贵州某鸡场大肠杆菌与葡萄球菌混合感染病原的分离鉴定

doi:10.16431/j.cnki.1671-7236.2017.05.030

《中国畜牧兽医》 ›› 2017, Vol. 44 ›› Issue (5): 1468-1476.doi: 10.16431/j.cnki.1671-7236.2017.05.030

贵州某鸡场大肠杆菌与葡萄球菌混合感染病原的分离鉴定

陈绍品^1,2, 温贵兰^1,2, 张升波^1,2, 伍昌华^1,2, 林汉卿^1,2, 范昌龙^1,2, 管国丹^1,2, 汪德生^1,2, 文明^1,2, 周碧君^1,2, 程振涛^1,2

1. 贵州大学动物科学学院, 贵阳 550025;
2. 贵州省动物疫病与兽医公共卫生重点实验室, 贵阳 550025

收稿日期:2016-11-01 出版日期:2017-05-20 发布日期:2017-05-18
通讯作者: 温贵兰, 汪德生 E-mail:524340732@qq.com;16525986@qq.com
作者简介:陈绍品(1993-),男,贵州毕节人,硕士生,研究方向:兽医病毒学及免疫学,E-mail:1334177986@qq.com
基金资助:
贵州大学博士基金（贵大人基合字201312号）；贵州省科技计划课题联合基金（黔科合LH字20147666号）；贵州大学“本科教学工程”项目（JKSP2013004）；贵州省科学技术基金（黔科合J字20152046号）；贵州省科技创新人才团队项目（黔科合人才团队20154016）

Isolation and Identification of Pathogen ofEscherichia coli andStaphylococcus Mixed Infection of Chichen in Guizhou Province

CHEN Shao-pin^1,2, WEN Gui-lan^1,2, ZHANG Sheng-bo^1,2, WU Chang-hua^1,2, LIN Han-qing^1,2, FAN Chang-long^1,2, GUAN Guo-dan^1,2, WANG De-sheng^1,2, WEN Ming^1,2, ZHOU Bi-jun^1,2, CHENG Zhen-tao^1,2

1. College of Animal Science, Guizhou University, Guiyang 550025, China;
2. Guizhou Institute of Animal Disease and Veterinary Public Health, Guiyang 550025, China

Received:2016-11-01 Online:2017-05-20 Published:2017-05-18

摘要/Abstract

摘要：

为探明贵州某鸡场患病鸡的细菌感染种类、耐药性及致病性等情况，本试验对送检的4只病鸡进行细菌分离培养，并对分离所得的细菌进行革兰氏染色镜检、生化试验、药敏试验、16S rDNA分子序列及动物感染试验等分析。结果显示，成功分离到了2株菌落形态不一的菌株，分别为革兰氏阴性杆菌与阳性球菌，根据分离地点和时间将其分别命名为GZHX2016-1及GZHX2016-2。GZHX2016-1为大肠杆菌，具有多重耐药性；GZHX2016-2为模仿葡萄球菌，血浆凝固酶阴性，对多种常用抗生素耐药。GZHX2016-1的16S rDNA与大肠杆菌（GenBank登录号：CP007442.1、CP014667、KT156725.1等）同源性达99.5%，GZHX2016-2与葡萄球菌（GenBank登录号：HM140412.1、AM944030.1、KM877513.1等）同源性达97.9%。GZHX2016-1与GZHX2016-2对小鼠都有致病性，GZHX2016-1的最小致死量为1.12×10⁸ CFU，GZHX2016-2为3.20×10⁷ CFU。综上所述，本试验成功从送检鸡中分离出1株致病性大肠杆菌和1株致病性血浆凝固酶阴性模仿葡萄球菌，该鸡场鸡群存在大肠杆菌与葡萄球菌混合感染的情况，且该鸡场感染细菌对多种抗生素耐药，应采取有效措施控制细菌在鸡群中的污染并限制抗生素在养鸡过程中的使用，以减少细菌耐药性的产生，保障鸡肉及鸡肉制品的食品安全。

关键词: 模仿葡萄球菌; 大肠杆菌; 生化鉴定; 16S rDNA

Abstract:

In order to analyze the variety, drug resistance and pathogenicity of bacterial of a chicken farm in Guizhou province, bacterial isolation culture was done from four sick chickens that were submitted by the farm. Gram staining, biochemical test, drug susceptibility test, 16S rDNA sequencing analysis and so on were carried out to identify the bacteria.The results showed that we isolated two strains of bacteria with different colony morphology that one of them was gram-negative bacilli,the other was gram-positive cocci, which were named as GZHX2016-1 and GZHX2016-2 according to the isolated location and time.GZHX2016-1 wasEscherichia coli (E. coli) with multiple drug, and GZHX2016-2 was Staphylococcus simulans which was negative on Staphylococcus coagulase test and was resistant to some antibiotics. The 16S rDNA sequencing of GZHX2016-1 was more close genetic relationship with E. coli (GenBank accession No.:CP007442.1, CP014667, KT156725.1, and so on), and the homology was 99.5%.The GZHX2016-1 was more close genetic relationship with Staphylococci (GenBank accession No.:HM140412.1, AM944030.1, KM877513.1, and so on), and the homology was 97.9%. GZHX2016-1 and GZHX2016-2 were pathogenic to mouse. The minimum lethal dose of the GZHX2016-1 was 1.12×10⁸ CFU of each mouse by intraperitoneal injection, and the minimum lethal dose of GZHX2016-2 was 3.20×10⁷ CFU.In summary, the experiment successfully isolated a strain of pathogenic E. coli and a strain of coagulase-negative Staphylococcus from chickens.The chicken farm was infected by E. coli andStaphylococcus,and the bacteria were resistant to multiple antibiotics. So, we should take effective measures to control the bacteria pollution in the chicken farm, and restrict the use of antibiotics to reduce the resistance bacteria, and ensure the food safety of chicken and chicken products.

Key words: i>Staphylococcus simulans; Escherichia coli; biochemical identification; 16S rDNA

中图分类号:

S858.31

陈绍品, 温贵兰, 张升波, 伍昌华, 林汉卿, 范昌龙, 管国丹, 汪德生, 文明, 周碧君, 程振涛. 贵州某鸡场大肠杆菌与葡萄球菌混合感染病原的分离鉴定[J]. 《中国畜牧兽医》, 2017, 44(5): 1468-1476.

CHEN Shao-pin, WEN Gui-lan, ZHANG Sheng-bo, WU Chang-hua, LIN Han-qing, FAN Chang-long, GUAN Guo-dan, WANG De-sheng, WEN Ming, ZHOU Bi-jun, CHENG Zhen-tao. Isolation and Identification of Pathogen ofEscherichia coli andStaphylococcus Mixed Infection of Chichen in Guizhou Province[J]. , 2017, 44(5): 1468-1476.

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贵州某鸡场大肠杆菌与葡萄球菌混合感染病原的分离鉴定

Isolation and Identification of Pathogen ofEscherichia coli andStaphylococcus Mixed Infection of Chichen in Guizhou Province

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