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Updated: Jan 30, 2026

Experimental Protocol for Examining Behavioral Response Profiles in Larval Fish: Application to the Neuro-stimulant Caffeine
Published on: July 24, 2018
2,4-Dichlorophenoxyacetic acid containing herbicide impairs essential visually guided behaviors of larval fish
Gavin K Dehnert1, William H Karasov2, Marc A Wolman1
1Department of Integrative Biology, University of Wisconsin - Madison, Madison, WI, USA.
Abstract:
Aquatic herbicides are used worldwide to eradicate nuisance and invasive plants despite limited knowledge of their toxicity to non-target organisms. 2,4-Dichlorophenoxyacetic acid (2,4-D) is a common active ingredient in commercial herbicide formulations, which triggers plant cell death by mimicking the plant-specific hormone auxin. Application practices of 2,4-D commercial herbicides typically coincide with yearly freshwater fish spawning periods. This practice exposes fish to xenobiotics at their vulnerable larval stages. The full impacts of 2,4-D on larval fish remains poorly understood, and hence, whether it may alter larval survival, larval behavior, fish populations, and ecosystem dynamics. In the present study, we exposed embryonic and larval zebrafish (Danio rerio) to the active ingredient 2,4-D (pure 2,4-D) or a 2,4-D containing commercial herbicide DMA4®IVM (DMA4) and evaluated morphology, survival, behavior, and nervous system function. At 2,4-D concentrations producing no overt morphological defects during embryonic or early larval stages, we observed reduced survival throughout a 21-day larval assay (4-8 ppm DMA4 and 0.75-4 ppm pure 2,4-D). Notably, prey capture, a behavior essential to survival, was reduced in 2,4-D-exposed larval zebrafish (4-8 ppm DMA4 and 0.75-4 ppm pure 2,4-D) and yellow perch (Perca flavescens) (4-20 ppm DMA4). In zebrafish, 8 ppm DMA4 exposure reduced prey capture when exposure was restricted to the period of visual system development. Consistent with these results, larval zebrafish exposed to 8 ppm DMA4 showed reduced neural activity within the optic tectum following prey exposure. Together, our results suggest that 2,4-D alters the development and function of neural circuits underlying vision of larval fish, and thereby reduces visually guided behaviors required for survival.
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