The ammonia oxidation of a mixed culture enriched from a wastewater treatment plant sludge was monitored by a DO probe and a nicotinamide adenine dinucleotide (phosphate) (NAD(P)H) fluorometer. Under fixed aeration, DO reflected ammonia oxidation effectively. According to the DO profiles and the total ammonia concentrations analyzed experimentally, the ammonia oxidation kinetics of the enriched culture was determined. The ammonia oxidation rate was found sensitive to pH, especially at low total ammonia concentrations. At high concentrations of total ammonia, the maximum ammonia oxidation rate occurred at pH 7.6+/-0.1. At low concentrations, the pH sensitivity intensified significantly and the apparent optimal pH shifted higher with decreasing total ammonia concentrations, because NH3 molecules were the true substrate for ammonia oxidation and more NH4+ ions were converted to NH3 molecules at higher pH. The ammonia oxidation kinetics was therefore developed according to the concentration of NH3 molecules, instead of the commonly used total ammonia concentration. The kinetics followed Michaelis-Menten behavior for both DO and NH3 concentration: the maximum rate was 16.7+/-0.7 mg NH3-N/(g TSS-h) and the Michaelis constants for DO and NH3 were (14+/-2)% air saturation and (4.4+/-0.4) x 10(-2) (mg NH3-N/L), respectively. It was also concluded from the study that with or without exogenous organic substances, the NAD(P)H fluorescence of the enriched nitrifying culture was undetectable. The fluorescence did not respond to addition or depletion of substrate (ammonia, glucose, or acetate), change between aerobic and anaerobic conditions, or even KCN addition to kill the culture.