Five chemicals with different modes of action were evaluated in laboratory studies to determine their acute toxicity (48-h median lethal concentration [LC50]) and behavioral effects on 30-d-old Japanese medaka (Oryzias latipes). The order of toxicity for these xenobiotics was permethrin > chlorpyrifos > 2,4-dinitrophenol (2,4-DNP) > strychnine > phenol. The 48-h LC50s were significantly different and ranged from 0.011 to 24.1 mg/L. In addition, chlorpyrifos and permethrin accumulated in the tissues of juvenile O. latipes. Observations of five behavioral/morphological responses, including changes in equilibrium, general activity, startle response, and morphology (e.g., hemorrhage and deformities) were used as indicators of sublethal toxicity. Each chemical, with the exception of 2,4-DNP, elicited a distinct behavior or set of behavioral responses. The behavioral toxicology bioassay may be valuable in comparing and predicting the mode of action of new or unknown toxicants in this species of fish.