牛病毒性腹泻-黏膜病病毒噬菌体单链抗体库的构建和筛选

为筛选牛病毒性腹泻-黏膜病病毒(Bovine viral diarrhea virus,BVDV)gP48蛋白的单链抗体,本研究利用噬菌体展示技术构建了gP48蛋白单链抗体克隆文库,通过微孔筛选法对抗体库进行3轮富集淘筛,利用ELISA技术测定单链抗体的亲和力,对亲和力较高的克隆进行基因测序分析和结合活性测定,旨在获得gP48蛋白的单链抗体。结果表明:1)成功构建了库容量为4.3×10⁷的噬菌体单链抗体库,其重组率为83.3%,经三轮筛选对噬菌体抗体库进行富集,富集倍数为1.43×10⁴;2)优化了间接ELISA方法,筛选到3株与BVDV gP48蛋白具有高亲和力的单链抗体以及其基因序列,通过IgBLAST分析显示,3株单链抗体均为鼠源IgG。综上,从gP48蛋白的单链抗体库中筛选获得3个具有高亲和力的单链抗体,可用于后续BVDV检测方法的建立。     

A spatial risk assessment of bighorn sheep extirpation by grazing domestic sheep on public lands

Bighorn sheep currently occupy just 30% of their historic distribution, and persist in populations less than 5% as abundant overall as their early 19th century counterparts. Present-day recovery of bighorn sheep populations is in large part limited by periodic outbreaks of respiratory disease, which can be transmitted to bighorn sheep via contact with domestic sheep grazing in their vicinity. In order to assess the viability of bighorn sheep populations on the Payette National Forest (PNF) under several alternative proposals for domestic sheep grazing, we developed a series of interlinked models. Using telemetry and habitat data, we characterized herd home ranges and foray movements of bighorn sheep from their home ranges. Combining foray model movement estimates with known domestic sheep grazing areas (allotments), a Risk of Contact Model estimated bighorn sheep contact rates with domestic sheep allotments. Finally, we used demographic and epidemiologic data to construct population and disease transmission models (Disease Model), which we used to estimate bighorn sheep persistence under each alternative grazing scenario. Depending on the probability of disease transmission following interspecies contact, extirpation probabilities for the seven bighorn sheep herds examined here ranged from 20% to 100%. The Disease Model allowed us to assess the probabilities that varied domestic sheep management scenarios would support persistent populations of free-ranging bighorn sheep.     

仔猪高死亡率腹泻病因探讨

2010年4月以来,全国各地先后出现高死亡率吮乳仔猪腹泻,以冬春寒冷季节多发、持续时间长、高死亡率为特征,日龄越小死亡率越高,用药物及生物制剂均无法有效控制,给养殖业带来巨大的经济损失。按照流行性腹泻病毒感染、霉菌毒素及中毒等采取相应的防治措施,效果均不明显。按中兽医整体观念、辨证论治,将两方面病因有机结合起来,分析该疾病的病理机制,采用中药复方进行临床验证,取得较好的防治效果,为该病的预防和治疗提供了新的思路与方法。     

奶牛四种流产性疾病的流行现状及疫苗研究进展

奶牛布氏杆菌病、传染性鼻气管炎、病毒性腹泻-黏膜病及新孢子虫病是导致我国奶牛流产发病比例升高的主要疾病,有效防控这四种疾病一直是世界关注的重点之一。本文对上述四种疾病的流行现状及疫苗研究进展进行了综述。     

2株猪流行性腹泻病毒的分离鉴定及ORF3基因的克隆与序列分析

为研究、防控广东地区猪流行性腹泻,首先通过RT-PCR法确定某猪场粪便样品中存在猪流行性腹泻病毒,随后利用Vero细胞进行盲传,最后经核酸类型鉴定、RT-PCR、病毒TCID50的测定、目的基因测序和序列分析等方法,确认分离到2株猪流行性腹泻病毒毒株,命名为PEDV GDKP-2013和PEDV GDKP2-2013,通过ORF3全基因核苷酸序列分析可知,GDKP-2013株与GDKP2-2013株与经典参考毒株的氨基酸序列一致性为 JX261936-CHGD-01,98.2%,98%;JQ039903-CH-GD-2011,98.6%,98.4%;AF353511-CV777,95.4%,95.1%;KC344845-STPED0810,95.1%,94.8%;EU054929-DR13,95.5%,95.2%;KF272920-USA-Colorado-2013,95.1%,94.8%,表明此2毒株与国内分离株的氨基酸同源性高,与韩国、泰国和美国分离株的氨基酸同源性要低一些,同时,与疫苗参考株的氨基酸同源性也相对较低;基因系统进化分析可知,GDKP-2013株与GDKP2-2013株主要处在Ⅰa分支,均与弱毒疫苗株（Ⅰb分支）相互之间遗传进化关系较远。     

励珂健喷剂对10日龄内仔猪腹泻和免疫力的影响

为研究励珂健喷剂（主要成分：杨树花提取物）对10日龄内仔猪腹泻和免疫力的影响,将18头PIC母猪随机分成3组,每组6头,分别为蒸馏水组、土霉素组和励珂健喷剂组,蒸馏水组仔猪1、3、5日龄时皮肤喷洒蒸馏水1 m L/头/d,土霉素组仔猪3、7日龄时分别肌肉注射20%长效土霉素注射液0.2 m L/头/d、0.5m L/头/d,励珂健喷剂组仔猪1、3、5日龄时皮肤喷洒励珂健喷剂1 m L/头/d。结果表明,励珂健喷剂可有效降低10日龄内仔猪的腹泻率和发病率,且励珂健喷剂组腹泻仔猪的治愈速度快于土霉素组和蒸馏水组。由此证明：励珂健喷剂对10日龄内仔猪腹泻有防治作用,并且还可以增强仔猪的免疫力。