Metabolic interactions in methanogenic and sulfate-reducing bioreactors.

In environments where the amount of electron acceptors is insufficient for complete breakdown of organic matter, methane is formed as the major reduced end product. In such methanogenic environments organic acids are degraded by syntrophic consortia of acetogenic bacteria and methanogenic archaea. Hydrogen consumption by methanogens is essential for acetogenic bacteria to convert organic acids to acetate and hydrogen. Several syntrophic cocultures growing on propionate and butyrate have been described. These syntrophic fatty acid-degrading consortia are affected by the presence of sulfate. When sulfate is present sulfate-reducing bacteria compete with methanogenic archaea for hydrogen and acetate, and with acetogenic bacteria for propionate and butyrate. Sulfate-reducing bacteria easily outcompete methanogens for hydrogen, but the presence of acetate as carbon source may influence the outcome of the competition. By contrast, acetoclastic methanogens can compete reasonably well with acetate-degrading sulfate reducers. Sulfate-reducing bacteria grow much faster on propionate and butyrate than syntrophic consortia.     

Anode and cathode materials characterization for a microbial fuel cell in half cell configuration

Microbial fuel cells (MFCs) are novel bioelectrochemical devices for spontaneous conversion of biomass into electricity through the metabolic activity of the bacteria. Microbial production of electricity may become an important source of bioenergy in future because MFCs offer the possibility of extracting electric current from a wide range of soluble or dissolved complex organic wastes and renewable biomass. However, the materials used in these devices are still not economic and researchers use different materials as cathode and anode in MFCs. This results in variable performance which is difficult to compare. We tested several commercially available materials for their suitability as anode in an acetate fed MFC. Besides, a novel non-platinized activated carbon (AC) based, gas porous air cathode was also tested. Both the anode and cathode were tested in a half cell configuration. Carbon cloth, graphite cloth and dynamically stable anode (DSA) served as ideal anode material with carbon cloth and graphite mesh reaching the open circuit voltage (OCV) of acetate oxidation (-500 mV vs. Ag/AgCl). The effect of increasing concentration of acetate on anode OCV was also investigated and results showed that on increasing the acetate concentration from 10 mM to 40 mM has no adverse impact on the anodic activity towards electrochemical oxidation of acetate. The AC cathode showed stable current (-1.2 mA/cm2) over a period of 100 days.     

Modelling the effect of the antimicrobial tylosin on the performance of an anaerobic sequencing batch reactor.

A laboratory-scale anaerobic sequencing batch reactor (ASBR) was fed a synthetic wastewater containing glucose to study the effects of the antimicrobial tylosin on treatment performance. Measurements of methane, volatile fatty acids, and COD concentrations suggested that the addition of 1.67 mg/L and 167 mg/l of tylosin to the synthetic wastewater inhibited propionate oxidizing syntrophic bacteria and aceticlastic methanogens. The latter is presumed to be an indirect effect. A modified version of the IWA Anaerobic Digestion Model No. 1 (ADM1) with extensions for microbial storage and hydrolysis of reserve carbohydrates, and tylosin liquid-solid mass transfer and inhibition adequately described the dynamic profiles observed in the ASBR.     

An extension of the Anaerobic Digestion Model No. 1 to include the effect of nitrate reduction processes.

Nitrate reduction processes were incorporated into the IWA Anaerobic Digestion Model No. 1 (ADM1) in order to account for the effect of such processes on fermentation and methanogenesis. The general structure of the ADM1 was not changed except for modifications related to disintegration and hydrolysis of complex organic matter and decayed biomass. A fraction of butyrate/valerate and propionate degraders was assumed to be the fermentative denitrifiers carrying out fermentation in the absence of N-oxides. Nitrate reduction proceeded in a stepwise manner to nitrite, nitric oxide, nitrous oxide and nitrogen gas using four substrates as electron and/or carbon source. The utilization of the four substrates and N-oxides was based on stoichiometry and kinetics. The inhibitory effect of N-oxides on the methanogens was accounted for by the use of non-competitive inhibition functions. Model simulations were compared with experimental data obtained with a batch, mixed fermenting and methanogenic culture amended with various initial nitrate concentrations.     

