Honey Bees Rely on Associative Stimulus Strength after Training on an Olfactory Transitive Inference Task

Transitive inference, the ability to establish hierarchical relationships between stimuli, is typically tested by training with overlapping premise pairs (e.g., A+B-, B+C-, C+D-, D+E-), which establishes a stimulus hierarchy (A > B > C > D > E). When subjects are tested with non-adjacent stimuli (e.g., B vs. D), a preference for B indicates transitive inference, while no preference indicates decisions based on stimulus associative strength, as B and D are equally reinforced. Previous studies with bees and wasps, conducted in an operant context, have shown conflicting results. However, this context allows free movement and the possibility to avoid non-reinforced options, thus reducing the number of non-reinforced trials. To address this, we examined whether honey bees could perform transitive inference using a Pavlovian protocol that fully controls reinforcement. We conditioned bees with five odorants, either forward- or backward-paired with a sucrose solution, across four overlapping discrimination tasks. In all experiments, bees showed no preference for B over D, choosing equally between them, regardless of the training schedule. Our results show that bees' choices were primarily influenced by stimulus associative strength and a recency effect, with greater weight given to the most recent reinforced or non-reinforced stimulus. We discuss these findings in the context of honey bee memory, suggesting that memory constraints may limit cognitive solutions to transitive inference tasks in bees