Subchronic impacts of 2,4-D herbicide Weedestroy®AM40 on associative learning in juvenile yellow perch (Perca flavescens)

•2,4-D in WAM40 exposure impairs associative learning in juvenile yellow perch•2,4-D in WAM40 did not impact hunger motivation, locomotion, or a visual startle response•Ecologically relevant concentrations of 2,4-D in WAM40 can impact essential behaviors Aquatic herbicides are commonly used to control a wide variety of invasive and nuisance plants. One common active ingredient used in commercial herbicide formulations in Midwestern states is 2,4-dichlorophenoxyacetic acid (2,4-D). Due to the stability of 2,4-D in aquatic environments, many non-target aquatic species experience prolonged exposure throughout critical developmental life stages that can affect essential behaviors. However, the impacts of 2,4-D exposure on learning behaviors in juvenile fish are poorly understood. Therefore, we conducted a series of experiments using a maze environment to determine the effects of a commercial 2,4-D amine salt herbicide formulation (Weedestroy®AM40; WAM40; at 0.00, 0.50, 2.00, and 50.00 mg/L 2,4-D acid equivalent (a.e.)) exposure on juvenile yellow perch's ability to perform a feed associated learning behavior. We observed a significant decrease in the ability of yellow perch to correctly complete the feed associated learning behavior within 200 s when exposed to WAM40 at 2.00 and 50.00 mg/L 2,4-D as compared to controls (p = 0.0002; p < 0.0001, respectively) and within 600 s when exposed to WAM40 at 2.00 and 50.0 mg/L 2,4-D as compared to the controls (p = 0.0107 and p < 0.0001). These data suggest that exposure to 2,4-D in WAM40 can both increase the amount of time it takes for yellow perch to complete a feed associated learning behavior and/or obstruct the behavior altogether. Further experiments showed no significant decreases in locomotion (p > 0.05), hunger motivation (p > 0.05), and a visually guided startle response (p > 0.05), in all treatment groups tested as compared to controls. This suggests that 2,4-D in WAM40 does not inhibit feed associated learning behaviors via interaction with these mechanisms. Altogether, the results indicate that the use of 2,4-D herbicides for weed control in aquatic ecosystems could present risks to cognitive functions that control essential behaviors of yellow perch.