The authors examined the effects of natural sediment physiochemical properties on the results of lab tests with the amphipod Hyalella azteca, the midge Chironomus tentans, and the oligochaete Lumbriculus variegatus. Ten-day exposures with the three species were conducted with 50 uncontaminated sediment samples from Lakes Erie, Huron, Superior, and Ontario, which differed markedly with regard to characteristics such as grain-size distribution, organic carbon content, and mineralogical composition. Tests were conducted both with and without the addition of exogenous food. Survival of Hyalella azteca, survival and growth of Chironomus tentans, and survival/reproduction and growth of Lumbriculus variegatus were significantly greater in tests in which the animals were fed vs. those in which they were not. Hence, there is significant potential for false positive results if Hyalella azteca or Chironomus tentans is not fed during sediment tests. Predictive modeling of the assay results in relationship to sediment physiochemical characteristics failed to reveal any additional factors that influenced survival of Hyalella azteca and Chironomus tentans, or reproduction and growth of Lumbriculus variegatus in tests in which the organisms were fed. However, linear modeling did suggest that growth of fed as well as unfed Chironomus tentans may have been influenced by grain-size distribution of the test sediments.