天山马鹿电刺激采精技术的研究

选用20头6～9岁的成年天山马鹿公鹿进行电刺激采精.采精前对公鹿采用鹿眠宝(2 mg/kg)、静松灵(3～5 mg/kg)、静松灵(2 mg/kg)+鹿眠宝(1 mg/kg)3种方法麻醉.结果表明:采用静松灵+鹿眠宝麻醉保定后,6头公鹿均可以正常采精,采出的精液品质可以用于人工授精.采用单独的静松灵麻醉保定后,6头公鹿电刺激采精除1头公鹿未能采下精液外,其它公鹿均能正常采精,其精液品质也可正常用于人工授精;单独采用鹿眠宝麻醉保定,7头公鹿电刺激仅3头能正常采精.     

亚洲黑熊电刺激采精及精液品质分析

采用电刺激法对 18头亚洲黑熊进行了 2 3次精液采集 ,并对所得精液进行了品质分析。亚洲黑熊每次射精量平均体积 (0 .2 5 9± 0 .185 )mL ,pH值 7.0± 0 .0 ,密度 (13.90± 8.19)× 10 8/mL ,活力 (75 .83± 14 .2 9) % ,运动状态 3.75±0 .2 7,畸形率 (2 3.0 9± 9.5 4 ) % ,顶体完整率 (80 .17± 13.2 6 ) %。盐酸氯氨酮与麻保静混合麻醉是一种适合黑熊电刺激采精的麻醉方法。     

抗冻保护剂的不同浓度对黑熊精液冷冻保存效果的研究

2001年6月至2002年6月，对吉林省延边白头山熊场的16只4－13岁的雄性黑熊进行了电刺激采精共72次，其中射精66次，占91.67％，并对精液进行了冷冻保存。黑熊的平均体重为225kg，射精量为0.3-5.0ml，精子活力为0.7-0.9。通过精液冷冻保存的预试，筛选出稀释液和抗冻保护剂，最后筛选出最佳抗冻保护剂的浓度。结果含4％甘油的稀释液对黑熊精液冷冻保存效果最好，冷冻后精子活力可达到0.4以上，已达到人工输精的要求指标。     

用低渗膨胀试验评价黑猩猩冷冻精子的膜完整性

精液品质评定的常见方法有精子活力、畸形率和顶体完整率等指标,但这些指标不能评定精子的真实受精能力。目前低渗膨胀试验作为检验精子膜功能的一种方法得到广泛认可,利用它可检验具有完整膜的精子是否具有生化活性。本试验通过对冷冻2只黑猩猩精液进行低渗膨胀来评定冷冻精子的膜完整性。试验结果显示,本研究中的黑猩猩的冷冻精子在低渗溶液中的膨胀率较高,表明试验黑猩猩的冷冻精液有较高的活力,精子具有较高的穿透率和受精能力。     

林麝电刺激采精及精液品质分析

采用电刺激采精法对 8头林麝采精 2 6次 ,并对所采得的精液进行了品质分析。结果表明 ,林麝每次射精量平均体积 0 3 5 6ml± 0 0 62ml,pH值 7 3 1± 0 2 74,精子平均密度为 ( 4 0 3 8± 0 62 4)× 10 8/ml ,精子平均活率为 78 75 %± 4 43 2 % ,畸形精子率为 14 5 7%± 2 0 3 1%。电刺激法采得林麝精液品质稳定 ,采精过程中用 846合剂麻醉保定 ,证明是一种适合林麝的人工采精方法     

鹌鹑采精方法与精液品质的研究

由于鹌鹑的精液量很少并且精子在体外存活的时间很短 ,仅有 (13± 4 )ｍｉｎ。鹌鹑采精方法的研究较少 ,笔者根据鹌鹑的生理特性摸索了一种鹌鹑按压式采精的方法 ,取得了较好的效果。同时由于未见有鹌鹑专用稀释液的资料 ,因此鉴于鸡与鹌鹑同属于鸟纲的不同属 ,既鸡属和鹌鹑属 ,而鸡的精液稀释液研究日趋完善 ,所以本试验根据鹌鹑精液生理生化指标 ,经查阅大量文献选用著名的鸡精液稀释液 ,ＢＰＳＥ液、Ｌａｋｅ液、ＢＶＰＥ液、5 .7%Ｇ液、5 .7%的葡萄糖液及 0 .9%氯化钠溶液 ,以期获得延长鹌鹑精液存活时间的缓冲稀释液。为此 ,于 2 0 0 2…     

