Mechanisms underlying the effect of chlorination and UV disinfection on VBNC state Escherichia coli isolated from hospital wastewater

The occurrence of viable but non-culturable (VBNC) bacteria in the wastewater system poses a huge threat to environmental and public health, in particular in hospital wastewater treatment system (HWTS). HWTS-oriented studies have been conducted to assess the effectiveness of chlorination and UV disinfection using indigenous bacteria. Results revealed that the VBNC Escherichia coli and ARGs remained persistent even at high chlorination (12 mg/L for 2.5 h) and UV doses (1000 mJ/cm²). The molecular mechanisms underlying chlorination-/UV-induced VBNC state in E. coli were explored through the transcriptomics and results suggested that most energy-dependent physiological activities (e.g., metabolism) have been suppressed in VBNC E. coli, while the pathogenicity-related genes varied insignificantly compared to the culturable cells, indicating that the VBNC E. coli could potentially display pathogenicity. Further Galleria mellonella model experiment has confirmed that although the disinfection-induced VBNC state made cells less infectious, these cells could regain their pathogenicity after resuscitation. This in vitro study can be used as a reference for studies on infections from VBNC bacteria and highlights the health risk due to VBNC pathogens in hospital effluents. There is a need to develop effluent standards specifically for healthcare facilities, and a stricter downstream disinfection strategy should be considered for the removal of VBNC cells and ARGs in the effluent.