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     

黄牛细管冷冻精液配种技术在基层推广中的应用

黄牛细管冷冻精液配种技术是继黄牛颗粒冻精之后牛冻改工作的一大进步,细管冻精是用直径小、管壁薄的无毒塑料管盛装精液后冷冻而成,由法式凯苏输精枪输精,与颗粒冻精比较,黄牛细管冷冻精液在冷冻时受冻均匀、解冻时受热一致、解冻方便、输精时不易污染、避免精液和液氮的直接接触、精子活力好,极大提高了黄牛冻精改良的受胎率,扩大了黄牛改良的覆盖面,可根据群众要求上门服务,比较方便基层养殖户。     

应用细管冷冻精液二次输精对提高母牛受胎率的效果

如何提高母牛的受胎率,是多年来畜牧兽医人员研究和探讨的一个问题。在近几年的黄牛冷配实践中,我们感觉到,把握授精时机和输精方法是问题的关键。由于不少配种员掌握不好发情鉴定,在一个情期内配种3—4次,这样不仅受胎率低,而且提高成本,并易造成子     

牛细管冷冻精液异地输精对受胎率的影响分析

本文对2011年青海省循化县积石镇采用细管冷冻精液异地人工输精的262头母牛进行了跟踪调查统计。结果表明,牛受胎数226头,使用细管冷冻精液304支,受胎率为86.26%。     

牛细管冷冻精液异地输精对受胎率的影响分析

2011年在循化县积石镇采用细管冷冻精液对当地262头母牛全部采用异地人工输精跟踪调查统计。结果表明：牛受胎数226头,空胎母牛36头,使用细管冷冻精液304支,受胎率达86．26％。其中积石镇受胎率86．71％,街子镇受胎率85．29％,查汗都斯乡受胎率85％,清水乡受胎率86．96％,文都乡受胎率85．71％。     

牛细管冷冻精液的使用技术

细管冷冻精液是指将经过技术处理后的精液盛装于塑料细管内经过超低温快速冷冻后浸泡在液氮(-195．8℃)中长期保存并可随时取用的冷冻精液。细管分为0．25mL和0．5mL两种,目前使用较为普遍的是0．25mL的细管,细管的直径为2mm,长为133mm,细管的一端封口为棉塞,棉塞中间为     

道赛特绵羊细管冷冻精液商品化研究

<正>无角道赛特产于澳大利亚和新西兰,是世界著名优良肉毛兼用品种,有早熟多胎、常年发情、背宽体高、生长发育快、肉用性良好、适应性和抗病性强等优良种质特性,是目前世界上生产羔羊最好的终端父系品种之一。我国的绵羊冷冻精液输精研究始于1974年,1981年进行了授精试验,母羊情期产羔率达到56.4%。     

肉羊细管冷冻精液的生产与应用试验

从2004年开始由内蒙古宏兴生物技术有限责任公司生产的肉羊细管冻精，先后在内蒙古乌兰察布市十个旗县市肉羊配种站上投放使用，并销售到山西大同、包头、固阳等地区。应用实验结果表明应用冻精平均情期受胎率70．1％以上，受到广大配种员和农牧民的认可和欢迎。现就肉羊细管冻精生产与应用试验情况总结如下：     

不同输精方法和输精次数对绵羊冷冻精液受精率的影响

选择小尾寒羊99只,采用过颈输精法输精,情期受胎率为42.31%;选择小尾寒羊59只,采用腹腔内窥镜子宫角输精法输精,情期受胎率为59.32%。子宫角输精法情期受胎率比过颈输精法提高17.01个百分点,差异极显著(P<0.01)。选择18只小尾寒羊,输精2次,受胎率为27.78%;选择12只小尾寒羊,输精3次,受精率为33.33%。后者比前者提高5.55个百分点,差异显著(P<0.05)。     

去精清对提高辽宁绒山羊细管冻精质量的影响

随着对山羊精液研究的不断加深,研究者发现山羊的精清中含有磷脂酶,类磷脂酶、磷酸酶等许多物质,当与卵黄接触后会使卵黄凝固,产生对精子有毒害和损伤作用的物质,致使精液酸度增高,造成精子死亡。卵黄作为山羊冻精稀释液的常用成分,来源广、价格便宜,其作用是多方面的,目前尚未发现有比较理想的物质可以完全代替卵黄。因此,要避免发生上述不良反应,就必须将精清中     

The Beltsville sperm sexing technology: high-speed sperm sorting gives improved sperm output for in vitro fertilization and AI