Effect of operating temperature on performance of microbial fuel cell

The performance of dual chambered mediator-less microbial fuel cell (MFC) operated under batch mode was evaluated under different operating temperatures, ranging between 20 and 55 degrees C, with step increase in temperature of 5 degrees C. Synthetic wastewater with sucrose as carbon source having chemical oxygen demand (COD) of 519-555 mg/L was used in the study. Temperature was a crucial factor in the performance of MFCs for both COD removal and electricity production. The MFC demonstrated highest COD removal efficiency of 84% and power density normalized to the anode surface area of 34.38 mW/m2 at operating temperature of 40 degrees C. Higher VSS to SS ratio was observed at the operating temperature between 35 and 45 degrees C. Under different operating temperatures the observed sludge yield was in the range of 0.05 to 0.14 g VSS/g COD removed. The maximum Coulombic and energy efficiencies were obtained at 40 degrees C, with values of 7.39 and 13.14%, respectively. Internal resistance of the MFC decreased with increase in operating temperature. Maximum internal resistance of 1,150 omega was observed when the MFC was operated at 20 degrees C; whereas the minimum internal resistance (552 omega) was observed at 55 degrees C.     

Mathematical model for microbial fuel cells with anodic biofilms and anaerobic digestion.

This study describes the integration of IWA's anaerobic digestion model (ADM1) within a computational model of microbial fuel cells (MFCs). Several populations of methanogenic and electroactive microorganisms coexist suspended in the anolyte and in the biofilm attached to the anode. A number of biological, chemical and electrochemical reactions occur in the bulk liquid, in the biofilm and at the electrode surface, involving glucose, organic acids, H2 and redox mediators. Model output includes the evolution in time of important measurable MFC parameters (current production, consumption of substrates, suspended and attached biomass growth). Two- and three-dimensional model simulations reveal the importance of current and biomass heterogeneous distribution over the planar anode surface. Voltage- and power-current characteristics can be calculated at different moments in time to evaluate the limiting regime in which the MFC operates. Finally, model simulations are compared with experimental results showing that, in a batch MFC, smaller electrical resistance of the circuit leads to selection of electroactive bacteria. Higher coulombic yields are so obtained because electrons from substrate are transferred to anode rather than following the methanogenesis pathway. In addition to higher currents, faster COD consumption rates are so achieved. The potential of this general modelling framework is in the understanding and design of more complex cases of wastewater-fed microbial fuel cells.     

Multi-walled carbon nanotubes as electrode material for microbial fuel cells

The microbial fuel cell (MFC) is a novel and innovative technology that could allow direct harvesting of energy from wastewater through microbial activity with simultaneous oxidation of organic matter in wastewater. Among all MFC parts, electrode materials play a crucial role in electricity generation. A variety of electrode materials have been used, including plain graphite, carbon paper and carbon cloth. However, these electrode materials generated only limited electricity or power. Recently, many research studies have been conducted on carbon nanotubes (CNTs) because of their unique physical and chemical properties that include high conductivity, high surface area, corrosion resistance, and electrochemical stability. These properties make them extremely attractive for fabricating electrodes and catalyst supports. In this study, CNT-based electrodes had been developed to improve MFC performance in terms of electricity generation and treatment efficiency. Multi-walled carbon nanotubes (MWCNTs) with carboxyl groups have been employed to fabricate electrodes for single-chamber air-cathode MFCs. The quality of the prepared MWCNTs-based electrodes was evaluated by morphology, electrical conductivity and specific surface area using a field emission scanning electron microscope, four-probe method and Brunauer-Emmerr-Teller method, respectively. The performance of MFCs equipped with MWCNT-based electrodes was evaluated by chemical analysis and electrical monitoring and calculation. In addition, the performance of these MFCs, using MWCNTs as electrodes, was compared against that using commercial carbon cloth.     

Thermal pretreatment of the solid fraction of manure: impact on the biogas reactor performance and microbial community.

Application of thermal treatment at 100-140 degrees C as a pretreatment method prior to anaerobic digestion of a mixture of cattle and swine manure was investigated. In a batch test, biogasification of manure with thermally pretreated solid fraction proceeded faster and resulted in the increase of methane yield. The performances of two thermophilic continuously stirred tank reactors (CSTR) treating manure with solid fraction pretreated for 40 minutes at 140 degrees C and non-treated manure were compared. The digester fed with the thermally pretreated manure had a higher methane productivity and an improved removal of the volatile solids (VS). The properties of microbial communities of both reactors were analysed. The specific methanogenic activity (SMA) test showed that both biomasses had significant activity towards hydrogen and formate, while the activity with the VFA - acetate, propionate and butyrate - was low. The kinetic parameters of the VFA conversion revealed a reduced affinity of the microbial community from the CSTR fed with thermally pre-treated manure for acetate, propionate and butyrate. The bacterial and archaeal populations identified by t-RLFP analysis of 16S rRNA genes were found to be identical in both systems. However, a change in the abundance of the species present was detected.     

