Oxygen intrusion into the anode chamber through proton exchange membrane can result in positive redox conditions in fed-batch, two chamber MFCs at the end of a cycle when the substrate is depleted. A slight increase in dissolved oxygen to 0.3 mg/L during MFC operation was not found to adversely affect power generation over subsequent cycles if sufficient substrate (acetate) was provided. Purging the anode chamber with air or pure oxygen for up to 10 days and 10 hrs also did not affect power generation, as power rapidly returned to previous levels when the chamber was sparged with nitrogen gas. When MFCs are connected in series, voltage reversal can occur resulting in a positive voltage applied to the anode biofilm. To investigate if this adversely affected the bacteria, voltages of 1, 2, 3, 4, and 9 V, were applied for 1 hr to the MFC before reconnecting it back to a fixed external load (1,000 Omega). A voltage of <2 V did not affect power generation. However, applying 3 V resulted in a 15 h lag phase before recovery, and 9 V produced a 60 h lag phase suggesting substantial damage to the bacteria that required re-growth of bacteria in the biofilm. These results indicate that charge reversal will be a more serious problem than oxygen intrusion into the anode chamber for sustained performance of MFCs.