Abstract
Honey bees (Apis mellifera L.) play vital roles as agricultural pollinators and honey producers. However, global colony losses are increasing due to multiple stressors, including malnutrition. Our study evaluated the effects of four pollen substitute diets (Diet 1, Diet 2, Diet 3, and Control) through field and cage experiments, analyzing 11 parameters and 21 amino acids. Notably, Diet 1 demonstrated significantly superior performance in the field experiment, including the number of honey bees, brood area, consumption, preference, colony weight, and honey production. In the cage experiment, Diet 1 also showed superior performance in dried head and thorax weight and vitellogenin (vg) gene expression levels. Canonical discriminant and principle component analyses highlighted Diet 1's distinctiveness, with histidine, diet digestibility, consumption, vg gene expression levels, and isoleucine identified as key factors. Arginine showed significant correlations with a wide range of parameters, including the number of honey bees, brood area, and consumption, with Diet 1 exhibiting higher levels. Diet 1, containing apple juice, soytide, and Chlorella as additive components, outperformed the other diets, suggesting an enhanced formulation for pollen substitute diets. These findings hold promise for the development of more effective diets, potentially contributing to honey bee health.