SDS—PAGE分离不孕奶牛宫颈黏液AsAb

免疫不孕严重影响奶牛正常繁殖,相对于其他不孕类型不容易被确诊,准确对孕牛与不孕牛之间宫颈黏液AsAb的差异进行定性和定量分析,对奶牛免疫不孕的治疗研究有重要的意义。本试验将浅盘凝集检测和实际生产结果相结合确定的不孕奶牛和可孕奶牛作为研究对象,对宫颈黏液中蛋白进行SDS—PAGE电泳并对其分离条件进行优选。结果表明,本试验建立了一种在8％分离胶上,以15p~L上样量,100V,电泳3～4h的SDS—PAGE分离奶牛宫颈黏液的有效方法：对浅盘凝集检测和实际生产记录相结合确定AsAb免疫性不孕奶牛的宫颈黏液,用SDS—PAGE分离得到一种分子量为22～25kDa蛋白,初步确定为AsAb轻链：发现100％（24／24）不孕奶牛的宫颈黏液中存在AsAb轻链,14．3％（2／14）可孕牛宫颈黏液中存在AsAb轻链,但表达量明显低于不孕牛。     

间接ELISA检测奶牛宫颈黏液ASA免疫不孕

为建立检测奶牛宫颈黏液ASA免疫不孕的间接ELISA方法,以精子膜蛋白为包被抗原,进行奶牛宫颈黏液ASA的间接ELISA的优化试验。对27头ASA阳性不孕和29头ASA阳性可孕奶牛进行了间接ELISA检测标准的确定试验;对137头奶牛的宫颈黏液ASA免疫不孕进行间接ELISA临床检测。奶牛宫颈黏液ASA的间接ELISA优化条件为:抗原包被量为5μg/mL,宫颈黏液稀释度为1:5,宫颈黏液反应时间1h,二抗反应时间1.5h;奶牛宫颈黏液ASA免疫不孕检测的判定标准为:当样品OD490nm值大于0.513时判定为ASA阳性免疫不孕,当样品OD490nm值小于0.410时可孕,当样品OD490nm值介于0.410~0.513时为ASA阳性疑似,重复试验的变异系数均小于10%。对临床137份宫颈黏液样品进行检测,发现13头ASA阳性免疫不孕奶牛都不能受孕,而98头阴性结果牛中有89头怀孕。结果表明,间接ELISA可以有效检测奶牛宫颈黏液ASA免疫不孕。     

猪源致病性大肠杆菌的分离鉴定及多价油苗的研制

从我省滇中地区11个县区所送检的66份可疑猪大肠杆菌病病料中分离到35株致病菌株。采用24种主要引起致病性大肠杆菌因子血清作平板凝集试验后鉴定出29株，其中O107 19株（1株含K88抗原）、O149 9株（4株含K88抗原）、O2 1株，占所分离致病菌株数百分比分别为54．0％（19／35）、25．7％（9／35）、2．9％（1／35）。从中挑战出含K88抗原菌株O107：K88、O149：K88与标准菌株K88、K99、F41、987P混合制备多价油苗，在以上分离地区应用1万头份，效果显著，平均保护率达70％以上，且免疫后猪只无不良反应。     

奶牛子宫内膜炎的临床诊断与防治

牛子宫内膜炎是子宫黏膜的黏液性或化脓性炎症。是奶牛最常见的产科病之一，是引起奶牛不孕的主要原因。据报道，每年美国因奶牛不孕症造成的经济损失近2．5亿美元；英国报导不孕牛中约95％是由于子宫内膜炎引起。根据报道，奶牛不孕症中，子宫内膜炎特别是慢性子宫内膜炎可占不孕牛的60．1％～92．O％。Barbu等（1980）报道，     

精子膜抗原的研究及其应用

为了研究精子膜抗原并研发避孕疫苗,试验从精子膜抗原结构和功能的角度出发(这是因为精子膜抗原不仅是精子发生、成熟过程的重要标志分子,与受精和胚胎的早期发育关系密切,而且其中部分抗原还可诱导机体产生相应的自身抗体,是导致免疫性不育的主要原因.)在研究精子膜抗原揭示受精过程和不育机制的基础上,将一些精子膜抗原作为候选成分来研...     

