Diet in the winter has a vital effect on the survival and condition of a honey bee (Apis mellifera) colony in the spring. The effect of supplementation of the diet with vitamin C (ascorbic acid) on the total antioxidant status (TAS), glutathione content, and activity of 4 antioxidative enzymes: superoxide dismutase (SOD), peroxidase (POX), catalase (CAT), and glutathione transferase (GST) of honey bee brood developing in the spring was studied. Twelve stages, from newly hatched larvae to emerging adult worker bees were studied, allowing changes in the antioxidant profile during brood development to be determined for the first time. It was shown that bees are more exposed to oxidative stress after emergence. In workers emerging in colonies after supplementation with vitamin C, higher contents of protein and glutathione, and higher activities of peroxidase, catalase, and glutathione transferase were observed. Vitamin C did not alter brood weight increase, and the level of protein in emerged workers was higher than in the control group. The mean of bee losses over winter were about 33% lower in colonies receiving vitamin C.