Spinal musculature plays a major role in spine stability, but its importance to spinal stiffness is poorly understood. We studied the effects of graded trunk muscle stimulation on the in vivo dynamic dorsoventral (DV) lumbar spine stiffness of 15 adolescent Merino sheep. Constant voltage supramaximal electrical stimulation was administered to the L3-L4 interspinous space of the multifidus muscles using four stimulation frequencies (2.5, 5, 10, and 20 Hz). Dynamic stiffness was quantified at rest and during muscle stimulation using a computer-controlled testing apparatus that applied variable frequency (0.46-19.7 Hz) oscillatory DV forces (13-N preload to 48-N peak) to the L3 spinous process of the prone-lying sheep. Five mechanical excitation trials were randomly performed, including four muscle stimulation trials and an unstimulated or resting trial. The secant stiffness (k (y) = DV force/L3 displacement, kN/m) and loss angle (phase angle, deg) were determined at 44 discrete mechanical excitation frequencies. Results indicated that the dynamic stiffness varied 3.7-fold over the range of mechanical excitation frequencies examined (minimum resting k (y) = 3.86 +/- 0.38 N/mm at 4.0 Hz; maximum k (y) = 14.1 +/- 9.95 N/mm at 19.7 Hz). Twenty hertz muscle stimulation resulted in a sustained supramaximal contraction that significantly (P < 0.05) increased k (y) up to twofold compared to rest (mechanical excitation at 3.6 Hz). Compared to rest, k (y) during the 20 Hz muscle stimulation was significantly increased for 34 of 44 mechanical excitation frequencies (mean increase = 55.1%, P < 0.05), but was most marked between 2.55 and 4.91 Hz (mean increase = 87.5%, P < 0.05). For lower frequency, sub-maximal muscle stimulation, there was a graded change in k (y), which was significantly increased for 32/44 mechanical excitation frequencies (mean increase = 40.4%, 10 Hz stimulus), 23/44 mechanical excitation frequencies (mean increase = 10.5%, 5 Hz stimulus), and 11/44 mechanical excitation frequencies (mean increase = 4.16%, 2.5 Hz stimulus) when compared to rest. These results indicate that the dynamic mechanical behavior of the ovine spine is modulated by muscle stimulation, and suggests that muscle contraction plays an important role in stabilizing the lumbar spine.