Although many areas in Denmark are intensively agricultured, the discharge of nitrate from groundwater aquifers to surface water is often lower than expected. In this study it is experimentally demonstrated that anoxic nitrate reduction in sandy sediment containing pyrite is a microbially mediated denitrification process with pyrite as the primary electron donor. The process demonstrates a temperature dependency (Q10) of 1.8 and could be completely inhibited by addition of a bactericide (NaN3). Experimentally determined denitrification rates show that more than 50% of the observed nitrate reduction can be ascribed to pyrite oxidation. The apparent zero-order denitrification rate in anoxic pyrite containing sediment at groundwater temperature has been determined to be 2-3 micromol NO3- kg(-1) day(-1). The in situ groundwater chemistry at the boundary between the redoxcline and the anoxic zone reveals that between 65 and 80% of nitrate reduction in the lower part of the redoxcline is due to anoxic oxidation of pyrite by nitrate with resulting release of sulfate. It is concluded that microbes can control groundwater nitrate concentrations by denitrification using primarily pyrite as electron donor at the oxic-anoxic boundary in sandy aquifers thus determining the position and downward progression of the redox boundary between nitrate-containing and nitrate-free groundwater.