Aging is a multifactorial process that occurs in all living organisms, including bees. One of the factors accelerating this process is stress caused in bees by Varroa destructor. The research aim was to compare antioxidant system activities in different tissues and in different fat body segments (sternite, tergite 3 and 5) in workers aging naturally (physiologically) and prematurely (affected by V. destructor). The CAT, GPx, GST, and SOD activities in naturally aging workers were higher in all the tissues/fat body segments and age groups compared to prematurely aging workers. These antioxidant activities increased with age, reaching a maximum at 21 (in tergite 3 and sternite) or 28 days of age (in the hemolymph and tergite 5) in naturally aging workers, and then decreased in the oldest ones (at 35 days of age). In the prematurely aging workers, the antioxidant activities in the fat body decreased along with age. The highest activities were identified in the fat body of tergite 5, which may suggest its role in detoxification processes. Our results are a starting point for a better understanding of the mechanisms related to oxidative stress, aging, and their correlation with the health and lifespan of bees.