Genetic Heterogeneity of Bovine Viral Diarrhoea Virus In Italy

The genetic characteristics, of 38 field isolates of bovine viral diarrhoea virus (BVDV) collected in 1999 from sick or healthy and persistently infected cattle of dairy farms situated in northern Italy, were investigated. A partial 5-untranslated region (5-UTR) sequence of each isolate was determined and a phylogenetic analysis was performed. All the isolates were classified as belonging to the BVDV-1 genotype and could be assigned to different BVDV-1 groups, namely BVDV-1b (n = 20), BVDV-1d (n = 6) and BVDV-1e (n = 10). Two remaining isolates could be classified as BVDV-1f and BVDV-1h, respectively. These results provided evidence for genetic heterogeneity of BVDV in Italy, and contribute to a better knowledge of the circulation of BVDV strains, and to their classification.     

A Universal `One-tube' RT-PCR Protocol for Amplifying Isolates of Bovine Viral Diarrhoea Virus

The increase in the knowledge of the genetic variability of BVDV and the identification of some of the genetic determinants of its pathogenicity require robust and practical tools for rapid molecular characterization of the various genotypes of this virus. This study was undertaken to develop a standard protocol for RT-PCR that allows the amplification of various parts of the genome of BVDV without the need for optimizing each individual reaction. The reaction set-up is very flexible because it consists of two pre-mixes. These are a master mix, with all the required reagents except the desired primers, which are the components of the second pre-mix and are therefore easily interchangeable between the different reactions. After adding any primer-containing pre-mix to the fixed master mix, a non-interrupted cycling protocol led to the generation of amplicons of up to 4 kbp in size in amounts sufficient for subsequent sequencing reactions. The method was applied to five different regions of the BVDV genome: (i) the well-known 5-UTR to differentiate genotypes I and II; (ii) the entire E2 gene, or an approximately 550 bp region within the E2 gene, in order to find the molecular equivalent of antigenic varieties; (iii) the entire structural protein coding region covering the Npro, capsid, E ^RNS, E1 and E2 genes; (iv) a 2.1 kbp region embracing the NS2/3 junction which is known to be cleaved in cytopathic biotypes of BVDV; and (v) the region covering the entire NS4B and NS5A/B genes. All six RT-PCRs were successfully applied using (i) primers with lengths of between 20 and 52 nucleotides, (ii) an aliquot of RNA extracted from either 10⁶ infected bovine embryonal lung cells or the same number of leukocytes from viraemic cattle, and (iii) all the genotype I and II strains of BVDV tested. The technique described was used to generate various Sindbis virus/BVDV recombinants. The correct processing of the amplicon-derived E2 glycoprotein of BVDV strain PT810 was demonstrated by its reaction with a monoclonal antibody in an immunofluorescence assay. Given the variety of RT-PCRs tested, we conclude that this universal protocol may be useful with other RNA viruses.     

Experimental Infection of Calves with Bovine Viral Diarrhoea Virus Type-2 (BVDV-2) Isolated from a Contaminated Vaccine

A non-cytopathic strain of BVDV-2 was isolated from a batch of live infectious bovine rhinotracheitis (IBR) vaccine, and inoculated intranasally into four 3-month-old calves. Severe signs of disease developed by days 4 and 6 in three of the calves, free of BVDV and antibodies to BVDV, that had been exposed to the virus. These calves survived the acute phase of the infection and progressively recovered. BVDV was consistently isolated, or the respective viral RNA was detected, in the buffy coats from blood samples collected starting from days 2 or 4 up to days 11 or 14 after the experimental infection. Viral RNA was also detected in sera from these infected calves until the presence in the serum of virus neutralizing antibodies was demonstrated. By contrast, the only calf having pre-existing neutralizing antibodies to BVDV at the start of the study was protected from the disease. No virus was detected at any time after experimental inoculation of this calf. Genomic characterization of the BVDV-2 isolated in cell cultures, or detected in sera from the experimentally infected animals, revealed 100% homology in the nucleotide sequence with the BVDV-2 detected as a contaminant of the live IBR virus vaccine. These findings provided evidence of the infective nature of the contaminant BVDV-2 and of its potential to generate disease outbreaks when inoculated into susceptible animals.     

