广州市生鲜鸡中肠炎沙门菌的监测及耐药基因的分子鉴定

doi:10.11843/j.issn.0366-6964.2018.02.018

畜牧兽医学报 ›› 2018, Vol. 49 ›› Issue (2): 388-395.doi: 10.11843/j.issn.0366-6964.2018.02.018

广州市生鲜鸡中肠炎沙门菌的监测及耐药基因的分子鉴定

孙朋浩¹, 邱荣超², 李书宁³, 郑双双¹, 杨伟聪¹, 张丽娜¹, 符颖¹, 詹泽强¹, 张建民¹, 廖明¹, 瞿孝云^1*

1. 华南农业大学兽医学院人兽共患病防控制剂国家地方联合工程实验室, 农业部人畜共患病重点实验室, 广东省动物源性人兽共患病预防与控制重点实验室, 广州 510642;
2. 河南牧业经济学院动物科技学院, 郑州 450011;
3. 云南省西双版纳州动物疫病预防控制中心, 景洪 666100

收稿日期:2017-08-03 出版日期:2018-02-23 发布日期:2018-02-23
通讯作者: 瞿孝云(1982-),女,主要从事动物传染病的研究,E-mail:qxy0926@126.com
作者简介:孙朋浩(1996-),男,山东青岛人,学士,主要从事细菌传染病学研究,E-mail:1220572895@qq.com
基金资助:
公益性行业（农业）科研专项（201403054）；广东省大学生科技创新培育专项（pdjh2016a0072）；国家自然科学基金（31402193）；国家肉鸡产业技术体系（cARS-41-G16）

Monitoring of Salmonella Enteritidis and Molecular Identification of Antibiotic Resistance Gene in Fresh Chicken of Guangzhou City

SUN Peng-hao¹, QIU Rong-chao², LI Shu-ning³, ZHENG Shuang-shuang¹, YANG Wei-cong¹, ZHANG Li-na¹, FU Ying¹, ZHAN Ze-qiang¹, ZHANG Jian-min¹, LIAO Ming¹, QU Xiao-yun^1*

1. National and Regional Joint Engineering Laboratory for Medicament of Zoonosis Prevention and Control, Key Laboratory of Zoonoses of Ministry of Agriculture, Key Laboratory of Zoonoses Control and Prevention of Guangdong, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China;
2. College of Animal Science, Henan University of Animal Husbandry and Economy, Zhengzhou 450011, China;
3. Yunnan Xishuangbanna Animal Disease Prevention and Control Center, Jinghong 666100, China

Received:2017-08-03 Online:2018-02-23 Published:2018-02-23

摘要/Abstract

摘要：

本研究旨在了解广州市鸡肉中沙门菌的流行情况、血清型分布及肠炎沙门菌耐药性与耐药基因的携带情况。对采集自广州市部分农贸市场的鸡肉样品，参照国标检测方法（GB4789.4-2016）进行分离培养并使用泰国S&A公司沙门菌属诊断血清进行血清分型；用Kirby-Bauer法测定肠炎沙门菌对16种抗菌药物的耐药性，用PCR方法检测肠炎沙门菌携带的耐药基因。结果显示：316份鸡肉样品中，阳性率为76.9%。共鉴定23种血清型，主要血清型为阿贡纳（Salmonella Agona，19.8%）、科瓦利斯（S.Kottbus，14.0%）、姆班达卡（S.Mbandaka，11.9%）、肯塔基（S.Kentucky，10.3%）、肠炎（S. enteritidis，7.8%）、布伦登卢普（S.Braenderup，7.4%）。耐药性结果显示：肠炎沙门菌对萘啶酸耐药率最高（100.0%），其次是磺胺复合物（79.0%）、氨苄西林（57.9%）、链霉素（36.8%）、四环素（21.0%）、庆大霉素（21.0%）、头孢噻肟（15.8%）、头孢他啶（5.3%），多重耐药率为68.4%。肠炎沙门菌中喹诺酮耐药决定区基因gyrA、gyrB、parC的突变率分别为94.7%、73.7%、15.8%，最常检出突变为苯丙氨酸-414→丝氨酸（73.7%），其次是天冬氨酸-87→甘氨酸（42.1%）、天冬氨酸-87→色氨酸（31.6%）、丝氨酸-83→色氨酸（21.1%）；喹诺酮类耐药质粒的检出率较低。β-内酰胺类耐药基因bla_TEM、bla_CTX-M的检出率分别为42.1%、10.5%。结果显示，广州市鸡肉中沙门菌的污染率较高，血清型复杂。肠炎沙门菌对萘啶酸、磺胺甲基异恶唑、氨苄西林等常用抗生素的耐药情况较为严重，多重耐药率高。肠炎沙门菌的喹诺酮耐药决定区基因突变率高，与肠炎沙门菌对喹诺酮类药物的耐药结果具有高度的一致性。

