禽波氏杆菌病研究进展

禽波氏杆菌病是由禽波氏杆菌引起的一种火鸡的传染病。本文综述了该病的病原特性,临床症状与病理变化,诊断方法,以及该病的治疗与预防。     

Back-calculation method shows that within-flock transmission of highly pathogenic avian influenza (H7N7) virus in the Netherlands is not influenced by housing risk factors

To optimize control of an avian influenza outbreak knowledge of within-flock transmission is needed. This study used field data to estimate the transmission rate parameter (β) and the influence of risk factors on within-flock transmission of highly pathogenic avian influenza (HPAI) H7N7 virus in the 2003 epidemic in The Netherlands. The estimation is based on back-calculation of daily mortality data to fit a susceptible-infectious-dead format, and these data were analysed with a generalized linear model. This back-calculation method took into account the uncertainty of the length of the latent period, the survival of an infection by some birds and the influence of farm characteristics. After analysing the fit of the different databases created by back-calculation, it could be concluded that an absence of the latency period provided the best fit. The transmission rate parameter (β) from these field data was estimated at 4.50 per infectious chicken per day (95% CI: 2.68–7.57), which was lower than what was reported from experimental data. In contrast to general belief, none of the studied risk factors (housing system, flock size, species, age of the birds in weeks and date of depopulation) had significant influence on the estimated β.     

Hemolytic activity of Trichomonas gallinae isolates does not correspond with clinical virulence

The hemolytic activity of 22 Trichomonas gallinae isolates was investigated using an 18 h erythrocyte hemolysis assay which has been shown to correlate with the clinical virulence of T. vaginalis. Absorbance of the assay supernatants was measured at 540 nm and expressed as percentage of complete hemolysis. Mean hemolytic activity of the T. gallinae isolates ranged from 3.5% to 53.4% and did not correspond with clinical virulence. The results of this investigation suggest hemolytic activity is not a useful in vitro virulence assay for T. gallinae.     

Multiple amino acid substitutions are involved in the adaptation of H9N2 avian influenza virus to mice

To explore adaptation of avian influenza virus to mice we previously performed serial lung-to-lung passages of the influenza A/Chicken/Jiangsu/7/2002 (H9N2) strain, resulting in the isolation of a variant influenza strain lethal for mice. We now report that virulence correlates with improved growth characteristics on mammalian cells and extended tissue tropism in vivo. Sequencing of the complete genomes of the wild-type and mouse-adapted viruses revealed 25 amino acid substitutions. Some were found to reiterate known substitutions in human and swine H9N2 influenza isolates. Functions affected include nuclear localization signals and sites of protein and RNA interaction, while others are known determinants of pathogenicity and host specificity such as the viral polymerase PB2 E627K substitution. These observations suggest that enhanced growth characteristics and modified cell tropism may contribute to increased virulence in mice. We conclude that multiple amino acid substitutions are likely to be involved in the adaptation of H9N2 avian influenza virus to mice.     

青海野生鸟类禽流感监测防控亟待关注解决的问题

近年来,青藏高原迁徙的野生鸟类不断发生高致病禽流感疫病,给生态安全和人们的健康带来了极大的威胁。如何面对和做好野生鸟类的疫源疫病监测与防控,本文通过青海省发生和开展的防控工作情况,就面临和亟待解决的问题,提出对策建议。     

Immunoadjuvant effects of bacterial genomic DNA and CpG oligodeoxynucleotides on avian influenza virus subtype H5N1 inactivated oil emulsion vaccine in chicken

