Performance and microbial community analysis of two-stage process with extreme thermophilic hydrogen and thermophilic methane production from hydrolysate in UASB reactors

The two-stage process for extreme thermophilic hydrogen and thermophilic methane production from wheat straw hydrolysate was investigated in up-flow anaerobic sludge bed (UASB) reactors. Specific hydrogen and methane yields of 89 ml-H₂/g-VS (190 ml-H₂/g-sugars) and 307 ml-CH₄/g-VS, respectively were achieved simultaneously with the overall VS removal efficiency of 81% by operating with total hydraulic retention time (HRT) of 4 days . The energy conversion efficiency was dramatically increased from only 7.5% in the hydrogen stage to 87.5% of the potential energy from hydrolysate, corresponding to total energy of 13.4 kJ/g-VS. Dominant hydrogen-producing bacteria in the H₂-UASB reactor were Thermoanaerobacter wiegelii, Caldanaerobacter subteraneus, and Caloramator fervidus. Meanwhile, the CH₄-UASB reactor was dominated with methanogens of Methanosarcina mazei and Methanothermobacter defluvii. The results from this study suggest the two stage anaerobic process can be effectively used for energy recovery and for stabilization of hydrolysate at anaerobic conditions.     

Enhancement of bioenergy production from organic wastes by two-stage anaerobic hydrogen and methane production process

The present study investigated a two-stage anaerobic hydrogen and methane process for increasing bioenergy production from organic wastes. A two-stage process with hydraulic retention time (HRT) 3 d for hydrogen reactor and 12 d for methane reactor, obtained 11% higher energy compared to a single-stage methanogenic process (HRT 15 d) under organic loading rate (OLR) 3 gVS/(L d). The two-stage process was still stable when the OLR was increased to 4.5 gVS/(L d), while the single-stage process failed. The study further revealed that by changing the HRT_hydrogen:HRT_methane ratio of the two-stage process from 3:12 to 1:14, 6.7%, more energy could be obtained. Microbial community analysis indicated that the dominant bacterial species were different in the hydrogen reactors (Thermoanaerobacterium thermosaccharolyticum-like species) and methane reactors (Clostridiumthermocellum-like species). The changes of substrates and HRT did not change the dominant species. The archaeal community structures in methane reactors were similar both in single- and two- stage reactors, with acetoclastic methanogens Methanosarcina acetivorans-like organisms as the dominant species.     

A new algorithm to characterize biodegradability of biomass during anaerobic digestion: influence of lignin concentration on methane production potential

We examined the influence of fibrous fractions of biomass on biochemical methane potential (BMP) with the objective of developing an economical and easy-to-use statistical model to predict BMP, and hence the biodegradability of organic material (BD) for biogas production. The model was developed either for energy crops (grass, maize, and straw) or for animal manures, or as a combined model for these two biomass groups. It was found that lignin concentration in volatile solids (VS) was the strongest predictor of BMP for all the biomass samples. The square of the sample correlation coefficient (R(2)) from the BMP versus lignin was 0.908 (p<0.0001), 0.763 (p<0.001) and 0.883 (p<0.001) for animal manure, energy crops and the combined model, respectively. Validation of the combined model was carried out using 65 datasets from the literature.     

Co-production of hydrogen and methane from potato waste using a two-stage anaerobic digestion process

Hydrogen and methane co-production from potato waste was examined using a two-stage process of anaerobic digestion. The hydrogen stage was operated in continuous flow under a pH of 5.5 and a HRT of 6h. The methane stage was operated in both continuous and semi-continuous flows under HRTs of 30 h and 90 h, respectively, with pH controlled at 7. A maximum gas production rate of 270 ml/h and an average of 119 ml/h were obtained from the hydrogen stage during the operation over 110 days. The hydrogen concentration contained in the gas was 45% (v/v), on average. The maximum and average gas production rates observed from methane reactor during the 74 days of semi-continuous flow operation were 187 and 141 ml/h, respectively, with an average methane concentration of 76%. Overall, 70% of VS, 64% of total COD in the feedstock were removed. The hydrogen and methane yields from the potato waste were 30 l/kg TS (with a maximum of 68 l/kg) and 183 l/kg TS (with a maximum of 225 l/kg), respectively. The total energy yield obtained was 2.14 kW h/kg TS, with a maximum of 2.74 kW h/kg TS.     

