Molecular Assessment of Genes Linked to Honeybee Health Fed with Different Diets in Nuclear Colonies

Honeybees are of economic importance not only for honey production, but also for crop pollination, which amounts to USD 20 billion per year in the United States. However, the number of honeybee colonies has declined more than 40% during the last few decades. Although this decline is attributed to a combination of factors (parasites, diseases, pesticides, and nutrition), unlike other factors, the effect of nutrition on honeybee health is not well documented. In this study, we assessed the differential expression of seven genes linked to honeybee health under three different diets. These included immune function genes [Cactus, immune deficiency (IMD), Spaetzle)], genes involved in nutrition, cellular defense, longevity, and behavior (Vitellogenin, Malvolio), a gene involved in energy metabolism (Maltase), and a gene associated with locomotory behavior (Single-minded). The diets included (a) commercial pollen patties and sugar syrup, (b) monofloral (anise hyssop), and (c) polyfloral (marigold, anise hyssop, sweet alyssum, and basil). Over the 2.7-month experimental periods, adult bees in controls fed pollen patties and sugar syrup showed upregulated Cactus (involved in Toll pathway) and IMD (signaling pathway controls antibacterial defense) expression, while their counterparts fed monofloral and polyfloral diets downregulated the expression of these genes. Unlike Cactus and IMD, the gene expression profile of Spaetzle (involved in Toll pathway) did not differ across treatments during the experimental period except that it was significantly downregulated on day 63 and day 84 in bees fed polyfloral diets. The Vitellogenin gene indicated that monofloral and polyfloral diets significantly upregulated this gene and enhanced lifespan, foraging behavior, and immunity in adult bees fed with monofloral diets. The expression of Malvolio (involved in sucrose responsiveness and foraging behavior) was upregulated when food reserves (pollen and nectar) were limited in adult bees fed polyfloral diets. Adult bees fed with monofloral diets significantly upregulated the expression of Maltase (involved in energy metabolisms) compared to their counterparts in control diets to the end of the experimental period. Single-Minded Homolog 2 (involved in locomotory behavior) was also upregulated in adult bees fed pollen patties and sugar syrup compared to their counterparts fed monofloral and polyfloral diets. Thus, the food source significantly affected honeybee health and triggered an up- and downregulation of these genes, which correlated with the health and activities of the honeybee colonies. Overall, we found that the companion crops (monofloral and polyfloral) provided higher nutritional benefits to enhance honeybee health than the pollen patty and sugar syrup used currently by beekeepers. Furthermore, while it has been reported that bees require pollen from diverse sources to maintain a healthy physiology and hive, our data on nuclear colonies indicated that a single-species diet (such as anise hyssop) is nutritionally adequate and better or comparable to polyfloral diets. To the best of our knowledge, this is the first report indicating better nutritional benefits from monofloral diets (anise hyssop) over polyfloral diets for honeybee colonies (nucs) in semi-large-scale experimental runs. Thus, we recommend that the landscape of any apiary include highly nutritious food sources, such as anise hyssop, throughout the season to enhance honeybee health.