Invasive red imported fire ants (Solenopsis invicta) create a serious threat to public safety, agriculture, biodiversity, and the local economy, necessitating early detection and surveillance, which are currently time-consuming and dependent on the inspector's expertise. This study marks an initial investigation into the potential of honeybees (Apis mellifera) to detect and discriminate the odor of S. invicta through the olfactory conditioning of proboscis extension responses. Deceased S. invicta were used as conditioned stimuli to ensure relevance to non-infested areas. The results showed that the bees rapidly learned to respond to deceased ant odors, with response levels significantly increasing at higher odor intensities. Bees exhibited generalization across the odors of 25 minor workers, 21 median workers, 1 major worker, and 1 female alate. When conditioned with deceased ant odors, bees effectively recognized live ants, particularly when trained on a single minor worker. Discrimination abilities varied by species and were higher when S. invicta was paired with Polyrhachis dives and Nylanderia yaeyamensis, and lower with S. geminata, Pheidole rabo, and Pheidole fervens. Notably, discrimination improved significantly with the application of latent inhibition. These findings suggest that trained honeybees have the potential to detect S. invicta. Further refinement of this approach could enhance its effectiveness for detection and surveillance.
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