正> 1984—1989年,我站在全胜多7个村、20个自然屯推广了马冷冻精液输精技术,取得了显著效果,近3年马匹冷冻精液输精受胎率已稳定达到鲜精水平。为提高全县马匹冷冻精液输精效果,我们在进一步总结该乡6年来马冷冻精液输精效果的同时,着重对影响马冷冻精液输精受胎效果的情期、冷冻精液解冻后活力和输精时间、输精月份和输精次数、解冻液、解冻方式等有关因素进行了分析调查。     

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。     

不同解冻方案对牛冷冻精液质量的影响

解冻是冷冻精子复苏的关键步骤,解冻方案的选择则是冷冻精子复苏的关键点。为筛选出适合目前我国规模化牧场冷冻精液的解冻方案,本研究分别采用6种解冻方案对冷冻精液进行解冻并观察其解冻活率。结果表明,随着解冻温度的升高和解冻时间的缩短,精子活率呈上升趋势,并且当解冻温度达到40℃,解冻时间为20s时,冷冻精子解冻后的活率达到最高。关于6种解冻方案对精液存活时间的影响研究结果表明,当采用38~39℃解冻30s方案时,精子存活6h的活率最好。     

牛精子冷冻损伤研究进展

人工授精技术是目前通过冷冻精液进行牛遗传改良而应用最为广泛的生物技术。虽然如此,由于精液产品的质量而导致人工授精的遗传影响被限制,在精液的冷冻－解冻过程中40%~50%的活精子失去完整性或功能受到损伤。作者在参阅国内外相关文献的基础上,通过阐述冷冻－解冻对精子细胞的损伤作用及机理、精浆对牛冷冻精液质量的影响、冷冻－解冻过程中精子的过氧化损伤,以及冷冻－解冻后精子相关基因与蛋白质的变化等,以期为牛高质量冷冻精液的研究提供一定参考,进一步提升冷冻精液质量。     

冷冻精液解冻后精子质量评估研究进展

随着人工授精技术的广泛应用,精液冷冻保存技术也迅速发展起来。冷冻精液解冻后精子的质量会直接影响其受精能力。本文综述了近年来国内外有关冷冻精液解冻后精子质量评估的传统方法及新方法的研究进展。     

猪冷冻精液的解冻及人工授精技术

在为猪场节约生产成本、提高猪种质量上,猪冷冻精液能够起到良好的作用。然而,由于冷冻精液具有不同于常温精液的特性,在用于人工授精前对其进行解冻、稀释等准备工作也有严格的要求。本文详细阐述了冷冻精液的准备工作,并着重介绍了解冻、稀释等关键环节的操作技术和步骤。     

猪冷冻精液与常温精液配种对比试验报告

为了探讨猪冷冻精液在现代养猪业的应用,以甘州区金园种猪场为试点,利用百钧达科技发展有限责任公司猪冷冻精液配套技术,分别对母猪的分娩率、总产仔数、产健仔数以及仔猪的初生重和21日龄断奶重等进行测定,探讨影响冷冻精液解冻和配种效果的因素,找出最佳解冻和配种方案。为生猪冷冻精液的应用提供科学依据。冷冻精液配种技术在养殖业是一项可行的先进技术,可在母猪繁殖场和种猪场推广使用。常温精液可在广大养殖户推广使用。     

冷冻速率和解冻温度对猪精液冷冻效果的影响

为优化冷冻和解冻方法,提高冷冻效果,本试验比较了不同冷冻速率(-100 ℃ 10 min、-120 ℃ 10 min、-140 ℃ 10 min)和不同解冻温度(37 ℃ 30 s、45 ℃ 30 s、52 ℃ 30 s、60 ℃ 30 s)对猪精液冷冻效果的影响。结果表明,采用-120 ℃熏蒸10 min,解冻后精子活力为0.36,质膜完整率和顶体完整率也优于其他2组,且差异显著(P<0.05)。采用37 ℃ 30 s方法解冻,精子活力、质膜完整率显著高于其他3组,顶体完整率也高于其他3组,但差异不显著(P>0.05),其畸形率最低和60 ℃ 30 s组差异明显(P<0.05),但与45 ℃ 30 s组和52 ℃ 30 s组差异不显著(P>0.05)。因此,采用-120 ℃平衡10 min冷冻,37 ℃ 30 s水浴解冻方法更为适合0.25 mL细管猪冻精解冻。     

Preservation of fowl semen in liquid nitrogen--an improved method

1. An improved method of storing fowl semen in liquid nitrogen is described. It uses glycerol as a cryopotectant and the degree of fertility obtained with semen stored for up to 2 1/4 years was very promising. 2. The essential features of the method are a slow freezing rate, a reduction of temperature to - 35 degrees C before the semen is plunged into liquid nitrogen and insemination within 15 min of thawing and removing the glycerol from the diluent. 3. It seems likely that further improvements in the technique may be expected through better control of the injection of liquid nitrogen into the freezing chamber, thereby minimising localised, rapid temperature changes around the ampoules during freezing. 4. Preliminary results suggest that it may prove possible to select males for the ability of their spermatozoa to withstand freezing. 5. The importance of recognising differences between males in the initial quality of semen and between females in the physiology of the reproductive organs in determining the fertilising ability of stored semen is discussed.     

