In 1972, the governments of Canada and the United States committed to restoring the physical, chemical, and biological integrity of the Laurentian Great Lakes under the Great Lakes Water Quality Agreement. Through this framework, the downstream-most section of Eighteenmile Creek, a tributary to the south shore of Lake Ontario in New York, was designated as an Area of Concern (AOC) because water quality and bed sediments were contaminated by past industrial and municipal discharges, waste disposal, and pesticide usage. Five beneficial use impairments (BUIs) have been identified in the AOC including the degradation of the “benthos”, or the benthic macroinvertebrate community. This investigation used sediment toxicity testing and macroinvertebrate community assessments to determine if the toxicity of bed sediments in the AOC differed from that of an unimpacted reference stream. Results from 10-day toxicity tests indicated that survival and growth of the dipteran Chironomus dilutus and the amphipod Hyalella azteca did not differ significantly between sediments from the AOC and reference area. Analyses of benthic macroinvertebrate community integrity and structure also indicated that macroinvertebrate communities, while impacted across most sites on both streams, were generally similar between the AOC and reference area. Despite these findings, the upstream-most AOC site consistently scored poorly in all analyses, which suggests that localized sediment toxicity may exist in the AOC, even if large scale differences between the AOC and a comparable reference stream are minimal.