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Genetic Diversity and Antibiotic Resistance of Escherichia coli Isolates from Different Leafy Green Production Systems

机译:不同叶类绿色生产系统中大肠杆菌分离物的遗传多样性和抗生素抗性

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Foodborne disease outbreaks linked to contaminated irrigation water and fresh produce are a public health concern. The presence of Escherichia coli isolates from irrigation water and leafy green vegetables in different food production systems (large commercial farms, small-scale farms, and homestead gardens) was investigated. The prevalence of antibiotic resistance and virulence in these isolates was further assessed, and links between water source and irrigated crops were identified using antimicrobial and genotypic analyses. Presumptive E. coli isolates were identified by matrix-assisted laser desorption ionization time-of-flight mass spectroscopy, and identities were confirmed by PCR using the uidA gene. Antimicrobial susceptibility was evaluated with the Kirby Bauer disk diffusion test; the presence of virulence genes was determined with enterobacterial repetitive intergenic consensus PCR assays. Of the 130 E. coli isolates from water (n = 60) and leafy green vegetables (n = 70), 19 (14.6%) were resistant to one antibiotic (tetracycline) and 92 (70.7%) were resistant to various antibiotics (including ampicillin, cefoxitin, and nalidixic acid). All E. coli isolates were susceptible to ceftriaxone and gentamicin. The virulence gene stx_2 was detected in E. coli isolates from irrigation water (8 [13.3%] of 60 isolates) and cabbages (3 [7.5%] of 40), but the virulence genes eae and stx_1 were not detected in any tested isolates from irrigation water and fresh produce samples. The prevalence of multidrug-resistant E. coli was lower in isolates from GLOBALG.A.P.-certified farms than in isolates from noncertified commercial and small-scale farms and homestead gardens. A link between the E. coli isolates from irrigation water sources and leafy green vegetables was established with phenotypic (antimicrobial) and genotypic (DNA fingerprinting) analyses. However, a link between virulence genes and the prevalence of antimicrobial resistance could not be established.
机译:与灌溉水和新鲜农产品污染有关的食源性疾病暴发是公共卫生问题。调查了在不同食品生产系统(大型商业农场,小型农场和宅基地花园)中从灌溉水和绿叶蔬菜中分离到的大肠杆菌的存在。进一步评估了这些分离株中抗生素耐药性和毒力的发生率,并使用抗微生物和基因型分析确定了水源与灌溉作物之间的联系。通过基质辅助激光解吸电离飞行时间质谱仪鉴定推定的大肠杆菌分离物,并使用uidA基因通过PCR确认身份。用Kirby Bauer纸片扩散试验评估抗菌药的敏感性;毒力基因的存在是通过肠细菌重复基因间共有PCR检测来确定的。从水分离出的130株大肠杆菌中(n = 60)和绿色蔬菜(n = 70)中,有19种(14.6%)对一种抗生素(四环素)有抗性,而92种(70.7%)对各种抗生素(包括氨苄西林,头孢西丁和萘啶酸)。所有大肠杆菌分离株均对头孢曲松和庆大霉素敏感。在灌溉水(60株菌株中的8 [13.3%])和大白菜(40株中的3 [7.5%])的大肠杆菌分离物中检测到毒力基因stx_2,但在所有测试菌株中均未检测到毒力基因eae和stx_1从灌溉水和新鲜农产品样品中提取。来自GLOBALG.A.P。认证农场的分离株的耐多药大肠杆菌的发生率低于未经认证的商业和小规模农场以及宅基地花园的分离株的流行。通过表型(抗微生物)和基因型(DNA指纹图谱)分析,建立了从灌溉水源分离的大肠杆菌和绿叶蔬菜之间的联系。但是,毒力基因和抗菌素耐药性的流行之间没有联系。

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