Percoll密度梯度离心分离牛精子初探

用2.9％柠檬酸钠将奶牛原精密度调到5×10~8/ml,取其3ml经26％Percoll洗涤(1700转,25分钟),置于pH7.0、渗透压300mmol/kg的9级Percoll梯度溶液上分离(1000转,15分钟)从第9层取得精子作去Percoll处理(2500转,20分钟),再用含甘油和卵黄的牛冷冻稀释液平衡滴冻,解冻后活力可达0.4左右。分离后精子的受胎率为52.2％(47/90),母犊率为44.2％(19/43),对照组母犊率为36.8％(21/57)。说明分离后精子可正常受胎,并有提高母犊率的趋势。     

离心方法对冷冻后民猪精子质量的影响

通过对比离心力、离心时间和离心次数与冻融后民猪精子质量的相关性,为改进民猪精液离心程序,提高冻精效果提供理论依据。试验分2组,第1组:将采集的新鲜民猪精液分组后分别在600×g,800×g,1 000×g及1 200×g离心力条件下离心10,15和20min;第2组:在800×g,离心15min条件下,分别离心1,3和5次;离心后的精子分别经过冷冻保存再融解过程后,检测不同离心条件下民猪精子冻融后的活率、顶体完整率、质膜完整率和线粒体膜电位的情况。结果表明,600×g(10,15,20min)及800×g(10,15min)离心力时,不同离心时间对精子质量影响不显著(P0.05)。1 000×g和1 200×g离心力时,随着离心力和离心时间的增加,精子质量明显降低,各指标组间差异显著(P0.05)。在800×g离心力,离心15min条件下,精子质量随离心次数增加而降低,各组间差异显著(P0.05)。因此,在本试验条件下,800×g离心力,离心15min,离心1次,可以获得较高的精子收集效率和冷冻后精子质量。     

不同精子密度仪测定猪精子密度的准确性比对试验

使用minitubeSDMI、分光光度计以及血细胞计数板手工计数法3种方法测定公猪精液密度,并将前两种仪器的测定结果与手工计数法进行准度和精度的比较以及配对T检验。结果表明,minitubeSDMl测定精子密度的准确性与精确性均高于分光光度计的测量结果,但这两种仪器与手工计数法的测定结果均存在显著差异。该结果提示我们。在这两种仪器的测定结果最好在样品量检测量较大的情况下用于精液密度的估计,而不能直接作为精液密度的检测结果。     

家兔结核菌素试验的研究

通过对家免结核菌素试验部位的形态学和组织学变化以及多次进行结核菌素试验的研究,查明了结核病家兔和健康家兔对多次注射结核菌素不可能产生脱敏和致敏反应,找到了可行的家兔结核菌素试验操作方法。     

兔出血症病毒提纯和多肽的比较研究

我们采用了甘油－酒石酸钿梯度离心、琼脂糖４Ｂ层析和ＤＥＡＥ３２离子交换层析等三种方法提纯兔出血症病毒（ＲＨＤＶ）进行比较，其结果发现ＤＥＡＥ３２离子交换层析效果最好，获得的病毒纯度最高，经ＳＤＳ－ＰＡＧＥ和Ｗ－Ｂｌｏｔ试验表明，ＲＨＤＶ只有一条蛋白多肽分子量为６１ＫＤ，截然不同于以往国内研究报道的ＲＨＤＶ具有３－６条结构多肽等结果，在国内属首次报道。     

醋糟饲喂家兔试验

醋糟是制醋的主要下脚料,据分析,它含粗蛋白6—10％,粗脂肪2—5％,无氮浸出物20—30％,粗灰分13—17％,粗纤维25—30％,钙0.25—0.45％,磷0.16—0.37％,营养比较丰富。但是,醋糟做为家兔饲料对增重的影响,国内尚少见报道。为探讨这一问题,我们进行了家兔日粮中添加一定比例醋糟的试验,以达到广辟饲料来源,降低饲养成本的目的。一、材科和方法 1、试验兔:选70～75日龄健康无病,胎次相同,体重相近的比利时生长兔五窝共30只,随机分成试验组(15只)和对照组(15只)。在试验期间分别投喂氯苯胍和敌     

