Activated sludge process is a widely used method to treat domestic and industrial effluents. The conditions of activated sludge wastewater treatment plant (AS-WWTP) are related to the morphological properties of flocs (microbial aggregates) and filaments, and are required to be monitored for normal operation of the plant. Image processing and analysis is a potential time-efficient monitoring tool for AS-WWTPs. Local adaptive segmentation algorithms are proposed for bright-field microscopic images of activated sludge flocs. Two basic modules are suggested for Otsu thresholding-based local adaptive algorithms with irregular illumination compensation. The performance of the algorithms has been compared with state-of-the-art local adaptive algorithms of Sauvola, Bradley, Feng, and c-mean. The comparisons are done using a number of region- and nonregion-based metrics at different microscopic magnifications and quantification of flocs. The performance metrics show that the proposed algorithms performed better and, in some cases, were comparable to the state-of the-art algorithms. The performance metrics were also assessed subjectively for their suitability for segmentations of activated sludge images. The region-based metrics such as false negative ratio, sensitivity, and negative predictive value gave inconsistent results as compared to other segmentation assessment metrics.