禽霍乱克隆89弱毒菌苗免疫原性及免疫持续期试验

禽霍乱克隆８９弱毒菌苗５×１０６个活菌对鸡即产生免疫，免疫剂量为１０×１０６个活菌，应用剂量为３０×１０６个活菌。应用剂量免疫后．用３０个强毒菌攻击，２４ｈ保护率８０％。免疫期１２５天，１００个以上强毒菌攻击，保护率６０％。     

赤水乌骨鸡MC1R基因多态性及其生物信息学分析

采用DNA混池及PCR产物直接测序技术,对赤水乌骨鸡、泰和乌鸡、兴义白鸡、兴义白鸡F2代、贵州黄快羽鸡、贵州黄慢羽鸡、罗曼蛋鸡和瑶山鸡8个鸡种群MC1R基因外显子区域进行多态性分析。结果显示:瑶山鸡与红色原鸡序列一致,无SNPs,赤水乌骨鸡的4个SNPs分别为G23A(Arg→His)、C69T、T212C、G274A,泰和乌鸡的8个SNPs分别为C69T、T212C、G274A、T398C(Leu→Pro)、G636A、T637C、A644C(His→Pro)、C834T,贵州黄快、慢羽鸡和兴义白鸡均有3个SNPs,分别为A427G(Thr→Ala)、G636A、T637C,罗曼蛋鸡的4个SNPs分别为T398A(Leu→Gln)、G636A、T637C、C834T,兴义白鸡F2代的4个SNPs分别为C69T、T212C、G274A、A644C(His→Thr);生物信息学分析发现,除G274A、T398C、T398A、A644C位点的稳定性增加外,其余位点的稳定性均有所降低;除G274A、T398C、T637C、A644C位点的整体多样性有所下降外,其余位点的均有所增加;除泰和乌鸡MC1R基因编码蛋白为不稳定蛋白外,其余种群的均属于稳定蛋白,且包括泰和乌鸡在内的7个鸡群MC1R基因编码蛋白均为疏水性蛋白和非分泌蛋白;赤水乌骨鸡、泰和乌鸡、兴义白鸡、兴义白鸡F2代和罗曼蛋鸡的MC1R蛋白三级结构均由α–螺旋、β–转角、无规则卷曲组成。     

Fowl adenoviruse-4 infection induces strong innate immune responses in chicken

Fowl adenovirus (FAdV), as the causative agent of hepatitis-hydropericardium syndrome (HHS), poses a significant threat to the poultry industry in China in recent years. In this study, we investigated the immunopathogenesis of a FAdV-4 strain HN/151025 in 60-day-old chickens. The virus was highly virulent in chickens, with a broader tissue tropism in chickens, causing 60 % mortality. Postmortem findings of dead chickens showed mild HHS and liver degeneration and necrosis. Importantly, FAdV-4 infection induced significant upregulation of genes encoding most toll-like receptors, some cytokines (interleukin-1β, 2, 6, 8, and 18, and interferon-γ), most of avian β-defensins, myeloid differentiation primary response protein 88, p38 mitogen-activated protein kinases, and inducible nitric oxide synthase, in tissues of infected chicken, especially in spleen and bursa of Fabricius. There was also a significant positive correlation between FAdV-4 genome load and the mRNA expression levels of most of these factors in specific infected tissues. The results indicated the potential role of these proteins in host immune response against FAdV-4 infection. However, overexpression of these proteins might contribute to tissue damage of FAdV-4 infected chickens, and eventually lead to chicken death.     

几种鸟类mtDNA限制性酶切图谱的比较研究

本文用５种限制内切酶（ＨｉｎｄⅢ，ＢｇＬＩ，ＰｓｔＩ和ＢａｍＨＩ）初步建立了头鹅、白鹅、火鸡和鹌鹑的肝脏线粒体ＤＮＡ的内切酶图谱。结果表明白鹅和狮头鹅的ｍｔＤＮＡ的酶切图谱极为相似。山鸡、火鸡和鹌鹑的ｍｔＤＮＡ限制性性内切酶图谱之间的差异是很大的，以上事实说明驯化中的鹅类的不同品种间ｍｔＤＮＡ差异不在。而鸡形目中的不同物种间的ｍｔＤＮＡ是有很大的差异的。     

