Pathogenesis of bovine viral diarrhoea-mucosal disease (BVD-MD) virus infection in tracheal organ cultures

Bovine foetal tracheal organ cultures were infected with two strains of BVD-MD virus and observed for 35 days. The effect of the virus was assessed by observing ciliary activity in gross tissue explants and histological changes in haematoxylin and eosin stained sections. Decrease in ciliary activity and mild epithelial degeneration were first observed at 4 days post infection (p.i.), the epithelial degeneration progressing to complete destruction by day 35 p.i. Viral titres in extracellular fluids rose sharply from day 4 p.i., reached peak levels (10⁵ TCID₅₀/ml) between 15 and 18 days p.i., and eventually declined but persisted at lower titres up to day 35 p.i., when observations were terminated.     

牦牛病毒性腹泻/粘膜病的防制研究

20世纪80年代以来，我国牦牛群中陆续发现牛病毒性腹泻／粘膜病(BVD／MD)，血清阳性率在30％～42．4％之间，病死率在30％左右，本研究先后从四川、西藏等地牦牛中分离出病毒，并对其进行各种生物学特征鉴定后，表明该病毒与标准毒属同一种，所不同的是四川牦牛病毒株属非致细胞病变型，即属NCP型。但回归本动物能复制出典型病例。目前尚无国产牛粘膜病疫苗用于生产。本研究依据猪瘟病毒与牛病毒性腹泻／粘膜病病毒具有交叉免疫性的原理，用猪瘟弱毒苗对牦牛病毒性腹泻／粘膜病进行预防，试验证明用猪瘟弱毒苗可以预防牛病毒性腹泻／粘膜病，且安全可靠，具有实用价值。     

Prevalence of antibodies to bovine virus diarrhoea-mucosal disease virus in Tanzanian cattle

A serological survey to determine the prevalence of antibodies to bovine virus diarrhoea-mucosal disease (BVD-MD) virus was conducted on 419 bovine serum samples originating from 18 of 20 regions (except Mwanza and Shinyanga) of the Tanzania mainland. The sera were a small proportion of samples collected for the appraisal of immune response to rinderpest vaccination. The survey indicated that the virus is prevalent in cattle populations and approximately 12% of sera tested contained demonstrable neutralising antibodies against BVD-MD virus.     

Implementation of two-step vaccination in the control of bovine viral diarrhoea (BVD)

Bovine viral diarrhoea (BVD) control/eradication programmes based on the test and removal of persistently infected cattle without use of vaccination were first introduced by the Scandinavian countries in the early 1990s. Within the last 10 years the programmes have proven to be very successful and have served as a blueprint for several other European regions. However, in areas with high cattle densities, intense animal trade and high BVD prevalence this control approach is risky, because there is a high probability that herds, which have been cleared of persistently infected (PI) animals and have become partly or fully susceptible to reintroduction of the virus, will come in contact with a BVD virus (BVDV) infected animal. A combination of the test and removal strategy with subsequent systematic vaccination of cattle could overcome this problem. The goals of vaccination in such a programme is protection against reintroduction of BVDV into herds free from PI cattle and foetal protection of pregnant animals accidentally exposed to the virus. Two-step vaccination is based on the use of inactivated BVDV-1 vaccine for priming followed by a live attenuated vaccine booster 4 weeks later. The immune response elicited by such a vaccination scheme has proven to be long lasting and foetal infection after challenge with BVDV-1 and BVDV-2 was prevented in pregnant animals 5 months after vaccination. These findings suggest that the implementation of a two-step vaccination in the initial phase of control programmes in addition to test and removal of PI animals in areas with high cattle densities and endemic BVD is practical and efficacious.     

A rapid, quantitative assay for titration of bovine virus diarrhoea-mucosal disease virus

An end point dilution microtitration assay is described that can be used for the titration of both cytopathic and non-cytopathic isolates of bovine virus diarrhoea-mucosal disease virus. Indirect immunofluorescence is used to detect infected MDBK cells in the wells of Terasaki plates. The virus titre is derived from the number of uninfected wells, using the Poisson distribution. The assay is simple, fast and economical. Titres of cytopathic virus determined by the microtitration assay and standard plaque assay are equivalent.     

