牛病毒性腹泻病毒(BVDV)抗原捕获ELISA检测方法的建立

用牛病毒性腹泻病毒(BVDV)单克隆抗体包被酶标板,以兔多克隆抗体作为夹心抗体,建立BVDV抗原捕获ELISA(AC-ELISA)检测方法,优化反应条件并对该方法的稳定性等指标进行了测试和评价。结果表明,单抗最佳包被质量浓度为5μg/mL,兔多抗血清最佳质量浓度为10μg/mL;单抗在4℃包被12~24 h,多抗在37℃作用1 h为双抗体的最佳反应条件;酶标抗体最适稀释度1∶10000,最适作用时间为1 h;采用1%BSA和1%明胶分别在抗体包被后和加入待检抗原反应后进行两次封闭效果好。用AC-ELISA方法检测临床采集的11份牛腹泻病料和12份健康牛组织样品,同时以病毒分离和RT-PCR检测方法做对比,3种方法符合率很高。研究表明AC-ELISA方法稳定性好,可用于BVDV的临床快速检测。     

鸭黄病毒SDbz株的分离与初步鉴定

鸭黄病毒病为近年来新发的鸭病,给养鸭业带来了极大的经济损失。为了深入研究本病的防制,本试验对鸭黄病毒(duck flavivirus,DFV)进行了分离鉴定。取疑似感染DFV的病鸭病料,经细菌分离初步排除细菌感染后,应用RT-PCR检测呈现DFV阳性,处理后将其接种到鸭胚成纤维细胞(DEF)和健康鸭胚上进行病毒分离传代。结果显示,在DEF细胞上第1代48 h就开始出现CPE,随着时间的延长CPE更加明显,通常在72~96 h产生典型CPE;接种鸭胚每一代均出现死亡,且死亡时间多集中于接种后60~72 h,死亡鸭胚胚体水肿、出血、发育不良、胚肝严重出血、肿胀或斑驳样坏死等病变。将病毒DEF细胞和鸭胚分离物应用血凝试验、毒价测定、病毒中和试验、RT-PCR及人工感染试验进行检测鉴定,证明所分离到的病毒为DFV,并将其命名为DFV SDbz株。     

Genetic variation for tolerance or resistance to barley yellow dwarf virus in durum wheat

Summary One durum wheat line (Triticum durum), cv. 82PCD476, with useful BYDV tolerance or resistance, was singled out of 5 152 lines evaluated between 1979 and 1986. A few other lines such as cv. Boohai and cv. 12th IDSN 227, slightly inferior to cv. 82PCD476, also showed some value. With an hybrid of cv. 12th IDSN227 with the susceptible cv. 84PCY-S531, broad-sense heritability values of 0.37–0.41 were obtained for symptoms and a heritability value of 0.55 was obtained for the total weight of spikes. The weight of spikes was considered as a good indicator of wheat tolerance to BYDV. Although BYDV resistance or tolerance genes are not very common in durum wheat, sources of heritable resistance could be found. However, the resistance ofT. aestivum to BYDV was superior to the one found inT. durum.Cintribution no. 323     

水稻抗条纹叶枯病基因Stv-bi的分子标记辅助选择

水稻条纹叶枯病是我国黄淮及长江流域粳稻区重要的病害。由于水稻条纹叶枯病的发病受外界条件影响较大,人工接种抗性鉴定比较困难,利用与抗病基因紧密连锁的分子标记进行标记辅助选择对提高抗性育种效率具有重要意义。来自籼稻抗源Modan的Stv-bⁱ是水稻育种中广泛应用的条纹叶枯病抗性基因。本研究设计了与Stv-bⁱ紧密连锁的SSR及STS分子标记,用3个抗条纹叶枯病混合群体F30718(圣稻13/镇稻88)、F50701(武优34/T022//圣稻806)、F60702 (V6/T022//镇稻88)进行分子标记检测和田间条纹叶枯病抗性鉴定,其结果的符合率分别为99.3%、87.7%和91.8%。表明这些分子标记可以用于条纹叶枯病抗性基因Stv-bⁱ分子标记辅助选择。     

