The composition of interstitial waters in Spain's Doñana National Park was assessed 4 years after a major pyrite slurry spill occurred from the Aznalcollar Mine. Metal and nutrient concentrations in pore waters from two of the most important watercourses traversing Doñana Park were measured: Guadiamar River (affected by the accident) and Partido Stream (unimpacted by the accident). Concentrations of dissolved constituents in interstitial waters varied according to land use in the two watersheds and to the effects of the mine spill. Levels of dissolved Co, Cu, Mo, Ti, and Zn were higher in pore waters from the Guadiamar River than in the Partido Stream, suggesting that concentrations of trace elements are still influenced by the spill. In contrast, concentrations of dissolved nutrients (NH4+, NO2-, NO3-, PO4(-3)) and some trace metals used in fertilizers (e.g. Al and Cr) were higher in the Partido Stream. Levels of dissolved As, Cs, DOC, Ge, Hg, Rb and V in the interstitial waters were equal in both watercourses. Metal concentrations in interstitial waters of the Guadiamar River floodplain were between 0.3 (As) and 16,000 (Zn) times lower than those previously reported in the river and groundwater a few weeks after the mine spill. Although metals in pore water appear to have reached levels characteristic of the area before the accident, concentrations are 60-150 times higher than those in pore waters from other regions. Metal:Al ratios in Doñana's pore waters suggest a transport of contaminants from the Iberian Pyrite Belt into Doñana Park.