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Active OHO (ordinary heterotrophic organism) plays a key role in the structure of steady state design models and kinetic simulation models with respect to organic removal and denitrification. However, the concentration of OHO is not determined clearly in mathematical models because it can not be measured accurately with engineering or microbiological techniques. Thus, above all, three quantifying methods in the view of engineering were developed and applied in this research. To quantify active OHO, both the 10-day and 20-day SRT Modified Ludzack-Ettinger (MLE) activated sludge systems were operated and provided their mixed liquor to perform modified batch test and batch aerobic digestion test. As measurement techniques, (1) steady state design method, (2) modified batch test and (3) batch aerobic digestion test were applied and examined. In this paper, application of mathematical models for measuring techniques is mainly examined. By fitting the experimental results into mathematical model, the concept of the quantifying techniques could be evaluated. The conclusion of this research, through comparing the experimental results with the estimated values from the models, are as followings; the results of (1) steady stated design method and (3) batch aerobic digestion were considerably stable. However, the results of (2) modified batch test were rather fluctuated. Thus, in the following experiment, the comparison between the three methods and the abnormal behaviors of microorganisms in modified batch test condition need to be deeply examined.