Analysis of microbial diversity in oligotrophic microbial fuel cells using 16S rDNA sequences

Molecular ecological techniques were applied to analyze the bacterial diversity of two oligotrophic microbial fuel cells (MFCs) enriched using river water or artificial wastewater (AWW) as fuel. Denaturing gradient gel electrophoresis (DGGE) of the PCR amplified 16S rDNA showed that different microbial communities were present in the two MFCs and these were different from the river sediment used to initiate the enrichment. Nearly complete 16S rDNA was amplified and sequenced. Over 80% of the clones were Proteobacteria. Betaproteobacteria were the dominant clones (46.2%) in MFCs fed with river water, and about 64.4% of the clones in MFCs fed with AWW were Alphaproteobacteria. Actinobacteria were found only in the MFC fed with AWW, and Deltaproteobacteria, Acidobacteria, Chloroflexi and Verrucomicrobia in the MFC fed with river water. Many clones were related to uncultured bacteria, some with homology less than 95%, indicating that many novel bacteria were enriched in the oligotrophic MFCs.     

Characterization of microbial fuel cells enriched using Cr(VI)-containing sludge

Microbial fuel cells (MFCs) were successfully enriched using sludge contaminated with Cr(VI) and their characteristics were investigated. After enrichment, the charge of the final 10 peaks was 0.51 C +/- 1.16%, and the anodic electrode was found to be covered with a biofilm. The enriched MFCs removed 93% of 5 mg/l Cr(VI) and 61% of 25 mg/l Cr(VI). 16S rDNA DGGE profiles from the anodic electrode indicated that beta-Proteobacteria, Actinobacteria, and Acinetobacter sp. dominated. This study is the first to report that electrochemically active and Cr(VI)-reducing bacteria could be enriched in the anode compartment of MFCs using Cr(VI)-containing sludge and demonstrates the Cr(VI) removal capability of such MFCs.     

Electricity generation coupled to oxidation of propionate in a microbial fuel cell

Propionate was used as fuel to enrich an electrochemically-active microbial consortium in a microbial fuel cell, and the bacterial consortium was analyzed by culture-independent methods including denaturing gradient gel electrophoresis (DGGE) of the 16S rDNA, and by fluorescent in situ hybridization (FISH). MFCs fed with propionate produced a current of 4.88 ± 0.1 mA stably on 100 mg propionate/l as COD within 3 weeks of the enrichment. When the MFCs were fed with H₂-saturated fuel containing propionate, the current dropped to 3.82 ± 0.07 mA. The maximum current generated was up to 8.8 mA when MFCs were fed with 200 mg propionate/l as COD. The DGGE of 16S rDNA showed that propionate-enriched MFCs have a different bacterial population from that enriched with acetate and from the inoculum used for enrichment. The major member (42%) of the consortium was an unidentified bacterium followed by γ, β, and δ-proteobacteria.     

一株耐盐产电菌Shewanella algae E-1的分离及其产电特性分析

【背景】产电微生物的种类和电化学活性机制对微生物燃料电池的产电性能有着重要的影响。【目的】从海水中分离获得一株耐盐产电微生物,研究其产电特性并鉴定种属信息。【方法】以取自南海的海水为接种液启动并运行阳极液中含有不同盐浓度的微生物燃料电池,从富集的阳极生物膜上分离得到一株纯培养的微生物菌株,命名为E-1。通过接种于阳极液中添加不同盐浓度的微生物燃料电池中对其产电特性进行分析,并利用形态学观察、Biolog分析和16SrRNA基因序列比对相结合的方法进行种属鉴定。【结果】菌株E-1在无外源添加和外源添加6.6%NaCl条件下产生的功率密度分别为51.69 m W/m2和26.56 m W/m2,这与其良好的耐盐能力相关。菌株E-1被鉴定为海藻希瓦氏菌(Shewanella algae),表现出多样的底物利用能力,生长的温度范围为25-40°C,pH范围为5.0-10.0。【结论】这是首次对Shewanella algae种内微生物产电性能及其在微生物燃料电池中应用的报道,丰富了产电微生物的多样性,菌株E-1能够在较高盐浓度条件下表现出良好的产电性能,为微生物燃料电池在海水资源化处理方面的应用提供新的实验材料。     

Electricity generation and microbial community response to substrate changes in microbial fuel cell

