For decades, the conversion of organic nitrogen to dinitrogen gas by heterotrophic bacteria, termed heterotrophic denitrification, was assumed to be the main pathway of nitrogen loss in natural ecosystems. Recently, however, autotrophic bacteria have been shown to oxidize ammonium in the absence of oxygen, yielding dinitrogen gas. This process, termed anammox, accounts for over 50% of nitrogen loss in marine ecosystems1–5. However, the significance of anammox in freshwater ecosystems has remained uncertain 6,7. Here, we use molecular and isotopic techniques to monitor anammox activity in sediments sampled fromlake riparian zones in North China. Laboratory incubations in the presence of ammonium or nitrate—at concentrations equivalent to no more than 10% of those detected in situ—yielded some of the highest potential anammox activities reported for natural environments to date. Potential rates of anammox peaked in sediments sampled from the interface between the land and the water, as did the abundance of annamox bacteria. Scaling our findings up to the entire lake system, we estimate that interfacial anammox hotspots account for the loss of 103 gNm-2 yr-1 from this lake region, and around one fifth of the nitrogen lost from the land–water interface.