Studying the influence of defect on the dynamic behavior of wood in order to detect the local heterogeneities is of great importance in non-destructive testing of wood. The natural heterogeneities in wood are oriented in a volume. However, one- dimensional models are still used in dynamic characterization of wooden beams. The aim of this study was to experimentally investigate the effects of the orientation and position of an artificial defect on the flexural vibration frequencies. Different batches of Fagus orientalis specimens were drilled in the radial direction at five positions along the specimen. Dynamic tests in free flexural vibration were performed on the specimens before and after drilling both in the longitudinal-radial (LR) and longitudinal-tangential (LT) bending plan. The behavior in free flexural vibration was found to be different depending on the position and orientation of heterogeneity. When the drilling axis lies in the bending plane (LR), the weakening of frequency was maximal at the location of an antinode of vibration. On the contrary, the frequency offset was maximal in the place of a vibration node when the drilling axis was orthogonal to the bending plane (L T).