Elucidating the effects of acute and chronic exposure to 2,4-Dichlorophenoxyacetic acid on fathead minnow (Pimephales promelas) innate immunity.

Aquatic herbicides, such as 2,4-dichlorophenoxyacetic acid (2,4-D) formulations, are commonly used for invasive species management throughout the United States. Ecologically relevant concentrations of 2,4-D can impair essential behaviors, reduce survival, and act as an endocrine disruptor; however, there is limited knowledge of its effects on the health of non-target organisms. Here, we investigate the acute and chronic exposure impacts of 2,4-D on adult male and female fathead minnow (Pimephales promelas) innate immune function. We exposed both adult male and female fathead minnows to three different ecologically relevant concentrations of 2,4-D (0.00, 0.40, and 4.00 mg/L) and took blood samples at three acute time points (6, 24, and 96 h) and one chronic time point (30 days). We found that male fatheads had higher total white blood cell concentrations when exposed to 2,4-D at the acute time points. For the females, only proportions of specific cell types were altered when exposed to 2,4-D at the acute time points. However, we did not observe any significant impacts of chronic exposure to 2,4-D on any innate immune responses for either males or females. Overall, this study is the first step in answering an important question for game fisheries and management agencies while providing insight to future studies that investigate the impacts of herbicide exposure to freshwater fish health and immunity.

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

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.