The examination of the moment exerted by the hamstrings during maximum isokinetic knee extensor tests is useful when comparing isokinetic strength and muscle activity patterns between children and adults. The purpose of this study was to examine the effect of antagonist moment of the hamstrings on the isokinetic moment of the knee extensors in pubertal children and to determine whether this effect is altered following a fatigue task. Eighteen healthy pubertal males [age 14.3 (0.5) years] performed 34 maximal isokinetic concentric efforts of the knee extensors at 60 degrees.s(-1). The average moment of force and electromyographic (aEMG) signal of vastus medialis (VM), vastus lateralis (VL) and biceps femoris (BF) at 11-30 degrees, 31-50 degrees, 51-70 degrees and 71-90 degrees of knee flexion were calculated for each repetition. The hamstrings antagonist moment was determined before and after the fatigue task by fitting the aEMG-moment relationship at different levels of muscle effort using second-degree polynomials. The percentage contribution of the antagonist moment to the resultant joint moment ranged from 7.1 % to 60.4 % throughout the range of motion, with the highest percentage observed close to full knee extension (11-30 degrees). The antagonist effect was significantly greater during concentric tests of the knee extensors compared to the corresponding eccentric tests ( p<0.05). Following the fatigue test, there was an overall decline of the resultant joint moment, but no changes in the predicted hamstrings moment were observed. These results indicate that when testing maximal knee extensor isokinetic strength in pubertal boys, activity of the hamstrings implies a reduction of the net extensor moment as compared to the isolated capacity of the knee extensors. However, this antagonist effect is not altered following the performance of an isokinetic fatigue knee extension task.