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Distinctions in hydrolysis and acidogenesis were examined for a series of anaerobic batch reactors inoculated with three different anaerobic mixed cultures (mesophilic, thermophilic and hyperthermophilic anaerobic sludge) and operated at the temperature of inoculum's origin and additionally at 70 °C. Hyperthermophilic temperatures led to increased hydrolysis rates during the start-up stage but a rapid drop in pH limited the overall hydrolysis efficiency, indicating the importance of pH control to sustain the high reaction rates at higher temperatures. No significant difference (P > 0.05) was observed among hydrolysis efficiencies obtained for different reactors which ranged between 27 ±â€¯3% and 40 ±â€¯14%. The highest fermentation yield of 0.44 g COD of fermentation products/g VSS-CODadded was obtained under thermophilic conditions, followed by mesophilic (0.33 g COD ferm. prod./g VSS-CODadded) and hyperthermophilic conditions (0.05-0.08 g COD ferm. prod./g VSS-CODadded). Fermentative performance was better at mesophilic and thermophilic conditions as indicated by improved production of volatile fatty acids (VFA). VFAs accounted for 60-71% of the solubilised matter at thermophilic and mesophilic conditions. Acetic acid formed the primary VFA (70%) at mesophilic temperatures, while butyric acid was the major VFA at thermophilic (60%) conditions. Hyperthermophilic conditions led to increased production of lactic acid, which comprised up to 32% of the solubilised matter. Overall, the results indicate that different operating temperatures may not significantly affect the substrate degradation efficiency but clearly influence the biotransformation pathways.

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As an alternative to applying the hydrothermal treatment to the raw algal feedstock before the anaerobic digestion (i.e. pre-treatment), one considered a post-treatment scenario where anaerobic digestion is directly used as the primary treatment while the hydrothermal treatment is thereafter applied to the digestate. Hydrothermal treatments such as wet oxidation (WetOx) and hydrothermal carbonization (HTC) were compared at a temperature of 200°C, for initial pressure of 0.1 and 0.82MPa, and no holding time after the process had reached the temperature setpoint. Both WetOx and HTC resulted in a substantial solids conversion (47-62% with HTC, 64-83% with WetOx, both at 0.82MPa) into soluble products, while some total chemical oxygen demand-based carbon loss from the solid-liquid phases was observed (20-39%). This generated high soluble products concentrations (from 6.2 to 10.9g soluble chemical oxygen demand/L). Biomethane potential tests showed that these hydrothermal treatments allowed for a 4-fold improvement of the digestate anaerobic biodegradability. The hydrothermal treatments increased the methane yield to about 200 LSTP CH4/kg volatile solids, when related to the untreated digestate, compared to 66 LSTP CH4/kg volatile solids, without treatment.

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Electricity generation in a microbial fuel cell (MFC) fed with carbon monoxide (CO) has been recently demonstrated; however, the microbial ecology of this system has not yet been described. In this work the diversity of the microbial community present at the anode of CO-fed MFCs was studied by performing denaturing gradient gel electrophoresis (DGGE) and high-throughput sequencing (HTS) analyses. HTS indicated a significant increase of the archaeal genus Methanobacterium and of the bacterial order Clostridiales, notably including Clostridium species, while in both MFCs DGGE identified members of the bacterial genera Geobacter, Desulfovibrio, and Clostridium, and of the archaeal genera Methanobacterium, Methanofollis, and Methanosaeta. In particular, the presence of Geobacter sulfurreducens was identified. Tolerance of G. sulfurreducens to CO was confirmed by growing G. sulfurreducens with acetate under a 100 % CO atmosphere. This observation, along with the identification of acetogens, supports the hypothesis of the two-step process in which CO is converted to acetate by the carboxidotrophic Bacteria and acetate is then oxidized by CO-tolerant electricigenic bacteria to produce electricity.

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This study demonstrates microbial electrolysis cell (MEC) scale-up from a 50mL to a 10L cell. Initially, a 50mL membraneless MEC with a gas diffusion cathode was operated on synthetic wastewater at different organic loads. It was concluded that process scale-up might be best accomplished using a "reactor-in-series" concept. Consequently, 855mL and 10L MECs were built and operated. By optimizing the hydraulic retention time (HRT) of the 855mL MEC and individually controlling the applied voltages of three anodic compartments with a real-time optimization algorithm, a COD removal of 5.7g L(R)(-1)d(-1) and a hydrogen production of 1.0-2.6L L(R)(-1)d(-1) was achieved. Furthermore, a two MECs in series 10L setup was constructed and operated on municipal wastewater. This test showed a COD removal rate of 0.5g L(R)(-1)d(-1), a removal efficiency of 60-76%, and an energy consumption of 0.9Whperg of COD removed.

