Wang et al. Biotechnology for Biofuels 2014, 7:9 http://www.biotechnologyforbiofuels.com/content/7/1/9 RESEARCH Open Access Microbial community structures differentiated in a single-chamber air-cathode microbial fuel cell fueled with rice straw hydrolysate Zejie Wang1,4, Taekwon Lee3,5, Bongsu Lim1, Chansoo Choi2* and Joonhong Park3 Abstract Background: The microbial fuel cell represents a novel technology to simultaneously generate electric power and treat wastewater. Both pure organic matter and real wastewater can be used as fuel to generate electric power and the substrate type can influence the microbial community structure. In the present study, rice straw, an important feedstock source in the world, was used as fuel after pretreatment with diluted acid method for a microbial fuel cell to obtain electric power. Moreover, the microbial community structures of anodic and cathodic biofilm and planktonic culturewere analyzed and compared to reveal the effect of niche on microbial community structure. Results: The microbial fuel cell produced a maximum power density of 137.6 ± 15.5 mW/m2 at a COD concentration of 400 mg/L, which was further increased to 293.33 ± 7.89 mW/m2 through adjusting the electrolyte conductivity from 5.6 mS/cm to 17 mS/cm. Microbial community analysis showed reduction of the microbial diversities of the anodic biofilm and planktonic culture, whereas diversity of the cathodic biofilm was increased. Planktonic microbial communities were clustered closer to the anodic microbial communities compared to the cathodic biofilm. The differentiation in microbial community structure of the samples was caused by minor portion of the genus. The three samples shared the same predominant phylum of Proteobacteria. The abundance of exoelectrogenic genus was increased with Desulfobulbus as the shared most abundant genus; while the most abundant exoelectrogenic genus of Clostridium in the inoculum was reduced. Sulfate reducing bacteria accounted for large relative abundance in all the samples, whereas the relative abundance varied in different samples. Conclusion: The results demonstrated that rice straw hydrolysate can be used as fuel for microbial fuel cells; microbial community structure differentiated depending on niches after microbial fuel cell operation; exoelectrogens were enriched; sulfate from rice straw hydrolysate might be responsible for the large relative abundance of sulfate reducing bacteria. Keywords: Microbial fuel cell, Microbial diversity, 454-pyrosequencing, Rice straw biomass Background its constituent sugars through acidic and/or enzymatic Microbial fuel cells (MFCs) are devices to produce elec- hydrolysis; the produced sugars can be further used as tric energy from organic matters and treat wastewaters substrate to produce organic acids or bioethanol [5,6]. in both anode and cathode chambers [1,2]. Pure organic Therefore, the sugars produced from the rice straw hy- compound, real wastewater, and biomass have been drolysate might be used as a useful fuel for power gener- successfully used as fuel for power generation in MFCs ation from MFCs. [3]. Rice straw is one of the most abundant biomasses, In MFCs, microbes play crucial roles in energy output mainly composed of cellulose, hemicellulose, and some and organic contaminants removal [7]. The ability of mi- lignin [4]. The hemicellulose can be easily degraded to crobes to transfer electrons in the anode can significantly affect the performance of MFCs. Anodic microbial com- munities were reported to be significantly related with the * Correspondence: [email protected] 2Department of Applied Chemistry, Daejeon University, Daejeon, South Korea types of substrates fed into the anode chamber [8]. For ex- Full list of author information is available at the end of the article ample, Acetobacterium species (sp.), Geobacter sp., and © 2014 Wang et al.; licensee BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Wang et al. Biotechnology for Biofuels 2014, 7:9 Page 2 of 10 http://www.biotechnologyforbiofuels.com/content/7/1/9 Arcobacter sp. were detected in the anodic biofilm fed with formate [9]; however, Enterobacter sp. was the dom- inant bacterial species in the MFC with glucose as sub- strate [10]. For air-cathode MFCs, biofilm was commonly formed on the water-facing side of the cathode. It was dis- covered that the formation of biofilm on the Pt-loaded air-cathode could decrease the power output due to the increased cathodic resistance and limited proton transfer rate [11]; however, recent research demonstrated that the biofilm formation on a bare air-cathode could enhance the electric power output from air-cathode MFCs [12]. The different research conclusions might be caused by dif- ferent air-cathode configurations. Moreover, the cathodic biofilm in a Pt-loaded air-cathode