快速凝集试验与补体结合试验检测牛边缘无浆体感染的效果对比

从１３８份血清样品的比较试验结果显示，快速凝集成试验（ＲＣＡ）比补结合试验（ＣＦ）检测边缘无浆体感染的敏感性高（８８．９％：８１．５％），假阴性率低（１１．１％：１８．５％），两者都具有良好的特异性和预测性，检测阳性符合率高，快速凝集试验对一次感染牛的阳性时间更长久（３０３天：９２天）。     

PCR检测感染牛血液中边缘无浆体DNA

以边缘无浆体表面保护抗原的ｍｓｐｌβ基因设计和合成的１对２０ｍｅｒ寡核苷酸作为引物，用耐热的Ｔａｇ－ＤＮＡ聚合酶经５０个循环扩增边缘无浆体模板，扩增产物直接用凝胶电泳检测，并用标准分子量确定。结果表明，只有用边缘无浆体ＤＮＡ模板进行扩增时，扩增产物才能生成，而以血液原虫，如牛巴贝斯虫、双芽巴贝斯虫、锥虫和牛白细胞ＤＮＡ作模板扩增，无扩增产物生成。ＰＣＲ检测边缘无浆体的灵敏度可达约３００个无浆体的ＤＮＡ。边缘无浆体ＰＣＲ适用于牛边缘无浆体病的病因鉴定和检测     

抗牛边缘无浆体MSP5单克隆抗体的制备

为制备抗牛边缘无浆体膜表面蛋白(MSP5)单克隆抗体(MAb),以原核表达的重组MSP5蛋白(rMSP5)免疫BALB/c鼠,应用常规杂交瘤技术获得2株能稳定分泌特异性MAb的杂交瘤细胞株(1D8和2F3).间接ELISA检测腹水效价分别为5.49×105和7.83×104,亚类鉴定其重链为分别为IgG2b和1gG2a,轻链均为K型;ELISA叠加试验表明这2株MAb识别的抗原位点相同或相近;Western blot结果显示2株MAb均能与rMSP5发生反应.特异性抗MSP5的MAb的获得,为牛边缘无浆体检测奠定了基础.     

实时荧光PCR检测牛边缘无浆体方法的建立及应用

根据牛边缘无浆体表面蛋白4的保守基因序列设计特异引物AMOC9/AMOC5、AMOC10/AMOC12和特异性探针MP,首次建立了牛边缘无浆体的实时荧光PCR检测方法,检测DNA的最低限度为200 fg。对中央无浆体、绵羊无浆体、牛巴贝斯虫、双芽巴贝斯虫、羊莫氏巴贝斯虫、山羊泰勒虫、温氏附红细胞体、东方巴贝斯虫、刚地弓形虫和伊氏锥虫进行检测,无荧光检测信号。本研究用所建立的方法检测采自江苏和哈尔滨的180份抗凝血（奶牛和肉牛）,其阳性率为8.9%。结果表明,建立的实时荧光PCR检测牛边缘无浆体的方法具有较高的特异性和敏感性,可用于牛边缘无浆体病的流行病学调查、检疫和监测。     

检测牛边缘无浆体抗体的竞争抑制ELISA方法的建立

为建立检测牛边缘无浆体(Anaplasma marginale)抗体的方法,本研究以牛A.marginale膜表面重组MSP5蛋白作为包被抗原,抗MSP5单克隆抗体(MAb)作为竞争抗体,建立一种用于检测牛A marginale抗体的重组MSP5蛋白竞争抑制ELISA(CI-ELISA)方法.经优化确定CI-ELISA的最佳反应条件为:抗原包被浓度为2μg/孔,封闭液为2%脱脂乳,MAb的稀释度为1:400,酶标二抗的稀释度为1:1000,阴性和阳性血清临界值分别为33%和40%;该方法具有良好的特异性和重复性;2 348份临床血清样品的检测结果表明,217份为阳性,阳性率为9.2%,与IDEXXA marginale抗体检测试剂盒的阳性符合率为95.3%,阴性符合率为100%.本实验建立的ELISA方法具有较高的特异性和重复性,可用于流行病学调查研究.     

