Orf virus circulation in cattle in Turkey

Orf virus (ORFV) causes contagious skin disease that mainly affects sheep and goats with zoonotic potential. However, there is not enough information about the association between ORFV and occurrence of skin disease in cattle. The present study describes outbreaks of ORFV infection in cattle in different provinces that are located in the Aegean, Central Anatolian and Mediterranean regions of Turkey. During the months of June and August 2017, vesicular fluid and scab samples were collected from cattle which had proliferative skin lesions. First, presence of lumpy skin disease virus (LSDV) and bovine herpesvirus 2 (BoHV-2, known as the causative agent of pseudo-lumpy skin disease) were investigated by real time PCR and PCR, respectively. Then, samples tested for the presence of parapoxviruses by PCR using primers specific to major envelope protein gene (B2L). Parapoxvirus DNA was detected in investigated samples whereas LSDV and BoHV-2 DNA were not detected. The analysis of the B2L gene sequences revealed that cattle were infected with ORFV. The isolates in the present study shared 100% sequence identity at the nucleotide and amino acid level when compared with previously characterised Turkish field ORFV isolates from goats in 2016. Results of the study show unusual infection of cattle with ORFV, and suggest that ORFV jumps the host species barrier from goats to cattle.     

New gene-based approaches for an AIDS vaccine

Vaccine approaches against AIDS have focused on inducing cellular immune responses, since many studies revealed the role of T cell responses in the control of human immunodeficiency virus or simian immunodeficiency virus (SIV) infections. The experimental infection of rhesus macaques with SIV or chimeric SHIV is routinely used as a model for AIDS. In such models, DNA immunization is a tool to elicit specific T cell responses and to study their protective efficacy. DNA immunogenicity in primates depends on parameters such as level of antigen expression, choice of the antigen among SIV proteins, use of fusion proteins, route of immunization, and addition of adjuvants. Recent results suggest that priming with DNA and boosting with attenuated recombinant viral vectors, each expressing corresponding SIV antigens, leads to improved specific immunity and, in some cases, affords protection against pathogenic challenge. After preclinical evaluations, DNA has entered clinical trials for a therapeutic or prophylactic gene-based AIDS vaccine.     

Development of Nucleic Acid Lateral Flow Immunoassay Detection Method for Foot-and-mouth Disease Virus

In this article, through the combination of nucleic acid probes and immune chromatography, a simple, sensitive and specific detection system——nucleic acid lateral flow immunoassay (NALFIA) for amplifing foot-and-mouth disease virus (FMDV) 3D RT-PCR products was established.An ultrasensitive nucleic acid biosensor (NAB) based on streptavidin-labeled gold nanoparticles dual labels and lateral flow strip biosensor (LFSB) were used in this system.The biotinylated goat anti-rabbit IgG was marked to the NC membrane as the alleged strip and the anti-digoxin antibody was labeled to the NC membrane to capture the digoxin probe.After assemblying gold-labeled strip and detecting RT-PCR products, the detection limit of NALFIA was 0.3×10^-3 to 3×10^-3 μg/μL.The NALFIA was compared with agar gel electrophoresis analysis, the results showed that the sensitivity of NALFIA was higher than agar gel electrophoresis.There was an excellent agreement between the two methods.NALFIA was a method with high sensitive, low cost and short time.In conclusion, this method provided a good alternative to detect FMDV.     

贵州高羊茅新品种(系)比较试验

2007-2010年对选育鉴定的高羊茅(Festuca arundinacea)新品系,按照完全随机区组排列设计方法,进行品种比较试验.结果表明,在株高30～40 cm时刈割测产,大方高羊茅每年的鲜草产量和干物质产量都比对照Fawn高许多,其中第2年的鲜草产量和干物质产量分别为87 825和15 360 kg/hm2,...     

马流感病毒双抗体夹心ELISA检测方法的建立

为建立一种快速、有效的检测马流感病毒(Equine influenza virus,EIV)的方法,以EIV中国分离株A/马/新疆/07(H3N8)制备的多克隆抗体为捕获抗体,原核表达的核蛋白(NP)制备的单克隆抗体为检测抗体,在国内首次建立了检测EIV的双抗体夹心ELISA方法.用该检测方法分别检测EIV、马动脉炎病毒、马疱疹病毒1型、马疱疹病毒4型和马乙型脑炎病毒阳性样品.结果表明,该ELISA方法具有良好的特异性;与常规检测EIV的血凝试验相比,其敏感性是后者的2.5～10倍;同时与H7N7亚型EIV有交叉反应.攻毒试验结果表明该方法可有效检测鼻腔分泌物中的EIV.该方法的建立为EIV的检测及早期防控提供了有效工具.     