Identification of Papillomaviruses in Scrapings from Bovine Warts by use of the Polymerase Chain Reaction

Bloch, N., Sutton, R.H., Breen, M. and Spradbrow, P.B., 1997. Identification of papillomaviruses in scrapings from bovine warts by use of the polymerase chain reaction. Veterinary Research Communications, 21 (1), 63-68.     

吉林某牛场牛病毒性腹泻流行病学调查及分析

为全面了解吉林省某规模化牛场牛病毒性腹泻(BVD)的流行情况,试验采集临床血清样品157份,粪便样品18份,肝脏、精液等组织样品17份,应用BVDV抗体检测试剂盒进行血清抗体检测,利用BVDV1型引物,采用纳米PCR方法对血清及临床样品进行BVDV抗原检测,对抗原阳性样品进行测序分析;将抗原阳性样品经研磨、稀释后用0.22μm滤膜过滤除菌,接入牛肾细胞(MDBK)进行病毒分离培养,盲传3代后再次进行抗原检测;采用免疫荧光技术检测病毒对MDBK细胞的侵染作用;利用Mega软件绘制系统进化树并进行同源性比对分析。结果显示,该牛场临床血清BVDV抗体阳性率为77.1%,血清抗原阳性率为12.1%,临床粪便等样品抗原阳性率为74.3%;病料接入细胞未观察到细胞病变,分离毒株PCR产物测序分析结果与样品抗原检测结果一致;免疫荧光检测结果显示,分离株毒液正常吸附于MDBK细胞中,有明显荧光反应;抗原测序分析显示,该牛场BVD主要流行毒株与BVDV JL-1株同源性高达99.0%,且均为BVDV 1型毒株。本研究对该牛场BVDV流行情况进行了全面调查,为开展净化工作奠定了基础。     

Possible Causes of Diabetes Mellitus in Cattle Infected with Bovine Viral Diarrhoea Virus

In cattle, we encountered insulin-dependent diabetes mellitus (IDDM) associated with bovine viral diarrhoea virus (BVDV) infection. To estimate the correlation between IDDM and BVDV infection, the distribution of BVDV in the pancreas and islet-cell antibody (ICA) were investigated. The distribution of BVDV in the pancreas was examined by in situ hybridization using two oligonucleotide probes that recognized the gp25- and p14-coding regions of the BVDV gene. ICA was examined by indirect fluorescence antibody assay using the sera from affected cattle and pancreata from normal cattle. In the pancreata of all BVDV-infected cattle, including IDDM-complicated cattle, oligonucleotide probe hybridized portions were recognized. In short, BVDV genes were detected not only in IDDM-complicated cattle but also in uncomplicated cattle. Moreover, there was no hybridized portion in the islet cells. In BVDV-infected and IDDM-complicated cattle, ICA was frequently detected. On the other hand, ICA was not detected in BVDV-infected and IDDM uncomplicated cattle. These results suggest that IDDM associated with BVDV infection is not a direct effect of BVDV on islet cells. Therefore, as BVDV did not induce IDDM in any cases, it appears that BVDV does not induce IDDM directly, but rather may be an autoimmune disease induced by autoantibodies against islet cells.     

