Sub-lethal concentrations of heavy metals induce antibiotic resistance via mutagenesis

The emergence of antibiotic resistance is a growing problem worldwide. Numerous studies have demonstrated that heavy metals facilitate the spread of bacterial drug-resistance in the environment. However, the actions and mechanisms of metals at relatively low sub-lethal levels (far below the minimal inhibitory concentration [MIC]) on antibiotic resistance remain unclear. In this study, we investigated the effect of sub-lethal levels of heavy metals [Ag(I), Zn(II), and Cu(II)] on antibiotic resistance and explored the underlying mechanisms. The results demonstrated that sub-lethal levels of metal ions increased the mutation rates and enriched de novo mutants that exhibited significant resistance to multiple antibiotics. The resistant mutants exhibited hereditary resistance after 5-day of sub-culture. Whole-genome analysis revealed distinct mutations in genes involved in multiple drug and drug-specific resistance, as well as genes that are not associated with antibiotic resistance to data. The number and identities of genetic changes were distinct for mutants induced by different metals. This study provides evidence and mechanistic insights into the induction of antibiotic resistance by sub-lethal concentrations of heavy metals, which may enhance the emergence of antibiotic resistance in various environments. More consideration and regulations should be given to this potential health risk for long-standing and harmful heavy metals.