Combined effects of volume ratio and nitrate recycling ratio on nutrient removal, sludge characteristic and microbial evolution for DPR optimization.

The optimization of volume ratio (VAn/VA/VO) and nitrate recycling ratio (R) in a two-sludge denitrifying phosphorus removal (DPR) process of Anaerobic Anoxic Oxic-Moving Bed Biofilm Reactor (A2/O-MBBR) was investigated. The results showed that prolonged anaerobic retention time (HRTAn: 1.25→3.75 hr) exerted favorable effect on chemical oxygen demand (COD) removal (57.26%→73.54%), poly-ß-hydroxyalkanoates (PHA) synthesis (105.70→138.12 mgCOD/L) and PO43- release (22.3→38.9 mg/L). However, anoxic retention time (HRTA) and R exhibited positive correlation with PHA utilization (43.87%-81.34%) and denitrifying phosphorus removal (DPR) potential (ΔNO3-/ΔPO43-: 0.57-1.34 mg/mg), leading to dramatical TN removal variations from 68.86% to 81.28%. Under the VAn/VA/VO ratio of 2:6:0, sludge loss deteriorated nutrient removals but the sludge bioactivity quickly recovered when the oxic zone was recovered. The sludge characteristic and microstructure gradually transformed under the dissolved oxygen (DO) control (1.0-1.5→1.5-2.0 mg/L), in terms of sludge volume index (SVI: 194→57 mL/gVSS), median-particle-size (D50: 99.6→300.5 µm), extracellular polymeric substances (EPS) (105.62→226.18 mg/g VSS) and proteins/polysaccharides (PN/PS) ratio (1.52→3.46). Fluorescence in situ hybridization (FISH) results showed that phosphorus accumulation organisms (PAOs) (mainly Cluster I of Accumulibacter, contribution ratio: 91.79%-94.10%) dominated the superior DPR performance, while glycogen accumulating organisms (GAOs) (mainly Competibacter, contribution ratio: 82.61%-86.89%) was responsible for deteriorative TN and PO43- removals. The optimal HRTA and R assembled around 5-6.5 hr and 300%-400% based on the PHA utilization and DRP performance, and the oxic zones also contributed to PO43- removal although it showed low dependence on DO concentration and oxic retention time (HRTO).