陇西县牛巴贝斯虫病的流行病学调查

将重组的牛巴贝斯虫棒状体相关蛋白(Bo-RAP-1)作为诊断抗原建立ELISA检测方法,对2016—2017年采自甘肃省陇西县的188份黄牛血清样品进行抗Bo-RAP-1特异性抗体检测。结果检出47份阳性,阳性率为25%,说明该地区牛群存在牛巴贝斯虫病的感染。     

识别牛巴贝斯梨形虫的特异性DNA探针

用质粒PuN121从墨西哥虫株M建立牛巴贝斯梨形虫DNA的基因族,并克隆于大肠杆菌上,选择几个与标记的牛巴贝斯梨形虫基因DNA杂交的重组质粒,进一步分析,发现PMu—B_1敏感性较高,能检测出25pg纯净的牛巴贝斯梨形虫DNA。10μl感染全血中300个梨形虫或0.00025％虫血症,PMu—B_1含有6.0kb牛巴贝斯梨形虫DNA插人物,该插人物不与双芽巴贝斯梨形虫、伊氏锥虫、恶性疟原虫、边缘边虫、微小牛蜱和奶牛DNA发生交叉反应,基因组DNA Southem印迹析,PMu—B_1能识别两种牛巴贝斯梨形虫地方株,即墨西哥虫株M和泰国TS_4株,因此,PMu—B_1探针可用于诊断牛和蜱巴贝斯梨形虫感染及识别牛巴贝斯梨形虫虫株。     

牛巴贝斯虫病的流行病学调查分析

牛巴贝斯虫病是媒介蜱传播的梨形虫纲巴贝斯科巴贝斯属的多种病原体寄生在动物红细胞、网状内皮细胞而所发的血液原虫病,牛巴贝斯虫病会对养牛场造成巨大的经济损失,其发病症状主要以血红蛋白尿、贫血、发热为主,由于当前对牛巴贝斯虫病的预防还没有比较好的实用方法,因此,加强牛巴贝斯虫病初期感染诊断有十分重要的作用。文章以某地区牛巴贝斯虫病感染情况为例,采用牛巴贝斯虫病间接ELISA检测方法,对牛巴贝斯虫病流行病学进行调查分析。     

牛巴贝斯虫TaqMan荧光定量PCR检测方法的建立

为建立一种灵敏、特异、快速的牛巴贝斯虫检测方法,针对牛巴贝斯虫Rap-1a基因设计引物进行PCR扩增,然后构建重组质粒制作标准品,经过优化反应体系、绘制标准曲线,建立了牛巴贝斯虫TaqMan荧光定量PCR方法,并进行灵敏度、特异性及稳定性检测,同时利用该方法对37份田间样品进行检测。结果显示：建立的牛巴贝斯虫TaqMan荧光定量PCR检测方法的标准曲线方程式为y=-3.362×Log（X）+43.32,相关系数R2=0.999,扩增效率为98.4%。该方法的灵敏度为1.0×102 copies/μL,是普通PCR（1.0×104 copies/μL）的100倍。该方法对牛双芽巴贝斯虫、大巴贝斯虫、卵形巴贝斯虫等7种常见的牛梨形虫病检测结果均为阴性,组内和组间重复试验的变异系数均小于2.5%,37份田间样品的阳性检出率为67.5%。结果表明,本试验建立的牛巴贝斯虫TaqMan荧光定量PCR灵敏度高,且特异、稳定,适用于牛巴贝斯虫的诊断,从而为其流行病学调查提供了快速有效的检测方法。     

一起牛双芽巴贝斯虫病的诊治

牛巴贝斯虫病是由巴贝斯虫属的原虫寄生于牛的血细胞和网状内皮系统引起的寄生虫病。临床主要特征为高热、贫血、黄疸和血红蛋白尿。在我国，引起牛巴贝斯虫病的病原体有3种，即双芽巴贝斯虫、牛巴贝斯虫和卵形巴贝斯虫。该病的发生呈一定的地区性、季节性。2009年7月23日我县某养牛户饲养黑白花母牛9头，其中4头突然发病，1头死亡，现将诊疗情况报告如下。     

