Parallel pheromone, metabolite, and lipid analyses reveal patterns associated with early life transitions and ovary activation in honey bee (Apis mellifera) queens

The honest signal hypothesis posits that queens emit pheromonal signals that convey information about fecundity, which workers use to make decisions around investing in direct or indirect fitness and queen acceptance. We used liquid chromatography-tandem mass spectrometry to measure queen retinue pheromone (QRP) components in relation to age, laying status, and likelihood of acceptance using a protocol that enables QRP to be measured concurrently with metabolomic and lipidomic analyses. Older queens ( >1 month) were more readily accepted by colonies than younger queens (10-12 d), regardless of the queen’s prior laying status, despite non-laying queens having significantly smaller ovaries at the time of introduction. Older queens produced higher levels of the QRP components 9(R)-HDA, LEA, and HVA compared to younger queens, with HVA also positively correlating with ovary mass. However, ovary mass is not an influential fertility metric for worker decision-making; therefore, the relationship between HVA and ovary mass is merely an honest signal of a non-influential metric. Metabolomic and lipidomic analyses showed that samples cluster strongly according to queen age and mating status, but not ovary mass. These data reveal some first indications of the importance of prostanoids in queen maturation and many other physiological changes occurring in the first month of a queen’s life.