河南部分地区乌鸡群鸭源鸡杆菌的分离鉴定及系统发育分析

doi:10.11843/j.issn.0366-6964.2018.11.017

摘要/Abstract

摘要：

为了解乌鸡中鸡杆菌的流行情况及其分离株的遗传进化特点，从河南信阳和南阳的两个乌鸡场采集棉拭子样品，进行鸡杆菌的分离和荧光定量PCR检测，对分离的菌株进行PCR鉴定、药敏试验和16S rRNA、rpoB及gyrB基因的PCR扩增，随后进行三个管家基因的序列测定分析，利用DNAStar中MegAlign程序计算分离株之间的相似性，利用MEGA5.0构建进化树进行遗传进化分析。结果显示：22只乌鸡中16只为鸭源鸡杆菌阳性，阳性率为72.7%；分离的4株鸡杆菌经鉴定均为鸭源鸡杆菌；4株鸭源鸡杆菌均为多重耐药菌株，只对头孢曲松、头孢噻肟、阿米卡星和妥布霉素等药物具有一定的敏感性；分离株与鸭源鸡杆菌的参考菌株相似性分别为99.1%~99.8%（16S rRNA）、85.4%~99.6%（rpoB）、92.7%~99.8%（gyrB）；在16S rRNA基因进化树中分离的4株鸭源鸡杆菌与参考鸭源鸡杆菌位于同一分支，在rpoB基因进化树中GAC193分离株与多杀性巴氏杆菌位于同一分支中，与其他鸭源鸡杆菌不在同一分支中，gyrB基因进化树中，GAC017与鸡杆菌基因种1在一个小的分支。结果表明：乌鸡中有鸭源鸡杆菌存在，并具有较高的检出率；从乌鸡中分离到4株鸭源鸡杆菌，这些菌株多重耐药现象严重；分离的部分鸭源鸡杆菌在rpoB和gyrB基因位点具有明显的遗传进化差异。

Abstract:

In order to understand the epidemic situation of Gallibacterium in silky and the genetic evolution characteristics of isolates, samples were collected for the isolation of Gallibacterium anatis and q-PCR identification from two silky farms in Xinyang and Nanyang, Henan province, then isolates were detected by Gallibacterium specific PCR followed by drug susceptibility test, and the PCR amplification and sequencing of the 16S rRNA, rpoB and gyrB genes. Finally, the MegAlign program was used for calculating the similarity between isolates of G. anatis, and MEGA5.0 for constructing evolutionary tree and genetic evolution analysis. Results were as follows:Of 22 silky chickens, 16 were found positive to G. anatis, the positive rate was 72.7%; the four Gallibacterium isolates were identified as G. anatis; the four isolates were all multidrug resistant, and were sensitive to ceftriaxone, cefotaxime, amikacin and tobramycin only; the homology between the isolates and the reference G. anatis species was 99.1%-99.8% (16S rRNA), 85.4%-99.6% (rpoB), and 92.7%-99.8% (gyrB), respectively. In the 16S rRNA gene evolution tree, the four isolates and the reference G. anatis were located in the same branch; the GAC193 isolate in the rpoB gene evolution tree were located in the same branch with Pasteurella multocida, not in the same branch with other G. anatis; in the gyrB gene evolution tree, GAC017 and G. genomosp.1 were in a small branch. The results showed that G. anatis were found in the silky chickens with a high detection rate, and four strains of G. anatis were isolated, which showing serious multidrug resistance. There are significant genetic differences in the rpoB and gyrB gene of some isolates of G. anatis.

中图分类号:

S852.61

皇甫和平, 杜振隆, 董青, 王宏魁, 孙彦婷, 郭宏伟, 姬向波, 贾含笑, 余萌薇, 卜亚歌. 河南部分地区乌鸡群鸭源鸡杆菌的分离鉴定及系统发育分析[J]. 畜牧兽医学报, 2018, 49(11): 2451-2460.

HUANGFU He-ping, DU Zhen-long, DONG Qing, WANG Hong-kui, SUN Yan-ting, GUO Hong-wei, JI Xiang-bo, JIA Han-xiao, YU Meng-wei, BU Ya-ge. Isolation,Identification and Phylogenetic Analysis of Gallibacterium anatis from Silky Population in Henan Province, China[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(11): 2451-2460.

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