The effect of substrate changes on the performance and microbial community of two-chamber microbial fuel cells (MFCs) was investigated in this study. The MFCs enriched with a single substrate (e.g., acetate, glucose, or butyrate) had different acclimatization capability to substrate changes. The MFC enriched with glucose showed rapid and higher power generation, when glucose was switched with acetate or butyrate. However, the MFC enriched with acetate needed a longer adaptation time for utilizing glucose. Microbial community was also changed when the substrate was changed. Clostridium and Bacilli of phylum Firmicutes were detected in acetate-enriched MFCs after switching to glucose. By contrast, Firmicutes completely disappeared and Geobacter-like species were specifically enriched in glucose-enriched MFCs after feeding acetate to the reactor. This study further suggests that the type of substrate fed to MFC is a very important parameter for reactor performance and microbial community, and significantly affects power generation in MFCs.     

Electrochemical treatment of graphite to enhance electron transfer from bacteria to electrodes

In this paper, graphite felts were continuously electrochemically oxidized to increase the current generation in microbial fuel cells (MFCs). The treated and untreated graphite felts were utilized as anodes in MFCs and current production was compared. The current production on electrochemically treated graphite felt anodes was about 1.13 mA, 39.5% higher compared with that of MFCs containing untreated anodes. The results demonstrated that the electronic coupling between graphite felt electrodes and electrogenic bacteria could be enhanced by electrochemical oxidization of the electrodes. Further study showed that the newly generated carboxyl containing functional groups from electrochemical oxidization were responsible for the enhanced electron transfer, due to their strong hydrogen bonding with peptide bonds in bacterial cytochromes.     

Improving energy accumulation of microbial fuel cells by metabolism regulation using Rhodoferax ferrireducens as biocatalyst

AIMS: To study the physiology and metabolism of microbial cells in the performance of microbial fuel cells (MFCs). METHODS AND RESULTS: A dual-chamber MFCs was constructed, and Rhodoferax ferrireducens was used as biocatalyst. To examine the physiology of microbial cells in the performance of MFCs, the anode media containing planktonic cells was replaced with fresh media in which KH(2)PO(4) and/or NH(4)Cl were excluded. The replacing of anode media containing planktonic cells with fresh media excluded of KH(2)PO(4) and NH(4)Cl made the coulombic yield remarkably increased by a factor of 68% (from 29.1 to 46.8C). The results showed that the electricity could be generated with cells in biofilms as biocatalyst, and coulombic yield was improved by limiting cell growth via removal of ingredients in anode media. By supplementation of glucose to the anode media when current declined to baseline, MFCs achieved about same platform current values immediately. MFCs could continue to produce electricity for about 30 h even after glucose was below detection. CONCLUSIONS: Biofilms and metabolism of glucose play important roles in the performance of MFCs. Coulombic yield of MFCs could be improved by regulating the media ingredients using the stable biofilms-electrode system. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first attempt to study the effect of ingredient compositions of anode media on the performance of MFCs. The observed results that MFCs continued to produce electricity after glucose was below detection was helpful to better understand the mechanism of microbial electricity production.     

Use of acetate for enrichment of electrochemically active microorganisms and their 16S rDNA analyses

A fuel cell-type electrochemical device has been used to enrich microbes oxidizing acetate with concomitant electricity generation without using an electron mediator from activated sludge. The device generated a stable current of around 5 mA with complete oxidation of 5 mM acetate at the hydraulic retention time of 2.5 h after 4 weeks of enrichment. Over 70% of electrons available from acetate oxidation was recovered as current. Carbon monoxide or hydrogen did not influence acetate oxidation or current generation from the microbial fuel cell (MFC). Denaturing gradient gel electrophoresis showed that DNA extracted from the acetate-enriched MFC had different 16S rDNA patterns from those of sludge or glucose+glutamate-enriched MFCs. Nearly complete 16S rDNA sequence analyses showed that diverse bacteria were enriched in the MFC fed with acetate. Electron microscopic observations showed biofilm developed on the electrode, but not microbial clumps observed in MFCs fed with complex fuel such as glucose and wastewater from a corn-processing factory.     