Methane production and methane consumption: a review of processes underlying wetland methane fluxes

Potential rates of both methane production and methane consumptionvary over three orders of magnitude and their distribution is skew.These rates are weakly correlated with ecosystem type, incubationtemperature, in situ aeration, latitude, depth and distanceto oxic/anoxic interface. Anaerobic carbon mineralisation is amajor control of methane production. The large range in anaerobicCH₄:CO₂ production rates indicate that a largepart of the anaerobically mineralised carbon is used for reduction ofelectron acceptors, and, hence, is not available for methanogenesis.Consequently, cycling of electron acceptors needs to be studied tounderstand methane production. Methane and oxygen half saturationconstants for methane oxidation vary about one order of magnitude.Potential methane oxidation seems to be correlated withmethanotrophic biomass. Therefore, variation in potential methaneoxidation could be related to site characteristics with a model ofmethanotrophic biomass.     

Eliminating methanogenic activity in hydrogen reactor to improve biogas production in a two-stage anaerobic digestion process co-digesting municipal food waste and sewage sludge

Laboratory scale two-stage anaerobic digestion process model was operated for 280 days to investigate the feasibility to produce both hydrogen and methane from a mixture feedstock (1:1 (v/v)) of municipal food waste and sewage sludge. The maximum hydrogen and methane yields obtained in the two stages were 0.93 and 9.5 mL/mL feedstock. To eliminate methanogenic activity and obtain substantial hydrogen production in the hydrogen reactor, both feedstock and mixed liquor required treatment. The heat treatment (100 °C, 10 min) for feedstock and a periodical treatment (every 2-5 weeks, either heating, removal of biomass particles or flushing with air) for mixed liquor were effective in different extent. The methane production in the second stage was significantly improved by the hydrogen production in the first stage. The maximum methane production obtained in the period of high hydrogen production was more than 2-fold of that observed in the low hydrogen production period.     

Processes involved in formation and emission of methane in rice paddies

The seasonal change of the rates of production and emission of methane were determined under in-situ conditions in an Italian rice paddy in 1985 and 1986. The contribution to total emission of CH₄ of plant-mediated transport, ebullition, and diffusion through the flooding water was quantified by cutting the plants and by trapping emerging gas bubbles with funnels. Both production and emission of CH₄ increased during the season and reached a maximum in August. However, the numbers of methanogenic bacteria did not change. As the rice plants grew and the contribution of plant-mediated CH₄ emission increased, the percentage of the produced CH₄ which was reoxidized and thus, was not emitted, also increased. At its maximum, about 300 ml CH₄ were produced per m² per hour. However, only about 6% were emitted and this was by about 96% via plant-mediated transport. Radiotracer experiments showed that CH, was produced from H₂/CO₂. (30–50%) and from acetate. The pool concentration of acetate was in the range of 6–10 mM. The turnover time of acetate was 12–16 h. Part of the acetate pool appeared to be not available for production of CH₄ or CO₂     

Batch anaerobic co-digestion of cow manure and waste milk in two-stage process for hydrogen and methane productions

Anaerobic co-digestion of cow manure (CM) and waste milk (WM), produced by sick cows during treatment with antibiotics, was evaluated in two-stage process under thermophilic condition (55 °C) to determine the effect of WM addition on hydrogen (H₂) and methane (CH₄) production potentials, volatile solids (VS) removal, and energy recovery. Six CM to WM VS ratios of 100:0, 90:10, 70:30, 50:50, 30:70, and 10:90 were examined using 1-L batch digesters. The WM VS ratio of 30 % was found to be the minimum limit for significant increases in specific H₂ and CH₄ yields, and VS removal as compared to digestion of manure alone (P < 0.05). The highest specific H₂ and CH₄ yields, VS removal and energy yield were 38.2 mL/g VS, 627.6 mL/g VS, 78.4 % and 25,459.8 kJ/kg VS, respectively, in CM:WM 30:70. Lag phases to H₂ and CH₄ productions were observed in CM–WM mixtures, increased with increasing the amount of WM in the feedstock and were greater than 72 h in CM:WM 50:50 and 30:70. The digestion system failed in CM:WM 10:90. The results suggest that CM:WM 30:70 was optimum, however, due to limited amount of WM usually generated and long lag phase at this ratio which may make the process uneconomical, CM:WM 70:30 is recommended in practice.     

