Serological characteristics of affected cattle during an outbreak of bovine enzootic encephalomyelitis caused by Akabane virus

During an outbreak of bovine enzootic encephalomyelitis caused by the Akabane virus (AKAV) in 2010, 210 serum samples were collected from the affected cattle, and serological investigations for the AKAV were performed using a serum neutralization test (SNT) and an enzyme-linked immunosorbent assay (ELISA). The seropositive rates for SNT and ELISA were 90.0 and 85.2 %, respectively. The titers of SNT (log₂) against the AKAV were higher than 4.0 in the highly affected cattle (80.0 %). This finding indicates that most affected cattle were infected with the AKAV and that strong immune responses against this virus were elicited in affected cattle. The strong immune response to the AKAV in cattle may provide insight into the occurrence of bovine encephalomyelitis caused by the AKAV.     

Genetic analysis of Akabane virus isolates from cattle in Korea

Bayesian Inference (BI) and Neighbor Joining (NJ) analyses of the phylogenetic relationships between the nucleotide sequences of the N gene of Akabane virus revealed an unclear topology among genogroups I–III, which was probably caused by genetic reassortment or recombination between these genogroups. In contrast, nucleotide and amino acid phylogenetic tree analyses of the M RNA segment agreed with the topologies obtained by using the BI and NJ methods. Therefore, distinct genogrouping of Akabane virus isolates should be performed using the M RNA segment. Four Korea isolates were classified into genogroup II together with Akabane virus strains isolated from all areas of Japan, including Okinawa Island. However, more nationwide isolates and more clinical data from Korean cattle farms will be required in the future to confirm the precise relationships between genotypes and pathogenicity.     

Elisa test for the serodiagnosis of Akabane virus infection in cattle

Summary A simple ELISA test has been developed for the detection of IgG and IgM antibodies to Akabane virus in bovine serum. The test is specific and its sensitivity higher than the serum neutralisation assay. Detection of IgM antibodies can serve as a rapid method of diagnosing primary infection with Akabane virus. The superiority of ELISA resides mainly in the rapidity of performance and that it can be performed with inactivated reagents at high dilutions of serum samples. Thus it might enable the surveillance of spread of infection in zones prone to be affected by insect-borne viral diseases.

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La Prueba Elisa Para El Serodiagnostico De La Infeccion En Bovinos Con El Virus De Akabane

Resumen Se desarrolló una prueba ELISA simple, para la detección en sueor bovino de anticuerpos IgG e IgM del virus de Okabane. La prueba es específica y la sensibilidad superior a aquella de la seroneutralización. La detección de anticuerpos IgM puede servir como un método diagnóstico rápido para detectar infecciones primarias causadas por el virus. La superioridad de la prueba ELISA reside principalmente, en la rapidez de ejecución de la misma y que ésta puede realizarse con reactivos inactivados, a altas diluciones de la muestra de suero. Esta prueba ELISA podría facilitar la vigilancia epidemioloógica de la extensión de la enfermedad, en zonas en donde las condiciones ecológicas facilitarían el establecimiento de infecciones transmitidas por insectos.

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Un Test Elisa Pour Le Serodiagnostic De L'Infection A Virus Akabane Chez Le Betail

Résumé Un test ELISA simple a été développé pour la détection dans le sérum bovin des anticorps IgG et IgM dirigés contre le virus Akabane. Le test est spécifique et sa sensibilité est supérieure à celle du test de séro-neutralisation. La détection des anticorps IgM peut servir de méthode rapide pour diagnostiquer une infection primaire par le virus Akabane. La supériorité de l'ELISA réside essentiellement dans la rapidité de son exécution et en ce qu'il peut être réalisé avec des réactifs inactivés, et a de hautes dilutions des échantillons de sérums. De la sorte, il permet la surveillance de la propagation de l'infection dans les zones aptes à être affectées par les maladies transmises par arthropodes.

     

Encephalitis caused by louping ill virus in a group of horses in Ireland.

An outbreak of LI infection in a group of free range horses is described. Three of 4 horses displayed signs of CNS disturbance and 2 of these died after illnesses ranging from 2-12 days duration. In both cases a variable degree of viral polioencephalomyelitis was observed. A virus antigenically indistinguishable from a reference strain of LI virus was isolated from the brain and cervical cord of a 3 y.o. draft mare. Serum samples obtained from 3 of the horses contained HI, CF, precipitating and neutralising antibodies to LI virus, with a rise in antibody titre being demonstrated in 2 animals.     