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This study demonstrated electricity generation in a thermophilic microbial fuel cell (MFC) operated on synthesis gas (syngas) as the sole electron donor. At 50°C, a volumetric power output of 30-35 mWL(R)(-1) and a syngas conversion efficiency of 87-98% was achieved. The observed pathway of syngas conversion to electricity primarily consisted of a two-step process, where the carbon monoxide and hydrogen were first converted to acetate, which was then consumed by the anodophilic bacteria to produce electricity. A denaturing gradient gel electrophoresis (DGGE) analysis of the 16S rDNA revealed the presence of Geobacter species, Acetobacter, methanogens and several uncultured bacteria and archaea in the anodic chamber.

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Electricity generation in a microbial fuel cell (MFC) using carbon monoxide (CO) or synthesis gas (syngas) as a carbon source has been demonstrated recently. A major challenge associated with CO or syngas utilization is the mass transfer limitation of these sparingly soluble gases in the aqueous phase. This study evaluated the applicability of a dense polymer silicone membrane and thin wall silicone tubing for CO mass transfer in MFCs. Replacing the sparger used in our previous study with the membrane systems for CO delivery resulted in improved MFC performance and CO transformation efficiency. A power output and CO transformation efficiency of up to 18 mW LR(-1) (normalized to anode compartment volume) and 98%, respectively, was obtained. The use of membrane systems offers the advantage of improved mass transfer and reduced reactor volume, thus increasing the volumetric power output of MFCs operating on a gaseous substrate such as CO.

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This study investigates the influence of anode and cathode size and arrangement on hydrogen production in a membrane-less flat-plate microbial electrolysis cell (MEC). Protein measurements were used to evaluate microbial density in the carbon felt anode. The protein concentration was observed to significantly decrease with the increase in distance from the anode-cathode interface. Cathode placement on both sides of the carbon felt anode was found to increase the current, but also led to increased losses of hydrogen to hydrogenotrophic activity leading to methane production. Overall, the best performance was obtained in the flat-plate MEC with a two-layer 10 mm thick carbon felt anode and a single gas-diffusion cathode sandwiched between the anode and the hydrogen collection compartments.

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This study demonstrates enhanced methane production from wastewater in laboratory-scale anaerobic reactors equipped with electrodes for water electrolysis. The electrodes were installed in the reactor sludge bed and a voltage of 2.8-3.5 V was applied resulting in a continuous supply of oxygen and hydrogen. The oxygen created micro-aerobic conditions, which facilitated hydrolysis of synthetic wastewater and reduced the release of hydrogen sulfide to the biogas. A portion of the hydrogen produced electrolytically escaped to the biogas improving its combustion properties, while another part was converted to methane by hydrogenotrophic methanogens, increasing the net methane production. The presence of oxygen in the biogas was minimized by limiting the applied voltage. At a volumetric energy consumption of 0.2-0.3 Wh/L(R), successful treatment of both low and high strength synthetic wastewaters was demonstrated. Methane production was increased by 10-25% and reactor stability was improved in comparison to a conventional anaerobic reactor.

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This work evaluates the impact of the external resistance (electrical load) on the long-term performance of a microbial fuel cell (MFC) and demonstrates the real-time optimization of the external resistance. For this purpose, acetate-fed MFCs were operated at external resistances, which were above, below, or equal to the internal resistance of a corresponding MFC. A perturbation/observation algorithm was used for the real-time optimal selection of the external resistance. MFC operation at the optimal external resistance resulted in increased power output, improved Coulombic efficiency, and low methane production. Furthermore, the efficiency of the perturbation/observation algorithm for maximizing long-term MFC performance was confirmed by operating an MFC fed with synthetic wastewater for over 40 days. In this test an average Coulombic efficiency of 29% was achieved.