Evaluation of hydrolysis and fermentation rates in microbial fuel cells

This study determined the influence of substrate degradation on power generation in microbial fuel cells (MFCs) and microbial community selection on the anode. Air cathode MFCs were fed synthetic medium containing different substrates (acetate, glucose and starch) using primary clarifier sewage as source of electroactive bacteria. The complexity of the substrate affected the MFC performance both for power generation and COD removal. Power output decreased with an increase in substrate complexity from 99 ± 2 mW m⁻² for acetate to 4 ± 2 mW m⁻² for starch. The organic matter removal and coulombic efficiency (CE) of MFCs with acetate and glucose (82% of COD removal and 26% CE) were greater than MFCs using starch (60% of COD removal and 19% of CE). The combined hydrolysis–fermentation rate obtained (0.0024 h⁻¹) was considerably lower than the fermentation rate (0.018 h⁻¹), indicating that hydrolysis of complex compounds limits current output over fermentation. Statistical analysis of microbial community fingerprints, developed on the anode, showed that microbial communities were enriched according to the type of substrate used. Microbial communities producing high power outputs (fed acetate) clustered separately from bacterial communities producing low power outputs (fed complex compounds).     

Comparison of anode bacterial communities and performance in microbial fuel cells with different electron donors

Microbial fuel cells (MFCs) harness the electrochemical activity of certain microbes for the production of electricity from reduced compounds. Characterizations of MFC anode biofilms have collectively shown very diverse microbial communities, raising ecological questions about competition and community succession within these anode-reducing communities. Three sets of triplicate, two-chamber MFCs inoculated with anaerobic sludge and differing in energy sources (acetate, lactate, and glucose) were operated to explore these questions. Based on 16S rDNA-targeted denaturing gradient gel electrophoresis (DGGE), all anode communities contained sequences closely affiliated with Geobacter sulfurreducens (>99% similarity) and an uncultured bacterium clone in the Bacteroidetes class (99% similarity). Various other Geobacter-like sequences were also enriched in most of the anode biofilms. While the anode communities in replicate reactors for each substrate generally converged to a reproducible community, there were some variations in the relative distribution of these putative anode-reducing Geobacter-like strains. Firmicutes were found only in glucose-fed MFCs, presumably serving the roles of converting complex carbon into simple molecules and scavenging oxygen. The maximum current density in these systems was negatively correlated with internal resistance variations among replicate reactors and, likely, was only minimally affected by anode community differences in these two-chamber MFCs with high internal resistance.     

Life-history strategies of soil microbial communities in an arid ecosystem

The overwhelming taxonomic diversity and metabolic complexity of microorganisms can be simplified by a life-history classification; copiotrophs grow faster and rely on resource availability, whereas oligotrophs efficiently exploit resource at the expense of growth rate. Here, we hypothesize that community-level traits inferred from metagenomic data can distinguish copiotrophic and oligotrophic microbial communities. Moreover, we hypothesize that oligotrophic microbial communities harbor more unannotated genes. To test these hypotheses, we conducted metagenomic analyses of soil samples collected from copiotrophic vegetated areas and from oligotrophic bare ground devoid of vegetation in an arid-hyperarid region of the Sonoran Desert, Arizona, USA. Results supported our hypotheses, as we found that multiple ecologically informed life-history traits including average 16S ribosomal RNA gene copy number, codon usage bias in ribosomal genes and predicted maximum growth rate were higher for microbial communities in vegetated than bare soils, and that oligotrophic microbial communities in bare soils harbored a higher proportion of genes that are unavailable in public reference databases. Collectively, our work demonstrates that life-history traits can distill complex microbial communities into ecologically coherent units and highlights that oligotrophic microbial communities serve as a rich source of novel functions.Subject terms: Microbial ecology, Community ecology     

Cascading influence of inorganic nitrogen sources on DOM production,composition, lability and microbial community structure in the open ocean

Nitrogen frequently limits oceanic photosynthesis and the availability of inorganic nitrogen sources in the surface oceans is shifting with global change. We evaluated the potential for abrupt increases in inorganic N sources to induce cascading effects on dissolved organic matter (DOM) and microbial communities in the surface ocean. We collected water from 5 m depth in the central North Pacific and amended duplicate 20 liter polycarbonate carboys with nitrate or ammonium, tracking planktonic carbon fixation, DOM production, DOM composition and microbial community structure responses over 1 week relative to controls. Both nitrogen sources stimulated bulk phytoplankton, bacterial and DOM production and enriched Synechococcus and Flavobacteriaceae; ammonium enriched for oligotrophic Actinobacteria OM1 and Gammaproteobacteria KI89A clades while nitrate enriched Gammaproteobacteria SAR86, SAR92 and OM60 clades. DOM resulting from both N enrichments was more labile and stimulated growth of copiotrophic Gammaproteobacteria (Alteromonadaceae and Oceanospirillaceae) and Alphaproteobacteria (Rhodobacteraceae and Hyphomonadaceae) in weeklong dark incubations relative to controls. Our study illustrates how nitrogen pulses may have direct and cascading effects on DOM composition and microbial community dynamics in the open ocean.     

