A Feasible Regulation Strategy for Conjugation of Antibiotic Resistance Genes Based on Different Bacterial Quorum Sensing Inhibition Methods

The dissemination of antibiotic resistance genes (ARGs) poses global environmental issues, and plasmid-mediated conjugation contributes substantially to the spread of ARGs. Quorum sensing (QS), an important cell–cell communication system that coordinates group behaviors, has potential as a feasible regulation pathway to inhibit the conjugation process. We examined the promoting effects of QS signal on conjugation, and this study is the first to report that QS inhibitors 2(3H)-benzofuranone and acylase I effectively repressed conjugation frequency of RP4 plasmid to 0.32- and 0.13-fold compared with the control respectively. The investigation of underlying mechanisms of QS inhibitors revealed a significant decrease in cellular contact and the formation of transfer channels. The downregulation of sdiA gene regulating the expression of QS signal receptor contribute to conjugation inhibition. Importantly, the expression of genes related to the formation of conjugative pili, which plays a role in plasmid mating bridge formation was downregulated, indicating QS inhibitors affect conjugation mainly through regulation of the mating pair formation system. Furthermore, 2(3H)-benzofuranone and acylase I achieved 84.07% and 66.05% inhibitory effect on plasmid spread in activated sludge reactors. Collectively, our findings demonstrate the feasibility of using different bacteria quorum quenching methods to control the spread of ARGs.