Agriculture and Agri-Food Canada, Guelph Food Research Center, 93 Stone Road West, Guelph, Ontario, N1G 5C9, Canada.
Agriculture and Agri-Food Canada, Pacific Agri-Food Research Centre, PO Box 5000, 4200 Highway 97, Summerland, British Columbia, V0H 1Z0, Canada.
Int J Food Microbiol. 2015 Dec 23;215:7-15. doi: 10.1016/j.ijfoodmicro.2015.07.029. Epub 2015 Jul 29.
Understanding the survival mechanisms used by Shiga toxin-producing Escherichia coli (STEC), including O157:H7 and non-O157 serotypes, is important for minimizing contamination of fresh produce and occurrence of foodborne outbreaks. Recent outbreaks linked to leafy green vegetables and sprouted seeds have prompted researchers to focus on investigating decontamination strategies. Several studies showed that hydrogen peroxide (H2O2) treatment has been effective in reducing pathogens on fresh produce. As such, the effect of hydrogen peroxide on stress-associated and virulence gene expression in six STEC isolates was investigated in this study. Logarithmic phase cells of E. coli O157:H7 (EDL933) and non-O157 serotypes, including E. coli O26:H11 (EC20070549), O103:H2 (EC19970811), O104:H4 (NML#11-3088), O111:NM (EC20070546) and O145:NM (EC19970355) were exposed to 2.5mM H2O2 for 40 min and gene expression was evaluated using quantitative real-time PCR. Different patterns of gene expression were observed in E. coli O157:H7 and non-O157 serotypes. Particularly, Shiga toxin gene stx2 was upregulated in O157:H7, but not in O104:H4. Moreover, stx1 was significantly upregulated in STEC O157:H7, but only slightly upregulated Stx1-positive non-O157 serotypes. However genes related to motility (fliC) and intimin gene (eae) were downregulated in most strains. Stress-associated sodA gene encoding manganese superoxide dismutase was significantly upregulated in all serotypes. The dps gene coding for non-specific DNA binding protein was upregulated in O145:NM, O111:NM, O103:H2 and O26:H11. However genes related to cold shock (cspC) and acid resistance (gadW) were significantly downregulated in all strains tested. The results of this study provide a basic understanding of the oxidative stress impact on survival and virulence of non-O157 serotype STEC strains.
了解产志贺毒素大肠杆菌(STEC),包括 O157:H7 和非 O157 血清型的生存机制,对于最大限度减少新鲜农产品的污染和食源性疫情的发生非常重要。最近与叶菜类蔬菜和发芽种子有关的疫情促使研究人员专注于调查去污策略。几项研究表明,过氧化氢(H2O2)处理在减少新鲜农产品上的病原体方面非常有效。因此,本研究调查了过氧化氢对 6 株 STEC 分离株应激相关和毒力基因表达的影响。对数期 E. coli O157:H7(EDL933)和非 O157 血清型,包括 E. coli O26:H11(EC20070549)、O103:H2(EC19970811)、O104:H4(NML#11-3088)、O111:NM(EC20070546)和 O145:NM(EC19970355)用 2.5mM H2O2 处理 40 分钟,并用实时定量 PCR 评估基因表达。在 E. coli O157:H7 和非 O157 血清型中观察到不同的基因表达模式。特别是,Shiga 毒素基因 stx2 在 O157:H7 中上调,但在 O104:H4 中没有上调。此外,stx1 在 STEC O157:H7 中显著上调,但在仅轻微上调 Stx1 阳性非 O157 血清型中上调。然而,大多数菌株的运动相关基因(fliC)和紧密素基因(eae)下调。所有血清型的应激相关 sodA 基因编码锰超氧化物歧化酶显著上调。编码非特异性 DNA 结合蛋白的 dps 基因在 O145:NM、O111:NM、O103:H2 和 O26:H11 中上调。然而,所有测试菌株的冷休克(cspC)和耐酸(gadW)相关基因显著下调。本研究的结果为非 O157 血清型 STEC 菌株的生存和毒力对氧化应激的影响提供了基本了解。