Identification of prevalent microbial communities in a municipal solid waste landfill.

To identify the microbial communities in Istanbul, Odayeri Municipal Solid Waste Landfill, leachate samples were collected from different sections at different stabilization phases. In identification of microbial communities in leachate samples, molecular techniques such as FISH, DGGE and cloning based on 16S rRNA and mcrA genes were used. As the chemical and microbiological compositions of the samples were compared, obvious correlations were found between the stability of the landfill section and abundance of active methanogens. On the other hand, there were considerable differences between acidogenic and mature leachate samples in DGGE profiles of archaeal and bacterial 16S rRNA genes. Moreover, in acidogenic leachate samples having BOD5/COD ratio of about 0.5 acetate utilizing Methanosarcina and Methanosaeta species were intensively detected in FISH. Although only very few H2-utilizing methanogens were identified with FISH analysis, most of the clones isolated from mature leachate samples clustered within H2-utilizing Methanobacteriales and Methanomicrobiales according to phylogenetic analysis of 16S rRNA and mcrA clones, respectively.     

Sulfide production and wastewater quality in pressure mains

An empirical model for predicting sulfide production in pressure mains (Hvitved-Jacobsen et al., 1988) was evaluated and modified based on results obtained from two intercepting pressure mains located in the Northern part of Jutland, Denmark. Mass balances in pipe influent and effluent were made for volatile fatty acids, VFA (formate, acetate, propionate and butyrate), dissolved COD, DOC and sulfide and biofilm surface rate for sulfide and organic matter were calculated. Relatively high sulfide formation rates were observed at low temperatures (5–12°C). The sulfide production rate strongly depended on wastewater quality in terms of VFA and dissolved carbohydrate concentration. Based on these two sets of observations — wastewater quality and temperature — the original empirical model was modified.     

Comparative 16S rRNA gene surveys of granular sludge from three upflow anaerobic bioreactors treating purified terephthalic acid (PTA) wastewater

The microbial communities from three upflow anaerobic bioreactors treating purified terephthalic acid (PTA) wastewater were characterized with 16S ribosomal RNA gene sequencing surveys. Universal bacterial and archaeal primers were used to compare the bioreactor communities to each other. A total of 1,733 bacterial sequences and 383 archaeal sequences were characterized. The high number of Syntrophus spp. and Pelotomaculum spp. found within these reactors indicates efficient removal of benzoate and terephthalate. Under anaerobic conditions benzoate can be degraded through syntrophic associations between these bacteria and hydrogen-scavenging microbes, such as Desulfovibrio spp. and hydrogenotrophic methanogens, which remove H(2) to force the thermodynamically unfavourable reactions to take place. The authors did not observe a relatively high percentage of hydrogenotrophic methanogens with the archaeal gene survey because of a high acetate flux (acetate is a main component in PTA wastewater and is the main degradation product of terephthalate/benzoate fermentation), and because of the presence of Desulfovibrio spp. (a sulfate reducer that scavenges hydrogen). The high acetate flux also explains the high percentage of acetoclastic methanogens from the genus Methanosaeta among the archaeal sequences. A group of uncultured bacteria (OD1) may be involved in the degradation of p-toluate (4-methyl benzoate), which is a component of PTA wastewater.     

Polyhydroxyalkanoates production by activated sludge in a SBR using acetate and propionate as carbon sources.

In this work, sludge was submitted to aerobic dynamic substrate feeding. Two sequencing batch reactors were operated, with acetate or propionate as carbon substrates. When acetate was used the system only produced a homopolymer of polyhydroxybutyrate (PHB). In order to maximize the PHB production, tests with different concentrations of acetate and ammonia were preformed. The best results (67.2% of PHB by cell dry weight) were obtained for 0.7 Nmmol/l of ammonia and 180 Cmmol/l of acetate. The PHB cell content was further improved by pulse addition of substrate, three times 60 Cmmol/l of acetate, reaching a value of 78.5%. Propionate can be used as a precursor for hydroxyvalerate. In conjunction with other substrates, it allows for the formation of copolymers, which present better processing properties on commercial applications. Tests with different concentrations of propionate and ammonia were performed. Under the operating conditions used, the maximum PHA accumulated inside cells was 34.8%, with 30 Cmmol/l of propionate and no ammonia.     

