Many emerging contaminants tend to be biologically active at very low concentrations, occur in water as part of complex mixtures, and impact biota in ways that are not detected using traditional toxicity tests (e.g., median lethal concentration). To evaluate emerging contaminants, the authors developed a method for detecting sublethal behavioral effects by quantifying the swimming behavior of Daphnia pulex, a model organism for studying aquatic toxicity. This optical tracking technique is capable of measuring many swimming parameters, 2 of which-cumulative distance and angular change-are presented. To validate this technique, 2 prototypical compounds that exhibit different modes of action as well as corresponding insecticides that are commonly found in surface waters were investigated. The acetylcholinesterase (AChE) inhibitor physostigmine was used as the prototypical compound for the large number of AChE inhibitor insecticides (e.g., chlorpyrifos). Nicotine was used as the prototypical compound for neonicotinoid insecticides (e.g., imidacloprid). Results demonstrate that this assay is capable of detecting sublethal behavioral effects that are concentration-dependent and that insecticides with the same mode of action yield similar results. The method can easily be scaled up to serve as a high-throughput screening tool to detect sublethal toxic effects of a variety of chemicals. This method is likely to aid in enhancing the current understanding of emerging contaminants and to serve as a novel water-quality screening tool.

Contaminant exposure in aqueous systems typically involves complex chemical mixtures. Given the large number of compounds present in the environment, it is critical to identify hazardous chemical interactions rapidly. The present study utilized a prototype for a novel high‐throughput assay to quantify behavioral changes over time to identify chemical interactions that affect toxicity. The independent and combined effects of 2 chemicals, diazinon (an insecticide) and 4‐nonylphenol (a detergent metabolite), on the swimming behavior of the freshwater crustacean Daphnia pulex were examined. Cumulative distance and change in direction were measured repeatedly via optical tracking over 90 min. Exposure to low concentrations of diazinon (0.125–2 µM) or 4‐nonylphenol (0.25–4 µM) elicited significant concentration‐ and time‐dependent effects on swimming behavior. Exposure to 0.5 µM 4‐nonylphenol alone did not significantly alter mean cumulative distance but did elicit a small, significant increase in mean angle, the measure of change in direction. When 0.5 µM 4‐nonylphenol was used in combination with diazinon (0.125–0.5 µM), it augmented the adverse impact of diazinon on the swimming behavior of Daphnia. Additionally, enhanced sensitivity to diazinon was observed in animals exposed to treated wastewater effluent for 24 h prior to a diazinon challenge. The present experiments demonstrate that exposure to 4‐nonylphenol and complex chemical mixtures (e.g., treated wastewater) can enhance the toxicity of exposure to the insecticide diazinon. Environ Toxicol Chem 2015;34:1145–1153. © 2015 SETAC