奶牛巴贝斯虫病的诊断与治疗

牛巴贝斯虫病是由巴贝斯属（Babesia）的多种寄生虫寄生于牛红细胞内所引起的血液原虫病。我国已报道的牛巴贝斯虫有3个种：双芽巴贝斯虫（B．bigemina）、牛巴贝斯虫（B．bovis）和卵形巴贝斯虫（B．ovata）。前2个种流行广泛,传播媒介为微小牛蜱和镰形扇头蜱,危害较大;后1个种只在河南局部地区发现,传播媒介为长角血蜱,危害较小。     

水牛巴贝斯虫病病原—牛巴贝斯虫体外培养的研究

本研究采用微气静相培养技术,对一株采自人工感染的去脾脏水牛犊的牛巴贝斯虫(Babesia bovis)进行了80天的连续体外培养·继代26次,累积增殖6.51×10~((?))倍;累积稀释2.19×10~(35)倍。培养24小时红细胞的染虫率为2.63±0.50％;48小时为7.18±1.39％;72小时为20.78±4 52％。最高红细胞染虫率达33.50％。体外培养的牛巴贝斯虫与采自病牛血液内的牛巴贝斯虫形态一致,说明已建立了水牛巴贝斯虫病病原——牛巴贝斯虫的体外培养。 对影响牛巴贝斯虫体外培养的几种因素进行的实验结果表明:培养液pH值显著地影响虫体的繁殖,最适的体外连续培养的pH为7.2;合适的血清浓度为40％;血清灭活后,支持牛巴贝斯虫生长繁殖的能力明显降低;脱纤血分离的血清和自然凝固血析出的血清用于培养的效果相同;RPMI-1640和TCM-199培养液支持牛巴贝斯虫生长繁殖的能力没有差异。     

牛巴贝斯虫病

牛巴贝斯虫病旧称为牛焦虫病,是由数种巴贝斯虫引起的一种需经硬蜱传播的牛的血液原虫病,以急性型为多见。     

新疆和静县和吐鲁番地区牛感染巴贝斯虫病的血清学调查

为了解双芽巴贝斯虫和牛巴贝斯虫在新疆疫区牛感染的状况,从吐鲁番市周边散养户、和静县部分散养户采集了273份牛(牦牛)血清。采用间接ELISA方法对血清样本进行牛巴贝斯虫和双芽巴贝斯虫抗体检测。结果显示:和静县部分散养户被检牦牛血清抗牛巴贝斯虫(B.bovis)抗体阳性率为18.68%(17/91);被检牦牛血清抗双芽巴贝斯虫(B.bigemina)抗体阳性率为9.89%(9/91)。吐鲁番市周边散养户被检奶牛血清抗牛巴贝斯虫(B.bovis)抗体阳性率为15.38%(28/182);被检奶牛血清抗双芽巴贝斯虫(B.bigemina)抗体阳性率为9.34%(17/182)。通过调查发现牛巴贝斯虫和双芽巴贝斯虫均有混合感染的现象,其中和静牦牛混合感染率为6.59%(6/91);吐鲁番奶牛混合感染率为8.24%(15/182);不同品种牛均可感染牛巴贝斯虫和双芽巴贝斯虫,其感染程度及感染率具有一定的差异。本次试验结果可为有效防治新疆疫区牛梨形虫病提供重要依据。     

牛巴贝斯虫lytB基因的克隆与序列分析

为了对牛巴贝斯虫lytB基因进行克隆与序列分析,试验以兰州株牛巴贝斯虫lyt B基因组为模板进行PCR扩增,将扩增得到的特异性产物克隆到p GEM-T Easy载体上,并对其进行PCR检测、酶切鉴定及序列测定分析。结果表明:试验克隆得到的864 bp基因片段与Gen Bank收录的牛巴贝斯虫lyt B核苷酸序列同源性为97.8%。说明试验成功克隆出牛巴贝斯虫lyt B基因,与Gen Bank收录的牛巴贝斯虫lyt B核苷酸序列具有高度的同源性。     

