The vibration response of the bridges under the moving vehicular load is of importance for engineers to estimate the serviceability of existing bridges and to design new bridges. This paper deals with the three dimensional vibration analysis of prestressed concrete bridges under moving vehicles. The prestressed bridges are modeled by four-node isoparametric flat shell elements with the transverse shearing deformation taken into account. The usual five degrees-of-freedom (DOFs) per node of the element are appended with a drilling DOF to accommodate the transformation of the local stiffness and mass matrices to the global coordinates. The vehicle is modeled as a single or two-DOF system. A single-span prestressed Tee beam and two-span prestressed box-girder bridge are studied as the two numerical examples. The effects of prestress forces on the natural frequencies and dynamic responses of the bridges are investigated.