De novo granulation of sewage-borne microorganisms: A proof of concept on cultivating aerobic granular sludge without activated sludge and effective enhanced biological phosphorus removal

Long start-up periods for granulating activated sludge and concerns on granular stability are the bottlenecks reported during implementation of novel aerobic granular sludge (AGS) technology in municipal wastewater treatment plants. Here, de novo granulation of sewage-borne microorganisms without using activated sludge (AS) inoculum was investigated in bench-scale sequencing batch reactors (SBR). Data showed that formation of AGS from sewage-borne microorganisms was rapid and first granules appeared within one week. Granulation was indicated by appearance of biomass particles (size >0.12 mm), high biomass levels (similar to 8 g/L) and superior settling properties (SVI30 min: 30 mL/g). Granulation process involved distinct stages like formation of aggregates, retention of aggregates, and growth of millimetre sized granules. Simultaneous COD, nitrogen and phosphorous removal was established within 10 days of start-up in the SBR without using AS inoculum. However, phosphorus removal became stable after 50 days of start-up. Total nitrogen (TN) and total phosphorus (TP) removals of 92% and 70%, respectively, were achieved from real domestic wastewater. Furthermore, addition of granular activated carbon (GAC) had improved both granulation and biological nutrient removals. Interestingly, phosphorus removal became quite stable within 10 days of start-up in the SBR operated with GAC particles. TN and TP removals were found to be higher at >98% and >94%, respectively, in GAC-augmented SBR. Removal of ammonia and phosphorus were mediated by nitritation-denitritation and enhanced biological phosphorus removal (EBPR) pathways, respectively. The bacterial diversity of AGS was lower than that of sewage. Quantitative PCR indicated enrichment of ammonia oxidizing bacteria, denitrifying bacteria and polyphosphate accumulating organisms during granulation. De novo granulation of sewage-borne microorganisms is a promising approach for rapidly cultivating AGS and establishing biological nutrient removal in sewage treatment plants.