Abstract Despite the extensive use of the neonicotinoid insecticide clothianidin, and its known toxicity to beneficial insects such as pollinators, little attention has been given to its fate under agricultural field conditions. The present study investigated the fate and toxicity of clothianidin applied every other year as a corn seed-coating at 2 different rates, 0.25 mg/seed and 0.50 mg/seed, in an agricultural field undergoing a corn–soybean annual rotation, and conservation tillage. Concentrations were measured in soil, surface runoff, infiltration, and groundwater from 2011 to 2013. Clothianidin was detected at low concentrations in soil and water throughout the 2-yr corn and soybean rotation. Low and no-tillage had little or no effect on clothianidin concentrations. Laboratory toxicity bioassays were performed on nontarget species, including Daphnia magna, Hyalella azteca, Chironomus dilutus, Pimephales promelas and Eisenia fetida. Risk quotients were calculated from clothianidin concentrations measured in the field and compared with the laboratory toxicity bioassay results to assess the environmental risk of the insecticide. The risk quotient was found to be lower than the level of concern for C. dilutus, which was the most sensitive species tested; therefore, no short-term environmental risk was expected for the species investigated in the present study. Environ Toxicol Chem 2014;9999:1–8. © 2014 SETAC