Vaccination strategies to conserve the endangered African wild dog (Lycaon pictus)

Pathogens such as rabies virus and canine distemper virus present a significant risk to the long-term survival of some endangered African wild dog (Lycaon pictus) populations. Vaccination of wild dogs is one approach that might reduce population extinction risks; however clear guidelines for how best to do this are lacking. Hence, we developed a mathematical model of wild dog population dynamics that integrates a pathogen transmission model and a vaccination process. Simulations indicated that the most effective vaccination strategy includes all age classes (when pups are 2-months old), prioritizes the breeding pair, and vaccinates at least 4 animals per pack included in the vaccination effort. In addition, populations for which an Allee effect, high rates of pathogen introduction, or low rates of recovery and immunity were simulated, required greater vaccination coverage (dogs/pack), to protect an equivalent number of packs compared to populations for which no Allee effect, low rates of pathogen introduction, or high rates of recovery and immunity were simulated. For populations in which some packs (high-risk) have a greater risk of pathogen exposure than others (low-risk), vaccinating both high- and low-risk packs, or only low-risk packs, is more effective than vaccinating only high-risk packs when pathogen introduction rates are high. Finally, model results suggest that vaccination of wild dog populations against pathogens, such as canine distemper virus, that do not cause 100% mortality and against which some wild dogs develop acquired immunity, may be unnecessary.     

体内外感染PRRSV处理下仔猪脾淋巴细胞增殖活性的变化

为了探讨体内或体外感染猪繁殖与呼吸综合征病毒（porcine reproductive and respiratory syndromevirus,PRRSV）的仔猪脾淋巴细胞在体外培养的增殖能力,本研究将PRRSV—GXA分离株经Mare-145细胞大量增殖培养,然后体内感染仔猪,接种病毒后分别于第11天、第14天和第21天从活体猪收获脾脏,分离脾淋巴细胞后用ConA和LPS于体外进行诱导增殖。研究结果显示第11天收获的T淋巴细胞增殖能力高于对照组,而B淋巴细胞增殖能力明显下降;第14天收获的T淋巴细胞和B淋巴细胞的增殖能力均极显著低于对照组;第21天收获的B淋巴细胞增殖能力略高于对照组,T淋巴细胞增殖能力略低于对照组。同时,用不同滴度的PRRSV体外感染PRRSV阴性的猪脾淋巴细胞,经ConA和LPS诱导增殖后,发现病毒滴度在10^3-10^6TCID50/mL范围内,能明显提高脾淋巴细胞的体外增殖能力;而病毒滴度在10^0-10^2TCID50/mL范围时,脾淋巴细胞增殖能力低于对照组。本研究结果说明PRRSV体内或体外感染对仔猪脾淋巴细胞增殖活性均有显著的影响,将为临床上PRRS的综合防治提供理论依据。     

3种大蒜中洋葱黄矮病毒的RT-PCR分子鉴定

洋葱黄矮病毒（Onion yellow dwarf virus,OYDV）是危害大蒜产量和品质的主要病毒之一。快速有效的病毒检测方法可为大蒜病毒病的研究和有效防御提供理论与技术资料。本研究利用反转录-聚合酶链式反应（RT-PCR）技术对甘肃＂成县迟蒜＂和山东＂鲁蒜王2号＂中OYDV外壳蛋白基因进行特异性扩增,并克隆到载体PMD18-TEasy Vector上进行核苷酸序列测定及分析。成功分离得到OYDV的两个DNA片段——GSCCS和SDLSW2,与GenBank中已报道的AB000837.1、AB000838.1和AJ409311.1等22个OYDV DNA片段的核苷酸相似性分别为81%～98%和81%～84%,氨基酸相似性分别为85%～99%和89%～94%,二者之间核苷酸相似性为84%,氨基酸相似性为89%。系统进化树显示不同DNA片段可聚为3个组群,GSCCS与中国山东金乡OYDV DNA片段同属一组,SDLSW2与其它16个OYDV DNA片段同属一组,表明OYDV在甘肃＂成县迟蒜＂和山东＂鲁蒜王2号＂中均存在,但具有一定差异。本实验确定了OYDV的基因变异程度,为进一步研究病毒进化和变异提供了有利条件。     

