Stereological estimation of structural change in floc matrix following weak ultrasonication

Abstract Ultrasonication is an energy intensive process used to enhance organics’ solubilisation and used in promoting methane yield from wastewater sludge. Applying limited ultrasound without full disruption of floc structure or cell walls can significantly enhance methane yield from sludge at reduced energy cost. This work probed the spatial distribution of extracellular polymeric substances, cells in the original wastewater flocs, and the yielded fragments following weak ultrasonication using a process of multiple staining and a confocal laser scanning microscope technique. The stereological information of the three-dimensional architecture of the original wastewater floc and ultrasonicated fragments was extracted and compared. The local structural changes following weak ultrasonication were presented for the first time, which should also correlate with the enhanced methanogenesis thereby produced.