Although zebrafish are gaining in popularity among researchers, the utilization of zebrafish models remains limited, due to a scarcity of well-established behavioral paradigms (Gerlai, 2015 Sison, Cawker, Buske, & Gerlai, 2006). The zebrafish displays a repertoire of complex behaviors and has recently emerged as an excellent vertebrate model for behavioral neuroscience studies (Gerlai, 2010). Thus, Argus is a novel, cost-effective, and customizable method for the analysis of adult zebrafish behavior that may be utilized for the behavioral quantification of both single and dyadic interacting subjects, but further sophistication will be needed for the proper identification of complex motor patterns, measures that a human observers can easily detect. In contrast, the manual coding of behavior in The Observer showed weaker correlations with the two tracking methods (EthoVision and Argus). We found that Argus and EthoVision extract similar absolute values and patterns of changes in these values for several behavioral measures, including speed, freezing, erratic movement, and interindividual distance. Next we computed and quantified the behavioral variables that characterize dyadic interactions between zebrafish. We first also performed an analysis of the movement of individual fish and compared the performance of the three software applications. All three software applications were originally designed to quantify the behavior of a single subject. In this article, we compare Argus with Noldus EthoVision and Noldus The Observer, to validate this new system. Argus includes a new, user-friendly, and efficient graphical user interface, instead of a command-line interface, and offers simplicity and flexibility in measuring complex zebrafish behavior through customizable parameters. Building on our previous work with the RealFishTracker (in-house-developed tracking system), we present Argus, a data extraction and analysis tool built in the open-source R language for behavioral researchers without any expertise in R. Promising lines of research, however, require the development and validation of software tools that will allow automated and cost-effective behavioral analysis. Zebrafish show great potential for behavioral neuroscience.
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