Wavelike distributions of infections by an introduced and naturally occurring root pathogen along wheat roots

Previously, we showed that copiotrophic and oligotrophic bacteria fluctuate as moving waves along roots. These waves probably originate as a result of growth and death cycles at any location where a moving nutrient source passed. In this study, we placed sclerotia of Rhizoctonia solani AG8 along growing roots of wheat and showed that the proportions of root sections from which R. solani was isolated fluctuated with distance from the root tip. Similarly, proportions of root sections from which naturally occurring Pythium spp. were isolated fluctuated with distance from the root tip. Fourier analysis showed that these fluctuations constituted significant waves. Cross-correlation analyses demonstrated that there were negative correlations between R. solani infections and colony forming units of copiotrophic bacteria at the time of inoculation at the same locations on the root (lag = 0 cm), indicating that infection by R. solani could have been inhibited by these bacteria. There was a positive correlation between Pythium infections and copiotrophic bacteria at a lag of 6 cm along the roots. It therefore appears that Pythium infection took place shortly after the initial peak in copiotrophic bacteria following the passage of the root tip.     

Enhanced Coulombic efficiency in glucose-fed microbial fuel cells by reducing metabolite electron losses using dual-anode electrodes

Glucose-fed microbial fuel cells (MFCs) have displayed low Coulombic efficiency (CE); one reason for a low CE is metabolite generation, causing significant electron loss within MFC systems. In the present study, notable electron loss (15.83%) is observed in glucose-fed MFCs due to residual propionate, a glucose metabolite. In order to enhance the low CE caused by metabolite generation, a dual-anode MFC (DAMFC) is constructed, which are separately enriched by dissimilar substrates (glucose and propionate, respectively) to effectively utilize both glucose and propionate in one-anode chamber. In the DAMFC, propionate ceases to exist as a source of electron loss, and thus the CE increased from 33 ± 6 to 59 ± 4%.     

Performance and microbial diversity of palm oil mill effluent microbial fuel cell

Aim: To evaluate the bioenergy generation and the microbial community structure from palm oil mill effluent using microbial fuel cell. Methods and Results: Microbial fuel cells enriched with palm oil mill effluent (POME) were employed to harvest bioenergy from both artificial wastewater containing acetate and complex POME. The microbial fuel cell (MFC) showed maximum power density of 3004 mW m^?2 after continuous feeding with artificial wastewater containing acetate substrate. Subsequent replacement of the acetate substrate with complex substrate of POME recorded maximum power density of 622 mW m^?2. Based on 16S rDNA analyses, relatively higher abundance of Deltaproteobacteria (88·5%) was detected in the MFCs fed with acetate artificial wastewater as compared to POME. Meanwhile, members of Gammaproteobacteria, Epsilonproteobacteria and Betaproteobacteria codominated the microbial consortium of the MFC fed with POME with 21, 20 and 18·5% abundances, respectively. Conclusions: Enriched electrochemically active bacteria originated from POME demonstrated potential to generate bioenergy from both acetate and complex POME substrates. Further improvements including the development of MFC systems that are able to utilize both fermentative and nonfermentative substrates in POME are needed to maximize the bioenergy generation. Significance and Impact of the Study: A better understanding of microbial structure is critical for bioenergy generation from POME using MFC. Data obtained in this study improve our understanding of microbial community structure in conversion of POME to electricity.     

产电菌群及电子受体对微生物燃料电池性能的影响

采用2种类型的微生物燃料电池--常规微生物燃料电池(S-MFCs,以生活污水作为产电菌群接种源、以硝酸盐作为电子受体）和改进后的微生物燃料电池（A-MFCs,以厌氧发酵液作为产电菌群接种源、以铁氰化物作为电子受体）,分析了产电菌群和电子受体的改进对微生物燃料电池产电性能的影响.结果表明：产电菌群和电子受体对MFCs驯化周期和运行周期具有显著影响,使驯化周期由S-MFCs的500 h缩短到A-MFCs的430 h,运行周期由S-MFCs的100 h增加到A-MFCs的350 h;改进后的微生物燃料电池使COD去除率提升了25%,使电压输出提高了约300%.选择合适的产电菌菌种和电子受体标准电极电势是微生物燃料电池性能提升的基础.     

