Overuse and overtraining models have implicated both metabolic and mechanical disturbances as contributors to muscle damage and performance decrement but have produced equivocal results. The purpose of the present study was to investigate the impact of rest interval between sets of stretch-shortening cycles (SSC) on static and dynamic muscle performance. Animals were randomly assigned to groups (N = 8 per group) of 10-s, 1-min, or 5-min rest between sets of isometric contractions (10-s, 1-min, or 5-min CON), or SSC (10-s, 1-min, or 5-min INJ). The dorsiflexor muscles were exposed in vivo to either seven sets of 10 SSC (500 degrees . s) or seven sets of isometric contractions. Performance was characterized by isometric exertions and positive, negative, and net work, at pretest, during the sets of SSC, and 48 h postexposure. The isometric force at 48 h after the 10-s and 5-min INJ groups were statistically different from the 1-min group (P < 0.05), whereas there was no difference in the CON groups. Negative work of the INJ groups were statistically lower at 48 h than pretest values (P < 0.05), whereas there was no change in positive work. Of the real-time parameters, there was a difference in minimum force and positive work (P < 0.05) with treatment with the 10-s INJ group being most affected. SSC conducted at shorter work-rest cycles resulted in a more profound isometric force decrement 48 h postexposure, and in real-time changes in isometric prestretch force and positive work. These results indicate that short rest intervals between athletic or vocational tasks of heightened physical exertion (i.e., high intensity) may adversely affect performance and increase injury susceptibility.