山东种公猪精液携带主要病原的检测

采用PCR技术,对2007年-2009年山东部分地区的38个猪场和人工授精站的生产公猪精液样品727份进行了猪瘟病毒(CSFV)、猪繁殖与呼吸综合征病毒(PRRSV)、伪狂犬病病毒(PRV)、猪圆环病毒2型(PCV-2)和猪细小病毒(PPV)5种主要病原的检测。结果检出CSFV、PRRSV、PRV、PCV-2和PPV的阳性数和阳性率分别为18份(2.48%)、27份(3.71%)、7份(0.96%)、33份(4.54%)、9份(1.24%),有7份样品为2种以上病原混合感染,其中以PRRSV+PCV-2混合感染最多。结果表明精液传播病毒仍是当前母猪繁殖障碍的重要原因之一,应重点加强对种公猪的疫病净化和公猪精液管理,从源头控制传染源。     

西门塔尔牛BVD预防和治疗试验

肉牛的呼吸道疾病(BVD)多因运输应激而引起的一种顽固性疾病,常因临床治疗效果不佳,饲养成本增加。为降低本病给牧场造成 损失,自2020 年03 月26 日起,针对华润集团开鲁隔离场 牧场购自伊胡塔交易市场的西门塔尔品种架子牛,使用德国勃林格公司育肥牛到场操作流程方案进行应激处理并跟踪各项数据,与单纯使用勃林格公司驱虫药害获灭做对比评估,通过药物成本、死淘情况、增重指标的对比来评估方案优劣及成本核算分析,结果显示：(1)同样品种的西门塔尔育肥牛,在采购进入育肥场前后,进行不同应激处理方案,相比较只做害获灭+FMD 疫苗处理的牛群,使用专克灵+害获灭+FMD 疫苗处理的牛群,平均每头牛的日增重差别可以达到0.3kg/d,根据目前市场行情条件下,育肥牛销售约32 元/kg 的价格,在入场30 日内,平均每头牛可以多获得239.6 元的更多增重育肥收入;(2)在入场后使用专克灵预防BRD,配合害获灭驱虫,相比较入场后只做驱虫处理,能获得更低的BRD 发病率(低28.7%),发病牛更短的治疗天数(少0.5 d),更佳的平均日增重(多300 g),能够带来更好的增重收益;(3)实验组与对照组差别为0.3kg即平均日增重差别300g 。该数据可以直观的说明,入场时,是否全群专克灵处理牛群,预防BRD,对于后续日增重的影响,差异极其明显。本试验为华润集团隔离场领导及牧场技术人员参考。     

Prevalence of Bovine Viral Diarrhoea Virus antibodies among the industrial dairy cattle herds in suburb of Mashhad-Iran

Mashhad is a major dairy production in Iran. The subject of this study was to survey the seroprevalence of Bovine Viral Diarrhea Virus (BVDV) infection using an indirect Enzyme-linked immunosorbent assay (ELISA) test in industrial dairy cattle herds in suburb of Mashhad-Iran. Totally, 141 serum samples were tested. None of the herds had been vaccinated against BVDV. Commercial indirect ELISA kit was used. The herds divided to 3 sizes as cow population. They were included: small, medium and large herds. Data were analyzed using Chi-square test. Ninety-seven (68.79%) cows were ELISA seropositive. However, the true BVDV seroprevalence was 72.25%. All of the herds were antibody positive against BVDV. The prevalence ranged from 66 to 100% within the herds. There were no significant differences between the presence of antibodies to BVDV and the herd size (P > 0.05). The prevalence in animals lower than 2 years old differed significantly with cows higher than 2 years old (P < 0.05). According to the results, it is concluded that it is likely the presence of persistently infection (PI) animal(s) within the herds in suburb of Mashhad-Iran, which is responsible for the presence antibody.     