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Electricity production from carbon monoxide (CO) is demonstrated in a single chamber microbial fuel cell (MFC) with a CoTMPP-based air cathode. The MFC was inoculated with anaerobic sludge and continuously sparged with CO as a sole carbon source. Volumetric power output was maximized at a CO flow rate of 4.8LLR(-1)d(-1) reaching 6.4mWLR(-1). Several soluble and gaseous degradation products including hydrogen, methane, and acetate were detected, resulting in a relatively low apparent Coulombic efficiency of 8.7%. Tests also demonstrated electricity production from hydrogen and acetate with the highest and fastest increase in voltage exhibited after acetate injection. It is hypothesized that electricity generation in a CO-fed MFC is accomplished by a consortium of carboxydotrophic and carbon monoxide - tolerant anodophilic microorganisms.

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The treatment of reconstituted whey wastewater was performed in a 400 L digester at 20 degrees C, with an anaerobic digestion step, followed by a step of aerobic treatment at low oxygen concentration in the same digester. In a first set of 48 cycles, total cycle time (T(C)) of 2, 3 and 4 days were tested at varying organic loading rates (OLR). The COD removal reached 89+/-4, 97+/-3 and 98+/-2% at T(C) of 2, 3 and 4 days and OLR of 0.56, 1.04 and 0.78 g COD L(-1) d(-1), respectively. The activity of the biomass decreased for the methanogenic population, while increasing by 400% for the acidogens, demonstrating a displacement in the predominant trophic group in the biomass bed. A second set of 16 cycles was performed with higher soluble oxygen concentration in the bulk liquid (0.5 mg L(-1)) during the aerobic treatment at a T(C) of 2 days and an OLR of 1.55 g COD L(-1) d(-1), with a soluble COD removal of 88+/-3%. The biomass specific activities showed a compartmentalization of the trophic group with methanogenic activity maintained in the biomass bed and a high acidogenic activity in the suspended flocs.

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Methane production was studied in a laboratory-scale 10 L anaerobic upflow sludge bed (UASB) reactor with periodic variations of the reactor temperature. On a daily basis the temperature was varied between 35 and 45 degrees C or 35 and 55 degrees C with a heating period of 6 h. Each temperature increase was accompanied by an increase in methane production and a decrease in the concentration of soluble organic matter in the effluent. In comparison to a reactor operated at 35 degrees C, a net increase in methane production of up to 22% was observed. Batch activity tests demonstrated a tolerance of mesophilic methanogenic populations to short-term, 2-6 h, temperature increases, although activity of acetoclastic methanogens decreased after 6 h exposure to a temperature of 55 degrees C. 16S sequencing of DGGE bands revealed proliferation of temperature-tolerant Methanospirillum hungatii sp. in the reactor.

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In this study, an ADM1-based distributed parameter model was validated using experimental results obtained in a laboratory-scale 10 L UASB reactor. Sensitivity analysis of the model parameters was used to select four parameters for estimation by a numerical procedure while other parameters were accepted from ADM1 benchmark simulations. The parameter estimation procedure used measurements of liquid phase components obtained at different sampling points in the reactor and under different operating conditions. Model verification used real time fluorescence-based measurements of chemical oxygen demand and volatile fatty acids at four sampling locations in the reactor. Overall, the distributed parameter model was able to describe the distribution of liquid phase components in the reactor and adequately simulated the effect of external recirculation on degradation efficiency. The model can be used in the design, analysis and optimization of UASB reactors.

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The electrolytical methanogenic/methanotrophic coupling (eMaMoC) process was tested in a laboratory-scale single-stage reactor for the treatment of tetrachloroethene (PCE)-contaminated waters. A water electrolysis cell was placed directly in the effluent recirculation loop for the supply of both O2 and H2 to the system: H2 serving as the electron donor for both carbonate reduction into CH4 and reductive dechlorination. The concurrent presence of O2 and CH4 could be used by the methanotrophs for co-metabolically oxidising the chlorinated intermediates left over by the anaerobic transformation of PCE. At a PCE inlet of 33-52 microM and a hydraulic residence time (HRT) of 5.6 days, PCE reductive dechlorination to dichloroethene (DCE) was over 95% with a maximum DCE mineralisation of 83%. Fluorescence in situ hybridisation with 16S rRNA probes related to type I and type II methanotrophic bacteria were utilised to localise the methanotrophic communities in the anaerobic/aerobic granules. It evidenced that with operational time, along with increasing oxygenation rate, methanotrophic communities were specifically colonising onto the outermost layer of the anaerobic/aerobic granule.

