In this study, the behavior, distribution and form dynamics of overloaded Cu(II) in anaerobic ammonium oxidation (anammox) granular sludge reactors were investigated. The performance and physiological characteristics were tracked by continuous-flow monitoring to evaluate the long-term effects. High Cu loading (0.24 g L(-1)d(-1)) exceeded sludge bearing capacity, and precipitation dominated the removal pathway. The Cu distribution migrated from the extracellular polymeric substances-bound to the cell-associated Cu and the Cu forms shifted from the weakly bound to strongly bound fractions over time. Pearson correlation and fluorescence spectra analyses showed that the increase in protein concentrations in the EPS was a clear self-defense response to Cu(II) stress. Two remediation strategies, i.e., ethylenediamine tetraacetic acid (EDTA) washing and ultrasound-enhanced EDTA washing, weakened the equilibrium metal partition coefficient from 5.8 to 0.45 and 0.34 L mg(-1)SS, respectively, thereby accelerating the external diffusion of the Cu that had accumulated in the anammox granules.
Keywords: Anammox; Copper inhibition; Extracellular polymeric substances; Metal fractions; Remediation strategy.
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