Locomotion behaviors are susceptible to disruption by a broad spectrum of chemicals and environmental stresses. However, no systematic testing of locomotion behavior defects induced by metal exposure has been conducted in the model organism of nematode Caenorhabditis elegans. In this study, the acute toxicity from heavy metal exposure on the locomotion behaviors was analyzed in nematodes. Endpoints of head thrash, body bend, forward turn, backward turn, and Omega/U turn were chosen to evaluate the locomotion behavioral defects. Our data suggest that the endpoints of head thrash, body bend, and forward turn will be useful for the evaluation of heavy metal toxicity in nematodes. The endpoint of head thrash could detect the toxicity from Cd, Co, Cr, Cu, Hg, and Pb exposures at a low concentration (2.5 micromol/L). The endpoint of body bend could be explored to evaluate the toxicity from all assayed heavy metal exposures at different concentrations, whereas the endpoint of forward turn will be more useful for the evaluation of heavy metal toxicity at high concentrations. Thus, endpoints of these locomotion behaviors establish a fast and economic way to assess the presence of acute toxicity from heavy metal exposure in nematode C. elegans.