Increased Tolerance to Heavy Metals Exhibited by Swarming Bacteria

Abstract:

Pseudomonas aeruginosa is a ubiquitous, Gram-negative bacterium that utilizes several different modes of motility to colonize surfaces, including swarming, which is the coordinated movement of cells over surfaces in groups. Swarming facilitates surface colonization and biofilm development for P. aeruginosa, and it is known that swarming behavior is influenced by changes in nutrient composition and surface moisture. To understand the fate and cycling of heavy metals in the environment, it is important to understand the interaction and toxicity of these metals upon bacteria. While previous studies have shown surface-attached bacterial biofilms to be highly resistant to heavy metal toxicity, little is known about the influence of heavy metals upon surface motile bacteria and developing biofilms. Using a combination of laboratory assays we examined differences in bacterial behavior in response to two metals, Cd and Ni. We find that surface swarming bacteria are able to grow on 4x and 2.5x more Cd and Ni, respectively, than planktonic cells (i.e., test tube cultures). P. aeruginosa was able to swarm in the presence ≤0.051mM Ni and ≤0.045mM Cd. To investigate the bioavailability of metals to bacteria growing under our examined conditions, we separated cell and supernatant fractions of P. aeruginosa cultures, and used ICP-MS techniques to measure Cd and Ni sorption. A greater percentage of Cd than Ni was sorbed by both cells and supernatant (which contains rhamnolipid, a surfactant known to sorb some metals and improve swarming). While we show that cell products such as rhamnolipid bind heavy metals (as expected) and should limit metal bioavailability, our results suggest at least one additional mechanism (as yet undetermined) that promotes cell survival during swarming in the presence of these heavy metals.