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A proof of concept was performed in order to verify if the coupling of anaerobic and aerobic conditions inside the same digester could efficiently treat a reconstituted whey wastewater at 21 degrees C. The sequencing batch reactor (SBR) cycles combined initial anaerobic phase and final aerobic phase with reduced aeration. A series of 24 h cycles in 0.5 L digesters, with four different levels of oxygenation (none, 54, 108 and 182 mgO2 per gram of chemical oxygen demand (COD)), showed residual soluble chemical oxygen demand (sCOD) of 683 +/- 46, 720 +/- 33, 581 +/- 45, 1239 +/- 15 mg L(-1), respectively. Acetate and hydrogen specific activities were maintained for the anaerobic digester, but decreased by 10-25% for the acetate and by 20-50% for the hydrogen, in the coupled digesters. The experiment was repeated using 48 h cycles with limited aeration during 6 or 16 hours at 54 and 108 mgO2gCODinitial(-1), displaying residual sCOD of 177 +/- 43, 137 +/- 38, 104 +/- 22 and 112 +/- 9 mgL(-1) for the anaerobic and the coupled digesters, respectively. The coupled digesters recovered after a pH shock with residual sCOD as low as 132 mg L(-1) compared to 636 mg L(-1) for the anaerobic digester. With regard to the obtained results, the feasibility of the anaerobic-aerobic coupling in SBR digesters for the treatment of whey wastewater was demonstrated.

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Anaerobic digestion model no.1 (ADM1) was used for tuning and performance analysis of the multi-model observer based estimator (mmOBE). The mmOBE was based on the variable structure model (VSM) of the anaerobic digestion model, which consists of several local submodels, each of which describes a typical process state. Depending on the hydraulic retention time, ADM1 simulated the methanogenic, organic overload, and acidogenic states of the process. These simulations allowed for optimising tunable parameters of the mmOBE. Owing to relatively slow process dynamics, a data acquisition interval as large as one day was sufficient to obtain acceptable accuracy. The simulations of mmOBE performance showed excellent rate of mmOBE convergence to ADM1 outputs. Moreover, mmOBE successfully estimated key kinetic parameters, such as maximal transformation rates of CODs, VFAs, and methane. These estimations can be used in the development of the advanced knowledge-based process system, which uses both available measurements and estimations of key kinetic parameters for extended diagnosis of failures and process trend analysis.

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Observer-based estimators (OBE) were used for estimation of state variables and kinetic parameters in an anaerobic digestion (AD) process. A simplified first-order model with time-varying kinetic parameters was used to design an OBE for kinetic parameter estimation. This approach was validated on a laboratory-scale anaerobic reactor equipped with a multiwavelength fluorometer for on-line measurements of COD and VFA concentrations in the reactor effluent. The proposed estimators provide continuous adjustment of kinetic parameters and can be used for predictions of state variables between samples acquisition and during sensor failure.

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In this work, rhodamine-related fluorescence was measured on-line at four reactor heights in order to study hydrodynamics within an upflow anaerobic sludge bed reactor. A linear dependence of the dispersion coefficient (D) on the upflow velocity was observed, while the influence of the organic loading rate (OLR) was insignificant. Furthermore, the Bodenstein number of the reactor loaded with granulated sludge was found to be position-dependent with the largest values measured at the bottom of the sludge bed. This trend was not observed in the reactor without sludge. Chemical oxygen demand (COD) and volatile fatty acid (VFA) concentrations were measured at the same reactor heights as in rhodamine tests using conventional off-line analytical methods and on-line multiwavelength fluorometry. Significant spatial COD and VFA gradients were observed at organic loading rates above 6g COD l(R)(-1)d(-1) and linear upflow velocities below 0.8m h(-1).

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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.

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Applicability of multi-wavelength fluorometry for anaerobic digestion process monitoring was investigated in a 3.5 L upflow anaerobic sludge bed (UASB) lab-scale reactor. Both off-line and on-line monitoring of key process parameters was tested. Off-line emission spectra were measured at an angle of 90 degrees to the excitation beam using a cuvette. On-line measurements were carried out using a fiber optic probe in the external recirculation line of the digester. Fluorescence spectra were correlated to available analytical measurements to obtain partial least square regression models. An independent set of measurements was used to validate the regression models. Model estimations showed reasonable agreement with analytical measurements with multiple determination coefficients (R2) between 0.6 and 0.95. Results showed that offline fluorescence measurements can be used for fast estimation of anaerobic digestor effluent quality. At the same time, the on-line implementation of multi-wavelength fluorescence measurements can be used for realtime process monitoring and, potentially, for on-line process control.

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