The objective of the study was to explore the impact of different operation strategies on the performance of the Oxic-Settling-Anaerobic (OSA) process. A sequencing batch reactor was selected as the aerobic (oxic) unit of the OSA system and it was operated as a dual mixing phase with step feeding in order to optimize simultaneous nitrogen removal and sludge minimization. For this purpose, the effect of COD/N ratio, filling pattern and the fraction of anaerobic period was investigated on the performance of a laboratory-scale OSA system fed with synthetic wastewater (peptone mixture) approximating the characteristics of sewage. In nine consecutive experimental stages, each sustaining different metabolic processes and biochemical reactions, the sludge reduction potential of the OSA system was not impaired, maintaining an average observed yield value of 0.18 g VSS/g COD, which was approximately half the level observed in the reference reactor. Similarly, the OSA scheme of operation did not affect or alter the fundamental mechanisms of biological nitrogen removal, where system performance could be modified and optimized the same way as in a conventional activated sludge configuration. Furthermore, the OSA system maintained a higher level of biomass concentration compared with the reference reactor. This observation confirmed the previous analyses that the continuous biomass inflow from the side stream anaerobic bioreactor was the reason to establish the microbial mechanism that resulted in a much lower sludge generation, regardless of the biochemical processes taking place in the reactor.