青海省乌兰县牧羊犬犬新孢子虫病的血清学调查

用犬新孢子虫的重组蛋白GST-NcSAG1t作为ELISA诊断抗原,对青海省乌兰县的部分牧羊犬进行了犬新孢子虫病的血清学调查。经过对所收集的80份牧羊犬血清的检测,共检出犬新孢子虫病阳性血清25份,阳性率为31.25%。结果表明青海省乌兰地区的牧羊犬中存在犬新孢子虫感染。     

牦牛病毒性腹泻/黏膜病和传染性鼻气管炎血清学调查

为掌握青海省大通种牛场和海晏县牦牛病毒性腹泻/黏膜病、传染性鼻气管炎的感染和流行情况,2010年3月至5月在大通种牛场和海晏地区采集252份牦牛血清样品,应用定量酶联免疫吸附试验（ELISA）对牦牛病毒性腹泻/黏膜病和传染性鼻气管炎的进行了血清抗体检测。结果显示,大通种牛场牦牛群中牛病毒性腹泻/黏膜病阳性率为23.42%,传染性鼻气管炎阳性率为65.45%;海晏县牦牛群中牛病毒性腹泻/黏膜病阳性率为19.86%,传染性鼻气管炎阳性率为4.96%。     

Application of an indirect ELISA to milk samples to identify cows with Mycoplasma bovis mastitis

An indirect ELISA was used to detect antibodies to Mycoplasma bovis in milk samples collected from a herd with M bovis mastitis. Antibodies were detected in samples from nine cows which had developed clinical M bovis mastitis. Milk from only three consistently antigen-negative cows tested positive for M bovis antibodies. These results indicate the potential value of the indirect ELISA for the detection of cows which have recently developed M bovis mastitis during the early stages of an outbreak.     

Serological survey of Babesia bovis and Babesia bigemina in cattle in South Africa

A total of 719 serum samples collected from clinically healthy cattle from eight provinces located in different districts of South Africa were examined by the indirect enzyme-linked immunosorbent assay (ELISA) and the standard indirect fluorescent antibody test (IFAT) to determine the serological prevalence of Babesia bovis and Babesia bigemina. The results showed that 35.3% and 39.7% of cattle were positive for B. bovis and 30% and 36.5% were positive for B. bigemina antibodies on ELISA and IFAT, respectively. Mixed infections were detected in 18.2% and 26.3% of the samples using ELISA and IFAT, respectively. Consequently, the ELISAs with recombinant B. bovis spherical body protein-4 (BbSBP-4) and B. bigemina C-terminal rhoptry-associated protein-1 (BbigRAP-1/CT) were proven to be highly reliable in the serological diagnoses of bovine babesiosis in South African cattle, as evidenced by the significant concordance rates when the results were compared to those of IFAT. Moreover, the serological prevalence was significantly different among the tested provinces, in which the ranges exhibited between 15% and 73% for B. bovis infection and between 13% and 54% for B. bigemina infection. High sero-positive rates were present in Mpumalanga and KwaZulu-Natal provinces, while the lowest rate was in the North West province. Our data provide important information regarding the current seroprevalence of bovine babesiosis in South Africa, which might be beneficial in developing rational strategies for disease control and management.     

A serological survey of bovine babesiosis in northern and eastern Zimbabwe

The geographical distribution of Babesia bovis and Babesia bigemina antibodies in communal herds in northern and eastern Zimbabwe was determined using the ELISA technique. The animals in different herds in the study region had different levels of natural exposure to B. bovis (mean 32%, range 0-79%) and B. bigemina (mean 52%, range 5-92%) infections. The majority of herds (90%) were endemically unstable for B. bigemina and 62% were unstable for B. bovis. Natural region 5 and Manicaland province had the highest seroprevalence of B. bovis infection, while natural region 5 and Masvingo province had the highest seroprevalence of B. bigemina infection.     

