Sludge bulking is a major problem that restricts the development of the activated sludge process. The microbial community responsible for sludge bulking varies depending on water quality and operational conditions. This study analysed the microbial community of bulking sludge in oxidation ditch with ultra-low sludge loading and long sludge retention time using high-throughput sequencing. The study found that the relative abundance of bacterial genus Saprospiraceae_norank was the highest in bulking sludge, reaching 13.39–28.83%, followed by Comamonadaceae_unclassified, Ardenticatenia_norank and Tetrasphaera, with the relative abundance of 4.59–11.08%, 0.52–16.60% and 0.17–8.92% respectively. In contrast, the relative abundance of bacteria that easily caused sludge bulking including Microthrix (0.54–2.47%), Trichococcus (0.32–1.71%), Gordonia (0.14–1.28%), and Thiothrix (0.01–0.06%) were relatively low. Saprospiraceae_norank was predominant and induced sludge bulking in oxidation ditch. The relative abundance of fungal genus Trichosporon was the highest in bulking sludge, reaching 16.95–24.98%, while other fungal genera were Saccharomycetales_unclassified (5.59–14.55%), Ascomycota_norank (1.45–13.51%), Galactomyces (5.23–11.23%), and Debaryomyces (7.69–9.42%), whereas Trichosporon was the dominant fungal genus in bulking sludge. This study reported that excessive Saprospiraceae_norank can induce sludge bulking for the first time, which provides important knowledge to control sludge bulking.

Activated sludge from wastewater treatment plants was seeded into a sequencing batch reactor (SBR) in which synthetic wastewater was used as the influent. The sludge was bulked by decreasing the concentration of dissolved oxygen (DO). By adding a 30 min step of anaerobic stirring after the water inflow, the sludge bulking was rapidly inhibited after 10 running cycles, and the sludge volume index (SVI) decreased from 222 to 74 mL·g⁻¹. The results of high-throughput sequencing showed that the relative abundance of bacteria Thiothrix, bacteria norank_o_Sphingobacteriales and fungi Trichosporon was increased by 6.3, 4.3 and 81.2%, after initial SBR stages, but these bacteria were inhibited by the addition of an anaerobic step, as their relative abundances decreased by 0.7, 0.8 and 14.7%, respectively. The proliferation of Thiothrix, norank_o_Sphingobacteriales and Trichosporon was the primary reason for the observed sludge bulking in the reactor. After the anaerobic step was added, the sludge extracellular polymeric substances (EPS) concentration was increased from 84.4 to 104.0 mg·(gMLSS)⁻¹ (grams of mixed liquor suspended solids). Thus, the addition of an anaerobic step can inhibit the growth of filamentous bacteria, increasing the sludge EPS concentration and promoting the precipitation of activated sludge.

Oxidation ditches (ODs) and membrane bioreactors (MBRs) are widely used in wastewater treatment plants (WWTPs) with bacteria and antibiotic resistance genes (ARGs) running through the whole system. In this study, metagenomic sequencing was used to compare the bacterial communities and ARGs in the OD and MBR systems, which received the same influent in a WWTP located in Xinjiang, China. The results showed that the removal efficiency of pollutants by the MBR process was better than that by the OD process. The composition and the relative abundance of bacteria in activated sludge were similar at the phylum and genus levels and were not affected by process type. Multidrug, fluoroquinolones and peptides were the main ARG types for the two processes, with macB being the main ARG subtype, and the relative abundance of ARG subtypes in MBR effluent was much higher than that in the OD effluent. The mobile genetic elements (MGEs) in the activated sludge were mainly transposons (tnpA) and insertion sequences (ISs; IS91). These results provide a theoretical basis for process selection and controlling the spread of ARGs.