Characteristics of hydrogen and methane production from cornstalks by an augmented two- or three-stage anaerobic fermentation process

This paper presents the co-production of hydrogen and methane from cornstalks by a two- or three-stage anaerobic fermentation process augmented with effective artificial microbial community. Two-stage fermentation by using the anaerobic sludge and DGGE analysis showed that effective and stable strains should be introduced into the system. We introduced Enterobacter aerogens or Clostridium paraputrificum into the hydrogen stage, and C. paraputrificum was proven to be more effective. In the three-stage process consisting of the improved hydrolysis, hydrogen and methane production stages, the highest soluble sugars (0.482 kg/kg cornstalks) were obtained after the introduction of Clostridium thermocellum in the hydrolysis stage, under the thermophilic (55 °C) and acidic (pH 5.0) conditions. Hydrolysates from 1 kg of cornstalks could produce 2.61 mol (63.7 l) hydrogen by augmentation with C. paraputrificum and 4.69 mol (114.6 l) methane by anaerobic granular sludge, corresponding to 54.1% energy recovery.     

Studies on potential effects of fumaric acid on rumen microbial fermentation,methane production and microbial community

The greenhouse gas methane (CH₄) contributes substantially to global climate change. As a potential approach to decrease ruminal methanogenesis, the effects of different dosages of fumaric acid (FA) on ruminal microbial metabolism and on the microbial community (archaea, bacteria) were studied using a rumen simulation technique (RUSITEC). FA acts as alternative hydrogen acceptor diverting 2H from methanogenesis of archaea towards propionate formation of bacteria. Three identical trials were conducted with 12 fermentation vessels over a period of 14 days. In each trial, four fermentation vessels were assigned to one of the three treatment groups differing in FA dosage: low fumaric acid (LFA), high fumaric acid (HFA) and without FA (control). FA was continuously infused with the buffer. Grass silage and concentrate served as substrate. FA led to decreases in pH and to higher production rates of total short chain fatty acids (SCFA) mediated by increases in propionate for LFA of 1.69 mmol d^?1 and in propionate and acetate production for HFA of 4.49 and 1.10 mmol d^?1, respectively. Concentrations of NH₃-N, microbial crude protein synthesis, their efficiency, degradation of crude nutrients and detergent fibre fraction were unchanged. Total gas and CH₄ production were not affected by FA. Effects of FA on structure of microbial community by means of single strand conformation polymorphism (SSCP) analyses could not be detected. Given the observed increase in propionate production and the unaffected CH₄ production it can be supposed that the availability of reduction equivalents like 2H was not limited by the addition of FA in this study. It has to be concluded from the present study that the application of FA is not an appropriate approach to decrease the ruminal CH₄ production.     

Hydrogen and methane production through two-stage mesophilic anaerobic digestion of olive pulp

The present study focused on the anaerobic biohydrogen production from olive pulp (two phase olive mill wastes, TPOMW) and the subsequent anaerobic treatment of the effluent for methane production under mesophilic conditions in a two-stage process. Biohydrogen production from water-diluted (1:4) olive pulp was investigated at hydraulic retention times (HRT) of 30 h, 14.5 h and 7.5 h while methane production from the effluent of hydrogenogenic reactor was studied at 20 d, 15 d, 10 d and 5 d HRT. In comparison with previous studies, it has been shown that the thermophilic hydrogen production process was more efficient than the mesophilic one in both hydrogen production rate and yield. The methanogenic reactor was successfully operated at 20, 15 and 10 days HRT while it failed when an HRT of 5 days was applied. Methane productivity reached the maximum value of 1.13 ± 0.08 L/L/d at 10 days HRT whereas the methane yield increased with the HRT. The Anaerobic Digestion Model no. 1 (ADM1) was applied to the obtained experimental data from the methanogenic reactor to simulate the digester response at all HRT tested. The ability of the model to predict the experimental results was evident even in the case of the process failure, thus implying that the ADM1 could be a valuable tool for process design even in the case of a complex feedstock. In general, the two-stage anaerobic digestion proved to be a stable, reliable and effective process for energy recovery and stabilization treatment of olive pulp.     

