Influence of the copper-induced viable but non-culturable state on the toxicity of Pseudomonas aeruginosa towards human bronchial epithelial cells in vitro

The viable but non-culturable (VBNC) state of the opportunistic bacterium Pseudomonas aeruginosa was previously shown to be induced by copper ions in concentrations relevant to those in drinking water plumbing systems. This decrease of bacterial culturability without loss of viability might have an influence on human health due to an underestimation of the actual contamination in drinking water systems. The aim of this study was to investigate the influence of culturable P. aeruginosa, viable but not culturable as well as culturable again after resuscitation from the VBNC state on human bronchial epithelial cells (BEAS-2B) in vitro. Cyto- and genotoxic effects of P. aeruginosa at different states were studied using trypan blue, MTT, xCELLigence as well as the micronucleus assay. While P. aeruginosa in the VBNC state did not have any cytotoxic or genotoxic effect on BEAS-2B cells, untreated (culturable) and resuscitated P. aeruginosa did show cell damage, including disruption of cell membranes, inhibition of mitochondrial activity and cell proliferation as well as DNA-damaging effects. We conclude from our study that P. aeruginosa after resuscitation from the VBNC state regains its viability and cyto-/genotoxicity and therefore might influence human health.