There were few attempts about the requirements of honeybee to vitamins. Our results highlighted the effectiveness of Phenoloxidase (PO) and Antioxidant (AO) activity in honeybee workers and vitamin types added with sugar solution as early and vitamins important indicators. There was a clear improvement in the antioxidant system in bee colonies that were provided with sugary feeding with vitamins mixture compared to other vitamins individually (A, B, C and E) and control ones. This suggests that honeybee workers need artificial feeding with mixed vitamins during winter season possess mechanisms that reduce their oxidative stress. Results suggest that vitamins in winter feeding for 14 weeks (from 1 December to 14 March) has the potential to improve colony development and health during overwintering, a period of high colony losses. The results showed that vitamins diet increased bee activity and colony health escpicially in the dearth period, which reduce varroa infection and caused the absence of European foulbrood compared to the control.