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Abstract Reproductive division of labor in insect societies is regulated through multiple concurrent mechanisms, primarily chemical and behavioral. Here, we examined if the Dufour’s gland secretion in the primitively eusocial bumble beeBombus impatienssignals information about caste, social condition, and reproductive status. We chemically analyzed Dufour’s gland contents across castes, age groups, social and reproductive conditions, and examined worker behavioral and antennal responses to gland extracts. We found that workers and queens each possess caste-specific compounds in their Dufour’s glands. Queens and gynes differed from workers based on the presence of diterpene compounds which were absent in workers, whereas four esters were exclusive to workers. These esters, as well as the total amounts of hydrocarbons in the gland, provided a separation between castes and also between fertile and sterile workers. Olfactometer bioassays demonstrated attraction of workers to Dufour’s gland extracts that did not represent a reproductive conflict, while electroantennogram recordings showed higher overall antennal sensitivity in queenless workers. Our results demonstrate that compounds in the Dufour’s gland act as caste- and physiology-specific signals and are used by workers to discriminate between workers of different social and reproductive status. 

Honey bees are social insects that live in large groups called colonies, within structures known as hives. The young adult bees stay within the hive to build nests and care for the young, while the older bees leave the hive to forage for food. Honey bees store food and other valuable resources in their hives, so they are often targeted by predators, parasites and ‘robber’ bees from other colonies. Therefore, it is important for bees to determine whether individuals trying to enter the nest are group members or intruders. While it is known that social insects use blends of waxy chemicals called cuticular hydrocarbons to identify group members at the entrance to the colony, it is not clear how members of the same colony acquire a similar blend of cuticular hydrocarbons. Some previous work suggested that in some ant species (which are also social insects), colony members exchange cuticular hydrocarbons with each other so that all members of the colony are covered with a similar blend of chemicals. However, it was not known whether honey bees also share cuticular hydrocarbons between colony members in order to identify members of a hive. Vernier et al. used chemical, molecular and behavioral approaches to study the cuticular hydrocarbons found on honey bees. The results show that, rather than exchanging chemicals with other members of their colony, individual bees make their own blends of cuticular hydrocarbons. As a bee ages it makes different blends of cuticular hydrocarbons, and by the time it starts to leave the hive to forage it makes a blend that is specific to the colony it belongs to. The production of this final blend is influenced by the environment within the hive. Thus, the findings of Vernier et al. indicate that honey bees guarding the entrance to a hive can only identify non-colony-member forager bees as intruders, rather than any non-colony-member bee that happens upon the hive entrance. Honey bees play an essential role in pollinating many crop plants so understanding how these insects maintain their social groups may help to improve agriculture in the future. Furthermore, this work may aid our understanding of how other social insects interact in a variety of biological situations.