A nitrogen (N) mass-balance budget was developed to assess the sources of N affecting increasing ground-water nitrate concentrations in the 960-km 2 karstic Ichetucknee Springs basin. This budget included direct measurements of N species in rainfall, ground water, and spring waters, along with estimates of N loading from fertilizers, septic tanks, animal wastes, and the land application of treated municipal wastewater and residual solids. Based on a range of N leaching estimates, N loads to ground water ranged from 262,000 to 1.3 million kg/year; and were similar to N export from the basin in spring waters (266,000 kg/year) when 80-90% N losses were assumed. Fertilizers applied to cropland, lawns, and pine stands contributed about 51% of the estimated total annual N load to ground water in the basin. Other sources contributed the following percentages of total N load to ground water: animal wastes, 27%; septic tanks, 12%; atmospheric deposition, 8%; and the land application of treated wastewater and biosolids, 2%. Due to below normal rainfall (97.3 cm) during the 12-month rainfall collection period, N inputs from rainfall likely were about 30% lower than estimates for normal annual rainfall (136 cm). Low N-isotope values for six spring waters (??15N-NO3 = 3.3 to 6.3???) and elevated potassium concentrations in ground water and spring waters were consistent with the large N contribution from fertilizers. Given ground-water residence times on the order of decades for spring waters, possible sinks for excess N inputs to the basin include N storage in the unsaturated zone and parts of the aquifer with relatively sluggish ground-water movement and denitrification. A geographical-based model of spatial loading from fertilizers indicated that areas most vulnerable to nitrate contamination were located in closed depressions containing sinkholes and other dissolution features in the southern half of the basin. ?? 2009 American Water Resources Association.