猪瘟病毒四川分离株E2基因的分子克隆、原核表达及免疫学活性分析

分子克隆猪瘟病毒(classical swine fever virus,CSFV)四川分离株E2基因,并对其进行原核表达及免疫学活性分析。PK-15细胞培养增殖CSFV四川分离株提取总RNA,运用RT-PCR扩增E2基因,定向克隆构建原核重组表达载体,转化E.coli Rosetta-gami-TM(DE3)plysS,IPTG诱导表达,SDS-PAGE检测蛋白表达,Western blotting分析表达蛋白的免疫学活性。试验结果表明,成功克隆了E2基因,共1119 bp,包含有编码E2蛋白的完整序列,编码373个氨基酸;成功构建了CS-FVE2基因完整阅读框、主要抗原区原核表达载体pET-E2(pe)、pET-mE2(pe);蛋白质电泳结果显示,成功表达出约45.32、28.49 ku两目的蛋白,而且表达的E2(pe)、mE2(pe)融合蛋白均能被CSFV阳性血清所识别,具有良好的免疫学反应活性。因此,本试验成功获得CSFV四川分离株E2基因,表达出具有生物学活性的E2(pe)、mE2(pe)融合蛋白。     

A new freeze-dried living virus vaccine against sheep-pox

A sheep-pox virus strain has been adapted and multiplied in primary lamb kidney cell cultures. The main characteristics of the strain have been verified in vitro after clones were isolated, and the results confirmed its identity. The safety and the potency of the strain have been investigated in sheep.

The inoculation of the strain to sheep was followed by a post-vaccinal reaction materialised by a nodule at the site of inoculation and an increase of temperature by about 1°C. No reactions adversely affecting pregnancy have been noted. Immunisation was demonstrated by an increase in the level of neutralising serous antibodies and protection against the pathogenic virus. The immunity tended to decrease during the second year after primovaccination and a yearly booster vaccination appeared to be necessary. Primovaccination of lambs over 2 months of age produced a better immunity, especially when the lambs were born from vaccinated ewes.

This strain forms the active principle of a freeze-dried vaccine containing no adjuvant of the immunity.     

Effects of bisphenol A,diethylhexyl phthalate and pentabrominated diphenyl ether 99 on steroid synthesis in cultured bovine luteal cells

Bisphenol A (BPA), diethylhexyl phthalate (DEHP) and pentabrominated diphenyl ether 99 (PBDE 99) are environmental toxicants belonging to the endocrine disrupting compounds (EDCs). They exert adverse effects on the various physiological systems, especially the reproductive system of humans and animals. The aim of this study was to investigate the effects of BPA, DEHP and PBDE 99 on progesterone (P4) synthesis in cultured bovine luteal cells. The bovine luteal cells isolated from the mid-luteal corpora lutea were exposed to different concentrations of BPA (1, 3, 10 and 30 µM), DEHP (1, 3, 10 and 30 µM) and PBDE 99 (0.1, 0.3, 1 and 3 µM) in a serum-free culture media for 48 and 96 hr. At 48 hr, the P4 level in the luteal cells decreased after treatment with all concentrations of BPA; 3, 10 and 30 µM of DEHP; and 3 µM of PBDE 99 compared to the control (p < .05). Treatment of cells with 3–30 µM of BPA, 1–30 µM of DEHP and 1–3 µM of PBDE 99 for 96 hr resulted in reduction in P4 synthesis (p < .05). However, lower concentrations of PBDE 99 (0.1 and 0.3 µM) increased P4 levels at 48 and 96 hr. Synthesis of P4 was lower at 96 hr compared to the 48 hr in the groups treated with BPA (30 µM), DEHP (1–30 µM), PBDE 99 (0.3–3 µM) and control group. Our results showed that BPA, DEHP and PBDE 99 are able to alter luteal steroidogenesis in bovine cells and can disrupt hormonal balance in the ovary. However, it is necessary to evaluate the exact mechanism underlying these effects in future studies.     

