** AWARDS ISSUED PRIOR TO JANUARY 20, 2025, WERE FUNDED UNDER PREVIOUS ADMINISTRATIONS AND MAY NOT REFLECT THE PRIORITIES AND POLICIES OF THE CURRENT ADMINISTRATION.** THIS PROPOSAL ADDRESSES AFRI'S PROGRAM PRIORITY A113: POLLINATOR HEALTH: RESEARCH AND APPLICATION. POOR NUTRITION AND LANDSCAPE CHANGES ARE TWO TOP FACTORS RESPONSIBLE FOR CAUSINGLOSSES OF OVER 40% OF MANAGED HONEY BEES (APIS MELLIFERA) IN THE U.S. ANNUALLY. HOWEVER, WHAT CONSTITUTES POOR NUTRITION FOR HONEY BEES IS CURRENTLY INADEQUATELY DEFINED. ADDITIONALLY, THE QUALITY AND QUANTITY OF NUTRITIONAL RESOURCES AVAILABLE TO HONEY BEES ACROSS DIFFERENT LANDSCAPES IS POORLY CHARACTERIZED. THE SHORTCOMINGS IMPEDE THE DEVELOPMENT OF SATISFACTORY FOOD SUPPLEMENTS. OUR OVERARCHING HYPOTHESIS IS THAT HONEY BEES TIGHTLY REGULATE THEIR INTAKE OF MULTIPLE NUTRIENTS USING A TWO-LEVEL PROCESS. FIRST, FORAGERS SELECTIVELY COLLECT POLLEN BASED ON ITS NUTRITIONAL CONTENT. NEXT, NURSE BEES SELECTIVELY FEED ON STORED POLLEN (BEE BREAD) TO BALANCE THEIR NUTRIENT INTAKE, WHICH OPTIMIZES THEIR PERFORMANCE. WE PREDICT THAT THE NUTRITIONAL CONTENT OF POLLEN VARIES ACROSS LANDSCAPES AND SEASONS, BUT BOTHFORAGERS AND NURSE BEES CAN ASSESS THESE CHANGES AND RESPOND APPROPRIATELY. WE HAVE THREE OBJECTIVES. (1) PERFORM A MULTIDIMENSIONAL NUTRIENT ANALYSIS OF POLLEN TO CHARACTERIZE THE NUTRITIONAL SPACE AVAILABLE FOR HONEY BEES IN THREE DIFFERENT LANDSCAPES (AGRICULTURAL, URBAN, AND RURAL) ACROSS SEASONS. (2) PERFORM A MULTIDIMENSIONAL NUTRIENT ANALYSIS OF BEE BREAD TO UNDERSTAND THE ROLE OF PRE-DIGESTIVE POLLEN PROCESSING. THIS WILL BRIDGE THE GAP BETWEEN NUTRITIONAL INPUTS (POLLEN) AND THE DIET AVAILABLE TO NURSES (BEE BREAD). (3) CHARACTERIZE THE LINK BETWEEN BEE BREAD FATTY ACID COMPOSITION, NURSE BEE FEEDING BEHAVIOR, AND COLONY-LEVEL PERFORMANCE. THE DATA GENERATED FROM EVERY OBJECTIVE WILL INFORM BEEKEEPERS ON PRACTICES THEY CAN CHANGE/IMPLEMENT TO IMPROVE COLONY HEALTH.