Tank-dependence of the functionality and network differentiation of activated sludge community in a full-scale anaerobic/anoxic/aerobic municipal sewage treatment plant

● Environmental parameters affected functional bacteria and network associations.● The structure and interactions of AS networks changed greatly within tanks.● Anoxic co-occurrence network was more unstable and easily influenced.● Composition of functional bacteria had a seasonal succession pattern.● Tetrasphaera was the major PAO in spring and winter leading a better P removal.Understanding the structures and dynamics of bacterial communities in activated sludge (AS) in full-scale wastewater treatment plants (WWTPs) is of both engineering and ecological significance. Previous investigations have mainly focused on the AS communities of WWTP aeration tanks, and the differences and interactions between the communities in anaerobic and anoxic tanks of the AS system remain poorly understood. Here, we investigated the structures of bacterial communities and their inter-connections in three tanks (anaerobic, anoxic, and aerobic) and influent from a full-scale WWTP with conventional anaerobic/anoxic/aerobic (A/A/O) process over a year to explore their functionality and network differentiation. High-throughput sequencing showed that community compositions did not differ appreciably between the different tanks, likely due to the continuous sludge community interchange between tanks. However, network analysis showed significant differences in inter-species relationships, OTU topological roles, and keystone populations in the different AS communities. Moreover, the anoxic network is expected to be more unstable and easily affected by environmental disturbance. Tank-associated environmental factors, including dissolved oxygen, pH, and nutrients, were found to affect the relative abundance of functional genera (i.e., AOB, NOB, PAOs, and denitrifiers), suggesting that these groups were more susceptible to environmental variables than other bacteria. Therefore, this work could assist in improving our understanding of tank-associated microbial ecology, particularly the response of functional bacteria to seasonal variations in WWTPs employing A/A/O process.