Production of H2 by sulphur-deprived cells of the unicellular cyanobacteria Gloeocapsa alpicola and Synechocystis sp. PCC 6803 during dark incubation with methane or at various extracellular pH

AIMS: To examine sulphur (S) deprivation in combination with the presence of methane (CH4) and changes in extracellular pH as a method to enhance in situ hydrogen (H2) generation during fermentation in the unicellular non-diazotrophic cyanobacteria Gloeocapsa alpicola and Synechocystis PCC 6803. METHODS AND RESULTS: The level of H2 production, measured using a gas chromatography, was determined in S-deprived cells of G. alpicola and Synechocystis PCC 6803 during fermentation. Starvation on S enhanced the rate of H2 production by more than fourfold in both strains. S-deprived cyanobacteria were able to maintain maximum rate of H2 production during at least 8 h of fermentation representing the entire dark period of a day. Increased H2 production was observed during dark anoxic incubation with a gas phase of 100% CH4 (up to four times) at lower pH of the medium (5.0-5.5). CONCLUSIONS: S-deprivation in combination with CH4, added or maybe produced by another micro-organisms, and changes in the pH of the media can be used to further increase the specific capacity of unicellular non-N2-fixing cyanobacteria to produce H2 during fermentation with the overall aim of applying it for outdoor photobiological H2 production. SIGNIFICANCE AND IMPACT OF THE STUDY: S-deprivation with respect to H2 production is well studied in the green algae Chlamydomonas reinhardtii while its application for H2 production in cyanobacteria is novel. Similarly, the stimulation of H2 generation in the presence of CH4 opens up new possibilities to increase the H2 production. Natural gas enriched with H2 seems to be a perspective fuel and may be an intermediate step on the pathway to the exploitation of pure biohydrogen.     

Hydrogen peroxide as an effecter on the inactivation of particulate methane monooxygenase under aerobic conditions

Particulate methane monooxygenase (pMMO), a copper-containing membrane protein, catalyzes methane hydroxylation under aerobic conditions. We found that the activity of pMMO was increased by catalase, implying that hydrogen peroxide (H₂O₂) is generated by pMMO with duroquinol, an electron donor for pMMO, and that the generated H₂O₂ inhibits pMMO activity. In addition, reversible inhibition of pMMO with H₂O₂ was observed upon treatment of pMMO with H₂O₂ followed by the addition of catalase, and H₂O₂ formation by pMMO with duroquinol was detected using a fluorescence probe. The redox behavior of type 2 copper in pMMO measured by the electron paramagnetic resonance revealed that H₂O₂ re-oxidizes the type 2 copper in pMMO reduced with duroquinol.     

联合生物制氢方法及发展趋势

为了在生物制氢过程中最大限度提高产氢量和产氢速率,增大底物的利用率以及更好地发挥菌种间的协同作用,联合生物制氢技术成为近年来人们关注的焦点。综述了目前国内外几种联合生物制氢方法的研究现状。并从产氢机理的角度对几种联合制氢技术进行了分析比较,重点强调光合发酵和暗发酵联合生物制氢技术具有广泛的发展前景,并指出其存在的问题和未来的发展趋势。     

Evaluation and modeling of biochemical methane potential (BMP) of landfilled solid waste: A pilot scale study

The main goal of this study was to present a comparison of landfill performance with respect to solids decomposition. Biochemical methane potential (BMP) test was used to determine the initial and the remaining CH₄ potentials of solid wastes during 27 months of landfilling operation in two pilot scale landfill reactors. The initial methane potential of solid wastes filled to the reactors was around 0.347 L/CH₄/g dry waste, which decreased with operational time of landfill reactors to values of 0.117 and 0.154 L/CH₄/g dry waste for leachate recirculated (R1) and non-recirculated (R2) reactors, respectively. Results indicated that the average rate constant increased by 32% with leachate recirculation. Also, the performance of the system was modeled using the BMP data for the samples taken from reactors at varying operational times by MATLAB program. The first-order rate constants for R1 and R2 reactors were 0.01571 and 0.01195 1/d, respectively. The correlation between the model and the experimental parameters was more than 95%, showing the good fit of the model.     

Predicting ultimate methane yields of Jatropha curcus and Morus indica from their chemical composition

In this study, all the components of Jatropha curcus and Morus indica were chemically characterized and their biochemical methane potentials (BMP) were determined. From the variables that showed strong influence on the ultimate methane yield (B_o) of J. curcus, a multiple regression Jatropha model was developed. This model comprised of total carbohydrates, protein, lipid, acid-detergent fiber (ADF), cellulose and ash in ADF as independent variables, with r² value of 0.943. The Jatropha model was validated on 7 samples of M. indica parts and wastes from silkworm rearing trays of this study and 13 samples of heterogeneous organic wastes of earlier studies, to judge the prediction quality. It was found that most of the predicted values differed by less than 15% of their experimental B_o.     

