关于牛冻精质量检验中的若干技术问题

关于牛冻精质量检验中的若干技术问题农业部牛冻精质量监测中心（南京２１００９５）金穗华牛的冷冻精液作为一种产品在畜牧业生产上广泛应用，因此产品质量必然要受到社会的监督以维护养牛业的正常发展。在国家发布的牛冷冻精液标准ＧＢ４１４３—８４中，对该产品质量有...     

谈牛冷冻精液质量检测技术问题

谈牛冷冻精液质量检测技术问题○黑龙江省家畜冷冻精液监测中心刘彩兰郝爱玲○黑龙江省家畜繁育指导站（哈尔滨１５００６０）施维民李晓东为保证牛冷冻精液（下称冻精）出厂产品的质量，除国家监测中心对各地冻精质量作定期抽检以外，各冻精生产单位和地方监测单位都要经...     

影响种公牛冻精生产若干因素的探讨

影响种公牛冻精生产若干因素的探讨王书芳，耿方阁（吉林省白城市家畜冷冻精液站１３７０００）随着应用牛冷冻精液进行人工输精技术的推广普及，种公牛精液数量的多少和质量的高低，对冻精生产及供应冷冻精液影响颇大。在实践中掌握和运用种公牛精液生产规律，充分发挥其...     

利用监控办法不断提高牛冻精产品质量

为适应现代化畜牧业生产发展的需要,更好地执行《牛冷冻精液国家标准》,建立了黑龙江省家畜冷冻精液监测中心。该中心主要任务是按国家标准监测全省家畜精液质量。中心成立以来,在监测全省冻精生产基础上,重点监测了黑龙江省家畜繁育指导站生产的牛冷冻精液产品。因为该站是我国生产牛冷冻精液主要大站     

对牛冷冻精液微生物检测培养基种类培养时间的探讨及控制细菌数的措施

牛冷冻精液在采集、稀释、分装及冷冻等生产过程中都有可能被污染，并且有大量资料表明许多细菌性及病毒性传染病都可能通过精液而传播，因此，对牛冷冻精液的微生物检查是冻精质量检测的重要指标之一。我国《牛冷冻精液国家标准》中明确规定：牛冷冻精液解冻后的精液，无...     

浅析影响牛冷冻精液质量的几个重要因素

<正>随着冷冻精液在我国养牛业生产中的广泛推广应用,牛冷冻精液质量显得至关重要。影响种公牛精液质量的因素很多,如营养、年龄、品种、季节、卫生状况、温度等。本文简要介绍了影响牛冷冻精液质量的几个重要因素,以期其对生产优质牛冷冻精液起到指导作用。1公牛个体因素1.1年龄种公牛的造精功能在17~42月龄最为旺盛,以后缓慢下降。2岁左右的种公牛精子耐冻性不稳     

种公牛采精过程中的操作细节

在牛的冻精制作生产上,精液的活力和密度是其重要指标,公牛精液的采集是第一个重要步骤,要提高冷冻精液活力和质量,采精过程中的每一个细节都要一丝不苟,严格把关.     

牛细管精液冷冻温度曲线与冻精质量的研究

牛冷冻精液质量是否优良与鲜精品质、稀释液种类、稀释方法、冷冻方法和解冻温度及速度等因素有关,而冷冻温度是影响冻精质量的重要因素之一。本文就冷冻时不同细管支数而引起冷冻温度变化对冻精质量的影响及其有关冷冻机理进行了研究,以利于提高细管冻精生产的质量和数量。     

高原低气压条件下不同冷冻温度对牛精液冻后活力的影响

通过评定不同冷冻温度条件下生产的公牛细管冻精活力,来确定高原低气压条件下较为理想的牛细管冷冻精液生产的冷冻温度,生产优质细管冻精。     

目前牛冻精的质量现状及要注意的技术问题

<正> 牛的冻精技术从七十年代起国内发展很快,对提高我国牛群质量起到了积极的作用;特别是1984年颁布了《牛冷冻精液国家标准》后对于冻精质量把关提供了依据。我国牛的冻精质量与先进国家相比还有距离,1986年在南京举行的全国公牛站冷冻精液质量评比的结果并对照国家标准如下表。 看来冻精质量并不十分理想,而各站间的差异较大,据近年的一些动态和调查,结合1987年在南京举办的《全国牛冷冻精液质量检定班》中总结归纳,目前一些主要影响冻精质量的因素有以下几个方     

