Insights into the stress response of Escherichia coli biofilms upon Plasma-activated water treatment.

Biofilms on surfaces pose quality and health risks, resisting traditional antimicrobial methods. Plasma-activated water (PAW) shows promise in biofilm elimination, yet its genetic impact remains unclear. We investigated Escherichia coli biofilm responses to PAW treatment, revealing significant gene expression changes and pathway activations. Knockout mutants of key genes confirmed their role in PAW response. This study enhances our understanding of PAW's anti-biofilm mechanisms and bacterial stress responses. RNA-seq was performed to compare transcriptomes of Escherichia coli biofilm during in situ 2 min sub-lethal PAW treatment with and without superoxide (by adding the scavenger Tiron to remove it). Four independent biological replicates of 48 h ATCC 25922 E. coli biofilms were generated, each comprising four technical replicates were pooled. Pooled biofilms were treated in situ for 2 min with four different treatment solutions PAW, PAW-Tiron, Bubble, and Bubble-Tiron.