狐传染性脑炎活疫苗抗体消长规律的研究

应用血清中和试验和血凝抑制试验对狐传染性脑炎活疫苗免疫狐进行抗体水平动态监测,结果表明,疫苗接种14 d抗体达到保护,21~30 d抗体水平达到高峰;免疫240 d抗体仍在保护值以上。对SN抗体和HI抗体进行动态分析表明,两者呈正相关,均可用于狐传染性脑炎活疫苗免疫效力监测。     

鹿源牛病毒性腹泻病毒核酸疫苗免疫实验研究

试验应用间接ELISA方法检测鹿源牛病毒性腹泻分离株CCSYD核酸疫苗体液免疫效果,应用淋巴细胞转化实验检测其细胞免疫效果,并与鹿源牛病毒性腹泻分离株CCSYD灭活苗和标准株C24V灭活苗进行比较。结果表明:鹿源牛病毒性腹泻分离株CCSYD核酸疫苗产生了较好的体液和细胞免疫应答,免疫前后免疫应答水平差异均显著(P<0.05)。     

二种佐剂对哈氏弧菌OMPC-溶藻弧菌LPS偶联疫苗的增效作用

制备哈氏弧菌SpGY020601外膜蛋白复合物(OMPC)和溶藻弧菌EpGS021001脂多糖(LPS)偶联疫苗,设置偶联疫苗组、偶联疫苗 ISA763佐剂组、偶联疫苗 β-葡聚糖佐剂(DEAE)组以及生理盐水对照组,并以相同程序免疫卵型鲳鲹。溶菌酶活性检测、微量凝集反应试验结果显示:各疫苗组血清中的溶菌酶含量无显著差异(P>0.05),但均远高于生理盐水对照组(P<0.05);ISA763佐剂组抗哈氏弧菌SpGY020601抗体滴度较其他组出现的早,抗溶藻弧菌EpGS021001抗体滴度较其他组持续时间长;β-葡聚糖佐剂组次之,其次是无佐剂组,对照组始终未检测到抗体。各组分为腹腔注射攻毒与浸泡攻毒,对EpGS021001的攻击,ISA763佐剂组相对保护率分别为90%和70%,高于β-葡聚糖佐剂组的85%和55%,以及无佐剂组的80%和55%;对SpGY020601的攻击,ISA763佐剂组相对保护率分别为95%和80%,高于或相当于β-葡聚糖佐剂组的95%和75%,以及无佐剂组的90%和75%。     

无乳链球菌Sip-GAPDH嵌合核酸疫苗的制备及免疫效果评价

为了研究无乳链球菌Sip-GAPDH嵌合核酸疫苗对罗非鱼链球菌病的免疫保护作用,本研究通过PCR和重叠延伸拼接技术(SOEing)获得融合基因Sip-GAPDH,连接至真核表达载体pcDNA3.1(+)制备DNA疫苗,通过背鳍肌肉注射方式免疫吉富罗非鱼,免疫后第7、28天取样,采用PCR及RT-PCR方法检测pcDNA-Sip-GAPDH在各个组织(包括肌肉、脑、头肾、脾脏、鳃和肝脏)中的表达情况。采用ELISA、Real-time PCR法和体外攻毒法分别分析各实验组不同时间血清抗体效价、免疫基因(IgM、IL-1β和CD8)表达变化规律和相对保护率,研究该嵌合性疫苗对吉富罗非鱼的保护效果。结果显示,实验组在免疫接种第7和第28天时,注射点周围的肌肉、脑、头肾、脾脏、鳃和肝脏均能检测到嵌合性质粒,实验组罗非鱼血清抗体效价均高于对照组并于21 d时达到峰值(1∶4 096);qPCR数据表明,实验组鱼体胸腺、头肾和脾脏的IgM、IL-1β和CD8基因的mRNA表达量均出现了上调,并且在胸腺、头肾和脾脏中的表达量分别在免疫后第12和第48 h达到峰值;免疫后42 d进行人工攻毒后计算免疫保护率为93.3%。以上研究结果表明,本研究构建的无乳链球菌Sip-GAPDH嵌合核酸疫苗在罗非鱼链球菌病防治中具有潜在的应用价值。     

Surface-exposed expression of Edwardsiella tarda EseB in live attenuated Vibrio anguillarum based on novel surface display systems

Live, attenuated Vibrio anguillarum strains can serve as vectors for the delivery of heterologous antigens for development of multivalent recombinant vaccines. Based on the outer membrane anchoring elements of V. anguillarum , we have previously constructed several efficient surface display systems Lpp-Omporf1, Lpp-OmpU, Lpp-Omp26La, Wza-Omporf1, Wza-OmpU and Wza-Omp26La. In this study, with these constructed surface display systems, a putative antigen protein EseB from pathogenic Edwardsiella tarda was successfully expressed on the surface of an attenuated V. anguillarum strain to get multivalent vaccine candidates. Further immune protection evaluation in zebra fish ( Danio rerio ) demonstrated that the V. anguillarum EseB-display strain AV/pW-26La-B could trigger full protection against V. anguillarum infection and early protection against E. tarda infection in the immunized fish. These results suggest that surface display of heterologous protective antigens in attenuated V. anguillarum could be used as a tool to develop potential V. anguillarum vector vaccine.     

