Prevalence and Molecular Characterization of Antimicrobial Resistance in Escherichia coli Isolated from Raw Milk and Raw Milk Cheese in Egypt
Prevalence and Molecular Characterization of Antimicrobial Resistance in Escherichia coli Isolated from Raw Milk and Raw Milk Cheese in Egypt
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DOI：
10.4315/0362-028x.jfp-17-277
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发表时间：
2018-02-01
影响因子：
2
通讯作者：
Yamasaki, Shinji
中科院分区：
文献类型：
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作者：
Ombarak, Rabee A.;Hinenoya, Atsushi;Yamasaki, Shinji
本研究的目的是检测从187份原料牛奶和埃及两种最受欢迎的奶酪样品中分离出的222株大肠杆菌的抗微生物药物耐药性，并对其相关基因进行表征。采用纸片扩散法检测分离的大肠杆菌对12种抗菌药物的敏感性。在222株大肠杆菌中，66株（29.7%）对一种或多种抗菌素耐药，其中一半耐药菌株表现为多药耐药表型（至少对三种不同药物类别耐药）。对四环素（27.5%）、氨苄西林（18.9%）、链霉素（18.5%）、磺胺甲恶唑-甲氧苄啶（11.3%）、头孢噻肟（4.5%）、卡那霉素（4.1%）、头孢他啶（3.6%）、氯霉素（2.3%）、萘啶酸（1.8%）、环丙沙星（1.4%）耐药。对磷霉素和亚胺培南均无耐药。检出四环素耐药基因tetA 53株，tetB 9株，tetD 1株，未检出tetC。检测到氨基糖苷类耐药基因strA、strB、aadA和aphA1分别为41株、41株、11株和9株。磺胺耐药基因sul1、sul2和sul3分别在7株、25株和3株中检测到。42株氨苄西林耐药菌株中分别检出bla（TEM）、bla（CTX-M）和bla（SHV）菌株40株、9株和3株，其中10株（23.8%）耐药菌株产生广谱β -内酰胺酶。对产内酰胺酶广谱大肠杆菌的bla基因进一步分型为bla（CTX-M-15）、bla（CTX-M-104）、bla（TEM-1）和bla（SHV-12）。在28株耐药菌株中也检测到1类整合子，并通过pcr -限制性片段长度多态性获得了3种不同的整合子模式。变异区测序结果显示，dfrA12/orfF/aadA2为4株，aadA22为2株，dfrA1/aadA1为1株。这些数据表明，耐抗生素大肠杆菌广泛分布在埃及的牛奶生产和加工环境中，并可能在传播对其他致病菌和共生菌的抗微生物耐药性方面发挥作用。
The goal of this study was to examine antimicrobial resistance and characterize the implicated genes in 222 isolates of Escherichia coli from 187 samples of raw milk and the two most popular cheeses in Egypt. E. coli isolates were tested for susceptibility to 12 antimicrobials by a disk diffusion method. Among the 222 E. coli isolates, 66 (29.7%) were resistant to one or more antimicrobials, and half of these resistant isolates showed a multidrug resistance phenotype (resistance to at least three different drug classes). The resistance traits were observed to tetracycline (27.5%), ampicillin (18.9%), streptomycin (18.5%), sulfamethoxazole-trimethoprim (11.3%), cefotaxime (4.5%), kanamycin (4.1%), ceftazidime (3.6%), chloramphenicol (2.3%), nalidixic acid (1.8%), and ciprofloxacin (1.4%). No resistance to fosfomycin and imipenem was observed. Tetracycline resistance genes tetA, tetB, and tetD were detected in 53 isolates, 9 isolates, and 1 isolate, respectively, but tetC was not detected. Aminoglycoside resistance genes strA, strB, aadA, and aphA1 were detected in 41, 41, 11, and 9 isolates, respectively. Sulfonamide resistance genes sul1, sul2, and sul3 were detected in 7, 25, and 3 isolates, respectively. Of 42 ampicillin-resistant isolates, bla(TEM), bla(CTX-M), and bla(SHV) were detected in 40, 9, and 3 isolates, respectively, and 10 (23.8%) ampicillin-resistant isolates were found to produce extended-spectrum beta-lactamase. Each bla gene of extended-spectrum beta-lactamase-producing E. coli was further subtyped to be bla(CTX-M-15), bla(CTX-M-104), bla(TEM-1), and bla(SHV-12). The class 1 integron was also detected in 28 resistant isolates, and three different patterns were obtained by PCR-restriction fragment length polymorphism. Sequencing analysis of the variable region revealed that four isolates had dfrA12/orfF/aadA2, two had aadA22, and one had dfrA1/aadA1. These data suggest that antimicrobial-resistant E. coli are widely distributed in the milk production and processing environment in Egypt and may play a role in dissemination of antimicrobial resistance to other pathogenic and commensal bacteria.