Using multi-parameter flow cytometry as a novel approach for physiological characterization of bacteria in microbial fuel cells

This work demonstrates, for the first time, the potential of using multi-parameter flow cytometry to monitor changes in the microbial cytoplasmic membrane integrity and polarization during microbial fuel cells (MFC) operation. Such information is crucial to follow the dynamics of bacteria colonization of the electrodes and their viability maintenance during electrical current production. Interestingly, the results show that during voltage production, the electrostatic gradients of the bacteria cytoplasmic membrane are disturbed, leading to depolarization of a subpopulation (where less than 40% of the cells were polarized). Once the voltage dropped, due to substrate limitation, several cells in the anode supernatant restored their polarized state. This process was reversible and observed over more than 4 cycles of fresh substrate addition. Similar power outputs induced similar membrane polarization results, regardless of the substrate used. The percentage of non-viable cells was maintained constant during current production. This study opens new opportunities to monitor cell behavior, and thus increase the knowledge of dynamic mechanisms responsible for current production at the individual cell level. This technique could be of great interest for the development of new MFC configurations and optimization of MFC operation conditions toward increased performance.

► Flow cytometry allows to characterize the physiological dynamics of bacteria in MFCs. ► The percentage of anode non-viable cells is maintained constant during each MFC fed batch cycle. ► The MFC environmental conditions disturb cell polarization of the membrane. ► During voltage production the percentage of polarized cells decreased to values between 40 and 20%. ► Similar power outputs induces similar cell physiological changes, regardless the substrate used.