家禽精液冷冻保存技术研究进展

家鸡精液冷冻保护是物种资源保存最廉价的方法。本文从家鸡精液的稀释液、冷冻保护剂、冻精形式、平衡时间、冷冻速率、解冻速率、精液质量检测方法等几个方面,综述了国内外研究进展,并对目前家鸡精液冷冻技术应用存在的问题进行了分析,提出了初步研究思路。     

提高牛人工授精覆盖面的试验研究

目的,为了使远离牛冻配点区域的能翻母牛及时配种,发挥良种牛生产性能,提高养牛经济效益,增加农户收入。方法,通过离冻配点不同路程、不同发情时间及冻精解冻温度进行统计分析。结果,结果表明,对远离冻配点区域的能繁母牛,在发情结束后4-6小时,解冻温度为35-37C°,子宫角深部输精是完全可行的。     

Myeloperoxidase in Equine Semen: Concentration and Localization during Freezing Processing

Myeloperoxidase (MPO) is a pro-oxidant enzyme contained in and released by neutrophils during degranulation or after lysis. Post-thaw semen contains MPO, and concentration of this enzyme is associated with decreased motility. The aim of this study was to determine kinetics of MPO concentration during freezing, its origin, and its impact on frozen-thawed semen. Forty ejaculates were used. Semen was frozen using the classical freezing procedure. MPO concentrations were assayed in fresh semen, after centrifugation, and after cooling down to 4°C. Post-thaw MPO assay results and spermogram characteristics were determined. MPO immunocytochemistry was performed on 4 different ejaculates at each step of freezing procedure. MPO concentration increased after cooling down to 4°C and thawing compared with fresh semen. As temperature decreased, MPO was higher or tended to be higher in post-thaw poor quality samples. Nonsperm cells showed various degrees of MPO immunostaining and were observed as epithelial cells with nuclear pyknosis and keratinization. MPO immunostaining increased in medium and decreased in nonsperm cells during freezing. Our study shows that MPO concentration in equine semen increases when temperature decreases. We hypothesize that nonsperm cells present in fresh semen could release MPO.     

Effects of liquid nitrogen vapor sensitization conditions on the quality of frozen-thawed dog spermatozoa

The freezing conditions for preparation of frozen canine semen by the plunging method were investigated with regard to the period of sensitization in liquid nitrogen (LN2) vapor and the height from LN2, and the semen qualities after thawing were compared with those of canine semen prepared by the simple freezer method previously reported by us. In the plunging method, 9 semen straws were prepared under the same conditions, horizontally kept at 5, 7, and 10 cm above the LN2 surface in a styrene foam box for 5, 10, and 15 min, and then plunged into LN2. The semen qualities immediately after thawing were high in the 7 cm/10 min (cooling rate: -4 to -22 degrees C/min) and 10 cm/15 min groups (cooling rate: -6 to -10 degrees C/min). On comparison of frozen semen prepared by the plunging method (7 cm/10 min) with frozen semen prepared by the simple freezer method, sperm motility and viability were significantly higher for the frozen semen prepared by the plunging method. The cooling rate in freezing was higher for the simple freezer method (cooling rate: -6 to -50.9 degrees C/min) than the plunging method. Based on these findings, horizontal placement of canine semen straws above LN2 to reduce the temperature at a slow cooling rate of about -10 degrees C/min, followed by plunging into LN2 after sensitization for 10-15 min, provides good semen qualities after thawing.     

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。     

黑猩猩精液冷冻和优选研究

对精液进行有效的冷冻保存和解冻优选是人工授精顺利开展的重要步骤。本研究在黑猩猩上做了尝试,分别用人工按摩采精、电刺激采精、附睾采精进行6次精液采集,所测精子活率为0.65～0.90,密度为1.69×10~（10）～6.64×10~（11）（个/L）。用自配精液冷冻液,采用CL-8800程序降温仪成功对所采集的精液进行冷冻保存。在冷冻后的10～400 d,分别对6次冻精解冻,并用人用精液优选试剂盒对其后3次进行解冻后优选。结果显示,解冻后精液活率为0.35～0.7;优选后精液活率为0.85～0.9,密度为4.5×10~（11）～5.0×10~（11）（个/L）,基本可以满足人工授精的需要。     

猪精液冷冻技术试验

从稀释液和解冻液的筛选，平衡时间、平衡温度、冷冻剂型的选择等方面进行试验。试验结果：猪精液冷冻后活率为３０％－６０％；解冻后的复苏率为５１．４２％－５８．７２％；保存３０天的冻精与保存３６０天的冻精受胎率无明显差异（Ｐ＞０．０１）；冻精保存期一年以上；输精受胎率达６３．６４％（６０％－６６．６６％），平均窝产仔６．１头。