We measured locomotory behaviors (distance traveled, speed, tortuosity of path, and rate of change in direction) with computer-assisted analysis in 30 day posthatch rainbow trout (Oncorhynchus mykiss) exposed to pesticides. We also examined cholinesterase inhibition as a potential endpoint linking physiology and behavior. Sublethal exposure to chemicals often causes changes in swimming behavior, reflecting alterations in sensory and motor systems. Swimming behavior also integrates functions of the nervous system. Rarely are the connections between physiology and behavior made. Although behavior is often suggested as a sensitive, early indicator of toxicity, behavioral toxicology has not been used to its full potential because conventional methods of behavioral assessment have relied on manual techniques, which are often time-consuming and difficult to quantify. This has severely limited the application and utility of behavioral procedures. Swimming behavior is particularly amenable to computerized assessment and automated monitoring. Locomotory responses are sensitive to toxicants and can be easily measured. We briefly discuss the use of behavior in toxicology and automated techniques used in behavioral toxicology. We also describe the system we used to determine locomotory behaviors of fish, and present data demonstrating the system's effectiveness in measuring alterations in response to chemical challenges. Lastly, we correlate behavioral and physiological endpoints.