Acetonitrile-Degrading Halophilic Aerobic Granular Sludge: De Novo Granulation, Acetonitrile Biodegradation, and Nutrient Removal Pathways

De novo granulation of autochthonous microorganisms of water and wastewater reduces the start-up periods for cultivating aerobic granular sludge (AGS) and enrichment of degrading strains. However, it has not been demonstrated using refractory carbon compounds. This work investigated the formation of AGS from the seawater microbiome and establishment of pollutant removal pathways by feeding acetonitrile as the sole carbon and nitrogen source. Use of acetonitrile at an organic loading rate of 0.124 kg/m3/day enabled rapid emergence of aggregates and then stable granules (size: 1.3 mm; SVI5: 68 mL/g) within two weeks. TOC removal accompanied by ammonium nitrogen release was consistent and stable at 93% during the 50 days of bioreactor operation. Formation of acetamide and ammonium indicated involvement of nitrile hydratase and amidase enzymes in acetonitrile biodegradation. Ammonium released during acetonitrile biodegradation was removed by partial nitrification and the nitrite denitrification pathway. However, incomplete ammonium removal led to accumulation of up to 120 mg/L NH4+-N by day 50. Phosphate was removed via the enhanced biological phosphate removal pathway. This study shows that de novo granulation permits cultivation of AGS via the de novo granulation approach for simultaneous biodegradation of refractory acetonitrile and biological nutrient removal under saline conditions.