Porcine circovirus type 2 decreases the infection and replication of attenuated classical swine fever virus in porcine alveolar macrophages

Recently, it has been noted that porcine circovirus type 2 (PCV2) infection adversely affects the protective efficacy of Lapinized Philippines Coronel (LPC) vaccine, an attenuated strain of classical swine fever virus (CSFV), in pigs. In order to investigate the possible mechanisms of the PCV2-derived interference, an in vitro model was established to study the interaction of LPC virus (LPCV) and PCV2 in porcine alveolar macrophages (AMs). The results showed that PCV2 reduced the LPCV infection in AMs and the levels of PCV2-derived interference were dose-dependent. The PCV2-derived interference also reduced the replication level of LPCV in AMs. The full-length PCV2 DNA and its fragment DNA C9 CpG-ODN were involved in the reduction of LPCV infection in AMs, whereas UV-inactivated PCV2 was not. In addition, a moderate negative correlation between the LPCV antigen-containing rate and IFN-γ production was observed, and had a dose-dependent trend with the level of PCV2-inoculation. The results of the present study may partially explain how PCV2 infection interferes with the efficacy of LPC vaccine.     

Drivers and risk factors for circulating African swine fever virus in Uganda, 2012–2013

We explored observed risk factors and drivers of infection possibly associated with African swine fever (ASF) epidemiology in Uganda. Representative sub-populations of pig farms and statistics were used in a case-control model. Indiscriminate disposal of pig viscera and waste materials after slaughter, including on open refuse dumps, farm-gate buyers collecting pigs and pig products from within a farm, and retention of survivor pigs were plausible risk factors. Wire mesh-protected windows in pig houses were found to be protective against ASF infection. Sighting engorged ticks on pigs, the presence of a lock for each pig pen and/or a gate at the farm entrance were significantly associated with infection/non-infection; possible explanations were offered. Strict adherence to planned within-farm and community-based biosecurity, and avoidance of identified risk factors is recommended to reduce infection. Training for small-scale and emerging farmers should involve multidimensional and multidisciplinary approaches to reduce human-related risky behaviours driving infection.     

Comparison of two real-time RT-PCR assays for differentiation of C-strain vaccinated from classical swine fever infected pigs and wild boars

Classical swine fever is one of the most important infectious diseases for the pig industry worldwide due to its economic impact. Vaccination is an effective means to control disease, however within the EU its regular use is banned owing to the inability to differentiate infected and vaccinated animals, the so called DIVA principle. This inability complicates monitoring of disease and stops international trade thereby limiting use of the vaccine in many regions. The C-strain vaccine is safe to use and gives good protection. It is licensed for emergency vaccination in the EU in event of an outbreak. Two genetic assays that can distinguish between wild type virus and C-strain vaccines have recently been developed. Here the results from a comparison of these two real-time RT-PCR assays in an interlaboratory exercise are presented. Both assays showed similar performance.     

Up-regulated expression of PD-1 and its ligands during acute Classical Swine Fever virus infection in swine

In order to investigate the relationship between the PD-1 pathway and impairment of immune responses with the CSFV infection, the mRNA expression of PD-1 and its ligands were evaluated by quantitative polymerase chain reaction (qPCR) during artificial CSFV infection. Simultaneously, expression of IL-2 and IL-10 mRNA were detected. The T cell proliferation and CSFV load in plasma were also measured. Results showed that the expression of PD-1 and its ligands mRNA were significantly increased (p < 0.01) in PBMC from 3 to 7 days post infection (dpi). Meanwhile the level of IL-10 was up-regulated (p < 0.01). The IL-2 mRNA was not obviously changed but it is significantly increased from 14 dpi. The T cell proliferation was notably decreased at 7 dpi. The CSFV load was also increased in plasma. Overall, our results suggest that the expression of PD-1 and its ligands were up-regulated and probably correlated with immune inhibition during acute CSFV infection.     

Re-emergence of Pigeon Fever (Corynebacterium pseudotuberculosis) Infection in Texas Horses: Epidemiologic Investigation of Laboratory-Diagnosed Cases

In recent years, Texas has seen a dramatic increase in the number of clinical cases of Corynebacterium pseudotuberculosis (pigeon fever) infection in horses. Equine pigeon fever cases at Texas Veterinary Medical Diagnostic Laboratories (TVMDL) were analyzed with the objectives of investigating the spatiotemporal distribution and seasonal and annual trends of pigeon fever infection in horses in Texas between 2005 and 2011 and identifying high-risk areas and create a risk map for pigeon fever in horses in Texas. The study population consisted of horses culture-positive for C. pseudotuberculosis between January 1, 2005, and December 31, 2011 at TVMDL. The Poisson model of scan statistics was fitted to identify disease clusters. Empirical Bayesian smoothing was performed with the crude incidence estimates, followed by the geostatistical method of kriging to delineate high-risk areas. Cases increased 10-fold between 2005 and 2011. The annual cumulative incidence ranged from 9.3 to 99.5 per 100,000 horses at risk. Two seasonal peaks in the number of cases were observed in June and in December. Scan statistics identified a primary cluster in central Texas in 2011 (P < .0001 and relative risk of 9.2). Isopleth risk mapping also delineated a high-risk area in central Texas. High-risk areas were also detected in the panhandle and northern Texas. The epidemiological investigation supported anecdotal reports that pigeon fever is re-emerging in the Texas horse population. This study provides a baseline for future investigations of pigeon fever in the Texas horse population and serves as a reference for the disease distribution for veterinarians and horse owners.     

