The nitrogen (N) removal potential of constructed wetlands is increasingly used to lower the N load from agricultural nonpoint sources to inland and coastal waters. To determine the removal efficiency and key factors limiting wetland N removal, N fluxes were studied in a small constructed wetland in Central Switzerland. With an isotope mass balance approach integrating the natural isotope signature of nitrate (NO3-), ammonium (NH4+), and particulate nitrogen (PN), the N transformations such as assimilation, mineralization, nitrification, and denitrification were quantified. On average, the wetland removed 45 g m-2 yr-1 N during the studied 2.5 years, corresponding to a removal efficiency of 27%. Denitrification contributed 94% to the N removal, while only 6% of the removed N accumulated in the sediments. Denitrification was most efficient during periods with an oxic water column overlying anoxic sediments, as NH4+ released during mineralization of sediment organic matter was completely nitrified and subsequently denitrified at the sediment−water interface. During water column anoxia, NH4+ accumulated in the water and fueled assimilation by duckweed and internal recycling. The NO3−N isotope signature in the wetland mainly reflected the mineralization of sediment organic matter and subsequent nitrification, while denitrification at the sediment−water interface produced no fractionationwastewater-rootzone contact. The experimental systems used an upflow hydraulic format to achieve this design objective