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Thousands of stream miles in the southern Piedmont region are impaired because of high levels of suspended sediment. It is unclear if the source is upland erosion from agricultural sources or bank erosion of historic sediment deposited in the flood plains between 1830 and 1930 when cotton farming was extensive. The objective of this study was to determine the source of high stream suspended sediment concentrations in a typical southern Piedmont watershed using sediment fingerprinting techniques. Twenty-one potential tracers were tested for their ability to discriminate between sources, conservative behavior, and lack of redundancy. Tracer concentrations were determined in potential sediment sources (forests, pastures, row crop fields, stream banks, and unpaved roads and construction sites), and suspended sediment samples collected from the stream and analyzed using mixing models. Results indicated that 137Cs and 15N were the best tracers to discriminate potential sediment sources in this watershed. The delta15N values showed distinct signatures in all the potential suspended sediment sources, and delta15N was a unique tracer to differentiate stream bank soil from upland subsurface soils, such as soil from construction sites, unpaved roads, ditches, and field gullies. Mixing models showed that about 60% of the stream suspended sediment originated from eroding stream banks, 23 to 30% from upland subsoil sources (e.g., construction sites and unpaved roads), and about 10 to 15% from pastures. The results may be applicable to other watersheds in the Piedmont depending on the extent of urbanization occurring in these watersheds. Better understanding of the sources of fine sediment has practical implications on the type of sediment control measures to be adopted. Investment of resources in improving water quality should consider the factors causing stream bank erosion and erosion from unpaved roads and construction sites to water quality impairment.