Start-up of sequencing batch reactor with <Emphasis Type="Italic">Thiosphaera pantotropha</Emphasis> for treatment of high-strength nitrogenous wastewater and sludge characterization

Biological treatment of high-strength nitrogenous wastewater is challenging due to low growth rate of autotrophic nitrifiers. This study reports bioaugmentation of Thiosphaera pantotropha capable of simultaneously performing heterotrophic nitrification and aerobic denitrification (SND) in sequencing batch reactors (SBRs). SBRs fed with 1:1 organic-nitrogen (N) and NH₄ ⁺-N were started up with activated sludge and T. pantotropha by gradual increase in N concentration. Sludge bulking problems initially observed could be overcome through improved aeration and mixing and change in carbon source. N removal decreased with increase in initial nitrogen concentration, and only 50–60 % removal could be achieved at the highest N concentration of 1000 mg L^?1 at 12-h cycle time. SND accounted for 28 % nitrogen loss. Reducing the settling time to 5–10 min and addition of divalent metal ions gradually improved the settling characteristics of sludge. Sludge aggregates of 0.05–0.2 mm diameter, much smaller than typical aerobic granules, were formed and progressive increase in settling velocity, specific gravity, Ca²⁺, Mg²⁺, protein, and polysaccharides was observed over time. Granulation facilitated total nitrogen (TN) removal at a constant rate over the entire 12-h cycle and thus increased TN removal up to 70 %. Concentrations of NO₂ ^?-N and NO₃ ^?-N were consistently low indicating effective denitrification. Nitrogen removal was possibly limited by urea hydrolysis/nitrification. Presence of T. pantotropha in the SBRs was confirmed through biochemical tests and 16S rDNA analysis.