The Beltsville sperm sexing technology is currently the only effective means of altering the sex ratio of offspring in livestock. The method is based on the flow-cytometric separation of X- and Y-chromosome-bearing sperm based on X/Y DNA content difference. It is an effective means of producing progeny of predetermined sex in cattle, swine, sheep, and laboratory animals. The method involves treating sperm with a DNA-binding fluorochrome, Hoechst 33342, and flow-cytometrically sorting them into separate X and Y populations that can subsequently be used for surgical intratubal or intrauterine insemination, deep-uterine insemination, regular artificial insemination in some cases, in vitro fertilization to produce sexed embryos for transfer, and intracytoplasmic sperm injection of ova. Skewed sex ratios of 85 to 95% of one sex or the other have been repeatably achieved in most species. The method has been used worldwide to produce several hundred morphologically normal animal offspring of the predicted sex. It has also been validated in the laboratory using DNA reanalysis of the sorted sperm populations and by fluorescence in situ hybridization and PCR of individual sperm. We developed a new orienting nozzle that we have fitted to both conventional and high-speed cell sorters that have been modified for sperm sorting. Recently we completed the adaptation of the new orienting nozzle to a Cytomation MoFlo high-speed cell sorter modified for sperm. This adaptation of the nozzle has increased the overall production rate of sorted X and Y sperm from about .35 million/h to 5 or 6 million sperm/h (each population). Calves have been born from cows artificially inseminated using conventional technique and sexed sperm. In addition, numerous litters of pigs have been born after transfer of embryos produced from X or Y sorted sperm.     

发情羊血清、肝素和BSA对绵羊冷冻-解冻精子体外获能的影响

分析了在受精液SOF中添加发情绵羊血清、肝素和牛血清白蛋白(BSA)对绵羊冷冻-解冻精液体外获能和受精的影响。结果表明:(1)使用含发情羊血清浓度为0、5%、10%和20%的受精液,分裂率分别是0、73.79%、73.25%和77.61%。加发情羊血清的3个试验组分裂率差异不显著。而未加血清的试验组分裂率为0,证明发情羊血清促进了绵羊冷冻-解冻精液的体外受精效果。(2)以含5%发情羊血清受精液为基础液,添加0IU/mL、5IU/mL和10IU/mL的肝素,受精后分裂率分别达86.64%、86.63%和75.53%。不加肝素组和5IU/mL肝素组分裂率差异不显著,而10IU/mL高浓度肝素组同其他两组比较,差异极显著(p<0.01)。说明含10IU/mL肝素的受精液,降低了绵羊体外受精后的分裂率。(3)用含5%发情羊血清受精液为基础液,以添加3mg/mLBSA为试验组,不加BSA为对照组,受精后两组分裂率差异不显著。发情绵羊的血清促进绵羊冷冻-解冻精子的体外受精效果。在受精液中添加发情绵羊血清进行绵羊冷冻解冻精子的体外受精,无需添加肝素和BSA。而且添加10IU/ml肝素,降低了受精后的分裂率。建议受精液中添加发情绵羊血清浓度在2%～10%较合适。     

辽宁绒山羊细管冻精配种受胎率试验

通过几年的研究,辽宁绒山羊细管冷冻精液质量明显提高,为了检验细管冷冻精液的应用效果,我们在辽宁绒山羊原种场进行了配种受胎试验。1材料和方法1.1 试验材料 配种用精液为我场自己研制生产的0.25ml细管冻精,解冻后平均活力在50.68％以上,顶体完整率58.24％,每管含有效精子数约2.5×107个。     

Optimization of the in vitro fertilization protocol for frozen epididymal sperm with low fertilization ability in Ban—A native Vietnamese pigs

The aim of the present study was to improve the penetration during in vitro fertilization (IVF) of a frozen lot of epididymal sperm with a notoriously low fertilization ability of a Ban boar which is a native Vietnamese breed by optimizing different parameters of the IVF system. In Experiment 1, we determined that Pig‐fertilization medium was superior medium to Tyrode's albumin lactate pyruvate‐polyvinyl alcohol medium for IVF and defined the optimum the sperm concentration (1 × 10⁶ sperm/ml). In Experiment 2, we clarified that partial removal of cumulus cells from cumulus‐oocyte complexes by hyaluronidase treatment before IVF enhances sperm penetration, whereas complete cumulus removal reduces penetration. Finally, in Experiment 3 the elevation of concentration of caffeine in Pig‐fertilization medium from 2 to 5 mmol/L and the prolongation of the co‐culture of gametes from 3 to 5 hr significantly increased the total penetration rate from 15.2% to over 50%. In conclusion, the combination of partial oocyte denudation, an elevated caffeine concentration in Pig‐fertilization medium and an extended interval of IVF with using an optimized sperm concentration was a potent way to improve the fertilization results for a frozen epididymal Ban sperm lot with low fertility.     

