Odorants that Induce Hygienic Behavior in Honeybees: Identification of Volatile Compounds in Chalkbrood-Infected Honeybee Larvae

Social insects that live in large colonies are vulnerable to disease transmission due to relatively high genetic relatedness among individuals and high rates of contact within and across generations. While individual insects rely on innate immune responses, groups of individuals also have evolved social immunity. Hygienic behavior, in which individual honeybees detect chemical stimuli from diseased larvae and subsequently remove the diseased brood from the nest, is one type of social immunity that reduces pathogen transmission. Three volatile compounds, collected from larvae infected with the fungal pathogen Ascosphaera apis and detected by adult honey bees, were identified by coupled gas chromatography-electroantennographic detection and gas chromatography-mass spectrometry. These three compounds, phenethyl acetate, 2-phenylethanol, and benzyl alcohol, were present in volatile collections from infected larvae but were absent from collections from healthy larvae. Two field bioassays revealed that one of the compounds, phenethyl acetate is a key compound associated with Ascosphaera apis-infected larvae that induces hygienic behavior.