Sludge ozonation is a promising technology for dealing with the increasing challenge of excess sludge treatment and disposal. However, nitrogen behavior during sludge ozonation and subsequent biological removal remains unclear. To clarify the feasibility and stability of oxidizing organic nitrogen (released during sludge ozonation) in the bioreactor (but not during ozonation), and the best operational conditions for sludge ozonation, nitrogen behavior was investigated by a long-term observation. The results showed that when inlet ozone concentration increased from 30 to 80 mg O3/L, and ozonation time decreased from 29 to 11 h, less soluble organic nitrogen was oxidized to ammonia (from 66.1 to 18.7% of soluble total nitrogen). This can reduce the operational costs of sludge ozonation. Furthermore, it is feasible to convert organic nitrogen to nitrate by biological processes because full nitrification was restored in three weeks after shock loading of organic nitrogen owing to the change in ozonation conditions. After combining sludge ozonation with the anaerobic/oxic process, the mass balance for nitrogen showed that nitrogen in the excess sludge decreased with increasing sludge reduction rate. The decreased nitrogen in the excess sludge was mainly transformed to nitrogen gas by denitrification, whereas nitrogen in the effluent did not increase noticeably.