东佛里生奶绵羊精液冷冻保存方法的研究

本研究旨在探讨东佛里生奶绵羊冷冻稀释液中添加维生素B₁₂以及不同冷冻保护剂对冷冻精液品质的影响，并筛选适合东佛里生奶绵羊的精液冷冻程序。通过假阴道法采集3只健康配种用东佛里生奶绵羊的精液，分为3组，分别为对照组(基础稀释液+6%甘油)、维生素B₁₂组(基础稀释液+6%甘油+0.05 mg/mL维生素B₁₂)、冷冻保护剂组(基础稀释液+5%乙二醇)。在-10～-100℃温区内设置3种降温速率(40、30、20℃/min)进行冷冻。结果显示：维生素B₁₂组冷冻精液的运动性能和畸形率较对照组没有显著变化；对照组冷冻精液的总活率、前向运动比例、平均路径速度、直线速度、曲线速度、直线性、鞭打频率、摆动性优于冷冻保护剂组(P<0.05)，畸形率低于冷冻保护剂组(P<0.05)。在冷冻程序的设计上，在-10～-100℃温区内以-40℃/min速率进行降温的冷冻程序A，其冷冻精液总活率、前向运动比例、平均路径速度、直线速度、曲线速度、直线性优于其他2个程序(P<0.05)，其畸形率最低(P<...     

稀释液中添加多不饱和脂肪酸(PUFA)对绵羊精液冷冻效果的影响

以葡3-3稀释液(葡萄糖-柠檬酸盐-卵黄)为基础,添加富含多不饱和脂肪酸(PUFA),且含较高比例DHA和EPA的深海鱼油作为冷冻绵羊精液的新型添加剂,进而研究PUFA对绵羊精子冷冻-解冻过程优良的保护作用.研究结果表明,绵羊精液冷冻稀释液中添加3%PUFA的处理组,同时添加1%VE(V/V)作为抗氧化剂,经冷冻-解冻对精子的损害较小,解冻后活率和顶体完整率分别比对照组1(0)显著提高6%和10%(P＜0.05),精清中LDH酶(乳酸脱氢酶)活力和GOT酶(谷-草转氨酶)活力比对照组1(0)显著降低65%和15%(P＜0.05).     

绵羊精液冷冻保存研究进展

绵羊精液保存技术目前虽然已进入生产应用阶段,但冷冻精液人工授精受胎率低仍是限制绵羊冷冻精液进行生产推广的重要因素。目前,对于绵羊精液冷冻的研究主要集中在冷冻稀释液组分和冷冻程序等方面。文章针对国内外有关绵羊精液冷冻保存稀释液、冷冻程序、解冻方法和冷冻损伤等研究进展作一综述,旨在对今后精液冷冻保存研究提供一定的参考。     

绵羊冷冻精液研究进展

随着肉羊产业化、工厂化的发展,冷冻精液人工授精技术亟待应用于生产中.绵羊精液冷冻保存技术虽然得到了快速发展,但目前仍处在试验研究或小范围的应用阶段.因此,绵羊精液冷冻技术还需要深入、系统的研究.文章从绵羊精液冷冻保存稀释液中的添加剂、冷冻速率、冷冻-解冻对绵羊冷冻精液品质的影响,精液输精对受胎率的影响等方面进行了综述,为精液冷冻保存研究提供一定参考.     

绵羊精液冷冻保存技术的研究进展

绵羊精液冷冻保存技术目前推广还比较困难,受胎率不高是其主要原因。目前,对于绵羊精液冷冻的研究主要集中在冷冻稀释液和冷冻程序两个方面,现就国内外有关精液冷冻稀释液和冷冻程序的研究进展做一概述,希望对今后有关此方面的研究具有一定的参考价值。     

