A noninvasive three-dimensional (3D) cardiac electrical imaging (3DCEI) approach, which can estimate the location of the initiation site (IS) of activation and the resultant 3D activation sequence (AS) from body surface potential maps (BSPMs), was validated in an intact large mammalian model (swine) during acute ventricular pacing. Body surface potential mapping and intracavitary noncontact mapping (NCM) were performed simultaneously during pacing from both right ventricular (RV) sites (intramural) and left ventricular (LV) sites (endocardial). Subsequent 3DCEI analyses were performed on the measured BSPMs. In total, 5 RV and 5 LV sites from control and heart failure animals were paced. The averaged localization error of the RV and LV sites were 7.0+/-1.1 mm and 6.6+/-1.9 mm, respectively. The endocardial ASs as a subset of the estimated 3D ASs by 3DCEI were consistent with those reconstructed from the NCM system. The present experimental results demonstrate that the noninvasive 3DCEI approach can localize the initiation site and estimate cardiac activation sequence with good accuracy in an in vivo setting, under control, paced and/or diseased conditions.