Severe malformations in calves associated with bovine viral diarrhoea (BVD) virus infection in a dairy cattle herd

During late may 2004, Some dairy cows at Al-Kharj area of central Saudi Arabia, gave birth to severely malformed calves which died, few hours to few days following birth. Samples were collected from the affected calves and their dams of virological and serological investigations. Bovine viral diarrhoea virus was detected by capture enzyme linked immuno-sorbent assay (ELISA) in the brains of affected calves. Serum antibodies were detected in the dams. The present study indicated that in spite of vaccination against BVD in the country, still severe affections of the disease are encountered. Further insight epidemiological studies to elucidate the BVD situation in Saudi Arabia is urgently needed.     

牛病毒性腹泻病毒E2基因在毕赤酵母GS115中的表达与鉴定

将牛病毒性腹泻病毒(Bovine viral diarrhoea-mucosal disease virus,BVDV)河北分离株HB-bd毒株E2基因去除跨膜区获得sE2基因,将sE2基因克隆入巴斯德毕赤酵母(P.pastoris)分泌型表达载体pPIC9K中,筛选培养后提取阳性重组质粒,经酶切和PCR鉴定,命名为pPIC9K-sE2。重组质粒pPIC9K-sE2经SalⅠ酶切后,电穿孔导入P.pastorisGS115基因进行整合,使外源基因sE2稳定地整合到P.pastoris染色体中,G418筛选得到阳性高拷贝转化子GS115-pPIC9K-sE2。经甲醇诱导表达后,sE2融合蛋白获得了表达,表达产物经SDS-PAGE、Western-blot和Dot-ELISA分析,确定其表达的sE2融合蛋白相对分子质量大小为37 000,且具有天然蛋白的抗原特异性。免疫活性研究证明,P.pastoris表达的sE2蛋白能刺激动物产生抗体。     

牛病毒性腹泻病毒E2基因在毕赤酵母GS115中的表达与鉴定

将牛病毒性腹泻病毒(BVDV)河北分离株HB-bd毒株E2基因去除跨膜区获得sE2基因,将sE2基因克隆入巴斯德毕赤酵母(P.Pastoris)分泌型表达载体pPIC9K中,筛选培养后提取阳性重组质粒,经酶切和PCR鉴定,命名为pPIC9K-sE2.重组质粒pPIC9K-sE2经Sal Ⅰ酶切线性化,电穿孔导入P.Pastoris GS115基因进行整合,使外源基因sE2稳定地整合到P.Pastoris染色体中,G418筛选得到阳性高拷贝转化子GS115-pPIC9K-sE2.经甲醇诱导表达后,sE2融合蛋白获得了表达,表达产物经SDS-PAGE、Western-blot和Dot-ELISA分析,确定其表达的sE2融合蛋白相对分子质量大小为37 000,且具有天然蛋白的抗原特异性.免疫原性研究证明P.Pastoris表达的sE2蛋白能刺激动物产生特异抗体.     

Pathogenesis of bovine viral diarrhoea-mucosal disease: distribution and significance of BVDV antigen in diseased calves

The distribution of bovine viral diarrhoea virus (BVDV) antigen in tissues of animals with acute and chronic bovine viral diarrhoea-mucosal disease (BVD-MD) was examined using improved indirect immunofluorescence and indirect immunoperoxidase staining methods on cryostat and paraffin wax tissue sections. In lymphoid tissues the antigen was principally located in cells belonging to the mononuclear located in cells belonging to the mononuclear phagocyte system and in other cells with antigen-retaining capacity. The distribution of the infected cells within a particular organ varied with the clinical stage. In the non-lymphoid organs the antigen was detected in cells of the mononuclear phagocyte system as well as in epithelial cells. An apparent time lag between initial antigen-detection and progression of pathological manifestations was noticed in the intestinal mucosa and in the keratinized epithelia of the upper digestive system and in the skin. Only limited involvement of blood vessels was observed in the tissues investigated. In the light of the mononuclear phagocyte system being an apparent common denominator for all the different tissues involved in the morphological alterations which characterise BVD-MD, a possible pathogenesis of BVDV-MD is discussed.     

应用酶联免疫吸附试验检测牛病毒性腹泻-黏膜病病毒

牛病毒性腹泻——黏膜病 (BVD- MD)是由牛病毒性腹泻病毒 (BVDV)引起牛的以黏膜发炎、糜烂、坏死和腹泻为特征的疾病。本病呈全球性分布 ,各养牛业发达国家均有流行。在自然条件下 ,可感染家养和野生的反刍兽 ,主要侵害 6～ 1 8月龄的幼牛 ,患牛表现为发病急 ,体温突然升高至 4