The electromyographic activity (EMG) of the anterior temporal (AT), masseter (M), trapezius (T) muscles and anterior aspect of the digastric (D) was measured in 50 subjects, during six seconds of maximum contraction, bilaterally with and without unilateral premature contacts and individually for each tooth. Special occlusal interferences were designed to assess muscular activity. Muscular activity was measured simultaneously by placing premature contacts on each tooth, under T-Scan monitoring. Premature contacts reduced EMG activity during maximum contraction of the AT, D and M muscles, the highest disruption is in the AT muscle, at the level of upper right 2nd molar, with a 56% reduction in activity. Conversely, there was an increase of activity of the T muscle in all teeth when placing artificial occlusal premature contacts, with the highest difference in the upper right 1st bicuspid. Therefore, occlusal interferences can cause neuromuscular disruptions, thus inducing important muscular discrepancy. Both the EMG and T-Scan monitoring can be considered suitable methods to use in daily dental practice to identify premature contacts and to measure EMG activity.