鸡源巴氏杆菌GZ-TZ2017株的分离及其16S rRNA序列分析

Isolation of chicken source Pasteurella multocida GZ-TZ2017 strain and its 16S rRNA sequence analysis

为了探明贵州某肉鸡养殖场突发疾病病因,试验通过解剖、细菌分离、染色镜检、药敏试验、生化试验、16S rRNA的PCR扩增及序列分析等方法对病原进行分析。结果表明:从病鸡内脏中分离得到1株革兰氏阴性、两极浓染的球杆菌,将其命名为GZ-TZ2017;该菌株的培养特性、菌体形态、生化试验特征均与已报道的巴氏杆菌相似;GZ-TZ2017菌株对米诺环素、氧氟沙星和诺氟沙星等抗生素较为敏感,对头孢拉啶、苯唑西林、卡那霉素、青霉素等抗生素耐药;16S rRNA PCR扩增得到大小为1 400 bp左右的目的片段,分离菌GZ-TZ2017与各地分离的多杀性巴氏杆菌的同源性极高,均在99.0%以上;GZ-TZ2017与多杀性巴氏杆菌属细菌属于一个分支,和重庆分离株PM-LK(GenBank登录号为KX579746)、安徽滁州分离株CZ-6(GenBank登录号为KY673151)在同一小分支上,属于禽巴氏杆菌。说明该肉鸡场鸡群感染了禽巴氏杆菌。

In order to find out the causes of sudden illness in a broiler farm in Guizhou , the pathogen was analyzed by pathological examination, bacterial isolation, microscopic examination, drug sensitivity test, biochemical test and PCR amplication and sequence analysis of 16S rRNA. The results showed that a Gram negative and two polar dense Bacillus from the viscera of the sick chicken was named GZ^TZ2017.The culture characteristics, the morphology of the bacteria and the physiological and biochemical characteristics of the strain were similar to the reported Pasteurella. GZ-TZ2017 strains were sensitive to minocycline, ofloxacin and norfloxacin, and resistant to cefradine, oxacillin, kanamycin and penicillin. 16S rRNA PCR amplified the target fragment about 1 400 bp. The isolated bacteria were very homologous to the isolated Pasteurella multocida, and all homology were above 99. 0%. GZ-TZ2017 belongs to one branch of the genus Bacteria with Pasteurella multocida, and the Chongqing isolator PM-LK( GenBank No. KX579746) and the Anhui Chuzhou isolator CZ-6( Gen Bank No. KY673151) on the same small branch, belongs to poultry Bacillus. It shows that the chicken colony in the chicken farm is infected with poultry Bacillus.