Spatial data provided by the U.S. Department of Agriculture National Resource Conservation Service representing implementation at the field-level for a selection of agricultural conservation practices were incorporated within a spatially referenced regression model to estimate their effects on nitrogen loads in streams in the Chesapeake Bay watershed. Conservation practices classified as “high-impact” were estimated to be effective (p = 0.017) at reducing contemporary nitrogen loads to streams of the Chesapeake Bay watershed in areas where groundwater ages are estimated to be less than 14-years old. Watershed-wide, high-impact practices were estimated to reduce nitrogen loads to streams by 1.45%, with up to 60% reductions in areas with shorter groundwater ages and larger amounts of implementation. Effects of “other-impact” practices and practices in areas with groundwater ages of 14 years or more showed less evidence of effectiveness. That the discernable impact of high-impact practices was limited to areas with a median groundwater age of less than 14 years does not imply that conservation practices are not effective in areas with older groundwater ages. A model recalibrated using high-impact agricultural conservation practice data summarized by county suggests effects may also be detectable using implementation data available at such coarser resolution. Despite increasing investment, effects of agricultural conservation practices on regional water quality remain difficult to quantify due to factors such as groundwater travel times, varying modes-of-action, and the general lack of high-quality spatial datasets representing practice implementation.