Bluetongue: history, global epidemiology, and pathogenesis

Bluetongue is an arthropod-transmitted viral disease of ruminants and certain other animals that was recognized and described more than 100 years ago in southern Africa. Bluetongue virus (BTV) infection of ruminants and vector Culicoides insects is enzootic throughout tropical and temperate regions of the world; however, there have been drastic recent regional alterations in the global distribution of BTV infection, particularly in Europe since 1998. Multiple novel BTV serotypes also have been detected since 1998 in the south-eastern United States, apparently encroaching from the adjacent Caribbean ecosystem, and novel serotypes of BTV have been identified recently in other historically enzootic regions of the world, including the Middle East and Australia. It has been proposed, but certainly not proven, that global climate change is responsible for these events. BTV infection of ruminants is often subclinical, but outbreaks of severe disease occur with regular frequency especially at the upper and lower limits of the virus' global range where infection is highly seasonal - occurring in the late summer and autumn. Bluetongue disease results from vascular injury, likely through a process analogous to that of human hemorrhagic viral fevers in which production of vasoactive mediators from virus-infected macrophages and dendritic cells results in enhanced endothelial paracellular permeability with subsequent vascular leakage and hypovolemic shock.     

Bluetongue virus serotype 26: Infection kinetics,pathogenesis and possible contact transmission in goats

The aim of this study was to assess the pathogenicity and infection kinetics of Bluetongue virus serotype 26 (BTV-26) in goats. Out of a group of six goats housed in insect free accommodation, five were experimentally infected with BTV-26 and one was kept uninfected as an in-contact control. Samples taken throughout the study were used to determine the kinetics of infection using a pan specific BTV real time RT-PCR assay and a group specific ELISA. The five infected goats did not show clinical signs of BTV, however high levels of viral RNA were detected and virus was isolated from the blood of all 5 goats. Antibodies against BTV were first detected between 7 and 11 dpi in all 5 experimentally infected goats. Interestingly at 21 dpi viral RNA was detected in, and virus was isolated from, the blood of the in-contact control goat, which also seroconverted. These results suggest that BTV-26 replicates to high levels in goats, causing no obvious clinical disease, suggesting that goats may be the natural host for this virus. Preliminary evidence also indicates that BTV-26 may be spread by contact transmission between goats, however a more detailed study is required in order to confirm this observation.     

Infectious salmon anemia virus: causative agent, pathogenesis and immunity

Infectious salmon anemia (ISA) virus (ISAV), an economically important new pathogen in marine aquaculture, is classified in the family Orthomyxoviridae, genus Isavirus. The main structural properties of this genus include enveloped virions 90-140 nm in diameter with surface projections of a combined receptor-binding hemagglutinin and receptor-destroying enzyme activity demonstrated to be an esterase, hence recently designated HE, and a genome composed of eight segments of linear, single-stranded, negative sense RNA ranging in length from 1.0 to 2.4 kb, with a total size of approximately 14.3 kb. The viral genome encodes at least ten proteins, of which nine are structural and one is non-structural. Examination of more than 160 ISAV isolates has led to the identification of two hemagglutinin subtypes of ISAV, one North American and one European. The immune response against ISAV after infection or vaccination does not provide full protection against the infection. The recent discovery of antibody-mediated uptake and replication of ISAV in macrophage-like fish cell lines suggests that Fc receptor-mediated antibody-dependent enhancement of the ISA virus infection might also occur in vivo, as the virus in Atlantic salmon (Salmo salar) targets endothelial cells lining blood vessels and macrophage-like cells. Cumulative mortalities in Atlantic salmon during natural ISA outbreaks and experimental infections range from 0 to 100%. ISAV causes fatal systemic infections in marine-farmed Atlantic salmon and asymptomatic infections in feral fish. Experimentally induced fatal clinical disease in rainbow trout (Oncorhynchus mykiss) has identified a correlate of virulence of ISAV that may explain its emergence as a fish pathogen.     

