A simplified roller bottle platform for the production of a new generation VLPs rabies vaccine for veterinary applications

Rabies is a neglected disease with an estimated annual mortality of 55,000 human deaths, affecting mainly low-income countries. Over 95% of these cases result from virus transmission through the bite of infected dogs and for this reason there is a real need for a cheap and effective rabies veterinary vaccine to be used in mass vaccination campaigns. In this work, we describe the establishment of a simple platform for the production of a virus-like particles based rabies vaccine using mammalian cells and roller bottles as culture system. Adherent cells were cultured during more than 15 days and VLPs were continuously produced and secreted to the culture supernatant. Immunogenicity and protective efficacy of VLPs were tested through rabies virus neutralizing antibody test and NIH potency test. These viral particles induced high titer of long lasting neutralizing antibodies and protected mice against active virus challenge. Therefore, this development represents a promising platform for the production of a new generation and virus-free rabies vaccine candidate for veterinary applications.     

猪链球菌2型灭活疫苗对小鼠的免疫效果评价

在前期筛选出2株毒力强、抗原性好、遗传稳定的猪链球菌2型(SS2)疫苗菌株(HF2、HF3株)并分别制成灭活疫苗(采用ISA 201 VG矿物油佐剂)的基础上,进一步与SS2商品化灭活疫苗(HA9801株,铝胶佐剂)同步免疫小鼠,利用间接ELISA、流式细胞术、免疫攻毒试验、细菌定植试验和病理组织学观察等方法测定小鼠血清中IgG抗体效价,细胞因子含量(IL-4、IL-10、IFN-γ、TNF-β、MCP-1),外周血中CD4⁺/CD3⁺、CD8⁺/CD3⁺ T细胞亚群含量,攻毒保护率,组织荷菌数和病理组织变化等指标。结果显示,二免7 d后,HF2、HF3灭活疫苗组和HA9801商品化灭活疫苗组的血清IgG抗体效价分别为1:25 600、1:12 800、1:25 600;HF2灭活疫苗组的IL-4、IL-10含量显著高于HF3灭活疫苗组和HA9801商品化灭活疫苗组,IFN-γ、TNF-β含量显著高于HF3灭活疫苗组,但MCP-1含量显著低于HF3灭活疫苗组;HF2和HF3灭活疫苗组的CD4⁺/CD3⁺ T细胞比率均显著低于HA9801商品化灭活疫苗组,但CD8⁺/CD3⁺ T细胞比率差异不显著;HF2、HF3灭活疫苗和HA9801商品化灭活疫苗对小鼠的攻毒保护率均为100%;HF2灭活疫苗组小鼠的肺、脾脏组织荷菌数显著低于HF3灭活疫苗组;HF2灭活疫苗组小鼠的肺、肾、脾脏病理变化较HF3灭活疫苗组和HA9801商品化灭活疫苗组轻微。综合结果表明,由SS2(HF2株)制备的ISA 201 VG佐剂灭活疫苗免疫小鼠后不仅可产生较强的免疫应答,还可完全抵抗强毒株的攻击。     

抗菌药联用对海藻希瓦菌的体外抗菌效果

为评价硫酸庆大霉素和头孢噻肟钠对水产致病性海藻希瓦菌SFH3的体外联合抗菌效果,以6种渔用抗菌药作为对照,比较测定硫酸庆大霉素、头孢噻肟钠对海藻希瓦菌SFH3的体外抗菌作用,在此基础上进一步采用试管二倍稀释法测定分析硫酸庆大霉素和头孢噻肟钠在不同复配比例下对海藻希瓦菌SFH3的体外联合抑菌作用,并通过时间-杀菌曲线法测定其体外联合杀菌效果。试验结果显示,硫酸庆大霉素和头孢噻肟钠对海藻希瓦菌SFH3的最小抑菌质量浓度为80 mg/L和160 mg/L,抗菌活性明显优于甲砜霉素、多西环素、噁喹酸、磺胺间甲氧嘧啶钠、磺胺甲噁唑、盐酸恩诺沙星等渔用抗菌药。硫酸庆大霉素和头孢噻肟钠在复配比例8∶2和7∶3时,对海藻希瓦菌SFH3的体外联合抑菌效果表现为协同作用,对海藻希瓦菌SFH3的最小抑菌质量浓度达40 mg/L。此外,时间-杀菌曲线表明,40 mg/L硫酸庆大霉素和80 mg/L头孢噻肟钠对海藻希瓦菌SFH3的体外联合杀菌作用表现为协同效果。本研究结果证实,硫酸庆大霉素和头孢噻肟钠联用有利于提升对海藻希瓦菌SFH3的体外抗菌效果,可为研发水产致病性海藻希瓦菌的潜在控制药物提供理论参考。     

High protection of mice against Brucella abortus by oral immunization with recombinant probiotic Lactobacillus casei vector vaccine,expressing the outer membrane protein OMP19 of Brucella species

Brucellosis is a zoonotic disease threatening the public health and hindering the trade of animals and their products, which has a negative impact on the economic development of a country. Vaccination is the most effective way to control brucellosis. The recombinant vector vaccines are promising candidates for immunization in humans and animals. In this study, the gene encoding OMP19 antigen was primarily amplified and cloned into an expression vector called pT1NX, and then transformed to L. casei cell via electroporation technique. The expression was confirmed using specific antibody against the recombinant protein via immunological screening tests such as western blot and immunofluorescence assay. Finally, recombinant L. casei was orally fed to mice and the results were further recorded, indicating that the mice group which received OMP19 through L. casei based vaccine represented a very good general and mucosal immune responses protective against challenges with virulent B. abortus 544 strain compared with negative control recipient groups. Therefore, the vaccine produced in this research plan can be a very good candidate for protection against brucellosis.     