春尺蠖核型多角体病毒人工扩繁技术研究初报

利用自制的扩繁饲养棚开展春尺蠖Apocheima cinerarius Erschoff幼虫饲养、接毒增殖、采集回收、加工配制、储藏保存等一系列技术研究,结果表明,室内配比成虫平均产受精卵153粒;孵化率为86.9%;幼虫存活率70%;接毒幼虫感病率92.9%;回收率82.1%;在-18～3℃条件下冷藏保存2 a的病毒制剂毒力仅衰减1%。     

Molecular characterization of Korean rabies virus isolates

The nucleoprotein (N) and glycoprotein (G) of 11 Korean rabies virus (RABV) isolates collected from animals diagnosed with rabies between 2008 and 2009 were subjected to molecular and phylogenetic analyses. Six isolates originated from domestic animals (cattle and dogs) and five were obtained from wild free-ranging raccoon dogs. The similarities in the nucleotide sequences of the N gene among all Korean isolates ranged from 98.1 to 99.8%, while those of the G gene ranged from 97.9 to 99.3%. Based on the nucleotide analysis of the N and G genes, the Korean RABV isolates were confirmed as genotype I of Lyssavirus and classified into four distinct subgroups with high similarity. Phylogenetic analysis showed that the Korean isolates were most closely related to the non-Korean NeiMeng1025B and 857r strains, which were isolated from rabid raccoon dogs in Eastern China and Russia, respectively. These findings suggest that the Korean RABV isolates originated from a rabid raccoon dog in Northeastern Asia. Genetic analysis of the Korean RABV isolates revealed no substitutions at several antigenic sites, indicating that the isolates circulating in Korea may be pathogenic in several hosts.     

Development of a novel diagnostic test for detection of bovine viral diarrhea persistently infected animals using hair

The purpose of this study was to determine whether manually plucked hairs might serve as an alternative sample for a quantitative real time polymerase chain reaction (qRT-PCR) testing. Twenty three, 1~3 week old, non-bovine viral diarrhea virus (BVDV) vaccinated calves, found to be positive for BVDV by immunohistochemical staining, were selected and hairs were manually plucked from the ear. qRT-PCR was performed on samples consisting of more than 30 hairs (30~100) and whole blood. All 23 animals were positive for the virus by qRT-PCR performed on the whole blood and when samples of more than 30 hairs were assayed. Additionally, qRT-PCR was performed on groups of 10 and 20 hairs harvested from 7 out of 23 immunohistochemical staining-positive calves. When groups of 20 and 10 hairs were tested, 6 and 4 animals, respectively, were positive for the virus.     

羊传染性脓疱病毒趋化因子结合蛋白（CBP）基因的克隆及表达分析

参照GenBank中ORFV的毒力因子CBP基因的核苷酸序列设计合成1对特异性引物,采用PCR方法扩增该基因片段,并克隆入原核表达载体pGEX-4T-1中,经PCR、酶切鉴定和测序正确后,转入E.coli BL21（DE3）感受态细胞中,经IPTG进行诱导表达,并通过生物信息学软件进行分析。SDS-PAGE结果可见大小...     

猴痘病毒PCR检测方法的建立

从GenBank上调取猴痘病毒的基因序列,经过分析,找出了猴痘病毒的特异性靶基因序列F3L片段,人工合成猴痘病毒MPV(AF380138)F3L基因片段(48048-48509bp)并插入质粒,作为病毒检测的模拟阳性模板。根据该序列设计并合成PCR引物,对模拟的阳性模板进行扩增,结果能扩增出与目的片段大小一致的条带。建立了PCR检测方法,经条件优化后,对鸡痘、禽痘、羊痘等14种相似病毒核酸进行PCR扩增,发现具有良好的特异性。PCR方法能扩增出0.3pg的阳性模板,显示建立的PCR方法具有高效、快速、特异、灵敏的特点,可用于口岸猴痘病毒的检疫。     

检测流感病毒的商业PCR试剂盒比对验证

利用8种不同流感病毒株及其核酸,结合自然感染组织样品与人工混合样,验证比较了8种商业PCR试剂盒的质量参数。结果表明,3号荧光PCR试剂盒漏检1株外,其他试剂盒均可以检测8种毒株,对其他类似病毒如猪繁殖与呼吸综合征病毒、猪伪狂犬病毒、猪传染性胃肠炎病毒、猪细小病毒和猪环状病毒核酸等无交叉反应,最高可检测到10-7稀释的目标核酸,重复性与再现性均为理想,在有效期内检测结果没有发生变化。