电刺激采精器在荷斯坦种公牛上的应用

本文介绍了德国米尼图电刺激采精器(Electro-Ejakulator mit sonde 2,5’’)的基本原理、工作方式,以及利用电刺激采精器采集荷斯坦种公牛精液的情况,并与假阴道法采集该牛精液的结果进行比较。与假阴道法相比较,利用电刺激采精器法获得的精液量显著提高,但精子密度却显著降低;而在原精活力、冻精活力、冻精产量方面无显著差异。利用电刺激采精器对无法以假阴道法采集精液的公牛进行精液采集,取得了良好效果。     

三种采精方法在种公牛上的应用效果分析

本文针对大额牛和婆罗门牛 ,对电刺激法、按摩法和假阴道法三种方法的采精效果进行了比较研究。结果表明按摩法、假阴道法的采精量差异不大 ( P>0 .0 5) ,但均明显高于电刺激法 ( P<0 .0 5) ;而就精液密度而言 ,假阴道法明显高于电刺激法 ( P<0 .0 1 )和按摩法 ( P<0 .0 1 ) ,而后两者间无明显差异 ( P>0 .0 5) ;而就精液活率而言 ,三种方法无明显差异 ( P>0 .0 5) ;同时还进一步说明通过三种采精方法所获精液制作成的冷冻精液颗粒品质无明显差异 ( P>0 .0 5)。通过综合比较认为假阴道法是最好的采精方法 ,而电刺激法有着其特殊用途。     

大熊猫精液的研究

在1999年3月至2000年6月，对卧龙自然保护区大熊猫研究中心的9只5.5～16.5岁的雄性大熊猫进行了17次电刺激采精，9只大熊猫的平均体重为108.1±15kg，睾丸体积为289.4±108.6cm3，刺激电压2～8V，射精电压4～8V，平均每次采精量为3.5±1.91ml，精子密度为995.2±782.6×106个/ml，每次采精总量为3907±2981.7×106个，精液平均活力为69.1±28.9%，活率为79.6±23.1%，运动状态为2.7±1.1，畸形率为39.4±32.2%，平均顶体正常率为85.8±31.2%，pH值为7.9±0.4。其中3只是能进行自然交配并繁殖了后代的种公兽，其体重为122.7±7.1kg，睾丸体积为381.7±64.8cm3，精液活力为77.3±5.8%，运动状态为3.0±0.5，精子畸形率为18.0±9.6%，顶体正常率为92.7±1.5%，pH值为8.0±0.1。大熊猫异常精子主要有顶体异常，卷尾，中段弯曲含胞浆小滴，中段弯曲无胞浆小滴，尾弯曲含胞浆小滴，尾弯曲无胞浆小滴，近端含胞浆小滴，远端含胞浆小滴8种。在2月～6月都能通过电刺激采集到精液，在4月份每次产精量最高，这正好与在这期间发情和交配的大熊猫数量最多相吻合。     

种鸡人工授精技术要点

正常精液为乳白色浓稠液体。如果精液呈现黄色，则混有粪便；如果呈粉红色，则混有血液；如果有乳白色的棉絮状物，则含有尿酸盐。这些异常的精液都会严重影响种蛋的受精率，应当弃用。     

Semen collection techniques.

Semen collection techniques in the stallion have evolved considerably over the last 70 to 80 years and are used today primarily for artificial insemination. Semen can be collected from stallions that are otherwise unable to breed, allowing continued use of valuable animals. There are many options for collection of semen from stallions that present with ejaculatory dysfunction (see the article by McDonnell elsewhere in this issue.) Although there are many advantages to the use of artificial breeding, the collector must understand each step of the collection procedure as well as stallion preferences and proper use of an artificial vagina and mount source so that a representative semen sample is collected.     

鸵鸟采精方法研究

对 1 0只体况较好的黑颈公鸵鸟进行假阴道法采精试验 ,结果获得优良品质的精液 ,平均精液量为 0 94mL ,精子活率平均为 0 88。试验表明 ,在对公鸵鸟进行一定时间的调教之后 ,采用假阴道法采精是简单可行、具有实践意义的方法     

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。     

Fertilizing Ability of Electro-Ejaculated Cryopreserved Semen in the Domestic Cat

