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There is evidence that movement-based microbreaks can improve the cardiovascular health of desk-based employees, but their effect on mood states is yet to be investigated. As daily work tasks can potentially result in the loss of physical and psychological resources, the objective of this study was to measure the effect of movement microbreaks during formal work time on mood states. In a randomized-controlled pilot study with repeated measures (baseline, post-test, washout) of self-reported job stress and mood states (fatigue and vigor), police officers (N = 43) were exposed to movement microbreaks during work hours. A multivariate significant difference between groups was noted after the intervention period. Further analysis revealed that the experimental group reported a latent reduction in job-related stress after the 3-months washout period. Although the study was conducted with a small sample, our preliminary findings suggest that interrupting sedentary work with movement microbreaks may have beneficial effects on employee mental health. The implications of movement microbreaks for mitigating work-related stress of first responders, including police, is discussed, along with directives for future research.