Improvement of a microbial fuel cell performance as a BOD sensor using respiratory inhibitors

Studies were made to improve the performance of a microbial fuel cell (MFC) as a biochemical oxygen demand (BOD) sensor. The signal from MFCs decreased in the presence of electron acceptors of higher redox potential such as nitrate and oxygen. The addition of azide and cyanide did not change the signal in the absence of the electron acceptors. The respiratory inhibitors eliminated the inhibitory effects of the electron acceptors on the current generation from MFCs. Similar results were obtained using oligotrophic MFCs fed with an environmental sample that contained nitrate. The use of the respiratory inhibitors is therefore recommended for the accurate BOD measurement of environmental samples containing nitrate and/or oxygen with an MFC-type BOD sensor.     

Degradation of pentachlorophenol with the presence of fermentable and non-fermentable co-substrates in a microbial fuel cell

Pentachlorophenol (PCP) was more rapidly degraded in acetate and glucose-fed microbial fuel cells (MFCs) than in open circuit controls, with removal rates of 0.12 ± 0.01 mg/Lh (14.8 ± 1.0 mg/g-VSS-h) in acetate-fed, and 0.08 ± 0.01 mg/L h (6.9 ± 0.8 mg/g-VSS-h) in glucose-fed MFCs, at an initial PCP concentration of 15 mg/L. A PCP of 15 mg/L had no effect on power generation from acetate but power production was decreased with glucose. Coulombic balances indicate the predominant product was electricity (16.1 ± 0.3%) in PCP-acetate MFCs, and lactate (19.8 ± 3.3%) in PCP-glucose MFCs. Current generation accelerated the removal of PCP and co-substrates, as well as the degradation products in both PCP-acetate and PCP-glucose reactors. While 2,3,4,5-tetrachlorophenol was present in both reactors, tetrachlorohydroquinone was only found in PCP-acetate MFCs. These results demonstrate PCP degradation and power production were affected by current generation and the type of electron donor provided.     

Microbial fuel cells meet with external resistance

The influence of external load on the composition of the anodic biofilm microbial community and biomass yield was investigated in a microbial fuel cell fed with glucose and domestic wastewater was used as source of electrogens. Denaturing gradient gel electrophoresis (DGGE) of polymerase chain reaction (PCR) amplified 16S rRNA gene fragments revealed distinct differences in anodic bacterial communities formed at the anode of each MFC operated under a different external load. These results implied that in an MFC, electrogenic bacteria were enriched under higher current densities, i.e., low external load, and were able to sustain better current and effluent quality. The influence of the external resistance applied to the MFCs during formation of the bacterial communities from sewage wastewater was shown to have no significant effect on power performance of the MFCs nor to have a significant influence on their anodic activity with both glucose and brewery wastewater as fuel. As expected, current generation, COD removal and the biomass yield were all directly influenced by the external load. Significantly, when operated under lower external load, the biomass yield in the MFC was less than that in conventional anaerobic digestion (i.e., control).     

The influence of acidity on microbial fuel cells containing Shewanella oneidensis

Microbial fuel cells (MFCs) traditionally operate at pH values between 6 and 8. However, the effect of pH on the growth and electron transfer abilities of Shewanella oneidensis MR-1 (wild-type) and DSP10 (spontaneous mutant), bacteria commonly used in MFCs, to electrodes has not been examined. Miniature MFCs using bare graphite felt electrodes and nanoporous polycarbonate membranes with MR-1 or DSP10 cultures generated >8W/m(3) and approximately 400muA between pH 6-7. The DSP10 strain significantly outperformed MR-1 at neutral pH but underperformed at pH 5. Higher concentrations of DSP10 were sustained at pH 7 relative to that of MR-1, whereas at pH 5 this trend was reversed indicating that cell count was not solely responsible for the observed differences in current. S. oneidensis MR-1 was determined to be more suitable than DSP10 for MFCs with elevated acidity levels. The concentration of riboflavin in the bacterial cultures was reduced significantly at pH 5 for DSP10, as determined by high performance liquid chromatography (HPLC) of the filter sterilized growth media. In addition, these results suggest that mediator biosynthesis and not solely bacterial concentration plays a significant role in current output from S. oneidensis containing MFCs.