Semen collection and AI in the cat are still not routine procedures. The correlation between semen quality and fertility under natural conditions is a relatively unknown field in the cat. In the present study, functional in vitro tests, such as the ability to bind and penetrate the zona pellucida or to fertilize in vitro, were used to determine fertilizing ability of sperm cryopreserved with a practical and efficient freezing protocol previously developed in our laboratory. Semen was collected by electroejaculation, evaluated for motility and diluted with Tris-glucose-citrate egg-yolk extender supplemented with Equex STM paste (0.5% v/v). After equilibration and loading into 0.25 ml straws, semen was frozen at 3.85 degrees C/min. Frozen-thawed semen was co-cultured with in vitro matured cat oocytes. Penetration rate was recorded 30 h after in vitro fertilization and cleaved zygotes were cultured in vitro until day 7. A correlation was found between sperm motility index (SMI) after thawing and semen fertilizing ability (p<0.05). In conclusion, it was demonstrated that the post-thaw motility quality, expressed as SMI, of spermatozoa frozen using the protocol mentioned above can be considered an index of the sperm ability to penetrate in vitro matured oocytes.     

CASE STUDY: Use of Modified Phosphate-Buffered Saline as a Component of Semen Extenders Allows the Use of Transported Semen from Two Stallions

Conception rates for mares bred with transported-cooled and fresh stallion semen were collected over a 4-yr period (1998–2002) for two stallions. Both stallions stood at a commercial breeding farm. Semen from both stallions was used immediately after collection on the farm and after 24 to 48 h of cold storage when transported to locations in the U.S. and Canada. Semen for insemination of mares located on the farm was extended with a commercially available skim milk glucose extender (SKMG). Spermatozoal motility following cold storage for spermatozoa diluted in SKMG extender was unacceptable. Thus, semen from both stallions was centrifuged, and spermatozoa were resuspended in SKMG supplemented with modified PBS. In a previous study, the percentage of motile spermatozoa increased following centrifugation and reconstitution of the sperm pellet in SKMG-PBS as compared with semen dilution in SKMG (Stallion A: 15% vs 47%; Stallion B: 18% vs 43%). In the current study, 22 of 25 (88%) and 3 of 4 (75%) mares conceived with transported-cooled semen from Stallions A and B, respectively. Conception rates for mares inseminated with transported semen did not differ (P>0.05) from those inseminated on the farm with fresh semen. These data illustrate that stallion owners can modify standard cooled semen processing procedures and semen extender composition to improve post-storage spermatozoa motility and to obtain acceptable fertility.     

Genetic Parameters and Breeding Values for Semen Characteristics in Hanoverian Stallions

The objectives of this study were to show whether semen traits of 30 Hanoverian stallions regularly used in AI may be useful for breeding purposes. Semen characteristics were studied using 15 149 ejaculates from 30 Hanoverian stallions of the State Stud Celle of Lower Saxony. Semen samples were collected between 2005 and 2009. Traits analysed were gel‐free volume, sperm concentration, total and motile sperm number and progressive motility. A linear multivariate animal model was employed to estimate heritabilities and permanent environmental variances for stallions. The same model was used to predict breeding values for all traits simultaneously. Heritabilities were high for gel‐free volume (h² = 0.43) and moderate for total number of sperm (h² = 0.29) and progressive motility (h² = 0.20). Gel‐free volume, sperm concentration and total number of sperm were genetically negatively correlated with progressive motility. The effect of the permanent environment for stallions accounted for 9–55% of the trait variance. The total variance among stallions explained 37–69% of the trait variance. The average reliabilities of the breeding values were 0.43–0.76 for the 30 Hanoverian stallions. In conclusion, the study could demonstrate large effects of stallions, routinely employed in a breeding programme, on semen characteristics analysed here. We could demonstrate that estimated breeding values (EBV) with sufficient high reliabilities can be predicted using data from these stallions and these EBV are useful in horse breeding programmes to achieve genetic improvement in semen quality.     

Effects of Incubation Temperature and Semen Pooling on the Viability of Fresh,Chilled and Freeze‐Thawed Canine Semen Samples