Appearance of slow-reacting and complement-requiring neutralizing antibody in cattle infected with Akabane virus

Slow-reacting complement-requiring neutralizing (NT) antibody was detected in sera from cattle 2 weeks after infection with Akabane virus. Bovine sera obtained 3 or 4 weeks after infection contained slow-reacting noncomplement-requiring NT antibody. The slow-reacting complement-requiring NT antibody was sensitive to 2-mercaptoethanol (2-ME), whereas the slow-reacting noncomplement-requiring NT antibody was resistant to 2-ME. The initial phase may represent the IgM response and the later phase a change to IgG. A NT test was developed in which virus-serum mixtures were incubated at 4 degrees C for 48 h and then with complement at 37 degrees C for 60 min; this gave an improved sensitivity over the previous incubation at 37 degrees C for 60 min.     

Immunisation of cattle with a recombinant togavirus-vaccinia virus strain

Genetic engineering techniques have been used to construct a vaccinia virus recombinant which contains and expresses togavirus (Sindbis) genetic information. Intradermal inoculation of this recombinant strain into calves caused a transient pock-type lesion at the site of inoculation and elicited the production of substantial levels of anti-Sindbis virus neutralising antibodies. These results suggest that recombinant vaccinia virus vaccines may have potential for use in veterinary medicine.     

Encephalitis caused by a Sarcocystis-like organism in a steer

Sarcocystis spp can cause poor growth, anorexia, fever, anemia, muscular weakness, nervousness, abortion, and even death in cattle. Lesions in the CNS of cattle infected with Sarcocystis spp are microscopic and relatively minor, compared with those in other organs. We report extensive encephalitis in a steer attributable to a Sarcocystis-like organism.     

The shedding of a virulent Kabete O strain of rinderpest virus by cattle

A Muguga substrain of the virulent Kabete O strain of rinderpest virus was demonstrated in the ocular, nasal, oral and rectal swabs collected from infected cattle. Ocular shedding was detected at the onset of viraemia and before the onset of clinical signs whilst virus shedding in nasal, oral and rectal discharges appeared at the same time as lesions. It is suggested that virus isolation from ocular and nasal swabs should be considered in the diagnosis of rinderpest in addition to the other methods currently employed, as virus was isolated from swabs collected from dead animals.     

A serological survey of Akabane virus infection in cattle and sheep in northwest China

Purpose

Akabane disease characterized mainly by fetal damage is a ruminant disease caused by insect-transmitted Akabane virus infection.

Methods

We investigated Akabane disease using serum neutralization tests in 446 blood samples collected from 187 cattle and 259 sheep of Xinjiang province, northwest China.

Results

(1) The overall prevalence rate of neutralizing antibody was 19.06?% (85/446), (2) the prevalence rates of Akabane disease in cattle and sheep were 20.32?% (38/187) and 18.15?% (47/259), respectively, (3) the disease prevalence rates were not significantly different between cattle and sheep, but significantly different among samples collected from different sampling months, (4) the disease was most prevalent in July when mosquitoes and culicoides were most active, and (5) the disease prevalence rates were significantly different between individuals with abortion experience and without abortion experience (P?<?0.05), suggesting that Akabane virus infection may significantly increase abortion risk in cattle and sheep.

Conclusions

To our knowledge, this is the first report confirming that Akabane virus infection is common in cattle and sheep of Xinjiang province, northwest China and providing useful epidemiological information for cattle and sheep abortion prevention and control.     

云南黄牛血液中阿卡斑病毒的分离鉴定及S基因序列分析

为了解阿卡斑病毒在云南师宗的流行情况及病毒的遗传特征,本研究在云南师宗设立哨兵动物,2014年5～11月每周1次连续采集血清、血液样品,血清用做AKAV抗体检测,血液在C6/36、BHK-21、MDBK和Vero细胞上进行病毒分离工作,用S基因特异引物对阳性分离物进行鉴定及序列分析。结果显示,15头哨兵动物共采集到28批420份血清、420份血液样品,其中有3头牛和1只羊感染AKAV,从这些样品中分离获得2株AKAV(编号09312D和09312H),它们在BHK-21和MDBK细胞上48 h产生明显CPE。2株新分离AKAV的S片段序列核苷酸同源性为100%,氨基酸同源性为100%,与所引用的9株其他不同地区分离的AKAV的S片段序列核苷酸同源性在83.8%～97.7%之间,氨基酸同源性在90.6%～99.6%之间,且新分离毒株处于同一进化小分支,具有高度亲缘性,形成了一个独立的次级分支,同时与其他2株中国AKAV分离株处于同一进化簇。结果表明,云南省师宗县为AKAV流行地区,病毒在当地活动频繁,新分离得到的2株病毒与其他AKAV毒株具有较高的核苷酸同源性和氨基酸同源性,提示亚洲太平洋地区AKAV病毒长期流行且关系密切,同时其氨基酸同源性高于核苷酸同源性也提示了AKAV S片段良好的氨基酸遗传稳定性。     

Duration of immunosuppression caused by a vaccine strain of infectious bursal disease virus

The depression of response of Brucella abortus strain 19 caused by an infectious bursal disease vaccine virus given to chicks at one day old was shown to persist for four weeks. Histological examination of the bursa of Fabricius showed a gradual repopulation by bursal lymphocytes, after initial damage, concomitant with the development of a humoral response.     