云南省牛病毒性腹泻-黏膜病血清流行病学调查

[目的]牛病毒性腹泻—黏膜病(bovine viral diarrhea-mucosal disease, BVD)引起牛腹泻、呼吸系统疾病、生殖功能障碍、免疫抑制、母畜流产、死胎等,对牛产业发展危害严重。牛病毒性腹泻—黏膜病病毒(bovine viral diarrhea-mucosal disease virus, BVDV)在我国大部分牛群中普遍存在,各地流行情况严重程度不同,本调查旨在调查云南省BVDV的流行情况。[方法]调查通过采用商品化的酶联免疫吸附试验(ELISA)试剂盒对BVDV感染和流行情况进行调查,采集了云南省4个州10个县(市)的部分规模牛场及散养户的牛血清样品456份。[结果]结果发现,云南省不同地区所检的牛血清样品总抗体阳性率为16.45%(75/456),抗原阳性率为0.53%(2/378),其中抗体阳性率以大理州洱源县60.0%(9/15)最高,而丽江市华坪县的血清样品中未检测到抗体阳性,大理州宾川县和丽江市宁蒗县的抗原阳性率较低,分别为7.69%(1/13)和4.00%(1/25)。云南省规模场牛的血清样品抗体总阳性率为27.27%(45/165),抗原阳性率为1.15%(1/87),散养户牛的分别为10.31%(30/291)和0.34%(1/291)。[结论]云南省的牛群中存在着BVDV感染的情况,其抗体抗原的阳性率水平跟地区有着紧密的联系,不同的养殖方式抗原抗体水平存在着一定差异,必须要加强云南省BVDV监测与防控,减少其造成的经济损失。     

PCR方法检测奶牛粪便中鼠隐孢子虫

从含有鼠隐孢子虫卵囊的奶牛粪便中,直接提取 Ｄ Ｎ Ａ 用作 Ｐ Ｃ Ｒ 模板,用 １ 对人工合成的寡核苷酸作为 Ｐ Ｃ Ｒ 引物,扩增大小为 ５４０ ｂｐ 的特异片段。 Ｐ Ｃ Ｒ 产物经电泳鉴定,表明可从含隐孢子卵囊的奶牛粪便标本 Ｄ Ｎ Ａ 抽提物中扩增出目的片段,而其他几种寄生虫及阴性对照均不能扩增出特异片段。本方法的敏感性最低可检测到含卵囊 ４００ 个／ｍ Ｌ 的样本,具有敏感性高、特异性强的特点。     

Characterization of Newcastle Disease Viruses Isolated from Chickens and Ducks in Tamilnadu,India

During 1993, outbreaks of Newcastle disease occurred on many farms in Tamilnadu, India. Six Newcastle disease virus (NDV) isolates were obtained from the chickens on five different farms and from the birds on one duck farm during outbreaks of the disease. All the isolates were characterized as velogenic, based on the mean death time, intravenous pathogenicity index, intracerebral pathogenicity index (ICPI), stability of haemagglutinin at 56°C, agglutination of equine erythrocytes, haemagglutination elution pattern and adsorption of haemagglutinin by chick brain cells. The isolate obtained from ducks resembled a group D strain, based on its ICPI and its reaction with a panel of monoclonal antibodies. The other five NDV isolates obtained from chickens were placed in groups B(1), C1(2) and D(2) on the basis of their binding patterns with the panel of monoclonal antibodies. In challenge experiments, it was found that LaSota vaccine provided 100% protection against each of these field isolates and against a local NDV strain obtained from the Institute of Veterinary Preventive Medicine, Tamilnadu, India, while unvaccinated chickens succumbed to challenge. The possible origin of epizootic viruses causing outbreaks in vaccinated flocks is discussed.     

Amplification of Bovine Viral Diarrhoea Virus Introduced into an In Vitro Embryo Production System Via Oocytes from Persistently Infected Cattle

