Contrasting distribution of antibiotic resistance genes and microbial communities in suspended activated sludge versus attached biofilms in an integrated fixed film activated sludge (IFAS) system

Abstract

Suspended activated sludge (AS) and carrier-attached biofilms simultaneously exist in an integrated fixed film activated sludge (IFAS) system. However, the differentiation of antibiotic resistance genes (ARGs) and microbial communities in different types of biofilms is rarely reported. In this study, successions of ARGs and microbial communities of AS and two types of suspended carrier-attached biofilms over seasons were investigated in the IFAS system of one municipal wastewater treatment plant. Results showed that substantial differences were found in the distribution pattern of ARGs, bacterial communities, and predicted microbial function between AS and attached biofilms. The relative abundances of all detected ARGs in AS were significantly higher than those in attached biofilms. ARGs with higher relative abundances generally existed in K3 carrier (surface area ≥ 800 m²/m³) attached biofilms than those in K1 carrier (surface area ≥ 450 m²/m³) biofilms. The relative abundances of ARGs were negatively correlated with temperature and biochemical oxygen demand (BOD₅) and positively correlated with ammonium nitrogen contents for AS but not for attached biofilms. No significant relationship was found between the extracellular polymeric substance (EPS) content and ARG abundance for all samples. Temperature, BOD₅, and ammonium nitrogen contents were closely connected to microbial communities. The Bray-Curtis distance of bacterial communities between two adjacent sampling seasons for AS was larger than those of two attached biofilms. Network analysis indicated that the AS network had more positive links and intense connections than the attached biofilm networks, potentially facilitating the dissemination of ARGs. The differential distribution of ARGs among the three types of samples was significantly correlated with the microbial co-occurrence network topological properties. Bray-Curtis distance and network analysis suggest that microbial community is more robust in attached biofilms than in suspended AS. This work provides a more in-depth understanding of ARGs and microbial community distributions in wastewater biofilms.

Keywords: Activated sludge; Antibiotic resistance gene; Attached biofilm; Integrated fixed film activated sludge; Microbial community.