Here, we present the results of a 2-year field trial aimed at testing the effect of overwintering on different feeds on the course of Nosema ceranae infection. In August 2015, four experimental bee colony groups were established. After the last honey harvest, each colony was provided with 20 kg of feed, either honey, sugar (3:2 solution in tap water), inverted syrup made of sucrose, or wheat starch syrup. Samples of live bees were collected from each beehive in August (before feeding), November, and May. The following year, feeding and sampling were performed in the same way. Bees were examined microscopically to estimate the percentage of Nosema-infected individuals in the sample and the spore number per bee. Fitness parameters were also measured in all colonies. In all hives, presence of N. ceranae was confirmed through polymerase chain reaction. Nosema apis was not detected in the apiary. Significant differences in nosematosis prevalence and/or intensity were observed between the experimental groups. For most parameters, best results were recorded in the group fed with honey. Worst fitness and highest nosematosis prevalence and intensity were found in colonies fed with wheat starch syrup.