Nutrient export from the Everglades Agricultural Area (EAA) has been implicated in causing sulfur (S) enrichment of Everglades wetlands. However, quantification of the S budget and transformations in EAA soils is inadequate. The objective of this study was to quantify various S fractions and investigate how elemental S amendment affects S dynamics in EAA soils. Reduced S compounds were not detected in soil before elemental S application. Organic S was the major form of S, comprising 87% of total S, followed by extractable SO4-S (13%). Extractable SO4-S for soils receiving 448 kg S ha(-1) was 36%, 131%, 201%, and 270% higher than for unamended soils at 2, 6, 9, and 13 months, respectively. Elemental S was significantly higher in soils receiving 448 kg S ha(-1) (482 mg kg(-1)) than in soils receiving 224 (111 mg kg(-1)) and 112 kg S ha(-1) (55 mg kg(-1)) and unamended soil (0 mg kg(-1)) at 2 months after S application. Similar to extractable SO4-S, elemental S significantly decreased during the growing season. Sulfur application did not affect the sulfatase activity, however, mineralizable S increased concurrently with S application rate, and the effects continued throughout the growing season. This result was largely attributed to the oxidation of the applied elemental S. Our results suggest that large-scale S application in the EAA soils is likely to increase SO42- concentrations in soils, which poses a potential risk of SO42- export to sensitive Everglades wetlands.