小尾寒羊精液冷冻技术研究

本试验对11只2～3岁小尾羊种公羊的精液品质、保存稀释液和冻精制作技术进行了研究。结果表明:小尾寒羊种公羊的射精量、精液品质、精液pH值、密度、色泽、活率和畸形率均属正常范围,优于无角道塞特和莎福克绵羊;4种稀释液中,A液低温保存效果优于B、C、D 3种稀释液,差异极显著(P<0.01)。B液作为小尾寒羊冷冻精液稀释效果较理想(P<0.01)。6％甘油作为小尾寒羊冷冻精液保护剂效果较理想。     

小尾寒羊精液保存技术研究

为了充分发挥种公羊的生产潜力,快速扩繁优良群体,本实验以取材方便的小尾寒羊为实验动物,对其精液采用室温、低温和细管法冷冻3种保存方法进行了研究。结果表明:在低温(4±0.5)℃条件下稀释保存小尾寒羊精子,显著优于室温(23±2)℃条件下的保存效果。其中采用II液低温保存绵羊精子的时间和活率(168 h,0.35)显著优于III液(72 h,0.30)I、V液(48 h,0.35)和I液(18 h,0.42)。而采用III液冷冻保存绵羊精液,其解冻后活率(0.532±0.004)极显著高于IV液(0.470±0.003)I、液(0.445±0.004)和II液(0.432±0.012)3种冷冻稀释液(P<0.01)。4种冷冻稀释液冷冻解冻后的精子,均能在室温下6 h内保持0.35以上的活率。     

绵羊精液冷冻保存技术的研究进展

随着现代科学技术的发展,中国畜牧业的规模化和集约化程度越来越高,绵羊的人工授精技术也进入了应用阶段,从中可以提高优秀种羊的利用率和提高种羊配种效益,降低饲养的费用,提高繁殖的质量,但由于受胎率低的原因,现在处于研究阶段或小范围内使用。文章分析介绍了绵羊精液稀释液中的成分、稀释方法、冷冻温度、冷冻-解冻方法对绵羊精液品质的影响等,对绵羊精液冷冻保存技术提供参考。     

绵羊精液冷冻保存和人工输精技术的研究

本文综述了绵羊冷冻精液稀释液的种类、细管冷冻精液的分装、平衡、解冻方法和人工输精技术对受胎率的影响     

不同稀释液及绵羊品种对冷冻精液保存效果的影响

本文利用计算机精子辅助分析仪和姬姆萨染色方法评价无卵黄稀释液OPTIXcell和有卵黄稀释液Optidyl冷冻保存湖羊、白头杜泊、黑头杜泊和澳洲白绵羊精液的效果。选用湖羊、白头杜泊、黑头杜泊和澳洲白绵羊的种公羊各5只,通过假阴道方法采集精液,合格精液分别用OPTIXcell和Optidyl稀释液稀释并冷冻。结果表明:Optidyl冷冻保存白头杜泊羊和澳洲白绵羊精液的精子在活精子比例(MR)、直线速度(VSL)、平均路径速度(VAP)、曲线轨迹的直线性(LIN)、空间平均路径的直线性(STR)和尾部鞭打频率(BCF)上优于OPTIXcell,同样的,湖羊在MR、曲线速度(VCL)、精子头侧摆幅度(ALH)、VSL和VAP上高于OPTIXcell无卵黄稀释液,而黑头杜泊羊使用OPTIXcell在MR、VSL、VCL、ALH和BCF的平均值上效果更好。无论哪个品种使用Optidyl有卵黄稀释液,其绵羊精子畸形率的比例都低于使用OPTIXcell无卵黄稀释液。综上,冷冻保存湖羊、白头杜泊羊、澳洲白绵羊精液适合采用Optidyl,而黑头杜泊羊精液适合采用OPTIXcell。     

Dietary fish and evening primrose oil with vitamin E effects on semen variables in cockerels

