Viable but non-culturable E. coli induced by low level chlorination have higher persistence to antibiotics than their culturable counterparts

- Low level chlorination could induce VBNC bacteria.
- The chlorination-induced VBNC bacteria showed broad-spectrum high persistence to 9 typical antibiotics.
- VBNC bacteria had transcript levels of rpoS that were higher than the corresponding transcript levels in culturable bacteria.
- Identified up-regulated resistance genes belonged to multi-drug efflux genes.
- Lower metabolic activity, higher efflux rate, might be responsible for broad spectrum persistence in VBNC cells.

Disinfectant used in drinking water treatment and distribution system can induce culturable bacteria, including various kinds of pathogenic bacteria, into viable but non-culturable (VBNC) state. The loss of cultural state, resuscitation and environmental persistence of VBNC bacteria will severely damage drinking water microbiological safety and thus pose a risk to public health. The manner in which chlorination treatment induced a VBNC state in Escherichia coli and the antibiotic persistence of VBNC bacteria was investigated. It was found that low dosage of chlorine (0.5 mg L−1) disinfection effectively reduced the culturability of E. coli and induced a VBNC state, after which metabolic activity was reduced and persistence to 9 typical antibiotics was enhanced. Furthermore, RT-qPCR results showed that stress resistance genes (rpoS, marA, ygfA, relE) and ARGs, especially efflux genes were up-regulated compared with culturable cells. The intracellular concentration was tested and found to be lower in VBNC cells than in actively growing E. coli, which suggested a higher efflux rate. The data presented indicate that VBNC E. coli are more persistent than culturable counterparts to a wide variety of antibiotics. VBNC E. coli constitute a potential source of contamination and should be considered during monitoring of drinking water networks.

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