Quorum sensing signaling distribution during the development of full-scale municipal wastewater treatment biofilms

Abstract

Acylated homoserine lactone (AHL)-mediated quorum sensing (QS) is ecologically important in multi-species systems in laboratory-scale studies; however, little is known about QS in the biofilm formation process in full-scale wastewater treatment plants, which is driven by multiple environmental variables. Here, a model integrated fixed-film-activated sludge system was employed in full-scale municipal wastewater treatment plant to investigate the AHL distribution during the biofilm development process in response to variable environmental factors. The whole biofilm development process can be divided into three phases: initial biofilm attachment process (week 1 to 3), biofilm development and mature phase (week 4 to 6), and biofilm detachment and reformation process (week 7 to 17). N-decanoyl-DL-homoserine lactone (C10-HSL) and N-dodecanoyl-DL-homoserine lactone (C12-HSL) presented high concentrations during the biofilm formation process, which was closely related with the biofilm initial attachment process. The AHL concentration in biofilms was higher than in activated sludge. During the initial attachment process, tryptophan and protein-like substances related to biological substance were strongly positively correlated with all detected AHL concentrations (p < 0.05). Three environmental variables (total nitrogen, pH, and Na⁺) were closely related to AHL distribution in municipal wastewater biofilms. High wastewater pH was found to contribute to a low AHL concentration. AHLs in the biofilm were significantly (p < 0.01) influenced by the concentration of Na⁺, and higher concentrations of Na⁺ (10.84-18.58 mg/L) in wastewater treatment plants potentially contribute to the biofilm formation processes. In addition, bacteria with nitrogen removal ability showed QS functionality. The results of this study indicate that AHL-based regulation of tryptophan and protein-like substances related to biological substance production was significantly influenced by the surrounding chemical environment, which has been underestimated in previous studies. AHL-mediated QS potentially suggests a novel solution for the advanced AHL-based regulation of the biofilm development processes.

Keywords: Acylated homoserine lactones; Biofilms; Full-scale municipal wastewater treatment plants; Integrated fixed film activated sludge; Quorum sensing; Time series analysis.