Abstract:

This experiment was conducted to investigate the prevalence of Salmonella, the distribution of serotypes and the antibiotic resistance of Salmonella enteritidis, and resistant genes they carried, in chicken of Guangzhou city. According to the national standard GB4789.4-2016, the chicken samples were collected. Salmonella serotypes were determined using diagnostic sera from S&A, Thailand. The antibiotic resistance of Salmonella enteritidis was determined by using the Kirby-Bauer method. The resistance gene of Salmonella enteritidis was detected by PCR. In 316 samples of chicken, the positive rate was 76.9%. A total of 23 serotypes were identified. The main serotypes include Salmonella Agona (19.8%), Salmonella Kottbus(14.0%), Salmonella Mbandaka (11.9%), Salmonella Kentucky(10.3%), Salmonella enteritidis (7.8%) and Salmonella Branederup (7.4%). The resistance rates of Salmonella enteritidis to nalidixic acid, peptidylsulfanilamide, ampicillin, streptomycin, tetracycline, gentamicin, cefotaxime and ceftazidime were 100.0%, 79.0%, 57.9%, 36.8%, 21.0%, 21.0%, 15.8% and 5.3%, respectively. The multiple antibiotic resistance rate was 68.4%. The mutation rates of gyrA, gyrB and parC were 94.7%, 73.7% and 15.8%, respectively. The most main mutations were Phe-414→Ser (73.7%), followed by Asp-87→Gly (42.1%),Asp-87→Trp(31.6%) and Ser-83→Trp(21.1%). The detection rate of quinolone-resistant plasmids was low. The detection rates of bla_TEM and bla_CTX-M were 42.1% and 10.5%, respectively. The pollution rate of Salmonella in chicken of Guangzhou city is high and the serotype is complex. Salmonella enteritidis are resistant to Nalidixic acid, peptidylsulfanilamide, ampicillin. Multiple antibiotic resistance is high. The mutation rate of the gene in the quinolone resistance-determining region is high, which is highly consistent with the resistance of Salmonella to quinolones.

中图分类号:

R378

孙朋浩, 邱荣超, 李书宁, 郑双双, 杨伟聪, 张丽娜, 符颖, 詹泽强, 张建民, 廖明, 瞿孝云. 广州市生鲜鸡中肠炎沙门菌的监测及耐药基因的分子鉴定[J]. 畜牧兽医学报, 2018, 49(2): 388-395.

SUN Peng-hao, QIU Rong-chao, LI Shu-ning, ZHENG Shuang-shuang, YANG Wei-cong, ZHANG Li-na, FU Ying, ZHAN Ze-qiang, ZHANG Jian-min, LIAO Ming, QU Xiao-yun. Monitoring of Salmonella Enteritidis and Molecular Identification of Antibiotic Resistance Gene in Fresh Chicken of Guangzhou City[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(2): 388-395.

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广州市生鲜鸡中肠炎沙门菌的监测及耐药基因的分子鉴定

Monitoring of Salmonella Enteritidis and Molecular Identification of Antibiotic Resistance Gene in Fresh Chicken of Guangzhou City

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