环境中抗生素耐药性的出现和传播对全球公共卫生构成了严重威胁。这是公认的非抗生素的压力,包括消毒剂,药物和有机污染物,在抗生素抗性基因(ARGs)的水平传播中起着至关重要的作用。尽管广泛存在非甾体抗炎药(NSAIDs),特别是在地表水中,它们对ARGs转移的贡献尚未得到系统探讨。此外,以前的研究主要集中在模型菌株上,以研究污染物是否促进ARGs的共轭转移,在非抗生素污染物的选择压力下,在天然水性环境中抗生素耐药细菌之间传播ARG的机制尚不清楚.在这项研究中,大肠杆菌(E.coli)将携带RP4质粒的K12作为供体菌株,太湖地区含有利福平抗性基因的维氏气单胞菌,和大肠杆菌HB101被用作受体菌株,以建立种间和种内共轭转移系统,研究酮洛芬应激下的共轭转移频率。结果表明,酮洛芬通过多种机制加速了ARGs的环境传播。酮洛芬通过增加细胞表面疏水性和减少细胞表面电荷来促进细胞与细胞的接触,从而减轻细胞间的排斥。此外,酮洛芬诱导活性氧(ROS)产生水平增加,激活DNA损伤诱导反应(SOS),增强细胞膜通透性,促进ARG在属内和属间系统中的传播。外膜蛋白的上调,氧化应激,SOS响应,交配对形成(Mpf)系统,和DNA转移和复制(Dtr)系统相关基因,以及对全球调控基因的抑制,所有这些都有助于酮洛芬治疗下更高的转移效率。这些发现为全面评估NSAIDs在天然水性环境中抗生素耐药性传播中的作用提供了预警。
The emergence and spread of antibiotic resistance in the environment pose a serious threat to global public health. It is acknowledged that non-antibiotic stresses, including disinfectants, pharmaceuticals and organic pollutants, play a crucial role in horizontal transmission of antibiotic resistance genes (ARGs). Despite the widespread presence of non-steroidal anti-inflammatory drugs (NSAIDs), notably in surface water, their contributions to the transfer of ARGs have not been systematically explored. Furthermore, previous studies have primarily concentrated on model strains to investigate whether contaminants promote the conjugative transfer of ARGs, leaving the mechanisms of ARG transmission among antibiotic resistant bacteria in natural aqueous environments under the selective pressures of non-antibiotic contaminants remains unclear. In this study, the Escherichia coli (E. coli) K12 carrying RP4 plasmid was used as the donor strain, indigenous strain Aeromonas veronii containing rifampicin resistance genes in Taihu Lake, and E. coli HB101 were used as receptor strains to establish inter-genus and intra-genus conjugative transfer systems, examining the conjugative transfer frequency under the stress of
ketoprofen. The results indicated that
ketoprofen accelerated the environmental spread of ARGs through several mechanisms.
Ketoprofen promoted cell-to-cell contact by increasing cell surface hydrophobicity and reducing cell surface charge, thereby mitigating cell-to-cell repulsion. Furthermore,
ketoprofen induced increased levels of reactive oxygen species (ROS) production, activated the DNA damage-induced response (SOS), and enhanced cell membrane permeability, facilitating ARG transmission in intra-genus and inter-genus systems. The upregulation of outer membrane proteins, oxidative stress, SOS response, mating pair formation (Mpf) system, and DNA transfer and replication (Dtr) system related genes, as well as the inhibition of global regulatory genes, all contributed to higher transfer efficiency under
ketoprofen treatment. These findings served as an early warning for a comprehensive assessment of the roles of NSAIDs in the spread of antibiotic resistance in natural aqueous environments.