Ultrastructure of frozen and thawed ram sperm

With the aid of a transmission electron microscope we studied the ultrastructure of frozen and thawed ram sperm. The cryoprotective agents do not completely prevent the occurrence of structural changes in spermatozoa. The sperm membrane system is affected. The greatest and most frequent damage is to the acrosome, the cell membrane and mitochondria are less injured. We observed regional differences in the damage to the cell membrane. It is most damaged above the acrosome, least in the postacrosomal area and in the principal piece of the tail. We found considerable variability in the changes among individual spermatozoa from the same ejaculate. The changes are not in the nature of mechanical damage as a result of the occurrence of ice microcrystals. We therefore think that the occurrence of structural changes is dependent on damage at molecular level and consists of a change in the physical-chemical properties of membranes.     

浅议人工草地放牧羊的疫病综合防治技术

为有效控制人工草地上全日制放牧羊只的疫病死亡率,根据当地家畜疫病的发生规律,采取加强饲养管理,搞好环境卫生、定期消毒,严格检疫制度,加大定期驱虫次数,有计划地免疫接种,实施药物预防,预防毒物中毒,及时隔离或扑灭传染病羊等技术措施,羔羊死亡率从34%下降到12%,成年羊的死亡率从8.6%下降到3.7%,提高了草地畜牧业的经济效益和社会效益.     

蛋白酶体在精子获能和受精中的作用

26S蛋白酶体是一个多亚基蛋白酶,在人、鼠、猪、牛、海鞘和棘皮动物受精研究中,表明它是一种特异性的靶蛋白。在研究哺乳和非哺乳动物物种受精过程中发现蛋白酶体的位置,蛋白酶体和精子移动有关,蛋白酶体与透明带(zona pellucida,ZP)结合促进顶体胞吐,精子泛素化和蛋白水解酶抑制剂有助于阻碍精子多精入卵,蛋白酶水解被抑制改变精子获能进程,精子表面结合腺苷三磷酸(adenosine triphosphate,ATP)的消耗阻碍猪和棘皮动物受精,蛋白酶体水解抑制剂阻碍精子穿过ZP,但是不改变哺乳动物精子和卵子结合的效率,不改变和精子有关的去泛素化活性和体外受精(in vitro fertilization,IVF)时单精受精和多精受精的平衡。总的来说26S蛋白酶体与人和动物受精的诸多事件有关,为精子和卵子结合机制的研究提供理论依据。     

Laparoscopic artificial insemination in sheep

The goal of any AI program is to create improved offspring, and the achievement of this objective will depend on the breeding value of the ram and ewe selected. Laparoscopic AI is being utilized in the sheep industry to extend the use of superior rams, and it offers the producer the opportunity to maximize the reproductive potential of superior sheep. Rapid genetic trait infusion of known superior stud rams into the flock is the primary economic benefit of laparoscopic AI. The success of laparoscopic AI depends on events and factors that interrelate in a complex way. Once the selection and preparation of the ewe have been accomplished, one of the more important steps in the program is the successful synchronization of the ewe to deliver the necessary ova to the site of fertilization at a specific time. One of the best methods of synchronization for laparoscopic AI is the use of a progesterone product for a controlled time period and the administration of PMSG upon its removal. Detecting the onset of estrus is critical, and the addition of sterile (e.g., vasectomized) males is helpful, even essential, to accurately determine when each ewe begins her estrus. The ram effect has been shown to stimulate ovulation and estrus. Ewes must be inseminated within a narrow window of time after the synchronization product is removed. Ewes should be inseminated in the order in which they begin to exhibit signs of behavioral estrus, but age, stage of lactation, duration of behavioral estrus, and breed must be taken into account when this order is established. Fresh-extended semen works well throughout this preferred time frame established for laparoscopic AI, but frozen semen gives best results when used near the end. Advancement in manufacturing technology today removes equipment as a variable factor. It is important, therefore, that the inseminator develop a level of expertise in laparoscopy to ensure maximum fertilization rates. If available, fresh-extended semen is preferred over frozen semen, using at least the minimal number of spermatozoa necessary for fertilization. Evaluation of the post-thaw frozen or fresh semen is necessary to determine motility, morphology, and concentration, all of which help determine the volume of the insemination dose. The minimum necessary for laparoscopic AI in fine-wooled breeds is 20 X 10(6) normal motile spermatozoa; however, the more seasonal and less fertile American sheep need approximately 40 to 50 X 10(6) normal motile sperm to achieve acceptable fertility rates.(ABSTRACT TRUNCATED AT 400 WORDS)