The objective of the present study was to investigate the impact of muscle length during stretch-shortening cycles on static and dynamic muscle performance. Animals were randomly assigned to either an isometric group (CON, N = 12), short muscle length group (S-Inj, 1.22 to 2.09 radians, N = 12), or long muscle length group (L-Inj, 1.57 to 2.44 radians, N = 12). The dorsi flexor muscles were exposed in vivo to either 7 sets of 10 stretch-shortening contractions (conducted at 8.72 radians/s) or 7 sets of isometric contractions of the same stimulation duration using a custom-designed dynamometer. Performance was characterized by multi-positional isometric exertions and positive, negative, and net work, at pre-test, 6 hours post, and 48 hours post-exposure. Real-time muscle performance during the stretch-shortening cycles was characterized by stretch-shortening parameters and negative, positive, and net work. The S-Inj group recovery (force difference) was similar to CON force differences at 48 hours, while the L-Inj group was statistically greater at 1.39, 1.57, and 1.74 radians than CON (p < 0.05). Negative work of the S-Inj and L-Inj groups (p <0.05), and net work (p <0.05) were statistically lower from CON at 48 hours post-exposure. Of the real-time parameters, there was a difference in cyclic force with treatment during the stretch-shortening cycles (p < 0.0001) with the L-Inj group being the most affected. Thus, longer ranges of motion result in a more profound isometric force decrement 48 hours post exposure, and in real-time changes in eccentric forces.