The purpose of this study was to determine whether or not embryos derived from in vitro fertilization of oocytes from persistently infected (PI) cattle would contain infectious virus. Three in vitro embryo production treatment groups were assessed: 1) oocytes and uterine tubal cells (UTC) free of bovine viral diarrhoea virus (BVDV) (negative control), 2) oocytes free of BVDV fertilized and cultured in media containing UTC obtained from PI heifers, and 3) oocytes from PI heifers fertilized and cultured in media containing UTC free of BVDV. The developmental media, UTC and embryos (individual or groups of five) were assayed for virus. Virus was not isolated from any samples in treatment group 1. As shown in previous studies, a proportion of embryo samples were positive for BVDV in treatment group 2. In treatment group 3, the virus associated with the oocytes contaminated the developmental media and infected susceptible co-culture cells used during fertilization and culture. In addition, 65% (11/17) of the degenerated ova from treatment group 3 had infectious virus associated with them. While none of the ova developed into transferable embryos, the study did confirm that use of oocytes from PI cows could lead to amplification of BVDV and cross contamination during in vitro embryo production.     

Comparison of the Prevalence and Incidence of Infection with Bovine Virus Diarrhoea Virus (BVDV) in Denmark and Michigan and Association with Possible Risk Factors

Based on 2 previous surveys on the occurrence of infection with bovine virus diarrhoea virus (BVDV) in Danish and Michigan dairy herds, the prevalence and incidence of the infection were compared. The presence of certain possible risk factors for the occurrence of infection in the 2 areas were summarized and it was investigated if any of these risk factors had significant effect on the presence of animals persistently infected (PI) with BVDV in the dairy herds. Information on the cattle population density in the 2 areas was obtained from statistical yearbooks. Further information for the individual farms on age distribution, housing of animals, herd size, pasturing and purchasing policy was gathered. The prevalence of PI animals was more than 10 times higher in Denmark as compared to Michigan. In herds without PI animals, the annual incidence of seroconversion as calculated from the age specific prevalence of antibody carriers varied in most age groups between 20–25% in Denmark and between 5–10% in Michigan. All investigated risk factors except for herd size were in favour of a lower prevalence of infection in Michigan. The use of having animals on pasture and at the same time having purchased more than 40 animals within recent 31/2–4 years were significantly associated with presence of PI animals in the dairy herds (p = 0.01) when tested by the Mantel-Haenszel χ². Using mul-tivariable logistic regression, the occurrence of PI animals was found to be significantly related to the study area (Michigan and Denmark) as well as to herd size and purchase intensity.     

江苏某奶牛场健康奶牛体内产志贺毒素大肠杆菌的分离鉴定及其对小鼠致病性的研究

通过对江苏省某奶牛场连续6个月的定群、定畜跟踪调查,获得产志贺毒素大肠杆菌(STEC)在该牛场分布的广泛性、持续性和血清型多样化的资料,并对一些重要血清型分离株作致病性的鉴定.基于本实验室已经建立的多重PCR方法对stx1、stx2、eaeA、ehxA共4个基因进行检测,对检测出的阳性样品,非O157 STEC采用多重PCR结合CT-SMAC平板的分离方法,而O157 STEC通过免疫磁珠结合O157显色平板的分离方法.结果表明,该奶牛场STEC的初筛率为16.1%(112/696),分离率为11.1%(77/696).分离株属于35种O血清型和60种O:H血清型.该场的优势血清型为O4、O26和O93,O157在该场存在,但并非优势血清型.77个分离株中,stx2基因的检出率为68.8%,远远高于其它毒力基因,如stx1(19.5%)、eaeA(11.7%1)和ehxA(20.8%).该场分离到一些O157和O26血清型的菌株,对小鼠具有较强的致病性.奶牛是STEC的天然宿主,可健康带菌.除了O157STEC外,非O157 STEC中一些高致病力菌株对人类的健康也存在威胁.     

