牛病毒性腹泻-粘膜病病毒地方株动物回归试验

【目的】自流产奶牛粪便及血液中分离得到的非细胞病变型牛病毒性腹泻-粘膜病病毒(BVDV),为了解分离毒株的特性,进行回归动物试验;【方法】将此分离毒回归2月龄健康犊牛,接毒25天后扑杀剖检,观察临床症状及病理变化。自犊牛体内采取脾、骨髓、肠淋巴结及血液接种MDBK细胞进行培养,分离病毒,进行细胞传代,将此细胞毒进行RT-PCR检测;【结果】犊牛表现为典型的急性病毒性腹泻症状和病变。自病料分离病毒经MDBK细胞盲传至第9代,均未出现细胞病变。将此细胞毒进行RT-PCR检测,扩增出相应的665bp的片段;【结论】将分离株接种易感犊牛以复制出BVD-MD病症,并从试验牛体内分离得到BVDV,从而进一步确证分离到的毒株属BVDV。     

腹泻抗原融合基因CTB-CS3对胡萝卜的遗传转化

CS3是CFA(产肠毒素型大肠杆菌引起腹泻时产生两类毒性因子之一)阳性株的保护性抗原。临床研究结果表明,以CS3作为疫苗可有效地防止由相关细菌导致的腹泻(董自正等,     

谷氨酰胺对断奶仔猪生长性能的影响

选取21日龄断奶杜大长仔猪60头(体重接近5 kg),按试验要求随机分成2组,每组设3个重复,每个重复10头.研究饲粮中添加1%谷氨酰胺对断奶仔猪生长的影响.结果表明,试验组在第9 d后腹泻率明显下降,而对照组在整个过程中的腹泻率较高,断奶8～11 d一直维持50%～60%的腹泻率,另外腹泻延续的时间较长,断奶12～15 d仍有20%～30%的腹泻率.断奶1～10 d,试验组料重比比对照降低了12.05%(P＜0.05).断奶11～20 d,试验组日增重比对照组提高27.75%(P＜0.05),试验组采食量、料重比与对照组相近.断奶10 d,试验组血清尿素氮水平比对照组低17.36%(P＞0.05),但对血清总蛋白和血清白蛋白水平无明显影响.断奶20 d,谷氨酰胺对血清尿素氮和总蛋白有一定影响,试验组血清尿素氮水平比对照组低4.26%(P＞0.05),血清总蛋白水平比对照组高18.70%(P＞0.05),谷氨酰胺对血清白蛋白水平无明显影响.饲粮添加谷氨酰胺对T3,T4水平无明显影响,但对生长激素水平有一定的提高作用.     

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.     

猪流行性腹泻病毒M蛋白单克隆抗体的制备及鉴定

为制备猪流行性腹泻病毒(PEDV)抗M蛋白单克隆抗体(MAb),本研究以截短表达的His-M重组蛋白免疫BALB/c小鼠;以截短表达的GST-M重组蛋白作为包被抗原,采用常规的淋巴细胞杂交瘤技术制备杂交瘤细胞,通过间接ELISA进行筛选,得到一株稳定分泌抗M蛋白MAb.MAb亚类鉴定为IgG2b型,轻链为к链,杂交瘤细胞培养上清和诱导的小鼠腹水抗体效价分别为1:3000和1:2×105.Western blot试验表明该MAb能够识别重组及天然的PEDV M蛋白.间接免疫荧光试验表明该MAb能够与PEDV感染的Vero E6细胞产生特异性免疫荧光.     

牛病毒性腹泻病毒纳米抗体文库的构建与鉴定

通过构建牛病毒性腹泻病毒(BVDV)驼源重链抗体文库,并鉴定抗体文库的多样性,为筛选抗BVDV特异性重链抗体奠定基础。利用灭活的BVDV免疫双峰驼,多次免疫后测定血清抗体滴度,采集免疫后的血液并分离其外周淋巴细胞,抽提RNA,用RT-PCR法扩增重链抗体可变区(VHH)片段;将其克隆至载体pCANTAB5E,电转大肠杆菌TG1获得VHH抗体基因库,检测抗体库库容量,随机挑取克隆测序验证抗体文库多样性。结果表明,所构建的抗体文库的库容量为4.32×105;测序和聚类分析表明,抗体文库多样性丰富。利用辅助噬菌体救援后,得到噬菌体展示文库,测定滴度达1.3×1012 cfu/mL。     

猪两种腹泻病毒混合感染的病原诊断

对我省流行的疑似猪病毒性腹泻进行了病原诊断，通过细菌培养、ＲＮＡ电泳和电镜形态观察等辅助诊断和免疫萤光特异性诊断，确诊该病病原为猪传染性胃肠炎病毒（ＴＧＥＶ）与猪流行性腹泻病毒（ＰＥＤＶ）的混合感染，混合毒代号为ＴＰＶ３。     

犊牛腹泻的病因、诊断及综合防治

介绍了犊牛腹泻的流行病学、发病原因及临床症状,并对该病的实验室诊断、治疗及综合预防措施进行了综述。     

Serotypes of Escherichia coli strains isolated from piglets with diarrhea in swine industrial farms

Serotypes of E. coli strains isolated from piglets, which died with symptoms of diarrhea in 9 swine industrial farms, were determined. Large numbers of serotypes (from 16 to 27) in individual farms were detected. The sets of serotypes from 9 investigated farms differed among each other significantly, depending on the farm and time of examination. It was found that more than one serotype of E. coli may exist in the pig body and contribute to the development of disease. The predominant serotypes, i.e. those comprising more than 10% of serologically determined strains, were found to exist in 6 of the investigated farms and not in the remaining ones. Among the predominant serotypes, particularly important seem to be strains with K88 antigen. For prophylaxis of piglet colibacteriosis in industrial farms in Poland two vaccines for sows are recommended: one containing the K88 antigen only and the other the following serotypes: 0149:K91,K88; 020:K57; 020:K83; 0157:K88; 01:K1; 0136:K78; 024:K?; 078:K80 and 0118:K? Strains belonging to these serotypes were the most prevalent in our strain collection.