Silver Sink Effect of Humic Acid on Bacterial Surface Colonization in the Presence of Silver Ions and Nanoparticles.

Silver nanoparticles (AgNPs) released from consumer products may enter the environment and possibly harm microbial communities. Prior research showed that surface-adherent AgNPs inhibit bacterial surface colonization, a precursor to biofilm formation, only when planktonic bacterial inoculum concentrations are less than a threshold level ( Wirth and co-workers, J. Colloid Interface Sci. 2016 , 467 , 17 - 27 ). This inoculum effect is due to a decrease in free silver ion concentration associated with sublethal binding to bacteria. Natural organic matter can be an additional silver sink in environmental systems. Using Pseudomonas fluorescens as a model biofilm-forming bacterium, we find significant increases in minimum bactericidal concentrations for AgNP suspensions and Ag+ in solution when adding humic acid (HA) to bacterial suspensions. When HA is present, planktonic bacteria survive and colonize AgNP-laden glass surfaces at lower bacterial inoculum concentrations than were needed for survival and colonization in its absence. This occurs despite the observed tendency of HA to inhibit colonization on bare glass surfaces when silver is absent. Results are interpreted through equilibrium Ag+ binding isotherms to HA and suspended bacteria. These results indicate that silver ion sinks may lessen AgNP impacts on natural microbial ecology relative to the disruption observed in pristine laboratory conditions.

Inhibition of bacterial surface colonization by immobilized silver nanoparticles depends critically on the planktonic bacterial concentration.

Immobilization of antimicrobial silver nanoparticles (AgNPs) on surfaces has been proposed as a method to inhibit biofouling or as a possible route by which incidental releases of AgNPs may interfere with biofilms in the natural environment or in wastewater treatment. This study addresses the ability of planktonic Pseudomonas fluorescens bacteria to colonize surfaces with pre-adsorbed AgNPs. The ability of the AgNP-coated surfaces to inhibit colonization was controlled by the dissolved silver in the system, with a strong dependence on the initial planktonic cell concentration in the suspension, i.e., a strong inoculum effect. This dependence was attributed to a decrease in dissolved silver ion bioavailability and toxicity caused by its binding to cells and/or cell byproducts. Therefore, when the initial cell concentration was high (∼1×10(7)CFU/mL), an excess of silver binding capacity removed most of the free silver and allowed both planktonic growth and surface colonization directly on the AgNP-coated surface. When the initial cell concentration was low (∼1×10(5)CFU/mL), 100% killing of the planktonic cell inoculum occurred and prevented colonization. When an intermediate initial inoculum concentration (∼1×10(6)CFU/mL) was sufficiently large to prevent 100% killing of planktonic cells, even with 99.97% initial killing, the planktonic population recovered and bacteria colonized the AgNP-coated surface. In some conditions, colonization of AgNP-coated surfaces was enhanced relative to silver-free controls, and the bacteria demonstrated a preferential attachment to AgNP-coated, rather than bare, surface regions. The degree to which the bacterial concentration dictates whether or not surface-immobilized AgNPs can inhibit colonization has significant implications both for the design of antimicrobial surfaces and for the potential environmental impacts of AgNPs.