The purpose of this research is to find out the effects of free ammonia concentration and dissolved oxygen on nitrification and nitrite accumulation in a biofilm airlift reactor. Free ammonia seriously inhibited the activity of nitrite oxidizers at the concentration higher than 0.1 mg NH3-N/L and it was very effective for nitrite accumulation. Dissolved oxygen limitation in the biofilm also caused nitrite accumulation. Long term inhibition decreased the growth rate for nitrite oxidizers, and ammonia oxidizers were the dominant nitrifiers in the wastewater nitrification. Selective accumulation of ammonia oxidizers in the biofilm could be another reason for nitrite accumulation. Free ammonia inhibited nitrite oxidizers immediately, and adaptation to free ammonia was not observed. Therefore, the optimum control of free ammonia and dissolved oxygen concentration is critical for nitrite accumulation and the strategy can be used for selective accumulation of ammonia oxidizers in a bioreactor system.