Age-specific olfactory attraction between Western honey bee drones (Apis mellifera) and its chemical basis

During the mating season, drones (males) of the Western honey bee (Apis mellifera) form congregations numbering thousands high in the air. Virgin queens arrive at these congregations after they have formed and mate on the fly with 15-20 drones. To explain the formation of drone congregations, a drone-produced aggregation pheromone has been proposed many years ago but due to the low accessibility of natural mating sites in bees, its study has progressed slowly. Recently, we used a walking simulator in controlled laboratory conditions to show that drones are indeed attracted by groups of other drones. Since these previous experiments were carried out with drones captured when flying out of the hive, it is currently unclear if this olfactory attraction behaviour is related to the drones' sexual maturity (usually reached between 9 and 12 days) and may thus be indicative of a possible role in congregation formation, or if it is observed at any age and may represent in-hive aggregation. We thus assessed here the dependency of drone olfactory attraction on their age. First, we performed behavioural experiments in the walking simulator to measure olfactory preferences of drones in three age groups from 2-3 to 12-15 days. Then, we performed chemical analyses in the same age groups to evaluate whether chemical substances produced by the drones may explain age differences in olfactory attraction. We show that honey bee drones are attracted by conspecifics of the same age when they are sexually mature (12-15 days old) but not when they are younger (2-3 and 7-8 days old). In parallel, our data show that drones' chemical profile changes with age, including its most volatile fraction. These results are discussed in the context of drone mutual attraction both within the hive and at drone congregations.