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The simultaneous nitrification and denitrification in step-feeding biological nitrogen removal process were investigated under different influent substrate concentrations and aeration flow rates.Biological occurrence of simultaneous nitrification and denitrifieation was verified in the aspect of nitrogen mass balance and alkalinity.The experimental results also showed that there was a distinct linear relationship between simultaneous nitrification and denitrification and DO concentration under the conditions of low and high aeration flow rate.In each experimental run the floe sizes of activated sludge were also measured and the results showed that simultaneous nitrification and denitrification could occur with very small size of floc.