Comparative Analysis of the Mechanism of Resistance to Silver Nanoparticles and the Biocide 2,2-Dibromo-3-Nitrilopropionamide.

Antimicrobials such as nanoparticles and biocides are used to control microbial growth. We used Escherichia coli to study the process of acquired resistance to silver nanoparticles (Ag-NP) and the industrial biocide DBNPA when grown in sub-MICs. We determined the MICs of these two antimicrobials against E. coli. We then performed an experimental evolution study where E. coli was grown in subinhibitory concentrations of the antimicrobials and transferred 10 times. We then tracked the changes in growth characteristics, lactate dehydrogenase (LDH) activity, reactive oxidative species (ROS) production, and the role of efflux pumps in conferring resistance. We also performed genome sequencing to determine the genetic basis for acquired resistance. Our results showed that E. coli could rapidly develop resistance to Ag-NP and DBNPA after growth in low concentrations of the antimicrobials. The expression of efflux pumps plays a vital role in both Ag-NP and DBNPA resistance. Multiple mutations occurred in the adapted strains that may confer resistance to both Ag-NP and DBNPA. Our study provides insights into mechanisms of adaptation and resistance to antimicrobials. Our results suggest that there are some shared mechanisms to resist nanoparticles and biocides as well as some key differences. The mechanism of resistance to Ag-NP might be related to flagellin production, while efflux pumps seem to be associated with resistance to DBNPA. This work provides a comparative study of the mechanisms of acquired resistance to these two types of antimicrobials.