牛边缘无浆体病原学研究进展

边缘无浆体病是由边缘无浆体引起的一种严重危害牛养殖业的传染病。本文概述了边缘无浆体病原形态学、分类学、体外培养以及种系发生关系的研究现状,并总结了近年来对边缘无浆体主要表面蛋白（MSPs）的基因调控机理和病原表面蛋白的研究进展。     

某羊场和田羊与策勒黑羊无浆体季节性感染PCR检测

为了解和田羊和策勒黑羊无浆体季节动态感染情况,分别于2015年冬至、2016年春分、夏至和秋分,在策勒县某牧业合作社采集和田羊和策勒黑羊血液样本共120份,应用PCR对无浆体的MSP4基因和16SrRNA基因进行扩增。结果发现,检测绵羊的无浆体总阳性率为99.2%(119/120),以绵羊无浆体(Anaplasma ovis)和嗜吞噬无浆体(A.phagocytophilum)混合感染为主,占阳性样本比例为61.3%(73/119),A.ovis和A.phagocytophilum占阳性样本比例分别为31.1%(37/119)和7.6%(9/119)。在不同季节,和田羊和策勒黑羊无浆体均普遍感染,差异不显著(P>0.05)。结果显示,新疆策勒县某羊场绵羊的无浆体病全年均可流行,应引起人们重视。     

长沙地区奶牛无浆体动物感染试验及其传播途径调查

采集长沙地区某奶牛场疑似患病牛鲜血及牛舍周围吸血类昆虫(牛虻、厩蝇和蚊子等),分别作镜检观察,腹腔接种至试验小鼠体内(0.5 mL/只)进行培养和观察.为进一步证明其传播途径,将12只试验兔分别放置于牛舍不同方位饲养1周,进行临床感染试验.结果表明,病牛鲜血涂片染色后在血液红细胞内观察到呈圆点状或球状的无浆体虫体,绝大部分位于红细胞边缘,少数位于中央,染虫率达30%,患牛抗凝血在试验小鼠体内进行培养后,第4天开始在小鼠红细胞内查到虫体,第17天达到感染高峰;吸血类昆虫血液涂片、染色后,其中385片在血液红细胞内观察到虫体,虫体查出率达77%;吸血昆虫内容物在试验小鼠体内的培养情况大致与患牛抗凝血情况相同;在临床感染1周后的第6天,7只兔在血液红细胞中查到虫体,兔感染率达75%.     

利用西杂牛和秦川牛做受体进行奶牛胚胎移植的对比试验

[目的]为了选择奶牛胚胎移植的理想受体。[方法]通过利用不同品种的本地黄牛～西杂牛84头和秦川牛77头做受体,进行荷斯坦奶牛胚胎移植效果的对比试验。[结果]西杂牛的同期发情率、受体可用率、受胎率都显著高于秦川牛（P〈0．05）,而且难产率低,奶水充足。[结论]表明西杂牛是奶牛胚胎移植的理想受体。     

新疆褐牛与同龄牦牛放牧增重对比试验

对10头30月龄的新疆褐牛（（??）及牦牛（（?））,进行了128天暖季草原放牧增重对比试验。结果表明:新疆褐牛30月龄的体重为184.7 ±20.2kg,是同龄对照组牦牛的138％。36月龄的体重为253.6±32.5kg,是同龄对照组牦牛的154.7％。日增重538.4±157.5g,是同龄对照组牦牛的227％。两者之间差异极显著（P<0.01）。每头褐牛比牦牛多收入403.65元,经济效益明显。     

A Sero-Epidemiological Study of Haemophilus Somnus Infection in Dairy Cattle

Akhtar, S., Farver, T.B. and Riemann, H.P., 1997. A sero-epidemiological study of Haemophilus somnus infection in dairy cattle. Veterinary Research Communications, 21 (4), 229-239Five repeated cross-sectional serological surveys of 790 dairy cattle in 4 dairy herds between December 1985 and February 1987 provided an opportunity to study the changes in the seroprevalence of Haemophilus somnus across the 5 surveys and with respect to some demographic and disease variables. The demographic variables included were age (heifers or cows) and farm, representing two groups of herds (two herds in each group, located in the Central and Northern Valleys of California). The serological status of cattle as either negative or positive against H. somnus, Campylobacter fetus and Leptospira hardjo were determined with enzyme linked-immunosorbent assays. Logistic regression analysis was used to compute maximum likelihood estimates of adjusted odds ratios and their respective 95% confidence intervals.The baseline risk of being H. somnus seropositive in the cattle observed at first sampling did not vary significantly during the study period after adjustment for the effects of covariates. Only at first sampling were cows about twice as likely to be H. somnus seropositive than heifers. At samplings 1 and 3, but not at 2 and 5, being in the herds of the Central Valley appeared protective. In contrast, at sampling 4 the cattle in herds in the Central Valley were about 7 times more likely to be H. somnus seropositive. C. fetus-positive cattle were about 3 times more likely to be H. somnus seropositive at sampling 1 only. The relationship between H. somnus status and L. hardjo was not significant during the study period.     

