两种检测精子活率方法的比较

精子活率是评价精液品质的一项重要指标,其在光学显微镜下的测定,操作简便、直观,仍为临床常规检查所应用。本试验就精子用伊红-苯胺黑染色法和中性红染色法两种精子活率的测定方法进行比较,以便选择一种快速又准确的方法。1材料与方法1.1材料1.1.1试剂:伊红Y、中性红染液均购自上海试剂三厂。伊红Y染液:用蒸馏水配制,浓度为0.15%;中性红染液:pH7.4PBS缓冲液配制,浓度为0.01%。1.1.2精液的采集和处理:精液采自宁夏隆德县某奶牛场。精液采集后放入37℃的恒温水浴箱内。借助光学显微镜将精液高倍稀释(以在光学显微镜下可以清楚看到精子轮廓…     

精子活率的简易检测方法

精子活率是评价精液品质的一项重要指标，采用肉眼估测法、加热法、伊红-苯胺黑染色法、中性红染色法，其在光学显微镜下的测定，操作简便、直观，仍为临床常规检查所应用。     

检测肉羊精子活率方法的比较

就精子加热法、伊红-苯胺黑染色法、中性红染色法3种精子活率的测定方法进行比较,以便找到一种快速又准确的方法。通过测定肉羊精液,结果表明,用以上3种方法测定精子活率,差异不显著。但在试验过程中发现,伊红染液浓度对精子活率有影响。中性红染色法能在光学显微镜下对精子进行形态描述与识别。     

二甲基亚砜对蓝狐精子活率和体外获能的影响

DMSO是一种有机溶剂,影响精子体外获能的孕酮、肝素、钙离子载体多溶于此溶剂,因此确定蓝狐精子最适合的DMSO浓度及其作用时间是十分必要的.试验将分离、优化的蓝狐精子置于BO液中,在38.5 ℃、5%CO2培养箱中进行获能培养,然后分别在6个浓度(0%、0.1%、0.2%、0.5%、1.0%、2.0%)的DMSO中和4个时间(15分钟、30分钟、60分钟、120分钟)进行考马斯亮蓝染色和伊红-苯胺黑染色,检测并统计精子顶体反应率、精子活率,利用SPSS11.0统计分析软件分析试验数据.试验结果显示:蓝狐精子在DMSO浓度为0.2%和120分钟时体外获能效果最好.     

家畜精子低渗色试验研究

根据活精子对特定染料不着色而死精子易于着色，以及活精子在低渗环境中出现膨胀，死精子不能膨胀的原理。应用伊红低渗溶液对犬，猪，牛，鸡活精子的代谢能力进行评定，同时用常规精子评定法进行评定。二者进行差异显著性检验，差异均不显著。说明伊红低渗溶液对犬，猪，牛，鸡精子生活能力的评定是一种行之有效的方法。而且该方法简便，判断客观，具有实用性。     

透明质酸对猪精子活率,获能,顶体反应及体外受精穿透率的影响

采用具有高繁殖力的种公猪的新鲜精液及冷冻精液，加入不同获能剂咖啡因及透明质酸，在３９．５℃、５％ＣＯ２培养箱中孵化培养。结果表明，透明质酸能够在体外条件下保持精子的活率，并维持在稳定的水平，鲜精为４２．６％～４７．８％，冻精为２０．４％～２５．６％。荧光剂Ｈｏｅｃｈｓｔ染色表明，透明质酸能够显著增加培养３６０ｍｉｎ后的活精子率。在无蛋白来源的ｍＢＯ培养液中，培养０～６０ｍｉｎ，精子发生自发的获能和顶体反应，而受获能剂影响，精子发生获能的时间带在各处理组之间显著不同：咖啡因处理组的获能时间带是６０～３６０ｍｉｎ，透明质酸处理组则是６０～１８０ｍｉｎ。体外受精试验表明，去除卵母细胞周围卵丘细胞时，透明质酸和咖啡因对精子穿透率的影响没有区别；而在卵丘细胞存在的前提下，咖啡因处理组的穿透率显著高于透明质酸处理组。     

弱酸对猪精子活率及产仔性别的影响

分别向猪精液中加入醋权、乳酸、草酸、柠檬酸,改变了精液的ＰＨ值,并测定精子的活率,当精液的ＰＨ值达到６．５,精子沃经达到６０％时,将此褒庇人发情母猪人工配种,结果当每ｍｌ精液中加入１％的醋酸０．１５ｍｌ（ｐＨ６．５）时,对母猪的产仔数影响不显著（Ｐ〉０．０５）,但与对照组相比可显著地提高仔母猪的比例（Ｐ〈０．０５）。     

性成熟程度、天气情况、外界温度对雄蜂精子总数和活精子比率影响的研究

本试验旨在采集到高质量的雄蜂精沌为蜜蜂人工授精教术理论的发展提供依据。本文较详细地研究了性成熟程度、天气情况、外界温度对采集到的高加索蜜蜂雄蜂、东北黑蜂雄蜂和意大利蜜蜂雄蜂的精子总数和活精子比率的影响,结果表明：（1）性成熟度一般的（即在稍微挤压下,雄蜂外生殖器就外翻的）雄蜂精子总数、活精子比率要比欠成熟的（即在稍微用力挤压下,雄蜂外生殖器外翻的）多。（2）晴天时采集到的雄蜂的精子总数、活精子比率要比阴天时多。（3）随着外界温度的升高,所采集的雄蜂个体精子总数增多;活精子比率也有增高的趋势。     

甲基紫催化光度法测定饲料中的微量碘

在磷酸介质中,微量碘离子与碘酸钾氧化反应能催化甲基紫褪色,而且褪色程度与碘离子的量有关,最大吸收波长在575nm,在0～2μg/ml范围内呈线性关系。测试方法检测限为0.062μg/ml,回收率为96.7%～99.3%。用于饲料中微量碘的测定,结果令人满意。     

