Insight into biofilm-forming patterns: biofilm-forming conditions and dynamic changes in extracellular polymer substances.

The microbial biofilm adheres to the surface of the carrier, which protects the pollutant-degrading bacteria and resists harsh environments; thus, research on biofilm-forming patterns will help promote the application of biofilms in wastewater treatment. Herein, univariate analysis and response surface methodology (RSM) confirmed that glucose and mannose at 3-5 g/L promoted biofilm formation. Notably, the microplate method demonstrated that compared to trivalent cations, divalent cations could more greatly enhance the activity (especially magnesium) of the biofilm matrix, and the period of biofilm formation in the three strains was divided into the following stages: initial attachment (0-10 h), microcolony (10-24 h), maturation (24-48 h), and dispersion (36-72 h). During maturation, large amounts of extracellular polysaccharides (EPs) and extracellular DNA (eDNA) were distributed in the extracellular and intracellular spaces, respectively, as observed by super-resolution structured illumination microscopy (SR-SIM). This study enhances the understanding of the characteristics and patterns of biofilm formation and can facilitate the application of biofilms in wastewater treatment.