Our aim was to determine the effect of n-3 (2%, wt/wt, fish oil rich diet) and n-6 (2%, wt/wt, evening primrose oil rich diet) fatty acid dietary supplementation and their combination with two concentrations of vitamin E (40 vs 200 mg/kg) on semen variables and on fatty acid and vitamin E profiles of spermatozoa in broiler breeders at 32, 42 and 52 weeks of age. The inclusion of fish oil in the cockerel diets increased the docosahexaenoic acid proportion in the sperm phospholipid fraction, which was almost threefold higher compared to the other two groups irrespective of vitamin E supplementation. In contrast, an increase in the proportion of total n-6 polyunsaturates, mainly 22:4n-6, was observed in the evening primrose oil group compared to the control only when the dietary content of vitamin E was increased to 200 mg/kg. Sperm concentration was decreased in the fish and evening primrose oil groups if vitamin E was 40 mg/kg, but such an effect was prevented in the fish, not the evening primrose oil group, by increasing the vitamin E to 200 mg. The proportion of motile spermatozoa was improved by the increased supplementation of vitamin E in all oil treatments.     

提高绵、山羊颗粒冻精品质的研究

用波德代羊和无角陶赛特羊品质良好的鲜精，研制出9个绵羊精液冷冻稀释液配方，同时对稀释、平衡、解冻、授配等方法和步骤进行相庆优化试验研究，获得冻精解冻后活力在0．45以上，冷冻保存409天的冻精解冻后活力在0．5以上，组织300只当地绵羊授配试验，30天的情期不返情率为65．57％。研制出波尔山羊（Boer)冻精的优化的稀释液配方和优化的冷冻工艺，授配试验结果表明母羊情期不返情率为65．77％。     

Evidence of excretion of Schmallenberg virus in bull semen

Schmallenberg virus (SBV) is a novel orthobunyavirus, discovered in Germany in late 2011. It mainly infects cattle, sheep and goats and could lead to congenital infection, causing abortion and fetal abnormalities. SBV is transmitted by biting midges from the Culicoides genus and there is no evidence that natural infection occurs directly between ruminants. Here, we could detect SBV RNA in infected bull semen using qRT-PCR (three bulls out of seven tested positive; 29 positive semen batches out of 136). We also found that highly positive semen batches from SBV infected bulls can provoke an acute infection in IFNAR^-/- mice, suggesting the potential presence of infectious virus in the semen of SBV infected bulls.     

不同稀释液对陶赛特肉羊细管冻精品质的影响

利用五种稀释液制作的细管冻精,通过精子项体完整率、畸形率和存活时间等指标的对比观察,筛选出最佳细管冻精稀释液。     

Development of a PCR to diagnose BLV genome in frozen semen samples

The sanitary and economic impact of BLV infection is associated with the interference in the international movement of cattle and their germ plasm. Although experimental data support the improbability that semen from BLV-positive bulls could infect recipient cows, restriction for commercialization of semen from infected animals is still present. The objective of this work was to standardize a PCR assay to diagnose the presence of BLV genome in frozen semen samples. The developed methodology involves the amplification of an internal fragment of gag gene. The limit of detection of this technique was six viral particles, using gag-PCR followed by hybridization analysis. Frozen semen samples from seropositive bulls were analyzed. It was possible to detect proviral DNA in 9 out of 173 samples. Additionally, a biological test in susceptible sheep was performed in order to evaluate the transmission of BLV genome by semen from seropositive animals. This data strongly suggest that semen from seropositive bulls that resulted negative by PCR can be used for artificial insemination (AI), accompanied by proper collection protocols. The development of this PCR assay constitutes a valuable diagnostic tool to determine the BLV-free status of frozen semen samples used for AI.     

Bluetongue virus in bovine semen: viral isolation

Vero cell cultures and embryonating chicken eggs were used for direct isolation of bluetongue virus from cattle blood and from semen samples. Cell culture and embryonating chicken eggs each were more effective than was the blood autograft inoculation of susceptible sheep with selected blood and semen samples. Evaluation of the cell culture technique indicated that the quality of the distilled water was the primary factor responsible for the increased sensitivity of the Vero cell cultures for the present blue-tongue viral isolations. Test results showed that urine was a poor specimen for viral isolation when assayed in chicken eggs. A comparison of tests for precipitating and complement-fixing antibodies to bluetongue virus indicated that the precipitin test was the more accurate of the two tests.     

