Pyocyanin production by Pseudomonas aeruginosa confers resistance to ionic sliver

Silver in its ionic form (Ag⁺), but not the bulk metal (Agͦ), is toxic to microbial life forms and has been used for many years in the treatment of wound infections. The prevalence of bacterial resistance to silver is considered low due to the nonspecific nature of its toxicity. However, the recent increased use of silver as an antimicrobial agent for medical, consumer, and industrial products has raised concern that widespread silver resistance may emerge. Pseudomonas aeruginosa is a common pathogen that produces pyocyanin, a redox toxin and a reductant for molecular oxygen and ferric (Fe³⁺) ions. The objective of this study was to determine whether pyocyanin reduces Ag⁺ to Agͦ, which may contribute to silver resistance due to lower bioavailability of the cation. Using surface plasmon resonance spectroscopy and scanning electron microscopy, pyocyanin was confirmed to be a reductant for Ag⁺, forming Agͦ nanoparticles and reducing the bioavailability of free Ag⁺ by >95% within minutes. Similarly, a pyocyanin-producing strain of P. aeruginosa (PA14) reduced Ag⁺ but not a pyocyanin-deficient (ΔphzM) strain of the bacterium. Challenge of each strain with Ag⁺ (as AgNO3) gave MICs of 20 and 5 µg/ml for the PA14 and ΔphzM strains, respectively. Removal of pyocyanin from the medium strain PA14 was grown in or its addition to the medium that ΔphzM mutant was grown in gave MICs of 5 and 20 µg/ml, respectively. Clinical isolates demonstrated similar pyocyanin-dependent resistance to Ag⁺.We conclude that pseudomonal silver resistance exists independently of previously recognized intracellular mechanisms and may be more prevalent than previously considered.