Assessment of physiological state of microorganisms in activated sludge with flow cytometry: application for monitoring sludge production minimization

Many sludge reduction processes have been studied for the minimization of sludge production in biological wastewater treatment. The investigations on most of these processes have monitored the increase of the soluble chemical oxygen demand, the sludge mass reduction, or the decrease of the floc size, but little information has been obtained on cell lysis and the change of the biological cell activity. However, employing any strategy for reducing sludge production may have an impact of microbial community in biological wastewater treatment process. This impact may influence the sludge characteristics and the quality of effluent. The objective of this study concerns the determination of the physiological state of activated sludge microorganisms during a sludge minimization process. A thermal treatment at 80 degrees C for 5, 20, 40 and 60 min was chosen in this study. Staining bacteria with CTC and SYTOX green was used to evaluate biological cell activity and viability of cell types contained in activated sludge, respectively. The monitoring of cell activity and viability was performed using flow cytometry (FCM) analysis before and after thermal treatment of activated sludge. Results indicated an increase in the number of permeabilized cells and a decrease in the number of active cells, subsequent to the thermal treatment. The study also confirms the potential of FCM to successfully evaluate the physiological heterogeneity of an activated sludge bacterial population. Moreover, the experimentally observed correlations between the FCM results and the organic matter solubilization in activated sludge samples during thermal treatment revealed that the increase in the soluble organic matter concentration was predominantly due to an intracellular material release. Identifying the increase in activated sludge hydrolysis requires a precise knowledge of the involved mechanisms, and this study indicated that the FCM, used in conjunction with specific probes, could be a useful tool.