缩宫素与氯化氨甲酰甲胆碱对奶牛胎衣不下的疗效观测

对自然分娩后8h内尚未排除胎衣的28头奶牛,分别应用缩宫素（14头,肌肉注射50IU／头）和氯化氨甲酰甲胆碱（14头,皮下注射25mL／头）进行治疗后。结果,用缩官素所治疗的14头奶牛中13例（92．85％）和氯化氨甲酰甲胆碱治疗的14例奶牛中11例（78．57％）均于12h内胎衣完全排出,其疗效显著（P〈0．05）。     

杜氏利什曼原虫NH36基因的克隆及其在大肠杆菌中的表达

应用RT—PCR技术扩增并克隆杜氏利什曼原虫（Leishmania donovani）NH36编码基因，经鉴定及序列分析，构建其原核重组表达载体，经IPTG诱导表达，SDS-PAGE和Western-blotting鉴定其表达产物。结果显示，获得的NH36开放阅读框为945bp，编码314个氨基酸残基，与GenBank上发表的国际标准株NH36编码基因序列以及氨基酸序列同源性分别为88．57％（837／945）和89．17％（280／314）。表达蛋白为36ku，经1mol／L IPTG诱导6h后的表达量最高；薄层扫描显示表达的蛋白占菌体蛋白总量的57％；该蛋白可被抗杜氏利什曼原虫的多克隆抗体血清识别。     

孕马血清对提高奶牛受胎率的效果观察

对屡配不孕奶牛在发情周期第11天肌注PMSG，第13天肌注氯前列烯醇，发情后输精．可有效提高情期受胎率，另对产后60天以上未配过的奶牛作同期发情处理，在第11肌注PMSG，第13天肌注氯前列烯醇，发情后输精，也明显可提高情期受胎率。结果显示：在屡配不孕牛组中A1组的受胎率40％（4／10），A2组的受胎率10％（1／10）；同期发情组中B1组的受胎率43．8％（7／16）．B2组的受胎率30．8％（4／13）。以上结果显示应用PMSG可明显提高奶牛情期受胎率。     

博达（PortaSCC）法测试奶牛隐性乳房炎试验初报

利用美国近年研究并应用于实践的博达（P0rtaSCC）奶牛隐性乳房炎测试技术对太原某奶牛场的奶牛隐性乳房炎进行了监测，试验显示，博达（PortaSCC）隐性乳房炎测试技术方法简便快捷、测定数据准确，具有较大推广应用价值；此奶牛场奶牛隐性乳房炎头阳性率为54．29％（57／105），乳区阳性率为26．03％（107／411）；3胎及以上奶牛阳性率达到70．18％（40／57）并极显著高于1-2胎（35．42％，17／48）（P〈0．01）；奶牛个体混合乳样体细胞与个体乳区乳样体细胞平均数呈直线相关，差异极显著（r=0．7418，P〈0．01）。     

浅谈奶牛蹄病与保健

蹄是奶牛重要的支往器官，肢蹄健康是奶牛高产、健壮的保证。奶牛蹄病是制约奶牛发挥优良性能的极为重要的因素。因蹄病而被迫淘汰，带来的经济损失，是每个牧场管理者十分棘手的问题。1发病情况笔者于1993、1994、1995、1996、1997连续5年对济南地区国有奶牛场进行调查统计，该病发病率分别为43．45％（478／1100）、57．77％（632／IO95）、27．43％（378／1378）、26．15％（369／1411）、14．81％（224／1512）。以上所指的蹄病主要包括腐蹄病，趾（指）间炎、蹄叶炎及外伤。5年中因蹄病而被淘汰的奶牛分别占病死、淘汰的70．26％（36…     

Oestrus during pregnancy in the cow

Forty-three oestruses were observed in 35 pregnant cows in one beef suckler herd and 17 dairy herds; at each oestrus the pregnant cow stood willingly to be mounted by another cow or bull. Such oestruses were observed at all stages of pregnancy, although more often between 121 and 240 days, occurred more than once per pregnancy and were also seen in successive pregnancies. On one farm where the dairy cows were observed for four 30 minute periods daily, oestrus was recorded in 5.7 per cent of pregnant cows. The behavioural signs associated with oestrus during pregnancy were indistinguishable from those of true oestrus in non-pregnant animals and although its duration was shorter (mean 5.6 hours), its intensity was comparable to that of the true oestrus. Pregnant cows showing oestrus were usually seen interacting with other oestrous cows in the sexually active group. Pregnant cows showing oestrus had a higher mean condition score (3.9 +/- 0.64) than control pregnant cows (3.0 +/- 0.36). Physiological changes in the genital tract normally associated with true oestrus were not observed in pregnant cows showing oestrus. There was no ovulation or metoestrous bleeding. The characteristics of cervical mucus, including ferning patterns, were similar to those of pregnant cows at the same stage of pregnancy. Hormonal changes associated with oestrus in non-pregnant cows were not observed in the pregnant cows exhibiting oestrus. Seven of nine pregnant cows at oestrus stood willingly to be mounted by a bull. On seven occasions, bulls exposed only to cervical mucus from pregnant cows showing oestrus did not display flehmen.(ABSTRACT TRUNCATED AT 250 WORDS)     

