猪瘟病毒基因组及基因工程疫苗研究进展

本文着重介绍了猪瘟病毒基因组结构组成、基因组产物和以基因组为研究对象开展的亚单位疫苗、活栽体疫苗、标记疫苗等新型基因工程疫苗的研究现状。     

重组轮状病毒多抗原表位的克隆、表达及植物表达载体的构建

为了研制基于表位的轮状病毒亚单位疫苗,用PCR的方法得到RV(Rotavirus RV)VP4的抗原表位编码序列,并人工合成了RV VP7的3个抗原表位编码序列,通用辅助性T淋巴细胞表位(PADRE)氨基酸序列和破伤风毒素(Tetanus toxin,TT)上的一个T细胞激活位点[1,2],各位点间以非极性的甘氨酸和脯氨酸分隔以保持各表位的相对独立性,使用搭桥法PCR(Over lapextension PCR)、酶切、连接等基因工程的方法将各个表位片段串联拼接在一起,组合成一条轮状病毒(RV)多表位抗原基因,将其转入原核载体质粒pTHioHis B,在大肠杆菌DH5a克隆,并在BL21中表达,表达蛋白经SDS-PAGE分析,相对分子量与预期结果相符;重组蛋白经轮状病毒快速检测试剂盒检测有抗原性。将此多表位抗原基因插入高效植物表达载体pE1802,为研制重组轮状病毒抗原表位亚单位疫苗、转基因植物口服疫苗做了前期准备。     

猪源马链球菌兽疫亚种灭活疫苗对小鼠的免疫效力评价

 【目的】评价中国不同地区猪源马链球菌兽疫亚种分离株抗原性的差异,为猪链球菌病疫苗的研制提供指导。【方法】取从国内10省市分离的10株猪源马链球菌兽疫亚种分离株,分别制备灭活疫苗,免疫12周龄ICR小鼠,用同源菌株和ATCC35246株攻击。【结果】以同源菌株攻击免疫小鼠,除CG-74-63株外,其余菌株的保护率均在90％以上;而以异源菌株ATCC35246进行攻击,免疫小鼠的免疫保护率为80%～100%。【结论】中国不同地区的猪源马链球菌兽疫亚种分离株的抗原性差异不大,单一菌株制备的疫苗具有保护性。     

A first step towards improving copepod cultivation using modelling: the effects of density,crowding, cannibalism,tank design and strain selection on copepod egg production yields

Copepods are the optimal live feed for hatcheries and improvement of cultivation techniques, to provide a constant food source, is crucial for the expansion of the industry. However, studies based on experimental work and real observations can be labour intensive and expensive. A simple model was developed based on the well‐known life history traits of Acartia tonsa to describe batch cultures and their productivity. Model results were compared to observations from real cultures. For maximizing egg production yields, the optimal stocking density of copepods should be adapted to the design (depth) of the culture tanks. At high densities, stress due to encountering conspecifics, as well as cannibalism of eggs by adults, limits egg production yields. Using this model, the potential selection efficacy of copepod strains was also evaluated in order to increase production yields. Selecting larger copepods increases the egg production per litre of culture, but decreases the optimal stocking density and the range of densities at which egg production yield is high, and vice‐versa. Selecting copepods that are less affected by stress due to conspecifics only affect production yields at very high adult densities. However, selecting copepods with a high Specific Growth Rate (SGR), or improving their SGR, was found to be an alternative which did not affect the optimal cultivation densities but improved egg production yields.     

