The global spread of antibiotic-resistant bacteria and their resistance genes is a critical issue that is no longer restricted to hospital settings, but also represents a growing problem involving environmental and food safety. In this study, we have performed a microbiological and genomic investigation of critical priority pathogens resistant to broad-spectrum cephalosporins and showing endophytic lifestyles in fresh vegetables sold in a country with high endemicity of extended-spectrum β-lactamases (ESBLs). We report the isolation of international high-risk clones of CTX-M-15-producing Escherichia coli, belonging to clonal complexes CC38 and CC648, and Klebsiella pneumoniae of complex CC307 from macerated tissue of surface-sterilized leaves of spinach, cabbage, arugula, and lettuce. Regardless of species, all ESBL-positive isolates were able to endophytically colonize common bean (Phaseolus vulgaris) seedlings, showed resistance to acid pH, and had a multidrug-resistant (MDR) profile to clinically relevant antibiotics (i.e., broad-spectrum cephalosporins, aminoglycosides, and fluoroquinolones). Genomic analysis of CTX-M-producing endophytic Enterobacterales revealed a wide resistome (antibiotics, biocides, disinfectants, and pesticides) and virulome, and genes for endophytic fitness and for withstanding acidic conditions. Transferable IncFIB and IncHI2A plasmids carried bla_CTX-M-15 genes and, additionally, an IncFIB plasmid (named pKP301cro) also harbored genes encoding resistance to heavy metals. These data support the hypothesis that fresh vegetables marketed for consumption can act as a figurative Trojan horse for the hidden spread of international clones of critical WHO priority pathogens producing ESBLs, and/or their resistance genes, to humans and other animals, which is a critical issue within a food safety and broader public and environmental health perspective.

中文翻译：

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新鲜蔬菜中产生广谱{beta}-内酰胺酶的主要病原菌的全球克隆的内生性生活方式：有利于人类殖民的特洛伊木马策略？

抗生素抗性细菌及其抗性基因的全球传播是一个关键问题，已不再局限于医院环境，而是一个日益严重的问题，涉及环境和食品安全。在这项研究中，我们对耐广谱头孢菌素的关键优先病原体进行了微生物学和基因组学研究，并显示了在广谱β-内酰胺酶（ESBLs）高度流行的国家出售的新鲜蔬菜中的内生生活方式。我们报告了CTX-M-15生产大肠杆菌的国际高风险克隆的分离，属于克隆复合物CC38和CC648，以及肺炎克雷伯菌菠菜，卷心菜，芝麻菜和生菜经表面消毒的叶子浸软组织中复合CC307的合成。无论是哪种物种，所有ESBL阳性分离株均能够内生定植于普通豆（菜豆）幼苗，对酸性pH表现出抗性，并且对临床相关抗生素（即广谱头孢菌素，氨基糖苷和氟喹诺酮类）。对产生CTX-M的内生肠杆菌的基因组分析表明，其抵抗力范围广泛（抗生素，杀生物剂，消毒剂和农药）和毒理素，以及内生适应性和耐酸性条件的基因。可移植的IncFIB和IncHI2A质粒带有bla _CTX-M-15基因，以及另外一个IncFIB质粒（名为pKP301cro）也包含编码对重金属的抗性的基因。这些数据支持这样一种假设，即销售食用的新鲜蔬菜可以作为具有象征意义的特洛伊木马，以隐藏世卫组织生产ESBLs和/或其抗性基因的关键世卫组织重要病原体国际克隆对人类和其他动物的隐性传播，这是至关重要的。食品安全以及更广泛的公众和环境健康角度来看问题。