由禽波氏杆菌和奇异变形杆菌共感染为主所致鸡胚发病死亡的生物学鉴定  

Biological identification of dead chicken embryos mainly co-infected with Bordetella avium and Proteus mirabilis

作　　者：郭繁霞[1] 朱瑞良[1] 刘冠华[1] 崔国林[1] 

机构地区：[1]山东农业大学动物科技学院山东省动物生物技术与疫病防治重点实验室,山东泰安271018

出　　处：《中国兽医学报》2013年第11期1668-1673,共6页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(30972183;31272595)

摘　　要：2010年,辽宁铁岭某种鸡孵化场出现孵化率低、死淘率高的现象。通过对送检病死鸡胚进行病原检测,最终检测出5株革兰阴性杆菌和3种病毒,其中禽波氏杆菌和奇异变形杆菌的检出率分别为76.44%、61.36%,两者混合感染率达51.83%,而其他病原菌(大肠杆菌、沙门菌、绿脓杆菌)和病毒(ALV、REV、CIAV)的检出率均较低。采用1对根据病原菌23SrRNA基因的共同保守序列设计的引物,以及根据临床常见致病菌16S～23SrRNA基因间隔序列(ISR)两端的16S及23SrRNA保守序列而设计的通用引物分别对分离菌进行PCR扩增。并分别测定所得片段的DNA序列。结果显示,所得DNA片段分别与GenBank中收录的登录号为HM545299的禽波氏杆菌和登录号为AY993943的奇异变形杆菌的相应核苷酸序列同源性达99.5%和98.7%。本研究最终确定禽波氏杆菌和奇异变形杆菌的混合感染是引起该场疫情发生的主要原因。In 2010,low hatching rate appeared and high death ratein chicken hatchery in Tieling. Five isolates of gram-nagative bacteria and three kinds of viruses were isolated from dead chicken embryos. They were diagnosed as multiple infected with Bordetella avium , Proteus miralilis, and some other pathogenic bacteria and viruses. The infection rate of Bordetellaaviura was 76.44%, the infection rate of Proteus miralilis was 61.36% ,and the combined infection rate of Bordetella avium and Proteus miralilis was 51. 83% ,The influence of some other pathogenies such as esche- richia coli ,salmonella ,ALV,REV were very low. A pair of primers which was designed according to the 23SrRNA of Bordetella avium ,Escherichia coli ,Salmonella ,Pseudomonas aeruginosa and staphylococcus was used. And we used another pair of primers which was designed according to the conserved sequences of 16S-23SrRNA. The sequences located in the 16S-23S rRNA gene inter- genic spacer region （ISR） of clinical common pathogenic bacteria. The bacteria strains were iden tiffed by PCR and the products were identified by the PCR length polymorphism comparison, the sequences obtained in this study were compared with the conserved sequences reported in Gen Bank by the software of DNAStar analyses. The result showed that the homology with reference sequence of gordetella avium（No. HM545299） was 99.5% ,and the the homology with reference sequence of Proteus miralilis（No. AY993943）was 98.7%. This study finally determined that the co-infected of Bordetella avium with Proteus miralilis was the main pathogenic bacteria of the epidemic.

关 键 词：禽波氏杆菌 奇异变形杆菌 分离 鉴定 共感染 16S-23S rRNA 

分 类 号：S852.61[农业科学—基础兽医学]