喜马拉雅旱獭弓形虫病血清学检测

用弓形虫的重组蛋白GST-SAG1作为ELISA诊断抗原,采用ELISA检测方法,对青海省祁连县喜马拉雅旱獭血清进行了弓形虫病的血清学检测。结果表明,被检92份喜马拉雅旱獭共检出阳性血清25份,阳性率约为27.17%。结果提示,青海省祁连县的喜马拉雅旱獭中存在弓形虫病的感染。     

Babesia spp. infection in Boophilus microplus engorged females and eggs in Sao Paulo State, Brazil

Babesia spp. infections were investigated in Bos taurus x Bos indicus dairy cows and calves and in Boophilus microplus engorged female ticks and eggs. Blood samples and engorged female ticks were collected from 25 cows and 27 calves. Babesia spp. was detected in ticks by microscopic examination of hemolymph of engorged female and by squashes of egg samples. Cattle infection was investigated in blood thin smears and by DNA amplification methods (PCR and nested PCR), using specific primers for Babesia bovis and Babesia bigemina. Merozoites of B. bovis (3 animals) and B. bigemina (12 animals) were detected exclusively in blood smears of calves. DNA amplification methods revealed that the frequency of B. bigemina infection in calves (92.6%) and in cows (84%) and of B. bovis in calves (85.2%) and in cows (100%) did not differ significantly (P > 0.05). Babesia spp. infection was more frequent in female ticks and eggs collected from calves (P < 0.01) than from cows, especially in those which had patent parasitemia. Hatching rates of B. microplus larvae were assessed according to the origin of engorged females, parasitemia of the vertebrate host, frequency and intensity of infection in engorged female tick, and frequency of egg infection. Hatching rate was lower in samples collected from calves (P < 0.01) than from cows, and in those in which Babesia spp. was detected in egg samples (P < 0.01).     

甘肃棘豆内生真菌种群多样性

将采集于青海省祁连县甘肃棘豆的茎和叶部表面消毒后分别置于黑暗和光照条件以及PDA和BDA 2种培养基上进行内生真菌分离纯化,采用形态学和分子生物学分析技术鉴定种属,并计算多样性指数。结果显示,629块植物样品中共分离到内生真菌433株,分属9个科15个属28种,不同的培养条件下优势菌属不同;多样性指数计算结果显示,香农-维纳指数（H）为1.99,均匀度指数（E）为0.60,辛普森指数（D）为0.257。结果表明,采集于青海省祁连县的甘肃棘豆内生真菌多样性比较丰富,可为今后深入探讨甘肃棘豆内生真菌遗传多样性奠定基础,并为产苦马豆素目标菌株的筛选,提供丰富的内生真菌后备菌株。     

Prevalence and molecular detection of Anaplasma marginale, Babesia bovis and Babesia bigemina in cattle from Puntarenas Province, Costa Rica

A study was conducted in 2008 to determine the prevalence of Anaplasma and Babesia infections in cattle in the Puntarenas Province of Costa Rica. Blood samples were taken from a total of 449 cattle during the month of March at 30 farms in the region of Espiritu Santu, Costa Rica. Commercially available enzyme-linked immunosorbent assays (ELISA) were used to determine presence of antibodies to Babesia bigemina and Anaplasma marginale, and real-time PCR was used to determine the presence of DNA from the disease-causing organisms. The ELISA results indicated that 87.5% of the cattle sampled were positive for antibodies to A. marginale, while 59.1% were positive for antibodies to B. bigemina. The real-time PCR results showed that 235 cattle were carrying A. marginale DNA (56.9%), 6 with B. bigemina DNA (1.34%), and 2 with B. bovis DNA (0.45%).     

Cross-sectional estimation of Babesia bovis antibody prevalence in cattle in two contrasting dairying areas in Tanzania

The crude prevalence of antibodies to Babesia bovis infection in cattle was estimated by serology using indirect ELISA during the period January to April, 1999. Sera were obtained from 1,395 dairy cattle (of all ages, sexes and breeds) on smallholder farms, the majority being kept under a zero grazing regime. The crude prevalence of antibodies to Babesia bovis was 6 % for Tanga and 12 % for Iringa. The forces of infection based on the age sero-prevalence profile, were estimated at six for Iringa and four for Tanga per 100 cattle years-risk, respectively. Using random effect logistic regression as the analytical method, the factors (variables) of age, source of animals and geographic location were hypothesised to be associated with sero-positivity of Babesia bovis in the two regions.