Vertical transmission pathway of viruses in honey bee colonies were investigated by molecular methods. The virus status of individual queens was evaluated by examining the presence of viruses in the queens’ feces and tissues including hemolymph, gut, ovaries, spermatheca, head, and eviscerated body. Except for head tissue, all five tissues as well as queen feces were found to be positive for virus infections. When queens in bee colonies were identified to be positive for certain viruses, the same viruses were detected in their offspring including eggs, larvae, and adult workers. The presence of viruses in the tissue of ovaries and the detection of the same viruses in eggs and young larvae suggest a vertical transmission of viruses from queens to offspring. To our knowledge, this is the first evidence of vertical transmission of viruses in honey bee colonies.