发情母牛粘液对采精公牛交配能力及精液品质影响的研究

[目的]研究发情母牛粘液对采精种公牛交配能力以及精液品质的影响。[方法]从采精种公牛中随机抽出10头,进行2次采精试验,第一次按照正常方法采精,第二次在台牛尾根附近涂抹粘液,记录交配能力,将2次采精所采得的精液进行检测。[结果]第二次采精交配能力明显高于第一次。精液量、精液密度、活率、有效精子数、冻后活率、生产冻精数分别为：5.76 mL、19.23亿/mL、0.72、13.4、0.385、400.5,通过T检验P〈0.01,差异显著。[结论]在台牛后躯涂抹发情母牛粘液刺激公牛使交配能力和精液品质显著提高。     

Usefulnes of cervical mucus of cattle for pregnancy tests

A qualitative fructose test was adapted for a pregnancy test in cattle. Cervical mucus was heated 1-2 minutes in a corn syrup-water solution according to the method of Hopwood. In mucus from 320 slaughtered cows there were 8.5% false positives among 118 nonpregnant and 5.0% false negatives among 202 pregnant cows. 6 distinct positives, 3 equivocal tests, and 2 definite negatives were recorded in 11 cases of intrauterine death with pseudopregnancy. The mucus from 56 live cows produced 9.7% false positives and 12.0% false negatives. Fructose contents were determined quantitatively in 96 cervical mucus samples: 60 pregnant cows averaged 148 mg%; 36 nonpregnant 34.9 mg% (p less than .005); and pregnant in the first 5 weeks 111 mg%. This method was 90% accurate in the first 5 weeks.     

精子生育相关性抗原对奶牛妊娠率影响研究

为研究精子生育相关性抗原对奶牛妊娠率的影响,测试了16头种公牛精子的生育相关性抗原,进行人工授精,比较奶牛的妊娠情况.结果显示:测试的16头种公牛精子生育相关性抗原,其中13头呈阳性,3头阴性,阴性率为18.75%.在用FAA阳性精液进行人工授精的520头奶牛中,有433头奶牛怀孕,妊娠率为83.26%.而FAA阴性授精的80头奶牛中,只有52头奶牛怀孕,妊娠率为65.0%(P<0.01).因此,奶牛人工授精FAA阳性精子妊娠率比FAA阴性精子高18个百分点,大大提高了奶牛的妊娠率.     

Antisperm antibodies in repeat‐breeding cows: Frequency,detection and validation of threshold levels employing sperm immobilization,sperm agglutination and immunoperoxidase assay

Antisperm antibodies have been found in repeat‐breeding(RB) cows, and those causing agglutination and/or immobilization of sperm are considered to be closely related to unexplained infertility. However, a standard protocol for identifying antisperm antibodies (ASA) in cattle is not validated. Therefore, an investigation was undertaken to evaluate sperm immobilization (SIT), sperm agglutination (SAT) and immunoperoxidase (IPT)assays for detection of ASA in serum and their respective threshold levels for confirmation. Animals (heifers, normally breeding, repeat‐breeding and pregnant animals) that were free from IBR, brucellosis and uterine infections (screened by clinical examination) were included in the study. Sperm agglutinating, sperm immobilizing and antisperm antibodies evaluated by respective assay were significantly higher (p < .05) in RB cows compared to other groups. The SIT assay was able to identify 61% of RB caused by ASA, more than those employing SAT and IPT. Furthermore, a dilution rate of 1:5 and 1:80 (confirms 59.0 and 57.0% RB+ve)were sufficient to diagnose ASA by SAT and IPT, respectively. Results indicate the presence of __12.6% clumped spermatozoa and __ 2.6%(cut‐off value) peroxidase‐positive spermatozoa at 1:5 and 1:80 dilutions diagnosed with SAT and IPT, respectively, may be considered as repeaters arising out of ASA. Furthermore, study also showed the presence of lower incidence of ASA positivity in other groups of animals (heifer<normal breeder<pregnant animals) compared to repeaters. Study results show that although IPT is more specific and accurate but SAT and SIT are comparatively simple and cost‐effective assays suitable for detecting ASA under field conditions and thus can be recommended for screening of repeaters.     

