Virulence genes and drug resistance characteristics of <i>Escherichia coli</i> in laying duck under the two different breeding modes

doi:10.3785/j.issn.1008-9209.2019.03.281

Journal of Zhejiang University (Agriculture and Life Sciences)

2020, Vol. 46

Issue (2): 254-262 DOI: 10.3785/j.issn.1008-9209.2019.03.281

Animal sciences & veterinary medicine

Virulence genes and drug resistance characteristics of Escherichia coli in laying duck under the two different breeding modes

Yan ZHAO¹(

),Junjie JIN¹,Minmin REN²,Fengxiang HOU¹,Suzhen LIU¹,Chengjun XUE³,Yingping XIAO²(

)

^1.Institute of Animal Sciences, Wenzhou Academy of Agricultural Sciences, Wenzhou 325000, Zhejiang, China
^2.Institute of Quality and Standard for Agricultural Products, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
^3.Cangnan County Animal Husbandry and Veterinary Bureau, Cangnan 325800, Zhejiang, China

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Abstract

This study mainly compared the virulence genes and drug resistance characteristics of Escherichia coli in laying ducks under the two different breeding modes: net bed with pool and net bed dry-system. Six farms including three net beds with pool and three net bed dry-systems were selected in Wenzhou, Zhejiang Province, and ten laying ducks were collected from each farm. The cotton swabs were used to collect microorganisms in the rectum forthe isolation of E. coli. The VITEK 2 COMPACT analysis system was used to identify the isolates and analyze the drug resistance profile. The virulence genes, drug-resistant genes and class Ⅰ integron genes were amplified by polymerase chain reaction. The results showed that the isolation rate of E. coli in laying ducks was 100%. The detection rates of virulence genes eae, ipa H, invE, aggR and pic of E. coli in the net bed dry-system were higher than those in the net bed with pool. For the drug-resistant phenotype, the proportion of strains with ≥3 resistance in E. coli in the net bed dry-system was 23.33%, while the proportion of strains with ≥3 in the net bed with pool was 10.00%. Similarly, the detection rates of the drug-resistant genes and the class Ⅰ integron genes in the net bed dry-system were also higher than those in the net bed with pool. Four of the 10 strains of class Ⅰ integron-positive E. coli carried the gene cassette insert, one of which derived from the E. coli class Ⅰ integron gene cassette of laying duck in the net bed with pool was dfrA12-aadA2, and the other three strains derived from the E. coli class Ⅰ integron gene cassette of laying duck in the net bed dry-system were dfrA1-aadA1. The above results indicate that the virulence gene carrying rate and drug resistance of E. coli in the intestinal tract of laying duck in the net bed dry-system are higher than those in the net bed with pool. It is suggested tha the changes of breeding mode may have some effect on the drug resistance characteristics of bacteria in waterfowls.

Key words： laying duck Escherichia coli drug-resistant phenotype drug-resistant gene virulence gene class Ⅰ integron

Received: 28 March 2019 Published: 22 May 2020

CLC:

S 834

Corresponding Authors: Yingping XIAO E-mail: yanzhao_wk@163.com;ypxiaozju@126.com

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	Yan ZHAO
	Junjie JIN
	Minmin REN
	Fengxiang HOU
	Suzhen LIU
	Chengjun XUE
	Yingping XIAO

Cite this article:

Yan ZHAO,Junjie JIN,Minmin REN,Fengxiang HOU,Suzhen LIU,Chengjun XUE,Yingping XIAO. Virulence genes and drug resistance characteristics of Escherichia coli in laying duck under the two different breeding modes. Journal of Zhejiang University (Agriculture and Life Sciences), 2020, 46(2): 254-262.

URL:

http://www.zjujournals.com/agr/10.3785/j.issn.1008-9209.2019.03.281 OR http://www.zjujournals.com/agr/Y2020/V46/I2/254

2种养殖模式下蛋鸭大肠埃希菌毒力基因和耐药特征分析

为比较分析在网床水面圈养与网床全旱养2种不同养殖模式下蛋鸭肠道中大肠埃希菌的毒力基因状况和耐药特征，分别在温州地区选择3家网床水面圈养和3家网床全旱养的蛋鸭场，每个蛋鸭场各选取10只蛋鸭，用棉拭子在直肠中采集微生物，用于大肠埃希菌分离，采用VITEK 2 COMPACT全自动细菌鉴定及药敏分析系统对分离株进行鉴定和开展耐药表型分析，并通过聚合酶链式反应（polymerase chain reaction, PCR）扩增毒力基因、耐药基因和Ⅰ型整合酶基因。结果表明：蛋鸭中大肠埃希菌的分离率为100%；网床全旱养蛋鸭肠道大肠埃希菌的毒力基因eae、ipa H、invE、aggR和pic检出率高于网床水面圈养蛋鸭。对于耐药表型，网床全旱养蛋鸭肠道大肠埃希菌耐药数≥3的菌株比例为23.33%，而网床水面圈养中耐药数≥3的菌株比例为10.00%。同样地，耐药基因和Ⅰ型整合酶基因的检出率也为网床全旱养高于网床水面圈养。10株Ⅰ型整合子阳性大肠埃希菌中4株携带基因盒插入片段，其中：1株来源于网床水面圈养蛋鸭，其大肠埃希菌Ⅰ型整合子基因盒插入的为dfrA12-aadA2；3株来源于网床全旱养蛋鸭，均为dfrA1-aadA1。网床全旱养蛋鸭肠道大肠埃希菌的毒力基因携带率和耐药性表型均高于网床水面圈养蛋鸭，说明养殖模式变化对水禽肠道微生物耐药性可产生一定的影响。

关键词： 蛋鸭, 大肠埃希菌, 耐药表型, 耐药基因, 毒力基因, Ⅰ型整合子

Table 1 Information of virulence gene primers

Table 2 Information of drug-resistant gene primers

Table 3 Information of β-lactamase gene primers

Fig. 1 Electrophoretogram of PCR products of eae geneM: DL5000 marker; NC: Negative control; 1-2: E. coli isolated from laying ducks under net bed with pool; 3-4: E. coli isolated from laying ducks under net bed dry-system; PC: Positive control.

Fig. 2 Detection percentage of virulence genes in E. coli

Fig. 3 Drug-resistant strains of E. coli in laying ducks under net bed with pool and net bed dry-system

Fig. 4 Detection percentage of drug-resistant genes of E. coli in laying duck under the net bed with pool and net bed dry-system

Fig. 5 Electrophoretogram of PCR products of CTX-M geneM: DL5000 marker; NC: Negative control; 1-2: Bacterial strains isolated from laying ducks under net bed with pool; 3-4: Bacterial strains isolated from laying ducks under net bed dry-system; PC: Positive control.

Fig. 6 Detection percentage of β-lactamase drug-resistant genes in ESBLs-positive E. coli

Fig. 7 Detection percentage of class Ⅰ integron in E. coli


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