In the experiment, we tested the effect of 30% CBD oil on the activity of the antioxidant system (superoxide dismutase, catalase, glutathione peroxidase, glutathione), the level of total antioxidant capacity, and the concentrations of ions (calcium, magnesium, and phosphorus) in honeybee workers in the hive test. For this purpose, we prepared hives containing all stages of the development of honey bees and started the experiment by adding 200 marked, one-day old bees to each colony (intended for hemolymph collection). In the test, we created three groups (two colonies per group): (1) Experimental with CBD oil mixed with sugar syrup (CSy); (2) experimental with CBD oil on textile strips (CSt); and (3) control with pure sugar syrup only (C). Every week, we collected hemolymph from the marked bees. In the experiment, all antioxidant enzyme activities were higher for the experimental groups CSy and CSt compared to group C. The highest concentrations/levels were obtained for the CSy group. Concentrations of calcium, magnesium, and phosphorus ions were also higher for the experimental groups compared to the C group (the highest concentration for the CSy group). We conclude that CBD oil positively contributes to stimulating the antioxidant system of honeybees.