Induction of Escherichia coli and Salmonella typhimurium into the viable but nonculturable state following chlorination of wastewater

We examined the effects of chlorine disinfection on Escherichia coli and Salmonella typhimurium in secondary-treated wastewater to determine whether such treatment might induce these bacteria into the viable but nonculturable (VBNC) state. In this state, cells lose culturability but retain viability and the potential to revert to the metabolically active and infectious state. To examine the effects of chlorination on cells in different physiological states, cells from the logarithmic and stationary phases, or nutrient starved, or grown in natural wastewater, were studied. Isogenic cells with and without plasmids were also examined. Whereas a mixture of free and combined chlorine, as occurs under typical wastewater disinfection, was found to be rapidly lethal to most cells, regardless of their physiological state or plasmid content, c. 10⁴ of the original 10⁶ cells ml⁻¹ did survive in the VBNC state. While we were not successful in resuscitating these cells to the culturable state, the presence of such nonculturable cells in treated wastewater offers a potential public health hazard.