The freshwater Anacostia River watershed (Maryland, DC, USA) was surveyed for the sources of bioavailable US Environmental Protection Agency (USEPA) Priority Pollutants and toxic metals by active biomontoring (ABM) using the freshwater Asiatic clam Corbicula fluminea. The Anacostia River is a 456 km(2) tributary of the tidal freshwater Potomac River that includes the city of Washington, DC where edible fish are highly contaminated with PCBs and chlordane. From 1999 to 2011, Corbicula were collected for ABM from a Potomac reference site and translocated in cages placed at 45 sites in the tidal and nontidal Anacostia watershed. Minimum clam mortality and maximum contaminant bioaccumulation was with 2-week translocation. The clam tissues (28-50) were combined at sites and analyzed by TestAmerica for 66 USEPA Priority Pollutants plus technical chlordane, benz(e) pyrene, and 6 metals (As, Cd, Cr, Cu, Fe, Pb). Tissue contaminants reflected water, not sediment, levels. To compare sites, all contaminant data above detection or reference were grouped as total metals (TMET), total polycyclic aromatic hydrocarbons (TPAH), total PCB congeners (TPCB), total pesticides (TPEST), and total technical chlordane (TCHL). Tidal Anacostia ABM found highest TPAH and TCHL upstream at Bladensburg Marina (MD) except for TCHL at site PP near the confluence. Five nontidal MD subtributaries (94% of flow) had 17 sites with bioavailable TPAH, TPCB, or TCHL 2 to 3 times higher than found at the toxic-sediment "hotspots" near Washington. The only TMET noted was Fe at 1 site. TPAH in MD subtributaries was highest near industrial parks and Metro stations. A naphthalene spill was detected in Watts Branch. TPCB (low molecular weight) originated upstream at 1 industrial park. Total technical chlordane (80% of TPEST) was 2 to 5 times the US Food and Drug Administration action in 4 nontidal tributaries where heptachlor indicated legacy chlordane dumpsites. Total technical chlordane fell to reference below a stormwater pond, suggesting transport via suspended sediment. Controlling the formation and movement of contaminated TSS in MD should enable the uncontaminated-sediment capping of Washington DC's toxic-sediment "hot-spots" that are presently considered responsible for fish contamination. Integr Environ Assess Manag 2016;12:548-558. © 2015 SETAC.