was observed to be cap- able of removing nitrogen, with enhanced removal effi- ciency due to the pre-accumulation of nitrifying biofilm [13]. The aforementioned results indicate that the cath- odic biofilm deserves further research. Therefore, the purpose of the present study was to evaluate the availability of diluted acid-treated rice straw hydrolystate as fuel for an air-cathode MFC. In addition, microbial analysis at high resolution level using 454 py- rosequencing was carried out to evaluate the effect of the rice straw hydrolystate and niches on the microbial diversity and community. Results and discussion Performance of the MFC After addition of the rice straw hydrolysate as an anodic solution, cell voltage was immediately increased with no lag time. Stable voltage increased from 177.6 ± 17.3 mV for chemical oxygen demand (COD) of 100 mg/L to 524.7 ± 3.2 mV for COD of 400 mg/L, in response to the decrease in anodic potential from −110.5 ± 21.6 mV to −508.7 ± 6.9 mV (Figure 1a and b). The results indicated that or- ganic matters produced from the hydrolysate could be easily utilized by anodic microorganism and release elec- trons, decreasing the anodic potential and consequently increasing the cell voltage [14]. The stable anodic potential was appropriately −300 mV (versus standard hydrogen Figure 1 Voltage response of the microbial fuel cell (MFC) to electrode), similar to that of −340 mV observed by Wang chemical oxygen demand (COD) under batch-mode operation. et al (a) Voltage output and (b) anodic potential as a function of COD . [15]. For relatively low COD concentrations (100 and concentration at a fixed external resistance of 500 Ω. Figures on the 150 mg/L), the cell voltage showed a large difference be- plot represent the COD concentrations (mg/L). tween two separate cycles; while the COD concentration increased to 200 mg/L, the voltage was well reproduced, suggesting that the saturated COD for voltage output was Coulombic efficiency (CE) indicates the ratio of total 200 mg/L. The saturated COD concentration in the electrons recovered as electric current from organic present study was far lower than that of the 1,000 mg/L matter. In this study, the CE was calculated in the range observed when wheat straw hydrolysate was used as fuel in from 8.5% to 17.9%, and the COD removal efficiency a dual-chamber MFC [16]. A lower saturated COD con- ranged from 49.4 ± 4.0% to 72.0 ± 1.7% (Figure 2). The centration indicated higher capacity of power production present CE was lower than that of wheat straw- [16] or from the fuel based on the same quantity. Furthermore, corn stover biomass-fuelled [17] MFCs, which were 15.5% the discharge time increased from 17.1 ± 1.2 hours to 49.8 ± to 37.1% and 19.3% to 25.6%, respectively; whereas it was 2.4 hours relying on the increased COD concentration. higher than that previously reported using real wastewater Wang et al. Biotechnology for Biofuels 2014, 7:9 Page 3 of 10 http://www.biotechnologyforbiofuels.com/content/7/1/9 Figure 2 Effluent chemical oxygen demand (COD) concentration and COD removal efficiency (RE) and coulombic efficiency (CE). The initial COD concentration was 100, 150, 200, and 400 mg/L with an external resistance of 500 Ω. as fuels, which for example, is less than 1% for fermented wastewater [18], and a maximum CE of 8% for starch pro- cessing wastewater [19]. For air-cathode MFCs, oxygen diffused to the anode chamber can aerobically consume substrates other than anaerobically generated electrons, which would be the reason for the low CE of air-cathode MFCs [20]. The maximum power density (Pmax) was determined as 137.6 ± 15.5 mW/m2 for COD of 400 mg/L, at a current density of 0.28 A/m2 (Figure 3a). It was further promoted to 293.3 ± 7.9 mW/m2 at a current density of 0.90 A/m2 when the solution conductivity was adjusted from 5.6 mS/ cm to 17 mS/cm through addition of NaCl. Consequently, the internal resistance (Rint) of the MFC was decreased from 714.4 ± 19.1Ω to 229.9 ± 14.5Ω. Moreover, conduct- Figure 3 Power density and electrode potentials as a function ivity adjustment enhanced the anodic performance, reliev- of current density. (a) Polarization curves, and (b) electrode ing the mass transfer limitation while limiting the cathode potentials for conductivity unadjusted (CUA) and adjusted (CA) performance (Figure 3b). solutions with hydrolysate of a COD concentration of 400 mg/L. The 2 conductivity was adjusted from 5.6 mS/cm to 17 mS/cm with The Pmax (293.33 ± 7.89 mW/m ) was larger than that of addition of NaCl. wheat straw hydrolysate (123 mW/m2). Based on the max- imum power of 1.08 mW (293.33 mW/m2) and the stable discharge time of 24 hours, 2.592 × 10-5 kWh of electric can meet the annual demand of 35.56 million people for power can be extracted from a COD concentration of electric power.