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Organic carbon is important in regulating ecosystem function, and its source and abundance may be altered by urbanization. We investigated shifts in organic carbon quantity and quality associated with urbanization and ecosystem restoration, and its potential effects on denitrification at the riparian-stream interface. Field measurements of streamwater chemistry, organic carbon characterization, and laboratory-based denitrification experiments were completed at two forested, two restored, and two unrestored urban streams at the Baltimore Long-Term Ecological Research site, Maryland, USA. Dissolved organic carbon (DOC) and nitrate loads increased with runoff according to a power-law function that varied across sites. Stable isotopes and molar C:N ratios suggested that stream particulate organic matter (POM) was a mixture of periphyton, leaves, and grass that varied across site types. Stable-isotope signatures and lipid biomarker analyses of sediments showed that terrestrial organic carbon sources in streams varied as a result of riparian vegetation. Laboratory experiments indicated that organic carbon amendments significantly increased rates of denitrification (35.1 +/- 9.4 ng N.[g dry sediment](-1).h(-1); mean +/- SE) more than nitrate amendments (10.4 +/- 4.0 ng N.[g dry sediment](-1).h(-1)) across streamflow conditions and sites. Denitrification experiments with naturally occurring carbon sources showed that denitrification was significantly higher with grass clippings from home lawns (1244 +/- 331 ng N.g dry sediment(-1).h(-1)), and overall unrestored urban sites showed significantly higher denitrification rates than restored and forest sites. We found that urbanization influences organic carbon sources and quality in streams, which can have substantial downstream impacts on ecosystem services such as denitrification.