In 1972, the USA and Canada agreed to restore the chemical, physical, and biological integrity of the Great Lakes ecosystem under the first Great Lakes Water Quality Agreement. In subsequent amendments, part of the St. Lawrence River at Massena, New York and segments of three tributaries, were designated as an Area of Concern (AOC) due to the effects of polychlorinated biphenyls (PCBs), lead and copper contamination, and habitat degradation and resulting impairment to several beneficial uses. Because sediments have been largely remediated, the present study was initiated to evaluate the current status of the benthic macroinvertebrate (benthos) beneficial use impairment (BUI). Benthic macroinvertebrate communities and sediment toxicity tests using Chironomus dilutus were used to test the hypotheses that community condition and sediment toxicity at AOC sites were not significantly different from those of adjacent reference sites. Grain size was found to be the main driver of community composition and macroinvertebrate assemblages, and bioassessment metrics did not differ significantly between AOC and reference sites of the same sediment class. Median growth of C. dilutus and its survival in three of the four river systems did not differ significantly in sediments from AOC and reference sites. Comparable macroinvertebrate assemblages and general lack of toxicity across most AOC and reference sites suggest that the quality of sediments should not significantly impair benthic macroinvertebrate communities in most sites in the St. Lawrence River AOC.