Abstract Background: In two experiments, two groups (pilots and non-pilots) were compared to assess the effect of pre-learning and career on the accuracy of distance estimation in virtual environment. Materials and methods: Ten pilots and ten non-pilots were involved in this study. In the first experiment, the distance estimation error of two groups was assessed when looking from up to down versus looking from down to up. In the second experiment, distance estimation error was evaluated in photopic versus mesopic lighting conditions. All measures obtained in three distances: 2.8 m, 10.35 m and 14.39m. Results: In up to down looking, pilots group had lower errors than non-pilots .The value of error estimation was significantly higher in mesopic lighting conditions compere to photopic conditions. Conclusion: In general, the distances estimation in virtual environment had no significant differences between pilots and non-pilots.