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。     

微嗜酸寡养单胞菌EFS1的产电性能及在污水处理中的应用

【背景】微生物燃料电池(Microbial Fuel Cell,MFC)作为一种新型的燃料电池资源,在产电的同时可应用于污水处理领域,达到资源最大化的目的。【目的】从MFC中分离获得一株可培养微生物,研究其产电特性及在污水处理中的微生物絮凝、重金属耐受、苯酚降解性能,为扩展产电菌资源库提供理论基础。【方法】利用WO_3纳米探针从MFC阳极中筛选获得一株具备产电和絮凝性能的菌株,命名为EFS1。运用循环伏安分析结合扫描电镜观测阳极电极;改变外电阻测定极化曲线和功率密度曲线。测定菌株的絮凝、重金属耐受及苯酚降解性能。【结果】经16S rRNA基因序列分析,结合形态学和生理生化鉴定菌株EFS1为微嗜酸寡养单胞菌(Stenotrophomonas acidaminiphila)。菌株EFS1具有稳定的产电周期,周期电压最高可达300m V,功率密度可达56.25m W/m~2;扫描电镜发现菌株存在直接接触电极及分泌电子中介体传递电子的方式;MFC内阻为1 000Ω左右。有氧条件下菌株的絮凝率可达到70%,存在电子受体的无氧环境中可达到80%;该菌株还具有良好的Cd~(2+)、Cu~(2+)、Mn~(2+)耐受性及苯酚降解性能,在48 h、2–4 mg/L时苯酚降解率达到了100%。【结论】研究验证了产电菌EFS1具备絮凝能力、重金属耐受、苯酚降解的可能性,为产电菌的开发及污水处理方面提供理论依据。     

好氧-厌氧混合污泥启动微生物燃料电池产电性能及微生物群落动态特征

【目的】为探讨好氧-厌氧混合污泥启动微生物燃料电池(Microbial fuel cell,MFC)产电性能以及MFC对微生物群落的选择作用,【方法】以乳酸为底物,应用不依赖于培养的微生物分子生物学技术解析单室MFC启动过程中微生物群落的组成和结构动态学特征。【结果】结果表明,MFC经过3个周期启动成功,最高输出电压230 m V。当MFC外电阻为1656Ω时,最大功率密度11.15 W/m3,电池运行稳定。混合污泥启动MFC以后,阳极生物膜微生物群落结构同种泥差异较大,且多样性降低。生物膜中微生物类群按丰度依次为β-变形菌纲(Betaproteobacteria)24.90%、拟杆菌门(Bacteroidetes)21.30%、厚壁菌门(Firmicutes)9.70%、γ-变形菌纲(Gammaproteobacteria)8.50%、δ-变形菌纲(Deltaproteobacteria)7.90%、绿弯菌门(Chloroflexi)4.20%以及α-变形菌纲(Alphaproteobacteria)3.60%。有利于生物膜形成与稳定的动胶菌属(Zoogloea)和不动杆菌属(Acinetobacter)序列丰度分别占生物膜群落的5.00%和3.90%,与MFC产电能力直接相关的地杆菌属(Geobacter)序列由混合污泥中的0.60%上升至阳极生物膜中的2.60%。【结论】本研究表明,MFC阳极生物膜在驯化过程中对污泥中的微生物进行淘汰和选择,最终驯化形成了有利于生物膜形成与稳定、有机物厌氧发酵与产电的微生物菌群。     

利用微生物电解池处理牛奶废水过程中产电菌落与产氢性能之间的关系

【目的】 研究微生物电解池(Microbial electrolysis cell, MEC)利用复杂有机物作为底物的运行特性, 对其在废水处理中的应用有着重要的意义。【方法】 以模拟牛奶废水为基质, 通过构建MEC反应器来考察在不同外加电压条件下产电菌群的性能。【结果】 当外加电压升高到1.2 V时, 最大电流密度可达到261 A/m3, 产氢速率可达0.048 m3 H2/m3 d, 分别比外加电压为0.4 V的情形提高了467%和700%。外加电压为1.2 V时, 系统对COD和蛋白质去除率可分别达59%和74%, 其中COD去除较之0.4 V的情形提高了22.5%。PCR-DGGE的分子生物学分析结果表明, 阳极生物膜中以Geobacter sp.作为优势菌, 说明在利用大分子有机物作为基质时产电菌与非产电菌的协同作用更为明显。【结论】 MEC能够利用牛奶废水作为燃料, 在实现高效降解的同时以产氢的形式进行能量产出, 这为MEC的实际应用提供了研究思路。     