This study investigated the immunoadjuvant effects of three types of bacterial genomic DNA and CpG oligonucleotides (CpG ODN) on the avian influenza virus (AIV) subtype H5N1 inactivated oil emulsion vaccine under two immunization strategies. The genomic DNA extracted from Escherichia coli O₂, Staphylococcus aureus, Streptococcus faecalis FQ68, and synthetic CpG ODN were used as adjuvants, and their effects on the AIV oil emulsion vaccine were examined in chickens. The results indicated that when administered separately from the vaccine, adjuvants induced lower haemagglutination inhibition (HI) titres and serum IgG titres but resulted in higher concentrations of IFN-γ and IL-10. In contrast, when combined with the oil emulsion vaccine prior to inoculation, CpG ODN induced higher HI, IgG titres and IFN-γ concentration but resulted in lower IL-10 concentration. These data suggest that, depending on the immunization approaches, adjuvants may exert distinct immune effects in chickens receiving AIV H5N1 oil emulsion vaccine: the prior incorporation of CpG ODN into the vaccine may augment both the humoral and Th1 type immune responses, while separate inoculation of adjuvants has not shown better adjuvanticity.     

表达H5N1亚型禽流感病毒HA基因的重组鸭瘟病毒的构建

利用PCR技术扩增出鸭瘟病毒(DPV)生长非必需区的TK基因(约2.5kb),将其克隆入pUC19载体获得载体puDTK。根据已知载体pcDNA-LacZ的序列设计了一对引物,PCR扩增出pcDNA-LacZ上含CMV启动子、多克隆位点、SV40及LacZ的完整的基因表达盒插入puDTK的TK上,获得质粒puDTCL。根据Genbank已发表的H5N1亚型禽流感病毒的血凝素(HA)基因序列,设计引物从T-HA质粒上扩增出HA基因,克隆到puDTCL的表达盒的多克隆位点NheI与ApaI之间,成功构建含LacZ及HA基因的转移载体质粒puDTCL-HA。将这载体质粒与DPV34F2疫苗毒共转染鸡胚成纤维细胞(CEF),经蓝斑克隆筛选和纯化,获得了遗传性状稳定的表达HA基因的重组鸭瘟病毒。     

加拿大应对禽流感的扑杀补偿政策及启示

自2003年以来,禽流感疫情在全球各地不断暴发,禽流感已成为国际社会共同面临的公共卫生问题。本文介绍了加拿大应对禽流感的扑杀补偿政策,尤其是其扑杀补偿的标准和补偿金额的计算方法,并从中得到改善我国禽流感扑杀补偿政策的有益启示,包括扑杀补偿政策法律化、补偿政策的激励相容性、补偿范围和标准及补偿金额的计算方法等五个方面。     

H9N2亚型禽流感病毒血凝素基因的克隆及原核表达

据GenBank收录的H9N2亚型禽流感病毒血凝素（HA）基因序列设计并合成引物,以H9N2亚型禽流感病毒RNA为模板,用R1PCR方法扩增了预计约1700bp的HA基因,将此扩增产物克隆进pMD18-T载体,采用限制性酶切及序列测定鉴定阳性重组克隆子结果表明HA基因长为1683bp。基于HA信号肽在表达中的负作用,研究通过基因工程手段缺失HA蛋白位于起始的信号肽的编码户列,获得了缺失HA蛋白信号肽的HA基因,将其亚克隆到pGEX-KG中,与GST融合表达。SDS-PAGE显示：融合表达的蛋白分子量乡为90kDa。     

74种鸟类线粒体基因组碱基组成及特征分析

 在鸟类线粒体基因组中,不同物种的线粒体碱基组成和特性存在明显的差异。截至2008年5月,GenBank细胞器基因组资源数据库共公布了74种鸟类线粒体全基因组数据。本研究利用已公布的鸟类线粒体基因组全序列分析其碱基组成特征。结果表明：(1)鸟类线粒体基因组密码子第二位的GC含量值波动范围十分狭窄,而密码子第三位的GC含量值波动范围很大。(2) 密码子第三位碱基中C的含量波动范围较大,为32.60%-50.70%。(3)线粒体基因组GC含量主要由密码子第三位的碱基C和T的变化引起。(4)密码子第三位碱基的GC含量与线粒体基因组的GC含量的变化存在相关性。这些结果,为今后鸟类线粒体基因组的深入研究提供借鉴和参考资料。