The purpose of this study was to determine the effects of vibration (VIB) on motor unit (MU) behavior of the vastus lateralis (VL) muscle during a 40% maximal voluntary contraction (MVC). Eleven healthy (age 21.3 ± 2.6 years) individuals participated in the study. Surface electromyography (EMG) data were recorded from the VL during isometric trapezoidal muscle contractions at 40% MVC. Firing events of single MUs and EMG amplitude were reported for the first, middle, and final seconds of a 12-second steady force segment at 40% MVC. VIB was applied at 55 Hz to the patellar tendon for 15 minutes before and continued throughout the remainder of testing (VIB) or remained off (CON). There were significant increases in MU firing rates during VIB in comparison to CON and no differences in EMG amplitude between VIB and CON. The VIB-mediated reduction in muscle spindle function altered MU behavior at 40% MVC. Muscle Nerve 55: 500-507, 2017.