A novel dropping nitrification–cotton-based denitrification reactor was developed for total nitrogen (N) removal from ammonium (NH4+)-contaminated groundwater. The nitrogen removal ability of the reactor was evaluated for 91 days. A 1 m long dropping nitrification unit was fed with synthetic groundwater containing 30 mg-NH4+-N/L at a flow rate of 2.16 L/d. The outlet of the dropping nitrification unit was connected to the cotton-based denitrification unit. The NH4+ present in the groundwater

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... completely oxidized (>90% nitrification efficiency) by nitrifying bacteria to nitrite (NO2–) and nitrate (NO3–) in the dropping nitrification unit. Subsequently, the generated NO2– and NO3– were denitrified (96%–98% denitrification efficiency) by denitrifying bacteria in the cotton-based denitrification unit under anoxic conditions. Organic carbons released from the cotton presumably acted as electron donors for heterotrophic denitrification. Nitrifying and denitrifying bacteria were colonized in higher abundance in the dropping nitrification and cotton-based denitrification units, respectively. The total N removal rate and efficiency of the dropping nitrification–cotton-based denitrification reactor for 91 days were 58.1–66.9 mg-N/d and 96%–98%, respectively. Therefore, the dropping nitrification–cotton-based denitrification reactor will be an efficient, sustainable, and promising option for total N removal from NH4+-contaminated groundwater.