Estimating the risk of polychlorinated biphenyls (PCBs) in aquatic systems requires evaluation of exposure, usually based on exhaustive chemical extraction of sediment and potentially exposed organisms and an assessment of toxicity. Remediation can then focus on areas where the exposure leads to the highest risk. Although effective, an approach that estimates exposure, which accounts for bioavailabilty, bioaccumulation, trophic transfer potential, and transport of materials within and out of the waterway, should serve as a more comprehensive environmental assessment. The current study examined exposure of PCBs in several different trophic levels within the Campus Lake ecosystem, Carbondale, Illinois. The source of contamination and the distribution of PCBs among ecosystem components demonstrated contamination within the aquatic portion of Campus Lake and transport out of the aquatic environment to the riparian area. Several media were collected including sediment, emergent insects, spiders, and three species of fish. Sediment extractions demonstrated that PCBs were localized to one small cove and this area served as the source for transfer of PCBs to both aquatic and terrestrial organisms. Single-point 24-h Tenax extractions formed the basis for evaluating the bioavailable component of the sediment-associated PCBs with strong correlations to laboratory-based bioaccumulation assays for oligochaetes and chironomid larvae. Stable isotope data suggested that the source of carbon to the food web was relatively constant. Food web samples of emerging insects, fish, and spiders revealed that the PCBs in Campus Lake were bioavailable and the pattern of the PCB signature among food web components followed typical food web processes. The PCB congener pattern was consistent between emergent midges and spiders demonstrating the transfer of PCBs from aquatic to terrestrial species. The PCB concentrations detected in emerging insects from the contaminated area were on average 25 times greater than those detected in emerging insects from reference sites outside the area of greatest sediment contamination. High PCB concentrations found in several species of fish suggested that despite the localized sediment contamination, fish throughout the lake were exposed. These levels also exceeded the fish consumption advisory criteria. The PCB pattern comparisons suggested that the contaminated sediment was the source of exposure throughout the food web. This approach identified the scope of exposure to organisms, demonstrated bioavailability, and provided a basis for future PCB remediation and subsequent monitoring of Campus Lake. In comparison to studies focused solely on limited sampling of fish for consumption advisory purposes, this approach demonstrated the importance of more comprehensive studies to examine the range of ecosystem exposure even from very limited contamination sources.