Sub-lethal Exposure to 2,4-Dichlorophenoxyacetic Acid Disrupts Nursing and Foraging Behaviors in Honey Bees

Abstract

A popular herbicide from the chlorophenoxy group, 2,4-Dichlorophenoxyacetic acid (2,4-D) effectively controls broadleaf weeds in agricultural environments. However, its application threatens honey bee habitats and has been implicated in colony collapse disorder (CCD) due to its toxic effects. While the general hazards of 2,4-D to honey bees are recognized, its specific impact on nursing and foraging behaviors remains poorly understood. This study quantified the lethal dose (LD₅₀) of 2,4-D for honey bees across developmental stages, finding LD₅₀ values of 104.1 μg/bee for newly emerged bees, 456.6 μg/bee for nurse bees, and 221.6 μg/bee for foragers. We further investigated sub-lethal effects on nursing and foraging, observing that exposure led to significant reductions in hypopharyngeal gland (HG) acini size, essential for brood care, and decreased expression of AmGr10, an amino acid receptor gene linked to nursing behavior. For foragers, sub-lethal 2,4-D exposure impaired gustatory responsiveness to key feeding stimuli, such as sucrose and glucose. This impairment corresponded with a decrease in AmGr1 expression, a taste receptor gene critical for resource detection. Additionally, affected foragers showed reduced olfactory learning and memory, likely due to decreased expression of the octopamine receptor AmOA1, essential for associative learning processes. These findings provide compelling evidence that sub-lethal abdominal exposure to 2,4-D disrupts both nursing and foraging behaviors by impairing physiological and cognitive functions, ultimately jeopardizing colony health and resilience.