Simultaneous sewage treatment and electricity generation in membrane-less microbial fuel cell

Long term performance of mediator-less and membrane-less microbial fuel cell (ML-MFC) was evaluated for treatment of synthetic and actual sewage and electricity harvesting. The anode chamber of ML-MFC was inoculated with pre-heated mixed anaerobic sludge collected from a septic tank. The ML-MFC was operated by feeding synthetic wastewater for first 244 days, under different organic loading rates, and later with actual sewage for next 30 days. Maximum chemical oxygen demand (COD) removal efficiency of 91.4% and 82.7% was achieved while treating synthetic wastewater and actual sewage, respectively. Maximum current of 0.33 mA and 0.17 mA was produced during synthetic and actual sewage treatment, respectively. Maximum power density of 6.73 mW/m(2) (13.65 mW/m(3)) and maximum current density of 70.74 mA/m(2) was obtained in this membrane-less MFC with successful organic matter removal from wastewater.     

Anaerobic biodegradability of Tween surfactants used as a carbon source for the microbial reductive dechlorination of hexachlorobenzene.

Three structurally-related, nonionic, polysorbate surfactants (Tween 60, 61, and 65) were used as the sole carbon source to sustain the microbial, sequential reductive dechlorination of hexachlorobenzene (HCB) in a mixed, methanogenic culture derived from a contaminated estuarine sediment. The surfactants were partially degraded and fermented to methane with no measurable accumulation of volatile fatty acids, indicating that methanogenesis was rapid relative to the rates of hydrolysis and acidogenesis. Addition of the methanogenesis inhibitor 2-bromoethanesulfonic acid resulted in acetate accumulation without impact on the sequential dechlorination of HCB. An anaerobic biodegradability assay was performed and the following data were obtained for the Tween 60, 61, and 65, respectively: 53, 62, and 62% COD destruction; 35, 57, and 48% COD to methane conversion; and 38, 38, and 45% COD to acetate conversion. These data suggest that the hydrophobic moiety (stearate) of the surfactants was preferentially degraded, most likely through beta-oxidation, to acetate and ultimately to methane and carbon dioxide. Between 38 and 47% of the initial surfactant COD remained after 46 d incubation, which most likely corresponds to the hydrophilic polyoxyethylene moiety. An anaerobic biodegradation pathway of the Tween surfactants is proposed.     

Activity of sulphate reducing bacteria according to COD/SO4(2-) ratio of acrylonitrile wastewater containing high sulphate.

This study was performed to evaluate the biodegradability of acrylonitrile wastewater, microbial inhibition effect of acrylonitrile wastewater on removal efficiency and the activity of sulphate reducing bacteria (SRB) according to COD/sulphate ratio. Acrylonitrile wastewater was hardly biodegradable in a biodegradability test, however, SRB activity was 57% for overall consumption of electron donor and it was relatively high value compared to 17% of reference test with glucose. COD removal of acrylonitrile wastewater was improved to 57% and 61% from 20% as the COD/sulphate ratio were 0.5 and 0.3 by sulphate addition to acrylonitrile wastewater. First order reaction rate constants k on organic removal of acrylonitrile wastewater were 0.001, 0.004 and 0.004 at each COD/sulphate ratio of 0.9, 0.5 and 0.3. Thus it was suggested that the activity of SRB was a significant factor for removing organics and sulphate simultaneously in acrylonitrile wastewater.     

Biohydrogen production by mesophilic fermentation of food wastewater.

Batch experiments were conducted to convert molasses wastewater (10-160 g COD/L) into hydrogen at 35 degrees C at various pH (4-8). The maximum hydrogen productivity (HP) and hydrogen production rate (HPR) reached 47.1 mmol-H2/g COD(re) and 97.5 mmol-H2/L-day, respectively, at a substrate concentration of 40 g COD/L and pH 6.0. The methane-free biogas contained up to 50% (v/v) of hydrogen. Fermentation at wastewater concentrations higher than 60 g COD/L required a long acclimation period (more than 20 h). Though the fractions were substrate concentration and pH-dependent, acetate and butyrate were the two main liquid fermentation products. A comparison of the HP and HPR data indicates that defining a hydrogen yield indicator to evaluate hydrogen generation efficiency should be taken into consideration in practical fermentation operations.     

The role of sulphate reduction on the reductive S decolorization of the azo dye reactive orange 14.

