Bacterium–Phage Interactions Enhance Biofilm Resilience during Membrane Filtration Biofouling under Oxidative and Hydraulic Stresses

Microbial interactions on membrane surfaces can facilitate biofilm formation and biofouling, which poses a significant challenge for pressure-driven membrane filtration systems. This multiomics study investigates the adaptive responses of bacterium-phage interactions under varying oxidative and hydraulic stress during membrane backwashing and their biological contributions to biofouling. Oxidative and hydraulic stress distinctly shaped bacteria and phage diversity and community composition. Under moderate oxidative backwashing (300 ppm of NaClO), diversity was maintained, with increased antioxidant enzyme activities, extracellular polymeric substance (EPS) production, and quorum sensing (QS) signaling, promoting bacterial resilience and biofilm formation. In contrast, excessive oxidative stress (600 ppm of NaClO) reduced bacteria and phage diversity, disrupted antioxidant responses, and increased microbial sensitivity. Hydraulic stress predominantly influenced viral diversity and co-occurrence network topology, favoring the expansion of broad host-range phages and lysogenic lifestyles under combined stresses. Phage-bacterium interaction analyses highlighted phages' adaptive preferences for hosts with high network centrality and broad ecological niches, which enhanced microbial interactions and resilience. Transcriptomic profiling demonstrated the early enrichment of genes associated with energy metabolism, ROS detoxification, and biofilm formation, followed by stabilization as biofilms matured. Phage-encoded auxiliary metabolic genes were involved in DNA repair, QS, and EPS biosynthesis, contributing to microbial adaptation through oxidative stress resistance and biofilm stabilization. Overall, these findings provide mechanistic insights into biofouling dynamics and highlight the need to optimize chlorine dosing to prevent suboptimal levels of microbial adaptation and biofouling.