一株土壤产电菌Clostridium sporogenes的分离及其产电性能

【目的】从土壤中分离获得产电菌纯菌株SE6,鉴定其种类并分析其产电性能。【方法】通过厌氧培养分离得到纯菌株,根据其形态、生理生化性质及16S r RNA基因测序分析确定其种属。以该菌株作为产电菌接种源,液体LB培养基和铁氰化钾溶液分别作为阳极液和阴极液,构建双室微生物燃料电池(Microbial fuel cells,MFCs),研究其产电能力;根据交流阻抗图谱,分析MFCs的内阻。应用循环伏安测试确定该菌株的胞外电子传递方式。并利用扫描电镜对阳极表面产电菌形态进行观察。【结果】菌株SE6的16S r RNA基因序列与Clostridium sporogenes有100%同源性,结合其形态特征和生理生化特性,确定其属于梭菌属(Clostridium)。SE6接种到MFCs中可以获得44.42 m W/m~2的最大功率密度。MFCs的阳极内阻、阴极内阻和欧姆内阻分别为(1488±193)Ω/cm~2、(0.92±0.01)Ω/cm~2和(20.69±1.76)Ω/cm~2。其循环伏安图谱显示体系中存在电化学活性物质且峰值电流随扫速升高线性增大。扫描电镜观察到阳极表面聚集附着着长度约1μm的杆菌。【结论】本研究成功从土壤中分离出具有一定产电能力的菌株C.sporogenes SE6,可直接将电子传递至阳极,其产电过程阻抗较大。     

微生物燃料电池中假单胞菌F026的产电性能研究

基于微生物燃料电池的反应装置,从污水处理厂曝气池的污泥中通过富集,筛选和基于16S rRNA基因序列的系统发育分析等手段驯化出1株高效产电假单胞菌F026。以F026为阳极产电菌制作微生物燃料电池,考察了底物种类、温度和p H值等因素对微生物燃料电池产电性能的影响。结果表明,F026最适合在以可溶性淀粉为底物,p H为中性偏碱性,温度在30~35℃的环境下生长。在此条件下,微生物燃料电池的最高电压达到500 m V,体积功率密度达到2 W/m3。     

pH值对微生物燃料电池处理生物废弃物的影响

以生物废弃物为底物,采用双室微生物燃料电池对生物废弃物进行处理,研究阳极液初始pH值对生物废弃物CODcr、TOC去除率的影响,以及对微生物燃料电池产电性能影响。实验结果表明,当阳极液pH值为6时,CODcr去除率达72.1%,TOC的去除率达44.5%,其输出电压最大为1.27 V,平均电压797 mV,最大功率密度达到136.6 mW/m2。     

异化铁还原梭菌Clostridium bifermentans EZ-1产氢与电化学特性

【目的】从黄河三角洲滨海湿地土壤中获得同时具备产氢、产电以及异化铁还原能力的多功能菌株。【方法】通过厌氧分离技术从黄河三角洲土壤中分离得到纯菌株,16S r RNA基因测序后与数据库已有序列进行比对。利用革兰氏染色及扫描电镜观察菌株形态,并用气相色谱(gas chromatography,GC)和液相色谱(high performance liquid chromatography,HPLC)检测其生理代谢底物和产物。通过添加不同形态铁氧化物检测该菌株Fe(Ⅲ)的还原能力。构建微生物燃料电池(Microbial fuel cells,MFCs)检测该菌株的电化学活性。【结果】16S r RNA基因序列比对发现其与双酶梭菌Clostridium bifermentans的相似性达97.99%。革兰氏染色结果显示为阳性菌。能够利用葡萄糖为底物发酵产生氢气、二氧化碳、乙酸和丁酸。Fe(Ⅲ)还原能力检测发现,其不仅可以还原柠檬酸铁(FeC_6H_5O_7)中可溶性的Fe(Ⅲ),还可以还原无定形铁水铁矿(FeOOH)和晶型纳米磁铁矿(Fe_3O_4)中的Fe(Ⅲ)。此外,经MFCs检测发现,该菌具有电化学活性,最大电流输出密度可达6.5 m A/m~2,且在0.15 V位置存在氧化峰。【结论】本研究从土壤中成功分离得到了一株同时具有产氢、产电以及异化铁还原能力的多功能梭菌菌株,保藏并命名为Clostridium bifermentans EZ-1。     