Risk for interspecies transmission of zoonotic pathogens during poultry processing and pork production in Peru: A qualitative study

Interspecies transmission of pathogens is an unfrequent but naturally occurring event and human activities may favour opportunities not previously reported. Reassortment of zoonotic pathogens like influenza A virus can result from these activities. Recently, swine and birds have played a central role as “mixing vessels” for epidemic and pandemic events related to strains like H1N1 and H5N1. Unsafe practices in poultry markets and swine farms can lead to interspecies transmission, favouring the emergence of novel strains. Thus, understanding practices that lead to interspecies interactions is crucial. This qualitative study aimed to evaluate poultry processing practices in formal and informal markets and the use of leftovers by swine farmers in three Peruvian cities: Lima (capital), Tumbes (coastal) and Tarapoto (jungle). We conducted 80 direct observations at formal and informal markets and interviewed 15 swine farmers. Processors slaughter and pluck chickens and vendors and/or processors eviscerate chickens. Food safety and hygiene practices were suboptimal or absent, although some heterogeneity was observed between cities and chicken vendors versus processors. Both vendors (76%) and processors (100%) sold the chicken viscera leftovers to swine farmers, representing the main source of chicken viscera for swine farms (53%). Swine farmers fed the chicken viscera to their swine. Chicken viscera cooking times varied widely and were insufficient in some cases. Non‐abattoired poultry leads to the sale of poultry leftovers to small‐scale swine farms, resulting in indirect but frequent interspecies contacts that can lead to interspecies transmission of bacterial pathogens or the reassortment of influenza A viruses. These interactions are exacerbated by suboptimal safety and hygiene conditions. People involved in these activities constitute an at‐risk population who could play a central role in preventing the transmission of pathogens between species. Educational interventions on hygiene and food safety practices will be important for reducing the risk of interspecies influenza transmission.     

我国不同地区麦蚜携带BYDV-GAV比率的差异

为了研究我国不同地区麦蚜携带大麦黄矮病毒麦二叉蚜麦长管蚜非专化性株系(BYDV GAV)比率的差异,采用RT-PCR技术,对BYDV-GAV的传毒介体麦蚜带毒情况进行检测.所用方法具有较高的灵敏度和特异性,测定样本用量可少至1/200头蚜虫;对采自我国主要麦区的蚜虫样本进行分子检测,山西、甘肃、青海、陕西11个小麦黄矮病重病区蚜虫样本带毒率为56％～91.5％,而河北、河南两省4个非重病区蚜虫样本带毒率为2.5％～33％.通过试验证实,我国不同地区麦蚜携带BYDV-GAV比率存在差异,小麦黄矮病重病区山西、甘肃、青海、陕西等地的麦蚜带毒率高,而非重病区河北、河南等地的麦蚜带毒率低.     

一例猪繁殖与呼吸综合征的RT-PCR诊断与防制

2008年3月,新乡市某猪场发生了以一种经产母猪和初产母猪流产、死产、产木乃伊胎、死胎及弱仔为特征的疾病。采集流产胎儿和死胎的肺、肝、脾脏等病料,制悬液,取上清,进行细胞培养,观察细胞病变。另根据猪繁殖与呼吸综合征病毒美洲株VR-2332 ORF7基因序列设计一对特异引物,提取流产胎儿的肺、肝等组织中PRRSV的RNA,经RT-PCR扩增,检测猪繁殖与呼吸综合征病毒为阳性,结合病史和临床症状,诊断为猪繁殖与呼吸综合征。通过对该猪场采取紧急接种疫苗、彻底消毒等综合防制措施,控制了该病的流行与蔓延。