The pesticide imidacloprid and the heavy metal copper provide some degree of protection to plants, while at the same time causing varying degrees of damage to bees. However, few studies have investigated the negative effects of imidacloprid and copper exposure on newly emerged bees (young bees), especially when both are present in a mix. In this study, young bees were exposed to sterile sucrose solutions containing imidacloprid (10 μg/L, 100 μg/L), copper (10 mg/L, 50 mg/L), or a mix of both (10 μg/L + 10 mg/L) for 5 days to assess their gut system and behavior, with survival and dietary consumption recorded over 21 days. We found that imidacloprid and copper reduced honeybee survival, dietary intake, and learning ability, decreased gut microbiota diversity, and caused metabolic disruptions. Notably, the mix of imidacloprid and copper had a synergistic negative effect. Correlation analyses revealed that the honeybee gut microbiota influences bee immunity and behavior by regulating metabolic pathways related to ascorbate, tryptophan, and carbohydrates. Our results demonstrate that imidacloprid and copper, either alone or in a mix, alter young bee health through a complex mechanism of toxicity. These findings highlight imidacloprid and copper's negative effects on young honeybees, offering insights for future pesticide and heavy metal impact research.Copyright © 2025. Published by Elsevier Inc.