Marked interspecific differences in the neuroanatomy of the male olfactory system of honey bees (genus Apis).

All honey bee species (genus Apis) display a striking mating behavior with the formation of male (drone) congregations, in which virgin queens mate with many drones. Bees' mating behavior relies on olfactory communication involving queen-but also drone pheromones. To explore the evolution of olfactory communication in Apis, we analyzed the neuroanatomical organization of the antennal lobe (primary olfactory center) in the drones of five species from the three main lineages (open-air nesting species: dwarf honey bees Apis florea and giant honey bees Apis dorsata; cavity-nesting species: Apis mellifera, Apis kochevnikovi, and Apis cerana) and from three populations of A. cerana (Borneo, Thailand, and Japan). In addition to differences in the overall number of morphological units, the glomeruli, our data reveal marked differences in the number and position of macroglomeruli, enlarged units putatively dedicated to sex pheromone processing. Dwarf and giant honey bee species possess two macroglomeruli while cavity-nesting bees present three or four macroglomeruli, suggesting an increase in the complexity of sex communication during evolution in the genus Apis. The three A. cerana populations showed differing absolute numbers of glomeruli but the same three macroglomeruli. Overall, we identified six different macroglomeruli in the genus Apis. One of these (called MGb), which is dedicated to the detection of the major queen compound 9-ODA in A. mellifera, was conserved in all species. We discuss the implications of these results for our understanding of sex communication in honey bees and propose a putative scenario of antennal lobe evolution in the Apis genus.

"Up" or "down" that makes the difference. How giant honeybees (Apis dorsata) see the world.

A. dorsata builds its large exposed comb high in trees or under ledges of high rocks. The "open" nest of A. dorsata, shielded (only!) by multiple layers of bees, is highly vulnerable to any kind of direct contact or close range attacks from predators. Therefore, guard bees of the outer layer of A. dorsata's nest monitor the vicinity for possible hazards and an effective risk assessment is required. Guard bees, however, are frequently exposed to different objects like leaves, twigs and other tree litter passing the nest from above and falling to the ground. Thus, downward movement of objects past the nest might be used by A. dorsata to classify these visual stimuli near the nest as "harmless". To test the effect of movement direction on defensive responses, we used circular black discs that were moved down or up in front of colonies and recorded the number of guard bees flying towards the disc. The size of the disc (diameter from 8 cm to 50 cm) had an effect on the number of guard bees responding, the bigger the plate the more bees started from the nest. The direction of a disc's movement had a dramatic effect on the attraction. We found a significantly higher number of attacks, when discs were moved upwards compared to downward movements (GLMM (estimate ± s.e.) 1.872 ± 0.149, P < 0.001). Our results demonstrate for the first time that the vertical direction of movement of an object can be important for releasing defensive behaviour. Upward movement of dark objects near the colony might be an innate releaser of attack flights. At the same time, downward movement is perceived as a "harmless" stimulus.

Comb construction in mixed-species colonies of honeybees, Apis cerana and Apis mellifera.

Comb building in mixed-species colonies of Apis cerana and Apis mellifera was studied. Two types of cell-size foundation were made from the waxes of these species and inserted into mixed colonies headed either by an A. cerana or an A. mellifera queen. The colonies did not discriminate between the waxes but the A. cerana cell-size foundation was modified during comb building by the workers of both species. In pure A. cerana colonies workers did not accept any foundation but secreted wax and built on foundation in mixed colonies. Comb building is performed by small groups of workers through a mechanism of self-organisation. The two species cooperate in comb building and construct nearly normal combs but they contain many irregular cells. In pure A. mellifera colonies, the A. cerana cell size was modified and the queens were reluctant to lay eggs on such combs. In pure A. cerana colonies, the A. mellifera cell size was built without any modification but these cells were used either for drone brood rearing or for food storing. The principal elements of comb-building behaviour are common to both species, which indicates that they evolved prior to and were conserved after speciation.