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Conventional microbial desalination cells (MDCs) can extract organic energy from wastewater for in situ utilization in saline water desalination, but are mostly unable to achieve enhanced treatment and desalination for one stream of wastewater. In this study, a multi-stage MDC (M-MDC) with two alternating anodes and cathodes was fabricated, and operated with domestic wastewater in two operational modes. In ACAC operational mode, with wastewater flowing serially from anode-1 → cathode-1 → anode-2 → cathode-2, the M-MDC realized current production of 11.4 mA and desalination efficiency of 52.4%, effluent chemical oxygen demand and total nitrogen removed 92.5% and 87.0% respectively, due to cooperative biological nitrification/denitrification and electrical migration. As a contrast in AACC mode, with wastewater flowing serially from anode-1 → anode-2 → cathode-1 → cathode-2, a higher current …