Four horizontal subsurface flow constructed wetlands (HSFCWs), named HSFCW1 (three-stage, without step-feeding), HSFCW2 (three-stage, with step-feeding), HSFCW3 (five-stage, without step-feeding) and HSFCW4 ( five-stage, with step-feeding) were designed to investigate the effects of dissolved oxygen (DO) and step-feeding on nitrogen removal. High removal of 90.9% COD, 99.1% ammonium nitrogen and 88.1% total nitrogen (TN) were obtained simultaneously in HSFCW4 compared with HSFCW1-3. The excellent TN removal of HSFCW4 was due to artificial aeration provided sufficient DO for nitrification and the favorable anoxic environment created for denitrification. Step-feeding was a crucial factor because it provided sufficient carbon source (high COD: nitrate ratio of 14.3) for the denitrification process. Microbial activities and microbial abundance in HSFCW4 was found to be influenced by DO distribution and step-feeding, and thus improve TN removal. These results suggest that artificial aeration combined with step-feeding could achieve high nitrogen removal in HSFCWs. (C) 2014 Elsevier Ltd. All rights reserved.