The long start-up time and facile biomass loss of aerobic granular sludge (AGS) impede its application for actual wastewater treatment. The present study investigates a novel assist-aggregation strategy based on Aspergillus tubingensis (AT) mycelium pellets to accelerate sludge granulation, and engineered Fe3O4 nanoparticles (NPs) were used to further enhance flocculent sludge (FS) aggregation. The AT mycelium pellets, modified by 0.5 g/L Fe3O4@SiO2-QC NPs (AT-V), had a more compact internal structure than the unmodified group (AT-I). The content of extracellular polymeric substances (EPS) and the zeta potential values were observed to increase from 39.86 mg/gVSS and −9.19 mv for AT-I to 69.64 mg/gVSS and 2.35 mv for AT-V, respectively. In optimized cultivation conditions, the aggregated sludge biomass of AT-V reached 1.54 g/g. An original AT-based AGS (AT-AGS) with a high biological activity (64.45 mgO2/gVSS·h as specific oxygen uptake rate) and enhanced velocity (58.22 m/h) was developed in only 9 days. The removal efficiencies of total nitrogen (TN) and total phosphorus (TP) of the AT-AGS were 12.24% and 16.29% higher than those of the inoculated FS under high feeding load. Additionally, the analysis of cyclic diguanylate (c-di-GMP) and con-focal microscope images implied that polysaccharide (PS) of EPS played an important role in maintaining the stability of the AT-AGS. Finally, the dominant functional species contributing to sludge aggregation and pollutants removal of the AT-AGS showed a larger richness and diversity than those of the inoculated FS. This study might provide a novel high-efficiency strategy for the fast formation of AGS.
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