猪流感病毒多表位融合蛋白对小鼠的免疫原性分析

作者拟利用表位多肽的抗原性及黏膜佐剂免疫增强作用,设计可通过黏膜途径免疫接种的猪流感病毒通用型疫苗.体外合成H1N1、H3N2亚型猪流感病毒表位抗原基因,C末端串联大肠杆菌热敏性肠毒素LTb基因,构建pET30(a)-ep-LTb表达载体,利用SDS-PAGE、Western blot分析重组融合蛋白表达及生物学特性,小鼠免疫试验分析融合蛋白免疫原性.SDS-PAGE检测表达蛋白相对分子质量约38 ku,主要以包涵体形式存在.重组蛋白可与抗His-tag抗体和CTB抗体发生特异性反应.ELISA及HI试验检测,经黏膜途径免疫的小鼠产生了针对重组表位模拟抗原蛋白及H1N1、H3N2亚型猪流感病毒的血清抗体及局部黏膜分泌型IgA抗体.利用猪流感病毒表位抗原与LTb基因串连获得的融合蛋白具有良好的免疫原性和反应原性,且在黏膜接种局部产生理想的分泌型IgA抗体,能有效阻断病原由黏膜局部感染,为研制猪流感病毒通用型疫苗奠定了基础.     

猪雄性生殖干细胞的分离培养及鉴定

本试验旨在探索猪雄性生殖干细胞(mGSCs)体外分离、培养的适宜条件,建立猪雄性生殖干细胞体外培养体系。采用两步酶消化法对新生小猪睾丸生殖干细胞进行了体外分离和初步的培养鉴定,并利用层黏连蛋白和明胶的不同贴壁特性,比较2种差易贴壁分选方法的富集效果,并对传代后的干细胞培养1周后进行碱性磷酸酶染色鉴定,通过免疫荧光技术检测培养细胞是否表达干细胞标志蛋白OCT-4。试验结果表明,层黏连蛋白更适用于猪生殖干细胞的富集、培养,细胞分选效率及增殖生长明显优于采用明胶分选的方法。培养的mGSCs拥有与小鼠mGSCs相同的形态、增殖及表达特征。鉴定结果显示,生长细胞克隆碱性磷酸酶染色呈阳性,支持细胞碱性磷酸酶染色呈阴性;培养的生殖干细胞克隆表达转录蛋白OCT-4,而饲养层支持细胞OCT-4抗体染色则呈阴性。结果表明培养的干细胞克隆仍保持较好的干细胞活性,保持正常的自我复制和分化潜能,初步建立了生殖干细胞培养体系。     

不同样品中犬输血传播性病毒的PCR检测比较与基因分析

根据NCBI上输血传播性病毒(Torque-Teno virus,TTV)犬基因组序列(Cf-TTV10,GenBank登录号:AB076002)设计嵌套引物,从犬血清与粪便中鉴定出TTV。199份犬血清样品中检出阳性率为14.6%(29/199),158份犬粪便样品的检出率为12.7%(20/158),通过基因分析与鉴定确定为同一型,验证了从粪便样品和血液样品的提取检测的犬TTV阳性率基本保持一致(P>0.05),为下一步进行TTV的传染特性和分子流行病学的研究提供了基础。     

Ⅰ型鸭肝炎病毒的分离与鉴定

鸭病毒性肝炎(duck virus hepatitis,DVH)是造成养鸭业严重经济损失的传染病之一。作者对松原市某鸭场送检的病死鸭进行病毒分离试验、雏鸭回归试验、DHV分离株ELD₅₀测定、雏鸭保护试验、鸡胚中和试验、鸭胚接种试验和电镜观察等,确诊该病为Ⅰ型鸭病毒性肝炎。试验所分离得到的Ⅰ型鸭肝炎病毒与标准毒株的回归试验病理特征存在一定差异,其基因序列的测定及分析还有待于进一步研究。     

The epidemiology and diagnosis of bluetongue with particular reference to Corsica

Bluetongue (BT) and/or BT viruses (BTV) have been identified in the Mediterranean basin and the Balkans each year from 1998 to 2002 and in particular BTV serotype 2 in the French Island of Corsica (2000 and 2001). In response to these virus incursions, the French Veterinary Authorities carried out epidemiological studies that included virological, serological and entomological analysis, and two vaccination campaigns performed in the winter of 2000/2001 and the winter and spring of 2001 and 2002. Rapid and reliable serotype differentiation is essential at the start of an outbreak to allow an early selection of vaccine to control the spread of the virus. Thus, molecular tools, that complement conventional methods, have been developed for early detection of infection, determination of the serotype, and differentiation between natural infection and vaccination. Serological results showed that the first vaccination campaign during the winter of 2000/2001 did not provide full protection for all sheep and during the summer of 2001, 335 sheep flocks in Corsica were again infected by BTV 2 (7-fold more that in 2000). Entomological studies have demonstrated that the only proven vector of the disease, Culicoides imicola, was present in the island in 2000 and that it has successfully established itself in Corsica. The safety and immunogenicity of the commercial South African vaccine were studied. Fourteen sheep were vaccinated and then observed for clinical signs. Blood, sera, spleen and lymph nodes were collected and analyzed, and the results confirmed the safety and potency of using this vaccine to protect sheep from clinical disease. As a result, an intensive vaccination campaign was performed during winter and spring 2001/2002. No cases of BT had been observed by the end of summer 2002, indicating that the vaccination campaign has been successful in protecting sheep from infection.