The objective was to provide an answer to “how to grow/survive in aggregative physiology” through evaluating the relation between physical stress and observed biomass characteristics. For that, a lab-scale sequencing batch reactor was operated at an anaerobic-aerobic mode and under altered hydraulic selection pressures of settling time (10–1 min) and hydrodynamic shear rates due to mechanical mixing (15.5–12.0 cm/s) and/or aeration (1.76–0.24 cm/s). Main physical stress experienced by the biomass was mechanical mixing, which resulted in extreme shearing conditions at the first operational stage (days 1–86), during which first granules formed but settling properties deteriorated and biomass was almost totally washed out. After relaxing the overall shear stress at the second stage, biomass formation accelerated, settling properties enhanced and granulation proceeded (days 86–136), until disturbance of the process at the last month of operation (days 136–163). Aggregative physiology-related parameters, being cell surface hydrophobicity and extracellular polymeric substances (EPS), followed increasing trends parallel to the progress of granulation, and then decreased upon disturbance of the process. There was an increase in the EPS production also during the first stage under extreme shear, while a substantial amount of biomass was present in the system. A direct correlation was also found between %hydrophobicity and EPS-composition expressed as ExoPN/ExoPS.

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