牛冷冻精液中前进运动精子数的研究

为提高我国种公牛的利用率提供理论基础，对牛冷冻精液相关标准的制、修订给出可靠的参考数据，本研究对国内近20年来牛冷冻精液的主要质量性状——每剂量中前进运动精子数进行了测定分析。结果表明，每剂量前进运动精子数平均为1342万个，其中最高的为3072万个，最低的为800万个；各年度间比较没有发现明显的变化规律，但是品种间（荷斯坦、西门塔尔、牦牛和水牛）差异显著（P〈0．05、P〈0．01）；经进一步分析测算，在基本不增加生产成本的情况下，目前牛冷冻精液的产量至少可以增加50％以上。     

两种氨酸对公牛精液冷冻效果研究

[目的]试验研究L—半胱氨酸、精氨酸对牛精液冷冻效果。[方法]基础液中分别添加不同浓度的L—半胱氨酸、精氨酸进行试验,再用L₉(3₄⁾正交表优化最佳配比,获得不同的稀释液配方,然后再进行对比试验,优选出使用效果最好的牛精液冷冻稀释液。[结果]牛精液冷冻稀释液添加0.125%、0.25%、0.375%的半胱氨酸;牛精液冷冻稀释液添加0.5%、0.75%精氨酸,牛冷冻精液解冻活力均得到提高。[结论] 结果表明,半胱氨酸、精氨酸可用于牛精液冷冻保存,能改善牛精液冷冻保存效果,对提高牛冷冻精液质量效果明显。     

HSP90表达与牛精子抗冻性研究进展

 热应激蛋白(HSPs)是生物体抵抗温度升高、氧化刺激和缺氧等环境变化时广泛合成的一组蛋白。论文简述了冷冻-解冻后精子蛋白质含量变化的研究进展以及热应激蛋白HSP90表达量对牛冷冻精液质量影响的机理,为提高牛精液冷冻品质提供理论参考。     

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。     

亚硒酸钠维生素E对牛冷冻精液品质的影响

本试验研究了亚硒酸钠维生素E对牛冷冻精液品质的影响。以常规Tris卵黄冷冻稀释液中分别添加2.5μL、5μL、7.5μL、10μL稀释后的亚硒酸钠维生素E(注射液)为试验组,以常规Tris卵黄冷冻稀释液为对照组,对解冻后的精液品质进行测定。结果显示添加5μL、7.5μL、10μL亚硒酸钠维生素E能够显著降低冷冻精液的精子畸形率,且10μL组顶体脱落的精子数量显著降低,精子顶体完整率上升。小剂量(<10μL)组改善了精子存活时间,使精子在体外37℃的存活时间延长至8.5h。     

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.     

Post-thaw viability of european bison (Bison bonasus) semen frozen with extenders containing egg yolk or lipids of plant origin and examined with a heterologous in vitro fertilization assay.