The U.S. Wholesale Market for Marine Ornamentals

The market for marine ornamentals encompasses all live marine species (fish and invertebrates) destined for the aquarium trade. In 1998, U.S. imports and exports were valued at approximately US $1.5 billion at the border (all dollar values cited in this paper are $US). In Florida, cultured freshwater and collected saltwater species accounted for $70 million and $4 million, respectively, in annual dockside revenue. To help explain Florida's marine landings and provide suggestions for future resource management and market campaigns that reflect the perceptions of the industry, a survey of wholesalers was initiated in 1999. Preliminary results indicate that there are several intermediate wholesale markets. Florida firms are closer to the supply as many also collect. Wholesalers in other states handle more freshwater product, are larger, and tend to import a larger share of their inventory. Consolidation is expected to continue, but niche markets for eco-friendly firms are expected to grow. In Florida, marketing strategies should focus on the observed higher survival rates and increasing supply of the popular invertebrate species. Many wholesalers are concerned about additional regulations that could reduce the number of collectors and thereby increase supply variability, a noted weakness of the Florida industry.     

鸡新城疫油佐剂灭活疫苗的研究

采用ＮＤ Ｌａｓｏｔａ株为种毒，福尔马林为灭活剂，７号白油为佐剂，吐温８０、司本８０为乳化剂，硫柳汞为防腐剂，高压匀浆泵为乳化工具，研制出ＮＤ油佐剂灭活疫苗。其剂型为双相型（Ｗ／Ｏ／Ｗ），呈稀薄乳状液，粒度为１ｕｍ左右，粘度≤１ｓ，通针性好。４￣８℃有效保存期１年。效力≥７０ＰＤ５０，最早产生免疫时间为１２天（１００％保护），生产中常用免疫剂量为２月龄内每只０．５ｍｌ，２月龄以上每只１．０ｍｌ皮下     

大批量注射花鲈疫苗方法的研究

研究了大批量注射疫苗中不同麻醉剂FA100的浓度、不同注射深度、不同注射部位和不同注射流程对体长(20.0±1.2)cm花鲈成活率的影响。试验结果表明,用麻醉剂与海水体积比为1∶2000的麻醉剂麻醉后,将鱼轻轻滑至容器壁腹腔注射,注射深度4 mm,成活率最高,可以避免鱼体表机械损伤,提高注射效率和成活率。利用此方法9 d共注射花鲈21万尾,成活率高达99.76%。     

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.     

长沙市生猪供应链垂直协作模式选择

从长沙市的生猪供应链具体情况入手,对长沙市生猪供应链内各个节点间的垂直协作形式进行了研究,探讨了其目前生猪供应链节点间的垂直协作模式,并提出"垂直一体化模式"和"上游供应商+规模猪场或养殖小区(合作社)+下游屠宰加工企业"2个生猪供应链的垂直协作模式,从而实现有效地将生猪供应链内部的各个成员有效地组织起来提高整体绩效。     

猪瘟兔化弱毒疫苗——半个世纪的回顾

 猪瘟是一种严重危害养猪业的毁灭性传染病。20世纪50年代中国首创了举世闻名的猪瘟兔化弱毒疫苗（即C株），随后创制了不同的疫苗制造工艺，如细胞培养苗、乳兔组织苗和牛体反应组织苗等。C株是一株非常安全的弱毒疫苗，对各种年龄和品种的猪都没有副作用，并且有良好的免疫效力，它能同时诱导体液免疫和细胞免疫应答，对不同基因型的猪瘟病毒株均能提供有效的免疫保护。免疫母猪通过母乳可对仔猪提供被动免疫保护，但过高水平的母源抗体会影响仔猪对C株疫苗的主动免疫应答。目前已经完成了包括C株及其亲本株在内的几十株猪瘟病毒的全基因组序列测定和注释，建立了猪瘟病毒的反向遗传操作系统，初步解析了猪瘟病毒主要基因的结构与功能，并构建了不同的反向遗传操作标记疫苗，赋予了C株疫苗新的生命和内涵。C株疫苗可以用于猪瘟的控制和根除，借助于C株疫苗密集接种和综合控制措施，有关国家有效地控制了猪瘟，甚至消灭了猪瘟。尽管如此，要在全球范围内根除猪瘟，今后依然有漫长的路要走，这可能有赖于对C株进行进一步改造和利用。