低扬程泵装置压力脉动特性

初步分析了低扬程泵及泵装置压力脉动的基本规律. 基于浙江姚江上游西排工程排涝泵站立式泵装置不同叶片角度和不同流量时的压力脉动特性模型试验结果,研究泵装置的叶轮室进口、叶轮室出口、导叶体出口和出水流道出口处的压力脉动规律. 从叶轮与导叶之间的水流条件、水泵运行工况等方面,对低扬程泵装置压力脉动进行了初步理论分析,提出了导叶体与叶轮间隙的大小、导叶体的叶片数对低扬程泵装置压力脉动特性的影响等需进一步研究的问题. 从水力因素和水泵制造等方面提出了避免低扬程泵装置压力脉动有害影响的对策:适当加大叶轮与导叶之间的间隙,避免水泵在偏离最优工况较多的条件下运行,提高水泵抗空化性能等;从避免产生共振的角度,提出适当加大叶轮叶片厚度、导叶叶片厚度和加强水泵的结构设计等措施.     

h指数及其衍生指标评估兽用疫苗领域单篇专利的适用性探索

随着禽流感、非洲猪瘟等疾病的出现,兽用疫苗的开发研制成为了动物防疫体系建设的重要一环,同时兽用疫苗领域国际竞争日益激烈,直接影响到我国相关企业的生存和发展,因此国家必须加强对兽用疫苗领域专利质量和专利保护的重视.本文以计量学中的h指数及其衍生指标为对象,研究其与传统专利价值评估指标的关系,通过相关性分析来筛选评估兽用疫苗领域高价值专利的指标.结果表明:h指数、g指数和hg指数这三个指标,与传统专利价值评估指标相关性强,且计算简便,相对于传统指标,能够快速准确地对兽用疫苗领域专利价值进行预估.     

商品化猪丹毒疫苗分类及特点

文章介绍了5种商品化猪丹毒疫苗的种类、毒株组成、抗原含量及相应免疫程序,为了解各种类型的猪丹毒商品化疫苗提供参考。     

Comparison of flagellin and an oil-emulsion adjuvant in inactivated Newcastle disease vaccine in stimulation of immunogenic parameters

The present study was designed to investigate the potential application of native (N) and recombinant (truncated modified [tmFliC] and full-length [flFliC]) flagellin proteins along with inactivated Newcastle disease virus (NDV). Fifty six SPF chickens were immunized twice with PBS (control), inactivated NDV (Ag), inactivated NDV/flFliC (AgF), inactivated NDV/tmFliC (AgT), inactivated NDV/N (AgN), commercial vaccine containing Montanide (Vac) and Vac/N (VacN), with a two-week interval. Blood was collected weekly and spleens were harvested after chickens were sacrificed. Interleukin-6 (IL-6) and tumor necrotic factor-α (TNF-α) gene expression in peripheral blood mononuclear cells were analyzed by Real-Time PCR. Antibody response was assessed by haemagglutination inhibition (HI). Cellular activity was quantified by MTT assay. Results showed that the most IL-6 and TNF-α gene expression was observed in AgF group (P < 0.01). The lowest gene expression among vaccinated groups was observed in Ag group for IL-6 and Ag and Vac group for TNF-α. The highest HI titer was observed in Vac, VacN, AgF and AgT groups. The AgF group showed the highest cellular activity (P < 0.01). In conclusion, flagellin-adjuvanted groups showed a pro-inflammatory effect and acted similarly to or better than the Vac group. Hence, flagellin can be proposed as a potential adjuvant for ND vaccine.     

油菜高亚麻酸育种探析

为促进通过提高油菜种子亚麻酸含量途径增加中国人群亚麻酸的摄入量,提高健康水平的工作进程。结合文献资料,在剖析开展油菜高亚麻酸育种的意义、可行性、难点及可利用的有利条件的基础上,提出油菜高亚麻酸育种策略与技术途径。在工作实践中,陕西省杂交油菜研究中心研究团队创建并完善"油菜高亚麻酸种质的筛选方法",利用该方法开展油菜高亚麻酸种质资源筛选和高亚麻酸育种取得良好成效:油菜种质资源的整体亚麻酸含量、具有较高亚麻酸含量种质的百分比率、亚麻酸含量最高值、亚麻酸含量15%的种质数量均明显增加;获得一批亚麻酸含量大于15%(最高含量达21%)的甘蓝型油菜非转基因种质,育成高亚麻酸的不育系和恢复系;甘蓝型油菜高亚麻酸杂交种‘秦杂油7号’通过国家品种登记;2个亚麻酸含量15%左右的杂交种正在申请品种登记。发现在A02、C02及A06染色体上存在多个与亚麻酸显著关联SNP位点。提出开展基础机理研究和分子设计育种创新体系建设,开展高亚麻酸种质资源创新、综合评价和利用研究,构建高亚麻酸育种体系,规化高效保优栽培技术路线,开展油菜籽加工技术、抗氧化研究,构建标准体系等6方面建议。     

野生牧草金荞麦与贵州省推广牧草栽培效益比较试验初报

金荞麦Poyigonum cymosum是一种药饲兼用的贵州野生植物资源,通过金荞麦与贵州省目前主要推广牧草的田间试验比较,研究金荞麦和推广牧草资源的产量、品质和饲用的不同特点。试验初步认为:金荞麦是一种高产优质的牧草资源,其产量与菊苣Cichorium intybus相似,比一般推广牧草如苜蓿Medicagosativa等高出2~3倍。其粗蛋白含量与菊苣相似,接近苜蓿。金荞麦的适口性好,耐瘠抗劣性强,易繁殖成活,有较大的推广潜力。