Community analysis of biofilms on flame-oxidized stainless steel anodes in microbial fuel cells fed with different substrates

Nweze Julius Eyiuche, Shiho Asakawa, Takahiro Yamashita, Atsuo Ikeguchi, Yutaka Kitamura, Hiroshi Yokoyama

The flame-oxidized stainless steel anode (FO-SSA) is a newly developed electrode that enhances microbial fuel cell (MFC) power generation; however, substrate preference and community structure of the biofilm developed on FO-SSA have not been well characterized. Herein, we investigated the community on FO-SSA using high-throughput sequencing of the 16S rRNA gene fragment in acetate-, starch-, glucose-, and livestock wastewater-fed MFCs. Furthermore, to analyze the effect of the anode material, the acetate-fed community formed on a common carbon-based electrode-carbon-cloth anode (CCA)-was examined for comparison. Substrate type influenced the power output of MFCs using FO-SSA; the highest electricity was generated using acetate as a substrate, followed by peptone, starch and glucose, and wastewater. Intensity of power generation using FO-SSA was related to the abundance of exoelectrogenic genera, namely Geobacter and Desulfuromonas, of the phylum Proteobacteria, which were detected at a higher frequency in acetate-fed communities than in communities fed with other substrates. Lactic acid bacteria (LAB)-Enterococcus and Carnobacterium-were predominant in starch- and glucose-fed communities, respectively. In the wastewater-fed community, members of phylum Planctomycetes were frequently detected (36.2%). Exoelectrogenic genera Geobacter and Desulfuromonas were also detected in glucose-, starch-, and wastewater-fed communities on FO-SSA, but with low frequency (0-3.2%); the lactate produced by Carnobacterium and Enterococcus in glucose- and starch-fed communities might affect exoelectrogenic bacterial growth, resulting in low power output by MFCs fed with these substrates. Furthermore, in the acetate-fed community on FO-SSA, Desulfuromonas was abundant (15.4%) and Geobacter had a minor proportion (0.7%), while in that on CCA, both Geobacter and Desulfuromonas were observed at similar frequencies (6.0-9.8%), indicating that anode material affects exoelectrogenic genus enrichment in anodic biofilm. Anodic community structure was dependent on both substrate and anode material. Although Desulfuromonas spp. are marine microorganisms, they were abundant in the acetate-fed community on FO-SSA, implying the presence of novel non-halophilic and exoelectrogenic species in this genus. Power generation using FO-SSA was positively related to the frequency of exoelectrogenic genera in the anodic community. Predominant LAB in saccharide-fed anodic biofilm caused low abundance of exoelectrogenic genera and consequent low power generation.