对实验性鸡霍乱病鸡组织中脂质过氧化物的初步分析

本文以TBA比色法测定了鸡霍乱病鸡肝、脾及十二指肠组织中脂质过氧化物(LPO)的含量。实验结果表明,禽巴氏杆菌可明显提高鸡肝、脾及十二指肠组织中LPO的水平(P<0.01),对照组与实验组LPO的水平分别为:肝358.1±48.01与548.41±49.84nmol(MDA)/g;脾412.61±63.51与521.62±45.13nmol(MDA)/g;十二指肠220.68±38.28与678.56±131.92nmol(MDA)/g。     

Ⅰ群禽腺病毒33K基因的原核表达与鉴定

33K蛋白是Ⅰ群禽腺病毒（fowl adenovirus groupⅠ,FAVⅠ）的非结构蛋白,在感染的过程中起重要作用。为了构建原核表达载体pGEX-33K作为诊断抗原,本研究根据GenBank中FAVⅠ33K基因序列,设计1对特异性引物。用PCR方法扩增目的片段,将其克隆至原核表达载体pGEX-4T-1,构建了重组质粒pGEX-33K。将其转化大肠杆菌感受态细胞DH5α,经IPTG诱导和SDS-PAGE分析,pGEX-33K表达的分子量为36.7KD,表达产物主要以可溶性的形式存在于菌体中;最佳IPTG诱导浓度为0.75mmol/L,最佳诱导时间为4h;经Western Blot鉴定,pGEX-33K重组蛋白可与FAVⅠ阳性血清发生特异性反应,为FAVⅠ抗体ELISA鉴别诊断试剂盒的研究奠定基础。     

Inhibition of electrolyte transport in isolated guinea-pig ileum by berberine

AIM: To investigate the effects of berberine (Ber) on ion transport and hypersecretion induced by cholera toxin(ChT) in ileum and its mechanism. METHODS: Ussing Chamber technique was used to measure the potential difference(PD), short-circuit current(SCC), and the resistance(R) in isolated guinea-pig ileum. The effects of Ber on PD, SCC and R in glucose Tyrode solution or glucose-free Tyrode solution were examined. The secretory diarrhoea model was made by ChT to investigate the effects of ChT and ChT+Ber on PD, SCC and R. RESULTS: (1)The PD and SCC were decreased in glucose Tyrode solution by the Ber added in mocosal side or in serosal side in ileum of normal guinea-pig and which was with secretory diarrhoea (P<0. 01, P<0. 01). 1. 0 mmol·L^-1 Ber in serosal face also decreased the R (P<0. 01). (2) In glucose-free Tyrode solution, the Ber in mucosal face did not decrease the PD, SCC and R, but 1. 0 mmol·L^-1 Ber in serosal face decreased the PD, SCC and R (P<0. 01, P<0. 01, P<0. 05). CONCLUSION: Ber not only inhibited electrogenic ion transport in normal guinea-pig ileum, but also reversed hypersecretion induced by ChT.     

禽霍乱蜂胶与氢氧化铝胶灭活疫苗免疫效果比较试验

利用蜂胶作为佐剂制成禽霍乱蜂胶佐剂灭活疫苗,将其与同批菌液制成的禽霍乱氢氧化铝胶灭活疫苗进行免疫效果比较试验,结果显示,用蜂胶佐剂制成的禽霍乱灭活疫苗免疫效果优于禽霍乱氢氧化铝胶灭活疫苗。     

猪群高热症的血清流行病学调查研究

采用血清学试验对贵阳市花溪区某种猪场的95份血清样本进行猪繁殖障碍性疫病血清学检测,以了解猪群免疫水平及疫病感染状况.结果显示,猪群O型口蹄疫（O-FMD）、猪瘟（HC）、猪繁殖与呼吸障碍综合征（PRRS）、猪伪狂犬病（PR）免疫合格率分别为100％ （33/33）、62.9％（39/62）、14.5％ （9/62）、61.3％ （38/62）,HC、PRRS和PR免疫合格率均未达到农业部要求;通过比较母猪免疫抗体与仔猪母源抗体水平相关性,发现HC与PR首免后抗体保护期维持较短,PRRS免疫不合格;猪群猪圆环病毒病（PC）、猪细小病毒病（PP）隐性感染抗体阳性率分别为70.5％ （67/95）、82.1％（78/95）,隐性感染情况严重.血清流行病学调查结果表明,该猪场应改进HC、PR免疫程序,立即对全群进行PRRS补免,并要高度重视PC和PP的防控。