Attempts at Preventing Further Spread of Bovine Virus Diarrhoea Virus (BVDV) Infection in 5 Danish Dairy Herds in which BVDV had been Isolated

In 5 herds in which bovine virus diarrhoea virus (BVDV) had been isolated, all animals were bled for virological and serological examination. After the herd blood test, follow up blood tests were made on calves born up to 6 months later in 1 herd, 9 months later in 1 herd and up to 12 months later in 3 herds. Persistently infected animals (PI animals) were removed and after a time period a small herd sample of 10 animals that were born after removal of the PI animals were examined for BVDV antibodies.At the herd blood test a total of 21 PI animals were detected. During the follow up period another 25 PI animals were born.Among animals in the small herd samples collected after removal of the PI animals, antibody positive animals were found in the 2 herds with the shortest follow up period. In the 3 herds with a 1 year follow up period there were no antibody carriers in the herd sample.It seems possible to prevent further spread of infection with BVDV if all animals in the herds as well as animals born during the following year are examined and PI animals removed.     

Polymerase chain reaction for detection of Ureaplasma diversum from urogenital swabs in cattle in Australia

Background Ureaplasma diversum has been associated with various reproductive problems in cattle, including granular vulvovaginitis, endometritis, salpingitis, early embryonic death, weak calves, decreased conception rates, balanoprosthitis, impaired spermatozoids and seminal vesiculitis in bulls. Methods This study briefly outlines the use of polymerase chain reaction (PCR) for the rapid detection of U. diversum directly from urogenital swabs collected from Australian beef cattle. Results The 16S ribosomal RNA gene sequences obtained from the PCR products of the clinical samples were closely related to U. diversum strain A417. Conclusion The present test enabled detection of the organism directly from clinical swabs collected from animals with or without lesions.     

牛十二指肠贾第虫的分子流行病学研究进展

贾第虫是一类在全球范围内广泛分布的寄生虫,包括7个虫种。其中的十二指肠贾第虫（Giardia duodenalis）是一种重要的肠道寄生虫,能感染人和大多数哺乳动物,可引起腹泻、营养不良和体重减轻等症状。分子分型工具的发展促进了贾第虫检测、基因分型和溯源的发展,大大改变了人们对贾第虫人兽共患潜力的理解。利用分子分型工具可将十二指肠贾第虫分为8种集聚体（A~H）,8种集聚体的宿主范围都存在差异,其中,集聚体A和B是人兽共患型。牛是十二指肠贾第虫的重要宿主,但目前对牛十二指肠贾第虫病的分子流行病学认识不够,其公共卫生意义也一直被忽视。本文汇总了国内牛十二指肠贾第虫的分子流行病学调查结果。结果发现,我国牛十二指肠贾第虫的感染是普遍存在的,其中,集聚体E为优势集聚体,集聚体A和B呈散发流行。近年来,一些国家出现了集聚体E感染人的报道,同时,集聚体A在牛中的感染率有上升的迹象,牛十二指肠贾第虫的人兽共患潜力正在逐步得到认识。     

Risk Factors Associated with the Occurrence of Bovine Tuberculosis in Cattle in the Southern Highlands of Tanzania

A study was conducted in the Southern Highlands of Tanzania to determine the prevalence of bovine tuberculosis and the risk factors associated with the occurrence of the disease in cattle of different categories and in different climatic zones. The overall prevalence of the disease was 13.2%, and 51% of the herds tested contained reactor cattle. Assessment of risk factors was based on comparisons of the reactivity of the cattle in the single comparative intradermal tuberculin test (SCITT). Older cattle were more affected by the disease than yearlings and calves (p<0.0001). There were significant differences between male and female cattle (p<0.05) and between cattle with exotic blood compared to indigenous Short Horn Zebu (SHZ) cattle (p<0.05). The castrated bulls, often used for draught power, were more frequently (p<0.01) affected than the entire bulls, mainly used for breeding. Reactivity to tuberculin did not appear to be influenced by the reproductive status of the animal. The reactivity to tuberculin of pregnant cattle was not significantly different from that of the rest of the cows (p>0.05). However, significantly more (14.6%) lactating cattle reacted in the SCITT than did non-lactating cows (12.0%) (p<0.05). There was a highly significant difference (p<0.001) between reactivity in the SCITT among cattle grazing in the hot and dry lower lands (14.0%) and that in those grazing in the cool and wet highlands (8.7%).