We studied the effects of mixed honeybee colonies of Apis mellifera and Apis cerana on the intraspecific and interspecific recognition of female brood stages in the honeybees A. cerana and A. mellifera by transferring brood combs between queenright colonies. In the intraspecific tests, significantly more larvae were removed in A. cerana than in A. mellifera, whilst significantly fewer eggs and pupae were removed in A. cerana than in A. mellifera. In the interspecific tests, A. cerana colonies removed significantly more larvae and pupae of A. mellifera than the same brood stages of A. cerana were removed by A. mellifera. We show there are highly significant differences in both intraspecific and interspecific brood recognition between A. cerana and A. mellifera and that brood recognition operates with decreasing intensity with increasing developmental age within species. This suggests that worker policing in egg removal is a first line of defense against heterospecific social parasites.

BACKGROUND: The honey bee is an important model system for increasing understanding of molecular and neural mechanisms underlying social behaviors relevant to the agricultural industry and basic science. The western honey bee, Apis mellifera, has served as a model species, and its genome sequence has been published. In contrast, the genome of the Asian honey bee, Apis cerana, has not yet been sequenced. A. cerana has been raised in Asian countries for thousands of years and has brought considerable economic benefits to the apicultural industry. A cerana has divergent biological traits compared to A. mellifera and it has played a key role in maintaining biodiversity in eastern and southern Asia. Here we report the first whole genome sequence of A. cerana. RESULTS: Using de novo assembly methods, we produced a 238 Mbp draft of the A. cerana genome and generated 10,651 genes. A.cerana-specific genes were analyzed to better understand the novel characteristics of this honey bee species. Seventy-two percent of the A. cerana-specific genes had more than one GO term, and 1,696 enzymes were categorized into 125 pathways. Genes involved in chemoreception and immunity were carefully identified and compared to those from other sequenced insect models. These included 10 gustatory receptors, 119 odorant receptors, 10 ionotropic receptors, and 160 immune-related genes. CONCLUSIONS: This first report of the whole genome sequence of A. cerana provides resources for comparative sociogenomics, especially in the field of social insect communication. These important tools will contribute to a better understanding of the complex behaviors and natural biology of the Asian honey bee and to anticipate its future evolutionary trajectory.

The honeybees Apis cerana and Apis mellifera are derived from the same ancestral base about two million years ago. With speciation and evolution, they have acquired many advanced living skills in common, but have also evolved very different living strategies due to different distributions. This thesis is an intensive study of the biology of the mixed-species colonies of these species, the aims of which were to investigate their behavioural relationships and uncover the evolutionary conserved features of their behaviours subsequent to speciation. The results show that the two species can form a stable society to perform normal tasks. First, workers of both species in the mixed-colonies could form the typical retinue behaviour to hetero-species queens, thus indicating that queen pheromones could be spread to and by both species. Secondly, both species did not show significantly different ovarian activation under hetero-species queens, suggesting that the queen pheromones more likely play a role of "honest signal" rather than a "repression" substance in the honeybee colonies. Thirdly, both species could mutually decode each other‘s waggle dances, with unexpectedly low misunderstanding; revealing that the dance language in a dark environment is quite adaptive for cavity-nesting honeybees. Fourthly, workers of both species could cooperate with each other in comb construction, although the combs they built contain many irregular cells. Interestingly, A. cerana workers could be stimulated by A. mellifera workers to perform this task, thus confirming self-organization theory in the colony. Fifthly, A. mellifera workers behaved more "defectively" in thermoregulation, but perhaps because A. cerana workers are more sensitive to changes in hive temperature. Given these differences in strategy, A. mellifera workers‘ performance might in fact reduce conflicts. Lastly, when faced with threats of predatory wasps, both species engaged in aggressive defence. Although they did not learn from each other‘s responses, species-specific strategies were adopted by each of them so that the defence of the mixed-colonies is very effective. I conclude that the two species can adapt to each other‘s efforts and task allocation is reasonably organized allowing mixed-species colonies to reach stability. These results suggest that all of the social behaviours discussed here were highly conserved following speciation. This thesis could provide some clues for the study of honeybee evolution from open-nesting to the transition of cavity-nesting.