The microbial communities developed from a mixed-species culture in up-flow and flow-through configurations of thermophilic (55 °C) microbial fuel cells (MFCs), and their power production from acetate, were investigated. The up-flow MFC was operated for 202 days, obtaining an average power density of 0.13 W/m3, and Tepidiphilus was the dominant transcriptionally-active microorganisms. The planktonic community developed in the up-flow MFC was used to inoculate a flow-through MFC resulting in the proliferation of Ureibacillus, whose relative abundance increased from 1 to 61% after 45 days. Despite the differences between the up-flow and flow-through MFCs, including the anode electrode, hydrodynamic conditions, and the predominant microorganism, similar (p = 0.05) volumetric power (0.11–0.13 W/m3), coulombic efficiency (16–18%) and acetate consumption rates (55–69 mg/L/d) were obtained from both. This suggests that though MFC design can shape the active component of the thermophilic microbial community, the consortia are resilient and can maintain similar performance in different MFC configurations. © 2019 Elsevier Ltd