Basic characteristics of European bison (Bison bonasus) semen were described and the efficacies of two extenders-Triladyl, containing egg yolk, and a synthetic extender, containing soybean lipids-were tested for semen cryopreservation. Seven ejaculates were collected by electroejaculation from a 10-yr-old, European bison bull. Each ejaculate was diluted at 37 degrees C to a final concentration of 200 x 10(6) sperm/ml with Triladyl or the synthetic extender. Extended semen samples were frozen according to a standard bull semen freezing protocol. After 2 wk of storage, one straw from each extender and ejaculate was thawed, and postthaw quality was evaluated by individual sperm motility and movement rate, numbers of sperm morphologic abnormalities and intact acrosomes, functional integrity of the sperm membranes determined by hypoosmotic swelling test (HOST), viability (live-dead, eosin-nigrosin stain), and a heterologous in vitro sperm penetration assay (SPA). A total of 600 in vitro-matured bovine oocytes were inseminated with 1 X 10(6) spermatozoa of Holstein semen frozen-thawed in Triladyl (control) or of European bison semen frozen in Triladyl or the synthetic extender. Nuclear status of the oocytes was determined after 18 h of sperm-oocyte coincubation. Extender had no effect on any evaluated parameters of semen after dilution and cooling (4 hr at 5 degrees C) or in postthaw individual motility, quality of movement, and sperm morphology. However, significantly (P < 0.05) higher numbers of spermatozoa with intact acrosomes, intact membranes (HOST), and viable sperm (P < 0.01) were in semen frozen in Triladyl than in the synthetic extender. Mean values for heterologous SPA for bull (control) and for bison semen frozen in the synthetic extender were very much alike-63.3+/-10.6% and 63.1 +/- 15.9%, respectively; bison semen frozen in Triladyl was lower, 43.0+/-24.2% but not significantly different. Cumulative results from a variety of viability assays of diluted/cooled and frozen-thawed semen, including the heterologous SPA, suggest that European bison semen can be successfully frozen in both extenders tested in this study.     

不同季节和气候条件对种公牛精液品质的影响的研究

[目的]研究不同季节气候条件对种公牛精液品质的影响。[方法]从采精种公牛中随机抽出5头,选取气温最高（7月份）最低（1月份）和适中温度（4,9月份）进行试验,分别对不同时期所采得的精液进行检测。[结果]在最高温度月份7月份（25℃）时5头种公牛的平均精液量为5.88mL,原精活率67.2%,精液密度16.64亿/mL,冻后活率33.8%,生产冻精数143.4剂,精子畸形率23%。1月份环境温度平均在-11.2℃时,5头种公牛的精液量平均为5.7mL,原精活率66.8%,精液密度13.3亿/mL,冻后活率37.4%,生产冻精数为275.8剂,精子畸形率17.8%。在4,9月份环境温度12℃时精液量为6.6mL,原精活率73.2%,精液密度16.92亿/mL,冻后活率41.8%,生产冻精数404.4剂,精子畸形率12.6%。精液量之间差异显著（P〈0.05）,原精活率之间差异显著（P〈0.05）,精子畸形率之间差异显著（P〈0.05）。冻后活率和冻精生产数差异不显著。[结论]环境温度过高或者过低都会影响精液品质,种公牛在适宜的温度下生产的精液品质也会更加优良,种公牛在温度较高的条件下生产精液品质比在低温条件下还要差,种公牛最适宜的生产温度为12℃～16℃。     

水牛性控精液微生物分析

研究针对目前分离水牛X和Y精子性别控制过程中,容易导致微生物快速繁殖进而污染性控精液的问题,对水牛新鲜及冷冻性控精液中的微生物进行分析研究,以期改进水牛分离精子保存方法。结果表明:新鲜采集的水牛精液、常规冷冻精液及分离后的冷冻精液中,平均菌落密度分别为10 619个/mL、293个/mL和18935个/mL,三者差异显著(P0.05)。新鲜精液检测到的菌落中,革兰氏阳性球菌占48%,阴性杆菌占41%,阳性杆菌占9%,霉菌占2%;性控冷冻精液中的菌落100%为革兰氏阴性杆菌。对性控冻精菌落药敏检测发现,菌落对妥布霉素和庆大霉素均呈现耐药性,而对丁胺卡那霉素以及药物环丙氟哌酸药物敏感性较强。本研究结果,为改进水牛分离精子稀释液以及冷冻保存液中的抗生素使用提供了有价值的参考。     

Inactivation of bovine herpesvirus 1 in semen using a hyperimmune egg yolk semen extender

Hyperimmune egg yolk semen extender was used for the inactivation of bovine herpesvirus (BHV-1) in experimentally infected bovine semen. As much as 5 x 10(4) TCID50/ml of virus was inactivated in semen as assayed by tissue culture. Moreover the hyperimmune egg yolk semen extender did not produce any adverse effect on the quality of the semen after being frozen/thawed in comparison with normal egg yolk semen extender (P > 0.05). The hyperimmune egg yolk semen extender is considered an important tool for containing the spread of BHV-1 from infected semen.