微生物燃料电池利用乳酸产电性能与微生物群落分布特征

【目的】为探讨以乳酸为基质的微生物燃料电池(Microbial fuel cell,MFC)产电性能以及微生物群落在阳极膜、悬浮液、阳极沉淀污泥中的分布特征,【方法】试验建立了双室MFC,以乳酸为阳极主要碳源,研究了反应器的启动过程及产电效能,同时以电镜和PCR-变性梯度凝胶电泳(Denaturing gradient gelelectrophoresis,DGGE)技术解析了微生物群落的空间分布特征。【结果】结果表明,反应器启动第7天时外电压达到0.56 V,当外阻为80Ω时,电流密度为415 mA/m2,MFC的功率密度达到最大值82 mW/m2。电镜观察发现大量杆菌附着在阳极表面,结合较为紧密;DGGE图谱显示阳极膜表面微生物与种泥最为相似,与阳极悬浮液、底部沉淀污泥中的主要菌群一致,条带序列与睾丸酮丛毛单胞菌(Comamonas testosteroni)和布氏弓形菌(Arcobacter butzleri)等最为相似。【结论】本研究表明以乳酸为基质MFC可产生较高的功率密度,阳极附着的优势菌与接种污泥来源密切相关。     

一株单环刺螠肠道电活性希瓦氏菌Shewanella marisflavi的生理学特性

【目的】探究以单环刺螠为代表的海洋环节动物肠道中电活性微生物的存在情况,并表征其生理学及电化学特性。【方法】采用平板划线法、16S rRNA基因测序技术分离纯化菌株并进行菌株鉴定。利用扫描电镜表征菌株形态。高效液相色谱(high performance liquid chromatography,HPLC)检测其无氧呼吸底物氧化及产物生成情况。通过菲啰嗪和甲醛肟显色法检测菌株的异化Fe(Ⅲ)和Mn(Ⅳ)还原能力。借助单室微生物燃料电池(single-chamber microbial fuel cells,SCMFCs)及循环伏安法检测菌株的电化学活性。【结果】从单环刺螠肠道中成功分离得到了一株兼性厌氧菌,16S rRNA基因序列比对结果显示该菌株与Shewanellamarisflavi的相似性达99.93%。扫描电镜结果显示其为杆状,长约2μm,宽度约为0.5μm。HPLC检测结果表明,该菌能以乳酸钠为电子供体,富马酸为电子受体进行无氧呼吸并伴随代谢产物乙酸钠和琥珀酸产生。菲啰嗪和甲醛肟显色法结果证实,该菌具有异化铁、锰还原能力。单室MFCs检测结果发现该菌的最大电流输出密度为146 mA/m2,循环伏安法检测结果显示该菌在0.14 V和–0.51 V位置处分别存在氧化峰和还原峰。【结论】本研究结果证实以单环刺螠为代表的海洋环节动物肠道中存在以Shewanella marisflavi UU-3-2为代表的电活性微生物。表明电化学活性微生物在海洋环节动物肠道中广泛存在。     

Electricity production from xylose using a mediator-less microbial fuel cell

Electricity generation integrated with xylose degradation was investigated in a two-chamber mediator-less microbial fuel cell (MFC). Voltage output followed saturation kinetics as a function of xylose concentration for concentration below 9.7 mM, with a predicted maximum of 86 mV (6.3 mW m(-2) or 116 mW m(-3)) and half-saturation constant (K(s)) of 0.29 mM. Xylose concentrations from 0.5 mM to 1.5 mM resulted in coulombic efficiencies and maximum voltage ranging from 41+/-1.6% to 36+/-1.2% and 55+/-2.0 mV to 70+/-3.0 mV respectively. Xylose degradation rate increased with increasing xylose concentration up to 9.7 mM and the predicted maximum degradation rate was 0.13 mM h(-1) and K(s) of 3.0 mM. Stirring by nitrogen in the anode chamber led to 99+/-2.3 mV maximum voltage (8.4+/-0.4 mW m(-2) or 153+/-7.1 mW m(-3)) and 5.9+/-0.3% coulombic efficiency at MFC running time 180 h, which were respectively 17+/-1.2% and 37+/-1.8%, higher than those without stirring. The COD removal under stirring was 22.1+/-0.3%, which was slightly lower than that of 23.7+/-0.4% under no stirring. However, stirring resulted in 59% lower xylose degradation rate. This work demonstrates that xylose can be used in the MFC for electricity production. Comparatively higher electricity generation and coulombic efficiency can be obtained by adjusting initial xylose concentration and applying stirring in the anode chamber.     

