Jesus Ortiz, BD; Felipe Rosa, BD; Imilce Rodriguez, PhD; Tugrul Giray, PhD

Introduction: Aging is a natural result of the accumulation of a wide variety of molecular and cellular damage over time, allowing a growing risk of disease and death. Intestinal permeability, passage of materials from gut lumen into circulation due to damage to epithelial cover of the gut is a good indicator of aging. The Smurf Assay (SA), first developed in the fly model, Drosophila melanogaster, allows an in vivo assessment of intestinal permeability based on food intake by the co-ingestion of a non-toxic blue dye (FD&C blue dye #1). Blue dye stays in the digestive tract in healthy individuals. The dye that enters the body therefore allows a direct quantification of gut health as a continuous phenotype. Here we report the development of the SA to be used in honey bees (Apis mellifera). SA could be used in honey bees if blue dye crosses the intestinal barrier of the aging bee, and bluish coloration will be obtained throughout the body. To our knowledge, this is the first study adapting SA in honey bees.

Methods: To address the main research question, firstly, the blue dye was evaluated for toxicity in forager bees at different concentration percentages, 0% (control), 0.1%, 0.5% & 2% blue dye, in 50% sucrose. With a total of 40-50 bees/box, for a total of 8 days, the number of dead bees per day was counted, and the count of Smurfs exhibiting this particular phenotype was recorded.

Results: Our results demonstrate that mortality does not increase with dye concentration, and it is not different across control vs treatment bees and is uniform across different concentrations. The emergence of Smurf bees commences after 24 hours (<1/100 bees). Over the 8 days, the number of deceased bees gradually increases, yet more than 50% remain alive within the cages. We report Smurf phenotype in dead and alive bees at the end of 8 days.

Conclusion: The future directions include SA applications to study adult development, gut health and nutritional quality, and gut health and cognitive performance in honey bees.

Acknowledgments: PR-LSAMP, Dr. Jose Agosto Rivera, Fanfan Noel & Yilmaz Berk Koru.