Bluetongue virus serotype 26: infection kinetics and pathogenesis in Dorset Poll sheep

Bluetongue virus serotype 26 (BTV-26) has recently been isolated from sheep in Kuwait. The aim of this study was to assess the pathogenicity and infection kinetics of BTV-26 in Dorset Poll sheep. Six sheep were experimentally infected with BTV-26 and samples taken throughout the study were used to determine the kinetics of infection using a pan specific BTV real time RT-PCR assay and two group specific ELISAs. Five of the six sheep showed mild clinical signs characteristic of bluetongue including conjunctivitis, reddening of the mouth mucosal membranes, slight oedema of the face and nasal discharge. Viral RNA was detected in 5 of the 6 sheep by real time RT-PCR, however the levels of viral RNA detected in the samples were lower and of shorter duration than seen with other field strains of BTV. Virus was isolated from the blood of infected animals at the peak of viraemia at around 9 dpi. Antibodies against BTV were first detected by 7 dpi using the early detection BTV ELISA and a little later (7-14 dpi) using a BTV specific competitive ELISA. Four of the five remaining sheep developed neutralising antibodies to BTV-26, measured by a serum neutralisation test (SNT), with titres (log(10)) ranging from 1.40 to 2.08.     

禽流感病毒的致病性和免疫性研究进展

介绍了禽流感病毒的病原、基因结构及其对禽类、人类的致病机理和危害，探讨了感染病毒后宿主的反应、机体的体液免疫和细胞免疫，在一定程度上概述了禽流感病毒的固有特点和规律，为进一步研究打下了基础。     

Bluetongue virus in bovine semen: viral isolation

Vero cell cultures and embryonating chicken eggs were used for direct isolation of bluetongue virus from cattle blood and from semen samples. Cell culture and embryonating chicken eggs each were more effective than was the blood autograft inoculation of susceptible sheep with selected blood and semen samples. Evaluation of the cell culture technique indicated that the quality of the distilled water was the primary factor responsible for the increased sensitivity of the Vero cell cultures for the present blue-tongue viral isolations. Test results showed that urine was a poor specimen for viral isolation when assayed in chicken eggs. A comparison of tests for precipitating and complement-fixing antibodies to bluetongue virus indicated that the precipitin test was the more accurate of the two tests.     

Bluetongue virus serotype 20: experimental infection of pregnant heifers

Three groups of 4 cows at 84 to 95 days, 100 to 160 days, and 170 to 180 days pregnant were inoculated both intradermally and subcutaneously with bluetongue virus serotype 20 (BTV20). Clinical observations and the viraemic and serological responses of the cows were followed for 9 to 17 weeks after inoculation. Viraemia developed in 9 of the 12 cows and was first detected 4 to 9 days after inoculation. Viraemia was detected for 4 to 21 days and in some animals only intermittently. The titre of the viraemia was obtained in 4 cows and ranged from detectable only, to 10(1) to 10(2.8) 50% tissue culture infecting doses per ml. Both serum neutralising and precipitating antibodies were detected in 11 of the 12 cows within 2 to 8 weeks after inoculation. No clinical responses were seen and one cow (516) did not develop a viraemia or produce detectable antibodies to the virus. The cows, calves and foetuses were necropsied following either parturition or slaughter between 200 and 270 days of pregnancy. No virus isolations were made from a wide range of tissues from the cows, calves or foetuses and no immunoglobulins or serum neutralising antibodies were detected in the serums of precolostral calves or foetuses at necropsy. No gross or histopathological lesions were seen in the cows, calves or foetuses, and there was no evidence that BTV20 crossed the bovine placenta or infected the foetus.     

Epidemiology,molecular virology and diagnostics of Schmallenberg virus,an emerging orthobunyavirus in Europe

After the unexpected emergence of Bluetongue virus serotype 8 (BTV-8) in northern Europe in 2006, another arbovirus, Schmallenberg virus (SBV), emerged in Europe in 2011 causing a new economically important disease in ruminants. The virus, belonging to the Orthobunyavirus genus in the Bunyaviridae family, was first detected in Germany, in The Netherlands and in Belgium in 2011 and soon after in the United Kingdom, France, Italy, Luxembourg, Spain, Denmark and Switzerland. This review describes the current knowledge on the emergence, epidemiology, clinical signs, molecular virology and diagnosis of SBV infection.     