The aim of this study was to investigate the impact of a broad range of sulphate concentrations (0-10g SO4(-2) L(-1)) on the reduction of an azo dye (reactive orange 14 (RO14)) by an anaerobic sludge. An increase in the sulphate concentration generally stimulated the reduction of RO14 by sludge incubations supplemented with glucose, acetate or propionate as electron donor. Sulphate and azo dye reductions took place simultaneously in all incubations. However, there was a decrease on the rate of decolorization when sulphate was supplied at 10g SO4(-2) L(-1). Abiotic incubations at different sulphide concentrations (0-2.5 g sulphide L(-1)) promoted very poor reduction of RO14. However, addition of riboflavin (20 microM), as a redox mediator, accelerated the reduction of RO14 up to 44-fold compared to a control lacking the catalyst. Our results indicate that sulphate-reduction may significantly contribute to the reduction of azo dyes both by biological mechanisms and by abiotic reductions implicating sulphide as an electron donor. The contribution of abiotic decolorization by sulphide, however, was only significant when a proper redox mediator was included. Our results also revealed that sulphate-reduction can out-compete with azo reduction at high sulphate concentrations leading to a poor decolorising performance when no sufficient reducing capacity is available.     

微生物燃料电池输出功率影响因素综述

微生物燃料电池(MFC)是一种通过微生物的催化作用将有机物中的化学能直接转化为电能的生物反应装置。本文在MFC产电原理的基础上,结合已有的研究结果综合叙述了微生物燃料电池中微生物的生物催化活性、产电底物、阳极材料及构造、阴极电子受体、膜材料及面积、电极间距和操作条件对MFC产电性能的影响。     

Microbial community analysis with a high PHA storage capacity.

Activated sludge was submitted to aerobic dynamic substrate feeding for the production of biodegradable plastics. Two sequencing batch reactors were operated with acetate or propionate as sole carbon substrates. With acetate a homopolymer of polyhydroxybutyrate (PHB) was obtained and with propionate a copolymer of hydroxybutyrate and hydroxyvalerate P(HB/HV) was produced. Three main morphotypes were identified in both sludges: two belong to the Alphaproteobacteria class and the third to the Betaproteobacteria class. Bacilli belonging to Betaproteobacteria were shown by FISH analysis, applied in combination with Nile Blue post-staining, to be the main responsible for PHAs storage. The latter were affiliated to Azoarcus genus within Betaproteobacteria.     

Syntrophic acetate oxidation in two-phase (acid-methane) anaerobic digesters

The microbial processes involved in two-phase anaerobic digestion were investigated by operating a laboratory-scale acid-phase (AP) reactor and analyzing two full-scale, two-phase anaerobic digesters operated under mesophilic (35 °C) conditions. The digesters received a blend of primary sludge and waste activated sludge (WAS). Methane levels of 20% in the laboratory-scale reactor indicated the presence of methanogenic activity in the AP. A phylogenetic analysis of an archaeal 16S rRNA gene clone library of one of the full-scale AP digesters showed that 82% and 5% of the clones were affiliated with the orders Methanobacteriales and Methanosarcinales, respectively. These results indicate that substantial levels of aceticlastic methanogens (order Methanosarcinales) were not maintained at the low solids retention times and acidic conditions (pH 5.2-5.5) of the AP, and that methanogenesis was carried out by hydrogen-utilizing methanogens of the order Methanobacteriales. Approximately 43, 31, and 9% of the archaeal clones from the methanogenic phase (MP) digester were affiliated with the orders Methanosarcinales, Methanomicrobiales, and Methanobacteriales, respectively. A phylogenetic analysis of a bacterial 16S rRNA gene clone library suggested the presence of acetate-oxidizing bacteria (close relatives of Thermacetogenium phaeum, 'Syntrophaceticus schinkii,' and Clostridium ultunense). The high abundance of hydrogen consuming methanogens and the presence of known acetate-oxidizing bacteria suggest that acetate utilization by acetate oxidizing bacteria in syntrophic interaction with hydrogen-utilizing methanogens was an important pathway in the second-stage of the two-phase digestion, which was operated at high ammonium-N concentrations (1.0 and 1.4 g/L). A modified version of the IWA Anaerobic Digestion Model No. 1 (ADM1) with extensions for syntrophic acetate oxidation and weak-acid inhibition adequately described the dynamic profiles of volatile acid production/degradation and methane generation observed in the laboratory-scale AP reactor. The model was validated with historical data from the full-scale digesters.