The present study evaluated the effect of activated carbon amendment in four freshwater sediments from the Great Lakes (North America) areas of concern with a wide range of sediment geochemical characteristics (0.83–5.1% total organic carbon) and polychlorinated biphenyl (PCB) concentrations (0.33–84.7 μg/g). The work focused on understanding the impact of activated carbon amendment on PCB aqueous partitioning, PCB desorption characteristics, and PCB biouptake in a freshwater oligochaete (Lumbriculus variegatus). The results showed that PCB aqueous equilibrium concentrations, rapid desorption fractions, and biouptake by the oligochaete were reduced after activated carbon amendment. Addition of activated carbon at a dose of 0.5-fold native organic carbon reduced PCB bioaccumulation by 42% for Niagara River sediment, 85% for Grasse River sediment, 74% for Milwaukee River sediment 1, and 70% for Milwaukee River sediment 2. A linear relationship was observed between log biota–sediment accumulation factor and the first 6-h desorption fractions for each PCB homologue for treated and untreated sediments. Waterlipid bioconcentration factors for PCB congeners were largely conserved after amendment with activated carbon. Our present results suggest that at steady state, changes in the aqueous PCB concentrations can be used to predict changes in PCB bioaccumulation in deposit-feeding organisms. Thus, use of advanced pore-water measurement techniques, such as solid-phase extraction passive samplers, may be suitable for long-term monitoring of treatment performance.