奶牛性控冻精和普通冻精授配效果对比试验

为了掌握性控冻精和普通冻精授配效果,采用性控冻精和普通冻精分组授配奶牛进行对比试验.性控冻精授配奶牛平均受胎率为89.2％;常规冻精授配母牛平均受胎率89.4％,差异不显著（P＞0.05）;性控冻精母犊率达到92.02％,普通冻精产母犊率51.63％,差异极显著（p＜0.01）.经对性控冻精所产母犊的体重、体尺进行测定,并与常规冻精所产母犊的生长发育指标进行比较,两者差异不显著.试验表明,奶牛性控冻精对实现规模化奶牛场的快速扩群和品质提升具有较高的实用价值.     

过瘤胃保护蛋氨酸对奶牛生产性能的影响及经济效益分析

本研究采用在生产条件下的对比饲养试验,通过研究日粮中添加本实验室研发的过瘤胃保护蛋氨酸产品对泌乳高峰期和泌乳中期高产荷斯坦奶牛产奶量和乳成分的影响,验证了产品的性能与使用效果,并对经济效益进行了分析。试验选择2～5胎之间、日产奶量30kg以上、处于泌乳高峰期的荷斯坦奶牛20头,随机分为两组,试验组每头每天添加本实验室研发的过瘤胃保护蛋氨酸产品75g。试验结果表明:日粮中添加过瘤胃保护蛋氨酸产品可明显提高荷斯坦奶牛的产奶量(3.0kg/头.d,P0.05)和极显著提高牛奶的乳蛋白含量(0.27个百分点,P0.01),使乳脂肪含量略有降低(0.13个百分点,P0.05),对乳糖含量和乳干物质含量影响很小。在高产奶牛泌乳高峰期日粮中添加过瘤胃保护蛋氨酸能够获得可观的经济效益(8.54元/头.d)。     

Relation between cervical mucus sperm toxicity and pregnancy in cows]

Cervical secretion was collected from 99 dairy cows of the Black Pied Lowland breed at an age of 24 to 109 months with a minimum performance of 4000 kg of milk annually in spontaneous oestrus 42 days post partum at the minimum. It was demonstrated by the radioimmunological determination of progesterone in skim milk that from all the breeding cows (n = 99) the oestral secretion was collected during an optimum insemination time (x = 0.19 ng.ml-1 progesterone). Of the statistical set of 99 cows, 55 (i.e. 56.57%) got in calf and 43 (i.e. 43.43%) remained barren. As found, the conception results were not influenced by the rank of the insemination after parturition and the conception capacity of the cows was not influenced by the length of time from parturition to the respective inseminations. It was demonstrated in our investigation that the conception capacity of the cows increases as the time of sperm survival prolongs in the cervical secretion in vitro. The highest conception capacity (80%) was recorded in the cows whose cervical secretion was free of spermiotoxic action. In such cases the motility of the spermatozoa in the oestral mucus was maintained at a temperature of 38 degrees C for longer than 300 minutes. On the other hand, it was demonstrated that the conception capacity of the cows declines with shortening time of sperm survival in the cervical mucus.     

以发情后期黏液带血的西杂牛为受体进行冷冻性控胚胎移植效果研究

在凉州区东河乡的王景寨等四个村,对农户饲养条件下,以西杂牛为受体,采用冷冻性控胚胎移植303头,怀孕162头,受胎率53.47％.其中：对自然发情的西杂繁殖母牛,以首次观察到黏液带血为0天,再加5天实施冷冻性控胚胎移植,受胎率54.84％;以发情盛期为0天,第7天进行移植的受体母牛受胎率为53.11％.经显著性测定二者差异不显著（P＞0.05）.移植初配受体母牛受胎率54.26％;移植经产受体母牛受胎率为53.11％.经显著性测定二者差异不显著（P＞0.05）.