Concentrations of perfluorinated chemicals (PFCs) were measured in surface waters and sediments from the Coosa River watershed in northwest Georgia, USA, to examine their distribution downstream of a suspected source. Samples from eight sites were analyzed using liquid chromatography‐tandem mass spectrometry. Sediments were also used in 28‐d exposures with the aquatic oligochaete, Lumbriculus variegatus, to assess PFC bioaccumulation. Concentrations of PFCs in surface waters and sediments increased significantly below a land‐application site (LAS) of municipal/industrial wastewater and were further elevated by unknown sources downstream. Perfluorinated carboxylic acids (PFCAs) with eight or fewer carbons were the most prominent in surface waters. Those with 10 or more carbons predominated sediment and tissue samples. Perfluorooctane sulfonate (PFOS) was the major homolog in contaminated sediments and tissues. This pattern among sediment PFC concentrations was consistent among sites and reflected homolog concentrations emanating from the LAS. Concentrations of PFCs in oligochaete tissues revealed patterns similar to those observed in the respective sediments. The tendency to bioaccumulate increased with PFCA chain length and the presence of the sulfonate moiety. Biota‐sediment accumulation factors indicated that short‐chain PFCAs with fewer than seven carbons may be environmentally benign alternatives in aquatic ecosystems; however, sulfonates with four to seven carbons may be as likely to bioaccumulate as PFOS.