This study assessed the effects of different incubation temperatures on semen viability and the influence of pooling on semen longevity. In experiment 1, semen samples were collected from five dogs, individually processed (individual semen: IS) and then aliquots from each male were pooled (pooled semen: PS). Semen samples (IS and PS) were diluted in a Tris‐glucose‐yolk extender and preserved as fresh (37 and 25°C) and chilled semen (4°C). Sperm motility and the percentages of sperm abnormalities and acrosome membrane integrity were assessed for 24 h. Storage at 25 or 4°C for the first 24 h yielded similar semen quality, but incubation at 37°C caused drastic reduction in sperm motility from 8 h of incubation onwards. In experiment 2, the semen was processed in the same way to that of experiment 1 and then preserved at 25 or 4°C until semen inactivation. Semen that was incubated at 25°C became completely inactive after 3–4 days of storage, while semen that was preserved at 4°C presented with more gradually decreased sperm motility (mean values of 40–60% for the first 8 days). In addition, the mixing of semen was only observed to influence the sperm quality of the samples stored at 4°C. In experiment 3, semen was collected from five dogs, pooled and frozen in liquid nitrogen; after thawing, it was preserved at 37, 25, 15 and 4°C, and the sperm quality was defined. The motility of the freeze‐thawed semen samples decreased quickly in the first 4 h after thawing, regardless of the preservation temperature of the thawed semen. This study confirmed that semen preserved at 37°C should be used within a maximum of 12 h, while the semen stored at 25°C shows acceptable quality for 24 h. Chilled semen presented highest most sustainable quality, especially when semen is processed as pooled semen.     

3种抗凋亡试剂在猪精液低温保存中的应用

The objective of the present study was to improve the low temperature preservation of boar semen and evaluate the effect of three anti-apoptotic drugs (puerarin, metoprolol tartaric and dichloroacetate) on keeping the quality of boar semen. Semen was collected from adult boars of proven fertility with the gloved-hand technique, and only the semen showing a minimum of 70% motile was used. Semen was diluted in extenders containing anti-apoptotic drugs (0,0.02,0.10,0.50 and 2.50 mmol/L puerarin, 0,0.02,0.10,0.50 and 2.50 mmol/L metoprolol tartaric acid, 0,0.04,0.20,1.00 and 5.00 mmol/L dichloroacetate). All sperm suspensions were stored at 5 ℃ and sperm motility was detected once a day. Six parameters (motility of extended soermatozoa,spermatozoal-survival effective time,spermatozoal-survival overall time, spermatozoal-survival index, the 72 h deformity rate and acrosomal integrity) were measured, six replicates were performed. The results showed that under the conditions of this study, a certain concentration of three drugs can extend the life-span of spermatozoa under the low temperature, and the optimal concentration of puerarin, metoprolol tartaric acid and dichloroacetate was 0.50, 0.50 and 1.00 mmol/L, respectively.     

中药王不留行对奶牛乳腺细胞增殖及泌乳的影响

为确定中药王不留行对奶牛乳腺细胞泌乳功能的影响,以王不留行为原料,通过水浸提得到王不留行提取液,测出王不留行提取液中增乳活性物质邻苯二甲酸二丁酯含量,设置不同浓度中药提取液进行细胞试验,观察其对细胞泌乳的影响,筛选出增乳效力最显著的药物浓度。结果2 mg/mL(生药浓度)药物组增乳效果最显著,提示中药王不留行可直接作用于奶牛乳腺上皮细胞促进泌乳。     

Stallion Semen Cooling Using Native Phosphocaseinate-based Extender and Sodium Caseinate Cholesterol-loaded Cyclodextrin-based Extender

The objective of this study was to compare semen parameters and embryo recovery rates of cooled stallion semen extended with INRA 96 or BotuSemen Gold. In experiment 1, 45 ejaculates from nine mature stallions were collected, assessed, and equally split between both extenders and then extended to 50 million sperm/mL. Then, the extended semen was stored in three passive cooling containers (Equitainer, Equine Express II, and BotuFlex) for 48 hours. In experiment 2, the same ejaculates extended in experiment 1 were cushion-centrifuged, the supernatant was discarded, and the pellets were resuspended at 100 million sperm/mL with their respective extender. Semen was then cooled and stored as in experiment 1. In both experiments, sperm motility parameters, plasma membrane integrity, and high mitochondrial membrane potential were assessed at 0, 24, and 48 hours post cooling. For experiment 3, 12 mares (n = 24 cycles) were bred with 48 hour–cooled semen from one stallion. Semen was processed as described in experiment 1. Mares had embryo flushing performed by 8-day post-ovulation. In experiment 1, BotuSemen Gold displayed superior total and progressive motility relative to INRA 96 (P < .05). There were no significant differences between the types of containers in any experiment. In experiment 2, INRA 96 and BotuSemen Gold extenders had similar total and progressive motility, but BotuSemen Gold had superior sperm velocity parameters at all timepoints. Embryo recovery was identical for both extenders (50%). Finally, the results obtained herein suggest that BotuSemen Gold is a suitable alternative to be included in semen cooling tests against INRA 96 in clinical practice.