Prolonged periods of standing and walking creates loading on the muscles of the back and legs causing pain and discomfort to many employees who stand on hard surfaces such as cement floors. Portable anti-fatigue mats are an ergonomic intervention that is thought to be a solution for musculoskeletal injuries. Thirteen healthy subjects ranging from 18-55 years of age were recruited for the study to represent a working population. Subject completed two 3 hour sessions, one with work boots, and the other with the combination of work boots and portable anti-fatigue mats. Subjects walked for a 3 hour period to simulate a prolonged walking period. Participants were instrumented with reflective marker to collect kinematic data and walked on the force plate to collect kinetic forces acting on the body. They also completed a discomfort questionnaire for “pre”, “during” and “post” exercise. Results showed no significant differences in mechanical variables, and showed significant differences in discomfort ratings for the ankle/foot and knee between conditions;​ while showing significant differences for the ankle/foot and low back over time.