山西猪瘟野毒株E_2基因序列测定与遗传变异多样性分析

为了解山西地区猪瘟病毒（CSFV）流行毒株的遗传变异情况,采用RT-PCR方法,2013年从山西部分地区分离出5株CSFV流行毒株,并进行了E2全基因扩增、克隆与序列测定,应用DNAStar分析软件对所测定的5株毒株与国内外参考毒株的相应序列进行了同源性分析,绘制系统发育进化树。结果表明：5株CSFV流行毒株之间E2基因核苷酸序列与所推导氨基酸序列的同源性分别为81.4%~100%和87.9%~100%,与CSFV石门毒株（Shimen株）的核苷酸与氨基酸的同源性分别为82.9%~94.8%和89.0%~94.9%,与CSFV兔化弱毒株（HCLV株）的核苷酸与氨基酸的同源性分别为81.6%~99.6%和87.9%~99.5%,与17株来自各国不同地区的CSFV参考毒株的核苷酸与氨基酸的同源性分别为81.5%~99.6%和86.3%~99.7%。经系统发育关系分析,4株属于基因2群,且705、713、725、729、734和738位氨基酸发生置换,另外1株属于基因1群。本研究揭示了山西猪瘟流行毒株的遗传变异多样性现状。     

某猪场猪瘟免疫程序的调整及免疫效果评价

为了更好地预防和控制猪瘟,找出适合四川某猪场的猪瘟免疫程序,笔者根据实际情况对免疫程序进行了调整。然后用ELISA和IHA法对免疫程序调整前后的不同日龄猪血清进行了抗体检测,结果显示:调整前用ELISA法检测到3日龄、25日龄、50日龄、90日龄和120日龄猪只的猪瘟抗体阳性率分别为80%、54.54%、77.78%、80%和95%,用IHA法测得的阳性率分别为100%、63.64%、77.78%、90%和95%;调整后用ELISA法测得的猪瘟抗体阳性率分别为100%、80%、77.27%、77.78%和95%,用正向IHA法测得的抗体阳性率分别为97.73%、94.44%、96.15%、100%和100%。调整前用ELISA和IHA测得的平均阳性率为77.46%、85.28%,调整后则变为86.01%、97.66%,虽然均符合农业部颁布的猪群猪瘟抗体阳性率应不低于70%的标准,但免疫程序调整后的抗体阳性率明显高于调整前,可见调整后的免疫程序更适合于该猪场。     

黄芪多糖对猪瘟活疫苗免疫应答的佐剂作用

为研究黄芪多糖对猪瘟活疫苗的佐剂作用,分别用0.5％黄芪多糖、疫苗专用稀释液、生理盐水（对照）稀释猪瘟耐热保护剂活疫苗免疫猪,测定免疫后不同时间段内血清中 IgG 及其亚型IgG1、IgG2、IgG3、IgG4和细胞因子IL 2、IL 6的含量。结果显示,与对照组相比,黄芪多糖作为疫苗稀释液可以显著提高血清中IgG及其亚型和细胞因子IL 2、IL 6的含量。与疫苗专用稀释液组相比,在免疫后第7、14、28天,黄芪多糖组IgG含量均提高,分别提高1.6％（P＞0.05）、6.4％（P＜0.05）、7.1％（P＜0.05）,IL 2含量分别提高12.1％（P＜0.05）、2.9％（P＞0.05）、15.3％（P＜0.05）,IL 6含量差异均不显著。表明,黄芪多糖作为疫苗稀释液对猪瘟活疫苗具有良好的佐剂作用。     

应用酵母双杂交方法筛选与猪瘟病毒Npro蛋白相互作用的宿主蛋白

为研究猪瘟病毒(CSFV)与宿主细胞间的相互作用,本研究采用酵母双杂交方法以猪瘟病毒Npro蛋白为诱饵,从猪外周血单个核细胞(PBMC) cDNA表达文库中筛选与之相互作用的蛋白,经共转化试验表明,共筛选到12个与Npro蛋白相互作用的宿主蛋白,应用Cytoscape 2.8.2软件绘制了蛋白质间相互作用关系图,根据GO分析结果,这些蛋白分别参与细胞代谢、细胞生长、免疫等过程,经双分子荧光互补(BiFC)试验表明筛得的部分宿主蛋白与CSFV Npro具有相互作用.本研究中筛选得到的蛋白质对CSFV复制的影响有待进一步深入研究.     

腺病毒/甲病毒复制子嵌合载体猪瘟疫苗rAdV-SFV-E2最早提供完全攻毒保护时间的研究

本实验室前期构建了腺病毒/甲病毒复制子嵌合载体猪瘟疫苗rAdV-SFV-E2,其免疫效力与商品化猪瘟C株疫苗相同,并具有鉴别检测特性.为进一步确定该嵌合载体猪瘟疫苗最早提供完全攻毒保护的时间,本研究将rAdV-SFV-E2免疫健康仔猪(n=5),免疫后于不同时间点采用猪瘟病毒(CSFV)强毒石门株进行攻毒,并检测其抗体应答、临床症状、病毒血症、病理学及组织病理学变化等.结果显示,rAdV-SFV-E2免疫后1周和2周,分别能对致死性CSFV强毒的攻击提供部分保护(3/5存活)和不完全保护(全部存活,但出现了短暂的发热和低水平的病毒血症);免疫后3周和4周,能够完全抵抗致死性CSFV强毒的攻击,表现为无病毒血症,无临床症状,无病理变化和组织病理学变化.该研究进一步证实rAdV-SFV-E2能够作为一种新型猪瘟标记疫苗,在猪瘟的紧急防控中能够起到一定的作用.