蓝舌病病毒VP7基因的原核表达

为获得蓝舌病病毒(BTV)VP7基因的原核表达蛋白,本实验采用RT-PCR方法扩增出了血清1型BTV的VP7基因,并克隆到原核表达载体pMAL-c2X中,构建了重组质粒pMAL-VP7.将重组质粒转化TBl感受态细胞,以0.5 mmol/L的IPTG进行诱导表达,SDS-PAGE电泳结果显示融合蛋白MBP-VP7以可溶形式存在,分子质量约为90 ku.以纯化的重组蛋白作为包被抗原,初步建立了检测BTV血清抗体的间接ELISA诊断方法,为今后进一步开展BTV诊断研究奠定了基础.     

Bluetongue virus in the French Island of Reunion

This paper records the results of a bluetongue virus (BTV) serological survey and reports the first isolation of BTV on the French Island of Reunion. In January 2003, the French Island of Reunion, located off the coast of Madagascar, reported an outbreak of disease in cattle that resembled clinical bluetongue (BT) in sheep. The suspected causal agent was isolated and identified as epizootic haemorrhagic disease of deer virus (EHDV). However, because of the similarity in the clinical signs to those of BT, a retrospective survey against BTV was carried out using sera collected in 2002. Results revealed the presence of antibody in all sera tested indicating that BTV has been resident on the Island since 2002, and probably earlier. Although up to July 2003 no clinical BT had ever been reported in sheep, BTV viral RNA was amplified by RT-PCR from a single sheep blood collected in February that year, which strongly suggested that BTV was currently circulating on the Island. Following a second outbreak of disease in August 2003, this time involving a flock of Merino sheep, infectious BTV was finally isolated, and identified by both traditional and molecular techniques as serotype 3. The nucleotide and amino-acid sequences of the RT-PCR products amplified for BTV segments 7 and 10 from the sheep blood collected in February and August from different areas of the Island, were sufficiently diverse as to suggest that they were of different origins and/or different BTV serotypes.     

蓝舌病病毒单克隆抗体的制备与生物学特性的鉴定

用纯化的蓝舌病病毒(BTV)免疫Balb/c小鼠,取免疫小鼠脾细胞与SP2/0骨髓瘤细胞进行细胞融合,经间接ELISA方法筛选,有限稀释法克隆,获得2株稳定分泌抗BTV特异性单克隆抗体的杂交瘤细胞株(1F5和4E5).其细胞培养上清ELISA效价分别为1:512和1:256,腹水ELISA效价分别为1:512 000和1:128 000.亚型鉴定表明,1F5和4E5分别为IgGl和IgG2a.ELISA结果显示,2株单克隆抗体仅与BTV反应,不与其他相关病毒反应,表明2株单克隆抗体特异性良好.2株单克隆抗体1F5和4E5的相对亲和力指数分别为5.14×106mol/L和6.71×106mol/L.这2株单克隆抗体的获得为建立BTV免疫学检测方法奠定了基础.     

Bluetongue virus seropositivity in sheep flocks in North West Frontier Province, Pakistan

The objectives of this study were to describe the prevalence and distribution of serum antibodies to Bluetongue virus (BTV) in a sample of 38 sheep flocks in northern areas of the North West Frontier Province of Pakistan and to identify demographic and productivity variables that are associated with BTV seropositivity. Blood samples were taken from a random sample of ewes in each flock in April 1995. The owners of the flocks were interviewed regarding some demographic, husbandry and productivity variables of the flocks on the day of blood sampling. A competitive enzyme-linked immunosorbent assay was conducted to test the serum samples for BTV group-specific antibodies. BTV seropositive reactions were obtained in 184 (48.4%) out of 380 tested sera, and in 89.5% (34/38) of the flocks. In the 34 seropositive flocks, the prevalences ranged from 12.5 to 100% (median = 47). A multivariable logistic analysis was carried out to study the influence of demographic and productivity variables on the BTV serological status of the sheep flocks. Abortion risk in the previous lambing season was mildly associated with the serological status of the flock (adjusted odds ratio = 1.16, P = 0.07). For the seropositive flocks, a linear multiple regression showed that distance travelled by the flock during transhumant movement was significantly associated with percent seropositivity (partial regression coefficient (± SE) = − 0.091 ± 0.045).