Antibodies to Akabane virus in horses,sheep and goats in Japan

High prevalences of neutralizing (NT) antibody to Akabane virus were obtained with horse (72%), sheep (38%) and goat (67%) serum samples collected in Chiba Prefecture, where outbreaks of abortion and congenital deformities caused by Akabane virus occurred among cattle. In these animal sera, titers of hemagglutination-inhibiting (HI) antibody to Akabane virus and of NT antibody were closely correlated.     

小反刍兽疫疫苗毒与野毒实时荧光RT—PCR鉴别检测方法的建立

为了建立鉴别检测小反刍兽疫疫苗毒与野毒的快速分子生物学检测方法,通过对自行测序的疫苗毒基因组序列及GenBank中登录的野毒基因组序列进行比对分析,设计了2套引物和TaqMan荧光探针,对实时荧光RT-PCR反应条件进行优化,建立了小反刍兽疫疫苗毒与野毒实时荧光RT-PCR鉴别检测方法。特异性试验证实,该检测方法只能检测到目的病毒核酸,表明其具有良好的特异性。灵敏性试验发现,检测疫苗株的最低检测限可达1．38mg／L的总RNA,检测野毒株的最低检测限为0．16mg／L的核酸。对同一样品进行重复性检测,检测的荧光扩增曲线阈值完全重舍,证明其重复性极好。从180份临床样品中,检测出2份疫苗病毒株阳性样品,其余均为阴性。结果表明,本研究所建立的实时荧光RT-PCR方法能对小反刍兽疫疫苗毒及野毒进行鉴别检测,具有特异性好、灵敏度高、重复性极好的优点,是开展小反刍兽疫疫情监测的有力工具。     

Sero-survey on Aino, Akabane, Chuzan, bovine ephemeral fever and Japanese encephalitis virus of cattle and swine in Korea

Vector-borne arboviruses produce mild to severe symptoms in domestic animals. Bovine ephemeral fever (BEF), Akabane, Aino, and Chuzan virus have been primarily attributed to reproductive disorders or febrile diseases in cattle, and Japanese encephalitis virus (JEV) is mainly associated with reproductive failures in swine. We investigated antibody titers from domestic swine against four bovine arboviruses (BEF, Akabane, Aino, and Chuzan virus) and from cattle against JEV in Korea. While the positive rates for Akabane and BEF were 37.4% and 15.7%, the positive incidence of Chuzan and Aino were relatively low, with positive rates of 3.04% and 0.4%, respectively, based on a virus neutralization assay. Antibody titers against more than one virus were also frequently detected in domestic swine. The incidence of JEV was 51.3% among domestic cattle. In addition, one positive case was detected in the thoracic fluids from 35 aborted calves, based on the hemagglutination inhibition test. Our results indicate that swine are susceptible hosts of bovine arboviruses without showing clinical symptoms in a natural environment. Moreover, we confirmed that JEV could be associated with reproductive failure in pregnant cattle, as were other vector-borne bovine arboviruses assessed in this study.     

赤羽病病毒核衣壳蛋白基因的表达纯化及活性鉴定

为获得赤羽病病毒（Akabane virus, AKAV）核衣壳蛋白重组抗原,经RT-PCR扩增了OBE-1株的S节段,将其插入到pMD 18-T载体,然后用带有酶切位点的引物从该重组载体亚克隆出表达核衣壳蛋白的N基因,用BamHⅠ和XhoⅠ双酶切后与同样处理的表达载体pET-28a（＋）进行连接,转化感受态细胞BL21（DE3）.将筛选出的阳性克隆进行测序、IPTG诱导表达和融合蛋白纯化及活性鉴定.结果表明,N基因含有1个全长702 bp的ORF,编码233个氨基酸,通过SDS-PAGE和Western-blotting分析,证实重组菌株可高效表达有免疫活性的分子质量约27 ku的可溶性融合蛋白,用薄层扫描仪分析,其最高表达量占可溶性菌体蛋白总量的58.5%.工程菌经超声波裂解高速离心后,上清液在ProBondTM Purification System（含Ni2＋）天然状态下纯化,可获得高纯度的融合蛋白.