Clinical usefulness of intradermal fluorescein and patent blue violet dyes for sentinel lymph node identification in dogs

The first lymph node receiving drainage from a specific anatomic region is referred to as the sentinel lymph node (SLN). This study sought to evaluate the intradermal use of two dyes, patent blue violet (PBV) and fluorescein (FL), for SLN mapping in the dog. Multiple intradermal injections were performed in five healthy dogs using two dyes, PBV in 0.9% NaCl and FL in solutions of 0.9% NaCl and 6% hetastarch. Skin flaps were raised and followed to the first area of discrete stain uptake. Areas of uptake were identified as lymph nodes grossly and by cytology. Identification of a SLN for each area of intradermal injection was accomplished for 98% of the injection sites. Intradermal injections of both PBV and FL dyes produce readily visible staining of lymphatic vessels and SLNs in healthy dogs and are sufficient to allow ready identification of these structures during postmortem dissection.     

Antibodies to West Nile virus and related flaviviruses in wild boar, red foxes and other mesomammals from Spain

Red foxes (Vulpes vulpes), wild boar (Sus scrofa) and Iberian pigs (Sus scrofa domestica) that are raised extensively outdoors, as well as other wild mesomammals from south central Spain and wild boar from Do?ana National Park (DNP), were tested for antibodies against related flaviviruses by ELISA and for antibodies against WNV by VNT. Mean flavivirus seroprevalence according to ELISA was 20.4±7.8% (21 out of 103) in red foxes, 12.6±2.8% (69 out of 545) in wild boars, and 3.3±2.7% (6 out of 177) in Iberian pigs. A stone marten (Martes foina) also tested positive. Flavivirus seroprevalence in wild boar was significantly higher in DNP, and increased with age. Haemolysis of the serum samples limited interpretation of VNT to 28 samples, confirming WNV seroprevalence in one red fox, four Iberian pigs and nine wild boars. ELISA positive, microVNT negative samples suggest presence of non-neutralizing antibodies against WNV or antibodies to other antigenically related flaviviruses. Despite the importance of wetlands for flavivirus maintenance and amplification, WNV/flavivirus seroprevalence in wild boar and red foxes was not associated to wetland habitats. This is the first report of exposure of red foxes to WNV. With view to use of the tested species as sentinels for flavivirus activity, limited exposure of Iberian pigs that would be available for regular sampling, low numbers of foxes collected and concentration of wild boar harvest in the winter season are major drawbacks.     

中性植酸酶在水产中的应用

本文综述了芽孢杆菌植酸酶的酶学性质、催化机制以及中性植酸酶在淡水鲤科鱼类饲料中的应用效果,并对其研究前景作了展望。     

Flow cytometric determination of the proportions of X- and Y-chromosome-bearing sperm in samples of purportedly separated bull sperm

A rapid assay for determining the proportions of X- and Y-chromosome-bearing sperm in semen samples would benefit research aimed at sex ratio control through sperm separation. It also would be of value for quality control should a separation technique be developed. Flow cytometric methods capable of measuring sperm DNA content precisely enough to resolve and quantify the X and Y populations in many mammalian species have been developed. They are effective for fresh and cryopreserved sperm of most domestic animals. Results are reported of flow cytometric analyses of bull sperm samples from seven commercial and academic sources after processing with procedures purported to separate the X and Y populations. In no case was enrichment of either sperm population observed. Breeding trials carried out by the sources of two of the sets of samples showed these procedures were ineffective in altering the sex ratio.     

Distribution of glycoproteins on feline testicular sperm, epididymal sperm and ejaculated sperm

Glycoproteins (GPs) are known to be involved in the phenomenon of sperm maturation and capacitation. In the present study, we investigated the attachment of GPs on sperm cell membrane during the process of feline sperm maturation from testicular sperm to ejaculated sperm by using 8 FITC-labeled lectins. The results showed that 3 types of GPs were presented on testicular sperm and 7 on caput epididymal sperm. Corpus and cauda epididymal sperm and ejaculated sperm had GPs detected by 8 FITC-labeled lectins used in the present study. This study demonstrates the part of the characteristic of GPs that are present on the feline sperm cell membrane during the process of sperm maturation.     

Evaluation of testicular toxicity and sperm morphology in rats treated with methyl methanesulphonate (MMS)

Methyl methanesulphonate (MMS), a potent alkylating agent and testicular toxicant, was orally administered to rats for 5 days at 40 mg/kg. During the recovery period of up to 5 weeks, males were evaluated for testicular toxicity and sperm morphology. The 5-week recovery period were designated as follows: Day 1 (the day after final treatment); Week 1, Week 2, Week 3, Week 4 and Week 5 (1, 2, 3, 4 and 5 weeks after final treatment). Morphologically abnormal sperm increased beginning in Week 3, peaked in Week 4 and declined slightly in Week 5. Histopathological examinations indicated retention of step 19 spermatids at stage IX from Day 1 through Week 3. Quantitative evaluation of spermatogenic cells indicated a decrease in the number of late pachytene spermatocytes and early spermatids on Day 1. TUNEL examination showed a significantly high frequency of apoptosis in the meiosis cells in Week 1. In the present study, genetic damage induced by treatment with MMS affected spermatogenesis and a wide variety of spermatogenic cells in the testis. Apoptosis in the course of meiosis seemed to be involved in the elimination process of genetically insulted germ cells, and this process seems to play an important role in eliminating and/or decreasing the germ cells with retention of spermatids and the potential to express morphologically abnormal spermatozoa.