The objective of this study is to estimate the soil N flux from the vadose zone to the aquifer of the Wilmot watershed (Prince Edward Island, Canada) for a typical three-year cropping rotation (barley-red clover-potato). A conceptual model estimates that 199-221 tons of N were yearly available for leaching at the watershed scale. A significant portion of this N amount was available for leaching at the end of the crop season representing 80-90% of the annual N balance. Drainage water nitrate concentrations were significantly higher after the potato-rotation year than during the crop season. Low nitrate concentrations were measured at spring thaw indicating that most of the nitrate available from the preceding potato crop season was likely leached at the end of fall or during winter. Early spring ionic exchange membrane sampling show a large availability of nitrate in soil possibly throughout winter as well, resulting from soil N mineralization and nitrification over the winter period. These findings are corroborated by the isotope natural abundance analysis of nitrate in groundwater implying that nitrifiers are significantly active during winter, as well as during the crop season, and that leaching of soil nitrates with seasonal signals takes place whenever recharge is occurring.