Analyzing the inhibitory effect of metabolic uncoupler on bacterial initial attachment and biofilm development and the underlying mechanism

Abstract

Metabolic uncouplers inhibit biofilm and biofouling formation in membrane bioreactor (MBR) systems, which have been considered as a potential biofouling control alternative. To better understand the inhibitory mechanism of uncoupler on biofouling, this study investigated the impact of the uncoupler 3, 3', 4', 5-tetrachlorosalicylanilide (TCS) on biofilm formation of B. subtilis in different development stages. Significant reductions in both the initial bacterial attachment stage and the subsequent biofilm development stage were caused by TCS at 100 μg/L. The motility of B. subtilis in semisolid medium was inhibited by TCS, which explicitly explained the reduction in initial bacterial attachment. Meanwhile, a reduction of extracellular polymeric substance (EPS) secretion owing to TCS suggested why biofilm development was suppressed. In addition, the fluorescent materials in tight-bound EPS (TB-EPS) and loose-bound EPS (LB-EPS) of Bacillus subtilis cultured in different TCS concentrations were distinguished and quantified by three-dimensional excitation-emission matrix (EEM) fluorescence spectroscopy coupled with parallel factor analysis (PARAFAC). The results of this study suggested that the biofilm inhibitory mechanism of the uncoupler was both a inhibition in bacterial motor ability and a reduction in EPS secretion.

Keywords: Bacterial attachment; Biofilm; EEM; EPS; Metabolic uncoupler.