The geochemistry of methane in Lake Fryxell,an amictic,permanently ice-covered,antarctic lake

The abundance and distribution of dissolved CH₄ were determined from 1987–1990 in Lake Fryxell, Antarctica, an amictic, permanently ice-covered lake in which solute movement is controlled by diffusion. CH₄ concentrations were < 1 υM in the upper oxic waters, but increased below the oxycline to 936 μM at 18 m. Sediment CH₄ was 1100 μmol (1 sed)⁻¹ in the 0–5 cm zone. Upward flux from the sediment was the source of the CH₄, NH₄ ⁺, and DOC in the water column; CH₄ was 27% of the DOC+CH₄ carbon at 18 m. Incubations with surficial sediments indicated that H¹⁴CO₃ ⁻ reduction was 0.4 μmol (1 sed)⁻¹ day⁻¹ or 4× the rate of acetate fermentation to CH₄. There was no measurable CH₄ production in the water column. However, depth profiles of CH₄, NH₄, and DIC normalized to bottom water concentrations demonstrated that a significant CH₄ sink was evident in the anoxic, sulfate-containing zone of the water column (10–18 m). The δ¹³CH₄ in this zone decreased from −72 % at 18 m to −76% at 12 m, indicating that the consumption mechanism did not result in an isotopic enrichment of ¹³CH₄. In contrast, δ¹³CH₄ increased to −55 % at 9 m due to aerobic oxidation, though this was a minor aspect of the CH₄ cycle. The water column CH₄ profile was modeled by coupling diffusive flux with a first order consumption term; the best-fit rate constant for anaerobic CH₄ consumption was 0.012 yr⁻¹. On a total carbon basis, CH₄ consumption in the anoxic water column exerted a major effect on the flux of carbonaceous material from the underlying sediments and serves to exemplify the importance of CH₄ to carbon cycling in Lake Fryxell.     

绿藻光合生物制氢技术进展

氢能作为可再生、环境友好的能源,已成为营造可持续发展的经济节约型社会的理想能源。绿藻因能利用光能分解水产氢,被称为最有应用前景的方法之一。本文将综述绿藻光合产氢的原理,介绍该生物制氢技术的研究现状和最新进展,并对其发展趋势做以展望。     

Immobilized microorganisms for hydrogen and ammonia production

Biological hydrogen and ammonia production are currently being studied as a means of lessening the dependence on non-renewable resources. Some of the systems considered show additional potential in various waste treatment processes. As the systems currently being examined employ a variety of organisms and metabolic pathways, the energetics of the various modes of hydrogen and ammonia production are examined in detail. The use of immobilized microorganisms may offer some advantages over free-living cultures. The present status of the work in this field employing immobilized microorganisms is reviewed, and areas for future research and the potential impacts of this technology are discussed.     

The response of Leuconostoc mesenteroides to low external oxidoreduction potential generated by hydrogen gas

AIMS: The physiological consequences of low external oxidoreduction potential in Leuconostoc mesenteroides were investigated. METHODS AND RESULTS: Leuconostoc mesenteroides was grown under two initial oxidoreduction potential conditions (Eh7: +200 mV and -400 mV) using nitrogen and hydrogen as reducing agents. Growth was affected by Eh7; the lag phase increased from 1 h at an initial Eh7 of +200 mV to 6 h at an initial Eh7 of -400 mV; the maximum specific growth rate at -400 mV was 68% of the one observed at +200 mV. The NADH/NAD+ ratio and (NADH + NAD+) pool were independent of the external Eh7. CONCLUSIONS: This study shows that changing the external oxidoreduction potential from +200 to -400 mV has a strong effect on the Leuc. mesenteroides physiology. The constancy of the maximum carbon and energetic fluxes (qglu, qATP) under the two Eh7 conditions accompanied by the decrease of YX/S and YATP suggested the existence of an uncoupling phenomenon, namely that some catabolized glucose and hence ATP was not associated with biomass production. SIGNIFICANCE AND IMPACT OF THE STUDY: This paper demonstrates the usefulness of taking into account, the effect of the oxidoreduction potential on the growth of Leuc. mesenteroides in the fermentation process.