哺乳动物精子分离技术研究进展

精子分离是控制动物性别简单而有效的方法之一,本文综述了X、Y精子分离的生物学基础,分离方法及其应用效果的研究进展,并阐述了精子分离技术存在的问题及其发展前景。     

家畜精子超弱化学发光的研究

利用液体闪烁计数仪测定绵羊精子的超弱化学发光,以研究它的机理和畜牧中的应用价值。结果表明,精子的发光与活力、呼吸、果糖分解、~(32)Pi摄入量和磷酸肌酸呈正相关(r=0.9806,P<0.01;0.9684,p<0.01;0.9882,p<0.01;0.9793,p<0.01;0.9962,p<0.01)说明了精子的发光与代谢存在有内在的联系,反映了精子的能量转化过程和精子细胞物理化学反应的信息,是评定精子质量很有价值的一个指标。     

Percoll法处理牛精子对体外受精胚胎发育的影响

牛冷冻精液解冻后,在Ｐｅｒｃｏｌｌ梯度液中４００ｇ离心２４ｍｉｎ有效地获得了５７．８７％±８．０％的精子回收率;对照组上游法精子回收率仅为１５．８７％±１．９％（Ｐ〈０．０５）。Ｐｅｒｃｏｌｌ和上游２种方法分离的精子在受精液中８,１８ｈ后的活率,分别为６０％,１０％和７０％,２０％,２种浓度Ｐｅｒｃｏｌｌ法分离的精子对不同成熟处理的卵母细胞体外受精子（ＩＶＦ）卵裂率的影响,精子最终浓度为２×１０＾     

利福昔明对家兔肠道菌群的影响

本试验将利福昔明按25、50、100 mg/kg 3个剂量组分别给40日龄家兔口服,连续用药3 d,并于首次用药后第4、7、10 d剖杀家兔,取盲肠和回肠内容物进行乳酸杆菌和大肠杆菌的选择性培养并计数,以此评价利福昔明对家兔肠道菌群数量的影响。结果显示：与对照组相比,利福昔明高、中、低剂量组的肠道大肠杆菌菌群数量减少,其中高剂量组（100 mg/kg）在首次用药后第7、10 d,对家兔回肠大肠杆菌的影响显著;与空白对照组相比,利福昔明高、中、低剂量组的肠道乳酸杆菌数量变化不显著。     

塞北兔血清蛋白遗传多态性研究

采用垂直板聚丙烯酰胺凝胶电泳法对２１６只塞北兔血清转铁蛋白（Ｔｆ）、前转铁蛋白（Ｐｒｔ）、后白蛋白（Ｐｏ）和白蛋白（Ａｌｂ）等４个蛋折位点的遗传多态性进行检测。结果表明,在塞北兔群体中,Ｔｆ和Ａｌｂ呈单态,Ｐｒｔ和Ｐｏ呈多态。Ｐｒｔ有６种表型,受３个共显等位基因控制;Ｐｏ有３种表型,受２个共显性等位基因控制。Ｘ＾２检验结果表明,塞北兔群体在Ｐｏ位点处于Ｈａｒｄｙ－Ｗｅｉｎ－ｂｅｒｇ平衡状态（Ｐ〉０     

Effect of Different Extenders and Seminal Plasma on the Susceptibility of Equine Spermatozoa to Lipid Peroxidation After Single-Layer Centrifugation, Through Androcoll-E

