Potential role of nanobubbles in dynamically modulating the structure and stability of anammox granular sludge within biological nitrogen removal process

Highlights: • Nanobubbles (NBs) generated by anammox granular sludge (AnGS) were investigated. • The existence and stability of NBs inside the AnGS system were discussed. • NBs changed local microstructure AnGS to avoid the blocking of gas channels by EPS. • NBs penetrated the cell clusters to form the nanopores in AnGS. • NBs played an important role in maintaining the stable porous structure of AnGS. The generation of visible macrobubbles considerably affects the structure and function of anammox granules in the anammox granular sludge (AnGS) system. However, the existence of nanobubbles (NBs) and their role in maintaining the AnGS structure and stability are unclear because of the complexity of the system and lack of effective analytical methods. In this study, methods for NB analysis and assessment of their effects were developed to investigate the formation and characteristics of NBs in an AnGS system and the effects of NBs on the properties and function of AnGS. The results indicated that dissolved gas supersaturation caused by AnGS generated NBs of 2.75 × 10⁸ bubbles/mL inside an AnGS reactor after running for 300 min at 30 °C. The increasing absolute value of the zeta potential of NBs with time indicated that the NBs in the AnGS system were gradually stable. The size of the stable NBs ranged from 150 nm to 400 nm. NB formation also increased the space and pressure between cells, leading to the breakage of the cell cluster and causing structural changes in granules. Changes in the local granular microstructure caused by NBs were favorable for the porous structure of granules to avoid granular disintegration and flotation caused by the excessive secretion of extracellular polymeric substances blocking gas channels. The formation and stability of NBs penetrating the cell clusters played a crucial role in the formation and stability of nanopores around or inside the cell clusters, further providing a basis for the formation of high-porosity structures and efficient mass transfer of AnGS.