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Prolonged sitting may involve several mechanisms that make it a risk factor for low back pain. The aim of this study was to investigate lumbar-pelvic kinematics and multifidus muscle (MF) activity during squatting and forward bending in relation to the sedentary behaviour of physically active students. Sixty-three students were divided into two groups according to the time spent in a sitting position during the day: ‘high’ (>7 h/day); ‘low’ (≤7 h/day). Lumbar-pelvic ratios, ranges of motion, angular velocities, and MF flexion-relaxation phenomenon were investigated. Data were obtained using the optical motion analysis system, and surface electromyography. The results indicated that lumbar-pelvic ratios during both tasks and velocity of lumbar spine during squatting were significantly greater in the ‘high’ than in the ‘low’ sitting group. Muscle activity showed no differences between groups. Prolonged sitting can be considered a factor that slightly, but statistically significantly influences the lumbar-pelvic kinematics in physically active people.