This study was conducted in an attempt to see whether single-layer centrifugation (SLC) increases the susceptibility of stallion spermatozoa to lipid peroxidation (LPO), in different extenders after removing all seminal plasma (SP). The susceptibility of stallion spermatozoa to LPO was studied before and after SLC. Each ejaculate was split, and aliquots extended with one of the three different extenders: INRA 96, Kenney's, or Equipro, and stored for 24 hours at 5°C (i). From the extended samples, an aliquot was kept as a control and the other was subjected to SLC through Androcoll-E. The selected spermatozoa were re-suspended in the appropriate extenders, without (ii) or with (iii) addition of 50% (v/v) pooled homologous SP for 24 hours at 5°C. Using ferrous sulfate as pro-oxidant, the susceptibility for LPO was flow-cytometrically assessed using the probe Bodipy^581/591-C₁₁. Sperm motility, monitored with a Qualisperm motility analyzer, increased after SLC treatment (P < .001). No significant correlations were found between motility and induced LPO with ferrous sulfate. The SP and extenders, per se, did not have a significant protective effect against LPO, but the interaction between SP and Kenney increased the susceptibility to LPO. However, the selected spermatozoa through Androcoll-E and the subsequent dilution in INRA had a significant protective effect against LPO (P < .05), especially when the oxidative insults were higher (80 μM).     

解冻温度和时间对牛精子活力的影响试验

[目的]为了找到较为理想的解冻温度与时间,为基层黄牛改良工作提供技术参考。在不同温度区、不同时间解冻牛细管冷冻精液,对解冻后的精子活力进行检测。[方法]采用50~90℃每隔10℃设一个温区解冻牛细管冻精,检测解冻后精液的活力。[结果]在50~90℃5个温度梯度下快速解冻不同时间后的精子活力均在0.35以上,其中90℃解冻3~4s,80℃解冻4~5s,70℃解冻4~6s,60℃解冻8~9s后的精子活力均大于0.4,显著高于其他时间解冻后的精子活力。[结论]不同温度、时间解冻后精子活力各异,其最佳解冻温度和时间为90℃解冻3~4s,80℃解冻4~5s,70℃解冻4~6s,60℃解冻8~9s。     

猪屎豆生产性能测定及其饲喂家兔的效果研究

为探讨猪屎豆作为家兔饲料的可行性及其开发利用前景,对猪屎豆进行了栽培试验和家兔饲养试验,结果：当年栽培的猪屎豆刈割3次,一年鲜草总产量为50420kg／hm。;经测定,猪屎豆的干物质中含粗蛋白21．0％、粗脂肪2．30％、粗纤维20．70％、粗灰分7．00％、水分10．20％、钙1．36％、磷0．48％;试验组与对照组兔绝对增重分别为4300g与4160g,差异不显著（P〉0．05）;对照组与试验组的饲料利用率分别为38．62％和40．56％,猪屎豆的饲料利用率高于对照组。结果表明：添加30％猪屎豆可以代替苜蓿草粉喂兔。有较大的开发利用价值。     

Effect of Storage Time and Temperature of Equine Epididymis on the Viability,Motion Parameters,and Freezability of Epididymal Sperm

The recovery of sperm from the epididymal cauda may be the last chance to obtain genetic material when sudden death or serious injuries occur in valuable stallions. However, the lack of technical knowledge regarding the storage and transportation of the epididymis often prevents the preservation of the sperm. Therefore, the aim of this study was to compare sperm parameters of sperm obtained immediately after orchiectomy with sperm recovered from epididymal cauda at different times after storage at 5°C and at room temperature (RT). For that, 48 stallions of different breeds were used. In group 1 (control group), eight stallions were used, and the harvest of the epididymal sperm was performed immediately after orchiectomy. In group 2, 40 stallions were used, which were divided into five groups according to the storage time of the epididymis after orchiectomy (6, 12, 18, 24, or 30 hours), making a total of eight stallions per group. One epididymis of each stallion was stored at 5°C, and the contralateral epididymis was stored at RT, both for the same period. The sperm parameters of total motility, progressive motility, progressive linear velocity, curvilinear velocity, percentage of rapid sperm, and plasma membrane integrity were evaluated in all the groups after sperm recovery, resuspension in a sperm freezing diluent, and thawing. In conclusion, the storage of the testis-epididymis complex at 5°C provided better preservation of epididymal sperm than the storage at RT, and regardless of the temperature, the progressive motility is the sperm parameter that is most sensitive to storage time.