牛结核病两种活体诊断方法检测结合病原学诊断的研究

本文报道应用两种活体诊断方法检测牛结核病的试验结果。2010～2011年度,应用PPD皮内变态反应方法分3批次对5156头奶牛和奶水牛实施监测,监测阳性数57头,阳性率1.11%。采集该57头牛抗凝全血,应用γ-干扰素酶联免疫吸附试验进行检测,检出阳性样品3份,阳性率5.26%。对14头第一批PPD皮内变态反应阳性牛进行病理检验和进行细菌分离培养和PCR检测,结果3份样品分离培养呈阳性,其中1份PCR鉴定为结核分枝杆菌,2份为非分枝杆菌。PCR方法检测的14份组织样品中,2份为结核分枝杆菌阳性,1份为牛结核分枝杆菌阳性。结合病理剖检和病原PCR诊断,分析比较PPD皮内变态反应试验和γ-干扰素酶联免疫吸附试验的敏感性和特异性,由于PPD纯度、非特异性反应、结果判定的主观性等因素,导致PPD皮内变态反应监测检出假阳性率较高。     

海晏县沼泽草场牛羊放牧情况及同盘吸虫感染调查

为了解海晏县沼泽草场放牧牛羊及同盘吸虫感染等情况而进行了本调查,结果为：沼泽草场放牧户占到全县总放牧户数的9.27%,放牧牛羊分别占到全县牛羊总数的10.93%和12.65%。77.78%的沼泽草场放牧户牛羊感染同盘吸虫,而牛群和羊群同盘吸虫感染率分别达到了59.26%和48.15%;牛羊群体混合感染率40.74%;牛羊个体同盘吸虫感染率分别为14.83%和17.04%。结果还表明专业技术人员等对同盘吸虫病了解不足,区域间感染率差别较大,牛羊感染率有上升的趋势,证明现在使用的驱除药物对同盘吸虫虫卵阳性率降低很不理想,今后对本病的预防知识普及宣传及技术培训、驱除药物研制和筛选、湿地椎实螺的防治十分迫切。     

牛奶中恩诺沙星和环丙沙星检测试纸的研制及性能测定

应用胶体金免疫层析技术建立了恩诺沙星和环丙沙星快速检测试纸,并对试纸条的检测限、特异性和稳定性等参数进行了确定。结果表明,该试纸条对牛奶样本的检测限为50μg/L,能够特异性地检测恩诺沙星和环丙沙星,与液相色谱-串联质谱方法检测的阴性样本符合率为97．8％。该试纸条在2℃～8℃或室温可保存12个月,可应用于恩诺沙星在牛奶中的快速筛查现场检测。     

Serological Diagnosis of Classical Swine Fever: A Comparison of a Modified Direct Complement Fixation Test with an Immunofluorescence Plaque Neutralization Test in the Diagnosis of Experimental Subclinical Infection

Antibody levels in post-infection sera from a pig inoculated with a low virulent strain of classical swine fever virus (Hannover 62) and in sera from two pigs inoculated with another low virulent strain (Spielbach 66) and from an in-contact pig were assayed by complement fixation and immunofluorescence using classical swine fever virus (ALD strain) and bovine virus diarrhoea virus (UG 59 strain) as antigens. The complement fixation test used was modified by addition of a preparation of porcine Glq to the complement and by mercaptoethanol treatment of the immune serum before use. The mercaptoethanol treatment of the immune serum resulted in complete elimination of a haemolytic prozone often seen with porcine immune sera.In the sera from the inoculated animals complement-fixing antibodies appeared earlier than neutralizing antibodies. A few weeks after inoculation there was a correlation between the presence of complement-fixing and neutralizing antibodies.During the entire observation period of 13 weeks it was not possible to demonstrate complement-fixing or neutralizing antibodies in serum from a pig exposed to infection by contact with the two pigs inoculated with the Spièlbach 66 strain of classical swine fever virus.     

Seroprevalence Investigation and Risk Factors Analysis of Chlamydia Infection in Dairy Cattle in Gansu and Ningxia Areas

The objective of the present investigation was to examine seroprevalence of Chlamydia in dairy cattle in Gansu and Ningxia areas, Northwest China, and to analyze the factors affecting Chlamydia infection in dairy cattle.Indirect hemagglutination assay (IHA) was used to detect antibodies against Chlamydia in 1 657 dairy cattle serum samples from Gansu and Ningxia areas.Epidemiological investigation and statistical methods were used to analyze the data.The results showed that the overall seroprevalence was 29.33%, logistic regression analysis was applied to evaluate the risk factors of Chlamydia infection in dairy cattle, and the results indicated that age and numbers of pregnancy of dairy cattle were not the significant risk factors (P>0.05), and were left out of the final model, however, region was considered as the main risk factor associated with Chlamydia infection (P<0.05).The highest titer was 1:1 024.In conclusion, the results of the present survey indicated the widespread of Chlamydia infection in Gansu and Ningxia areas.In order to ensure the economic benefit of dairy farming, we should pay more attention to Chlamydia infection in dairy cattle, and integrated control strategies and efficient management measures should be implemented to prevent and control Chlamydia infection in dairy cattle.