Twenty-four Bacillus strains predominantly outgrown in a night soil treatment system were isolated and characterized. Under various culture conditions, cell interactions took place among them and cell population changed. Maximum removal of NH4+-N and cell production by the isolates occurred under the conditions of 30% DO and C/N ratio of 8. Five dominant isolates were identified to be species of Bacillus cereus, Bacillus subtilis and Bacillus licheniformis with similarities of 78-94%. Additions of 0.8% peptone and 0.3% yeast extract to a basal medium influenced the growth of isolates and the removal of NH4+-N in flask culture. Metal ions such as Ca2+, Fe2+ and Mg2+ had a similar effect. The specific growth rates of the five isolates were found to be in a range of 0.43-0.55 h(-1). During the flask experiment of nitrogen removal under aerobic growth conditions, active nitrification by the isolates occurred largely in 1h with a decrease of COD and alkalinity reduced to only 74.6% of theoretical value. From the nitrogen balance, the percentage of nitrogen lost in the flask culture was estimated to be 33.0%, which was presumed to convert to N2 gas. This conversion of ammonia to N2 without formation of nitrous oxide under aerobic growth conditions was confirmed by GC analysis. From all the results, it has been found that the Bacillus strains were able to occur simultaneously aerobic nitrification/denitrification and the B3 process using the Bacillus strains seemed to possess some economic advantages.