Fabrication of stainless steel mesh gas diffusion electrode for power generation in microbial fuel cell

This study reports the fabrication of a new membrane electrode assembly by using stainless steel mesh (SSM) as raw material and its effectiveness as gas diffusion electrode (GDE) for electrochemical oxygen reduction in microbial fuel cell (MFC). Based on feeding glucose (0.5 g L(-1)) substrate to a single-chambered MFC, power generation using SSM-based GDE was increased with the decrease of polytetrafluoroethylene (PTFE) content applied during fabrication, reaching the optimum power density of 951.6 mW m(-2) at 20% PTFE. Repeatable cell voltage of 0.51 V (external resistance of 400 Ω) and maximum power density of 951.6 mW m(-2) produced for the MFC with SSM-based GDE are comparable to that of 0.52 V and 972.6 mW m(-2), respectively obtained for the MFC containing typical carbon cloth (CC)-made GDE. Besides, Coulombic efficiency (CE) is found higher for GDE (SSM or CC) with membrane assembly than without, which results preliminarily from the mitigation of Coulombic loss being associated with oxygen diffusion and substrate crossover. This study demonstrates that with its good electrical conductivity and much lower cost, the SSM-made GDE suggests a promising alternative as efficient and more economically viable material to conventional typical carbon for power production from biomass in MFC.     

Electricity generation using p-nitrophenol as substrate in microbial fuel cell

The cell voltage and degradation rate of p-nitrophenol (PNP) were monitored in a two-chambered microbial fuel cell (MFC) system. Degradation metabolites in the anode solution of MFC were analyzed by gas chromatography–mass spectrometry (GC–MS). PNP was used as substrate by the MFC that was inoculated with anaerobic sludge. The results showed that electricity output increased with the PNP concentration increased, the MFC displayed a maximum power density of 1.778 mW m⁻² and a maximum PNP degradation rate of 64.69% when PNP was used as a sole substrate. However, the cell voltage and the PNP degradation rate with sodium acetate (402.3 mV and 95.96%) were higher than those fed with glucose (341.9 mV and 83.51%) when glucose and sodium acetate were used as a substrate, respectively. Furthermore, GC–MS analysis showed that the PNP was biodegraded completely after 142 h in the MFC. These results demonstrate that PNP can be used for electricity generation in MFC for practical applications of wastewater treatment.     

A comparison of air and hydrogen peroxide oxygenated microbial fuel cell reactors

In this study, a two-compartment continuous flow microbial fuel cell (MFC) reactor was used to compare the efficiencies of cathode oxygenation by air and by hydrogen peroxide. The MFC reactor had neither a proton-selective membrane nor an electron transfer mediator. At startup, the cathodic compartment was continuously aerated and the anodic compartment was fed with a glucose solution. An increase of electrical power generation from 0.008 to 7.2 mW m(-2) of anode surface with a steady-state potential of 215-225 mV was observed within a period of 12 days. The performance of the air-oxygenated MFC reactor progressively declined over time because of biofilm proliferation in the cathodic compartment. Oxygenation of the cathodic compartment using 300 mL d(-1) of 0.3% hydrogen peroxide solution resulted in a power density of up to 22 mW m(-2) (68.2 mA m(-2)) of anode surface at a potential of 340-350 mV. The use of H2O2 for oxygenation was found to improve the long-term stability of the MFC reactor.     

一株海洋产电菌Shewanella sp. S2的筛选和产电分析

以厦门白城海域的潮间带表面沉积物为菌种来源筛选得到一株具有电催化活性的菌株S2,该菌株的16S rRNA和gyrB基因发育树与Shewanella oneidensis MR-1同支,相似性分别为98.5%和87%,葡萄糖、木糖、半乳糖等碳源利用及最佳生长的NaCl浓度与S.oneidensis MR-1有显著差别,因此初步鉴定为Shewanella属菌株,命名为Shewanella sp.S2。初步研究了菌株S2产电活性,在以乳酸作为碳源产电时,电压最高为150mV,相应的电流密度为66.1mA/m2。     

Microbial fuel cells demonstrate high coulombic efficiency applicable for water remediation

Microbial fuel cells (MFCs) convert biomass into electricity by the metabolic activity of microorganisms and are also used for remediation and water treatment. Power output was compared for a dual chambered membrane MFC using either E. coli or two Yamuna river samples, Yamuna (before the Sangam region)--slow flow (sample 1) and Sangam region--fast flow (sample 2). E. coli and the two river water samples 1 and 2 gave a maximum voltage of 779, 463 and 415 mV respectively. Using E. coli the maximum power density obtained with a 100 omega resistor was 220.66 mW/cm2 and the highest power generated 6068.41 mW. The results demonstrate E. coli, river sample 1 and river sample 2 have a comparable coulombic efficiency of 85.2, 71 and 77% respectively when using 0.4% sucrose as substrate. The decrease in chemical oxidative demand of all river water samples using MFC technology demonstrates efficient remediation of inland water.     