动物油脂中DNA的提取及牛、羊源性成分的PCR检测

用异硫氰酸胍法提取鱼油、加入牛肉DNA的鱼油、加入山羊肉DNA的互油和牛油中的DNA。18S rDNA片段以及牛、羊源性成分的PCR扩增结果表明，建立的动物油脂DNA提取方法是可行的。用建立的DNA提取方法提取3份送检鱼油的DNA，牛、羊源性成分的PCR检测结果均为阴性。     

Effect of Docosahexaenoic Acid on Quality of Cryopreserved Boar Semen in Different Breeds

During the cryopreservation process, the level of polyunsaturated fatty acids, especially docosahexaenoic acid (DHA), in the sperm plasma membrane decreases significantly because of lipid peroxidation, which may contribute to sperm loss quality (i.e. fertility) of frozen–thawed semen. The aim of this study was to investigate the effect of supplementation of DHA (fish oil) in freezing extender II on frozen–thawed semen quality. Semen from 20 boars of proven motility and morphology, were used in this study. Boar semen was split into four groups, in which the lactose–egg yolk (LEY) extender used to resuspend the centrifuged sperm pellet was supplemented with various levels of fish oil to reach DHA level of 1X (group I, control, no added fish oil), 6X (group II), 12X (group III) and 18X (group IV). Semen solutions were frozen by using a controlled rate freezer. After cryopreservation, frozen semen was thawed and evaluated for progressive motility, viability by using SYBR‐14/Ethidiumhomodimer‐1 (EthD‐1) staining and acrosome integrity by using FITC‐PNA/EthD‐1 staining. There was a significantly higher (p < 0.001) percentage of progressive motility, viability and acrosome integrity in DHA (fish oil) supplemented groups than control group. Generally, there seemed to be a dose‐dependent effect of DHA, with the highest percentage of progressive motility, viability and acrosome integrity in group‐III. In conclusion, supplementation of the LEY extender with DHA by adding fish oil was effective for freezing boar semen as it resulted in higher post‐thaw plasma membrane integrity and progressive motility.     

DMF effects on frozen gander semen

1. The objective of the present study was to determine if the age of semen donors affects the susceptibility of spermatozoa to freezing and whether DMF (dimethyl formamide) inseminated with freeze-thawed gander semen decreases fertility. 2. Semen was collected 3 times a week by dorsal-abdominal massage from two groups of White Italian ganders: 3 and 2 years-old. Both samples were diluted, mixed with DMF to a final concentration of 6% (v/v), pre-frozen and transferred into LN2. 3. Twice a week, the freeze-thawed semen was used for insemination of two groups of geese at a dose of 4 to 16 million live morphologically normal spermatozoa. One group was inseminated immediately after thawing, the 2nd with semen from which the DMF was removed. 4. Donor age had no effect on the spermatozoa's aptitude for freezing. The differences in quality and quantity of fresh and freeze-thawed semen produced by 3 or 2 year-old ganders were not significant. 5. The presence of DMF in the inseminated freeze-thawed semen did not affect the reproductive efficiency of spermatozoa. The fertility rate obtained with semen inseminated either with or without the cryoprotectant averaged 92.9% and 87.2% respectively. The hatchability of set eggs was 81.1% and 79.9% and, the hatchability of fertile eggs amounted to 87.3% and 89.4%.     

Cryopreservation of gander semen

1. The effect of dimethylacetamide (DMA) and dimethyl sulphoxide (DMSO) on the cryopreservation of gander semen were investigated. An improved survival rate of spermatozoa after freeze-thawing was obtained when semen was frozen by a fast-freezing procedure on dry ice with 9% DMA as the cryoprotectant. 2. Gander semen, which was frozen during mid season, was tested for fertilising ability in different times of the season. The percentage of fertility during d 3 to d 9 after 2 consecutive inseminations was 68% to 95%, depending on the date of artificial insemination.