蜂胶佐剂对异育银鲫嗜水气单胞菌灭活疫苗的免疫增强试验

本文就蜂胶佐剂对鱼类细菌性疫苗的免疫增强作用进行了首次评价。试验以异育银鲫嗜水气单胞菌灭活疫苗为对象,比较了蜂胶佐剂、铝胶佐剂和无佐剂三种疫苗对银鲫的免疫效果。经抗体监测表明,免疫接种后第2周,蜂胶佐剂组银鲫已检测到血清抗体,随后抗体效价迅速上升,第4周达到峰值2^5：铝胶佐剂组和无佐剂组第3周检测到抗体．铝胶佐剂组第5周达到峰值2^5,无佐剂组第4周达到峰值2^4。银鲫免疫后第5周进行攻毒保护试验,发现蜂胶佐剂组、铝胶佐剂组和无佐剂组的免疫保护率分别为77．3％．65．0％和56．3％。试验结果表明,蜂胶佐剂能有效增强异育银鲫细菌性灭活疫苗的免疫保护力。     

山莨菪碱提高嗜水气单胞菌灭活疫苗浸泡免疫鲫的效果

为研究山莨菪碱作为佐剂在细菌灭活疫苗浸泡免疫中的作用,将山莨菪碱和嗜水气单胞菌全菌灭活疫苗联合浸泡免疫异育银鲫,首次浸泡免疫7d后加强免疫1次.通过实时荧光定量PCR检测第2、4、7、11、14和21天脾脏中IgM、IL-1β、C3、C-凝集素以及溶菌酶的mRNA表达量,并在第21天进行同源菌株活菌攻击实验.结果显示,山莨菪碱组第4天IgM和IL-1 β的表达量达到最高值,分别为28.3和332.7;而无佐剂疫苗组第11天IgM和IL-1β达到最高值,分别为56.1和791.8.山莨菪碱组的补体C3、C-凝集素以及溶菌酶的表达量都显著高于无佐剂组和对照组,且表达持续时间长.活菌攻击实验表明山莨菪碱组的相对免疫保护率可达80.0％,显著高于无佐剂组的56.0％.结果表明,山莨菪碱与嗜水气单胞菌灭活疫苗共同浸泡免疫银鲫,可以增强脾脏中IgM、IL-1β、C3、C-凝集素以及溶菌酶基因表达,提高银鲫相对免疫保护率.     

草鱼出血病的免疫学研究进展

由于草鱼病毒性出血病发病率高,且尚未找到有效防治的药物,至今一直成为草鱼养殖发展中"瓶颈"问题的根本原因。因此,采取免疫学途径防治草鱼出血病是解决此问题的关键之所在。综述了采用草鱼出血病疫苗、干扰素及免疫增强剂进行草鱼出血病防治的研究现状;提出口服型免疫增强剂是生产上防治草鱼出血病最为有效的途径之一。     

Exploring the efficacy of vaccine techniques in juvenile sablefish,Anoplopoma fimbria

Wild sablefish, Anoplopoma fimbria, are a valuable commercial species whose populations are declining. Fortunately, sablefish are excellent species for commercial aquaculture. Sablefish raised under high‐density conditions in the marine environment require the use of efficacious vaccines to control disease. Sablefish impacted by disease in net pens may have poor flesh quality and high mortality during grow‐out. As a result, disease can cause financial hardship for sablefish aquaculture operators. The efficacy of a multivalent vaccine preparation for sablefish, administered either by intraperitoneal (i.p.) injection or by immersion, against atypical and typical Aeromonas salmonicida, the causative agents of atypical and typical furunculosis, respectively, was examined. A. salmonicida can affect sablefish at any age and size. Consequently, an efficacious vaccine that can be appropriately and optimally administered to all life stages is desirable. Sablefish vaccinated by immersion at ~1.5 or ~4.5 g with a whole‐cell multivalent vaccine were not protected against either typical or atypical A. salmonicida. Factors that may have contributed to the ineffectiveness of the immersion vaccine are discussed. By contrast, the relative per cent survival (RPS) or potency of the whole‐cell multivalent vaccine injected i.p. in juvenile sablefish at ~50 g against typical and atypical A. salmonicida was 94.3% and 81.7% respectively. The high RPS values indicated that the vaccine successfully initiated an immune response in sablefish upon a second encounter with the pathogen.     

草鱼呼肠孤病毒疫苗的研发与应用

正随着水产养殖产业的不断扩大,我国已成为世界上最大的水产品生产消费和进出口国家。2000年至2013年,水产养殖产量以每年5%~6%稳步增长,在2013年,中国水产养殖产量达4.542×10~7t,占全球水产养殖总产量的60%以上~[1]。根据世界粮农组织数据显示,2015年我国水产养殖总量达7.430×107 t。这些数据表明我国水产养殖业不仅保障了中国水产品的市场供应,也对世界水产品供