鳜源致病性维氏气单胞菌的鉴定及药敏试验

【目的】通过对从患病的鳜(Siniperca chuatsi)肝脏中分离得到的一株优势菌WJ2014-1进行鉴定,旨在确定病因并筛选出敏感药物,为今后鳜维氏气单胞菌(Aeromonas veronii)的防治提供参考。【方法】从患病鳜肝脏分离致病菌,通过对其生理生化特征与16S r RNA基因序列分析进行鉴定,并通过纸片扩散法进行药敏试验。【结果】用菌株WJ2014-1进行人工回归感染试验后,鳜发病症状与自然发病症状相似。根据该菌株的形态特征、生化特性、16S r RNA基因序列分析结果鉴定为维氏气单胞菌。该菌株对复方新诺明、强力霉素、罗红霉素、头孢噻肟、哌拉西林等21种抗生素敏感,对氯霉素、诺氟沙星、萘啶酸等9种抗生素耐药。【结论】分离得到的菌株WJ2014-1对鳜有致病性,生产中可选用复方新诺明、强力霉素、罗红霉素等药物进行防治。     

Influence of anodic biofilm growth on bioelectricity production in single chambered mediatorless microbial fuel cell using mixed anaerobic consortia

The effect of anodic biofilm growth and extent of its coverage on the anodic surface of a single chambered mediatorless microbial fuel cell (MFC) was evaluated for bioelectricity generation using designed synthetic wastewater (DSW) and chemical wastewater (CW) as substrates and anaerobic mixed consortia as biocatalyst. Three MFCs (plain graphite electrodes, air cathode, Nafion membrane) were operated separately with variable biofilm coverage [control; anode surface coverage (ASC), 0%], partially developed biofilm [PDB; ASC approximately 44%; 90 days] and fully developed biofilm [FDB; ASC approximately 96%; 180 days] under acidophilic conditions (pH 6) at room temperature. The study depicted the effectiveness of anodic biofilm formation in enhancing the extracellular electron transfer in the absence of mediators. Higher specific power production [29mW/kg COD(R) (CW and DSW)], specific energy yield [100.46J/kg VSS (CW)], specific power yield [0.245W/kg VSS (DSW); 0.282W/kg VSS (CW)] and substrate removal efficiency of 66.07% (substrate degradation rate, 0.903kgCOD/m(3)-day) along with effective functioning fuel cell at relatively higher resistance [4.5kOmega (DSW); 14.9kOmega (CW)] correspond to sustainable power [0.008mW (DSW); 0.021mW (CW)] and effective electron discharge (at higher resistance) and recovery (Coulomb efficiency; 27.03%) were observed especially with FDB operation. Cyclic voltammetry analysis documented six-fold increment in energy output from control (1.812mJ) to PDB (10.666mJ) operations and about eight-fold increment in energy from PDB to FDB (86.856mJ). Biofilm configured MFC was shown to have the potential to selectively support the growth of electrogenic bacteria with robust characteristics, capable of generating higher power yields along with substrate degradation especially operated with characteristically complex wastewaters as substrates.     

Characterization of Fluoroglycofen Ethyl Degradation by Strain <Emphasis Type="Italic">Mycobacterium phocaicum</Emphasis> MBWY-1

A fluoroglycofen ethyl-degrading bacterium, MBWY-1, was isolated from the soil of an herbicide factory. This isolated strain was identified as Mycobacterium phocaicum based on analysis of its 16S rRNA gene sequence and its morphological, physiological, and biochemical properties. The strain was able to utilize fluoroglycofen ethyl as its sole source of carbon for growth and could degrade 100 mg l⁻¹ of fluoroglycofen ethyl to a non-detectable level within 72 h. The optimum temperature and pH for fluoroglycofen ethyl degradation by strain MBWY-1 were 30°C and 7.0, respectively. Five metabolites produced during the degradation of fluoroglycofen ethyl and were identified by mass spectrometry as {5-[2-chloro-4-(trifluoromethyl) phenoxy]-2-nitrophenylacyl} hydroxyacetic acid, acifluorfen, 5-[2-chloro-4-(trifluoromethyl) phenoxy]-2-nitrobenzoate, 5-[2-chloro-4-(trifluoromethyl) phenoxy]-2-hydroxyl, and 3-chloro-4-hydroxyl benzotrifluoride. Identification of the metabolites allowed to propose the degradation pathway of fluoroglycofen ethyl by strain MBWY-1. The inoculation of strain MBWY-1 into soil treated with fluoroglycofen ethyl resulted in a higher fluoroglycofen ethyl degradation rate than in uninoculated soil regardless of whether the soil was sterilized or nonsterilized.