Indole Signaling Enhances Biofilm Formation and Quorum Sensing in Sequencing Biofilm Batch Reactors

Indole is a heterocyclic organic compound that acts a signaling molecule to regulate biofilm formation (BF) by microorganisms. However, the potential roles of indole in BF among microbial communities in wastewater treatment systems remain unknown. In this study, the effects of indole signaling on BF and quorum sensing (QS) of the complex microbial communities in sequencing batch biofilm reactors were systematically explored. The results showed that indole at 200 mg/L increased BF along with concentrations of homoserine lactones. High-throughput sequencing revealed notable differences in the BF capacity of microbial communities between the indole-treated and control groups. The indole-related metabolic and QS functions of the genera in the biofilm were annotated based on the QS-related protein database and previous studies. Relative abundance analysis showed that the QS-related genera, such as Acinetobacter, that produce indole oxygenase were significantly enriched in the indole-treatment group. In addition, bioinformatics analyses revealed that indole at 200 mg/L enhanced BF, QS, and protein efflux. Based on a phylogenetic molecular ecological network, QS was positively correlated to indole treatment of Burkholderia, suggesting that indole promotes QS and BF. This study provides new insights into the effects of indole on BF of microbes in wastewater treatment plants.