Bacteria-Inspired Synthesis of Silver-Doped Zinc Oxide Nanocomposites: A Novel Synergistic Approach in Controlling Biofilm and Quorum-Sensing-Regulated Virulence Factors in Pseudomonas aeruginosa

Multidrug-resistant <i>Pseudomonas aeruginosa</i> infections pose a critical challenge to healthcare systems, particularly in nosocomial settings. This drug-resistant bacterium forms biofilms and produces an array of virulent factors regulated by quorum sensing. In this study, metal-tolerant bacteria were isolated from a metal-contaminated site and screened for their ability to synthesize multifunctional nanocomposites (NCs). Rapid color changes in the reaction solution evidenced the biotransformation process. The potent isolated <i>Bacillus cereus</i> SASAK, identified via 16S rRNA sequencing and deposited in GenBank under accession number MH885570, facilitated the microbial-mediated synthesis of ZnO nanoparticles and silver-doped ZnO NCs. These biogenic nanocomposites were characterized using UV-VIS-NIR spectroscopy, FTIR, XRD, zeta potential, HRTEM, FESEM, and EDX analyses. At a sub-MIC concentration of 100 µg/mL, 2% Ag-ZnO NCs effectively inhibited virulent factor production and biofilm formation in <i>P. aeruginosa</i> without affecting bacterial growth. Notably, there was a significant reduction in violacein pigment (96.25%), swarming motility, and pyocyanin concentration (1.87 µg/mL). Additionally, biofilm formation (81.1%) and EPS production (83.9%) using <i>P. aeruginosa</i> were substantially hindered, along with reduced extracellular protease activity, as indicated by zone formation (from 2.3 to 1.8 cm). This study underscores the potential of Ag-ZnO NCs as promising agents for combating quorum sensing-mediated virulence in chronic infections caused by multidrug-resistant <i>P</i>. <i>aeruginosa.</i>