Metallic nanoparticles induced antibiotic resistance genes attenuation of leachate culturable microbiota: The combined roles of growth inhibition, ion dissolution and oxidative stress

The dissemination and propagation of antibiotic resistance genes (ARGs) is an emerging global health concern, and the potential effects of nanomaterials on ARGs fates have drawn much attention recently. In the current study, the effects of metallic nanoparticles on ARGs occurrence of leachate culturable microbiota were investigated by four typical metal and metal oxide nanoparticles (Cu, Zn, CuO, and ZnO). The ARGs diversity was remarkably decreased during the cultivation and enrichment of leachate microbiota, and their abundances decreased for 1.4-3.2 orders of magnitude. The presence of nanoparticles facilitated the ARGs attenuation, and the magnitude of effects depended on types of nanoparticles and ARGs. Metal oxide nanoparticles caused more remarkable effects than metal nanoparticles. Mechanism analysis indicated that bacterial growth was inhibited, and the dissolved metal ions from nanoparticles partially contributed to nanoparticles decreasing ARGs. Flow cytometry experiments further confirmed that nanoparticles could enter bacterial cells, and then induce excessive reactive oxygen species (ROS) generation and increase membrane permeability. Finally, the possible mechanisms were put forward, and the structural equation models (SEM) differentiated the contribution of different factors shaping ARGs. The dissolved metal ions and growth inhibition caused by nanoparticles decreased ARGs transfer frequencies via exerting excessive metal stress and lowering population density. On the other hand, nanoparticles were incorporated into the cells, and then induced the generation of ROS, which might facilitate ARGs horizontal transfer via increasing membrane permeability, or decrease ARGs via the damage of genomic and plasmid DNA. Therefore, nanoparticles could affect ARGs fates via several ways, and combined effects finally determined the ARGs variations.