Production of Bio-Energy from Pig Manure: A Focus on the Dynamics Change of Four Parameters under Sunlight-Dark Conditions

This study investigated the effect of sunlight-dark conditions on volatile fatty acids (VFAs), total ammonium nitrogen (TAN), total alkalinity (TA) and pH during pig manure (PM) digestion and then the subsequent influence on biogas yield of PM. PM₁ and PM₂ were performed in a transparent reactor and a non-transparent reactor, respectively. Two sets of experiments were conducted with a temperature of 35.0±2.0 °C and a total solid concentration of 8.0% to the digestion material. The dynamic change of the four parameters in response to sunlight-dark conditions resulted in variations of the physiological properties in the digester and affected the cumulative biogas production (CBP). PM₁ obtained higher CBP (15020.0 mL) with a more stable pH and a lower TAN concentration (1414.5 mg/L) compared to PM₂ (2675.0 mL and 1670.0 mg/L, respectively). The direct path coefficients and indirect path coefficients between the four parameters and CBP were also analyzed.     

Microbial processes influencing methane emission from rice fields

Irrigated rice fields are an important source of atmospheric methane. In order to improve our understanding of the controlling processes, we measured in situ CH₄ emission and CH₄ oxidation in an Italian rice field in 1998 and 1999, and studied CH₄ production in soil and root samples. The CH₄ emission rates were correlated with diurnal temperature variations and showed pronounced seasonal and interannual variations. The contribution of CH₄ oxidation to total CH₄ flux, determined by specific inhibition with difluoromethane, decreased from 40% at the beginning to zero at the end of the season. The stable carbon isotopic composition of the emitted CH₄ also decreased. The CH₄‐oxidizing bacteria probably became limited by nitrogen as indicated by the seasonal decrease of NH₄⁺. Thus, CH₄ oxidation had little effect on CH₄ emission. Methane production on rice roots was relatively constant over the season. Methane production in soil slowly increased after flooding and was highest in the middle of the season. Pore water concentrations of CH₄ showed a similar seasonal pattern. In 1999, CH₄ production increased later in the season and reached lower rates than in 1998. An additional drainage in 1999 resulted in higher ferric iron concentrations, higher soil redox potentials and lower acetate concentrations. As a result, acetate‐utilizing methanogens were probably out‐competed by iron‐reducers so that a larger percentage of [2–¹⁴C]acetate was converted to ¹⁴CO₂ instead of ¹⁴CH₄. The residual CH₄ production was relatively low and was mainly due to H₂/CO₂‐dependent methanogenesis. Experiments with radioactive bicarbonate and with methyl fluoride as specific inhibitor showed that the theoretical ratio of 7:3 of methanogenesis from acetate vs. H₂/CO₂ was only reached later in the season when total CH₄ production was at the maximum. In conclusion, our results give a mechanistic explanation for the intraseasonal and interannual differences in CH₄ emission.     

Bacterial community analysis in upflow multilayer anaerobic reactor treating high‐solids organic wastes

A novel anaerobic digestion configuration, the upflow multi‐layer anaerobic reactor (UMAR), was developed to treat high‐solids organic wastes. The UMAR was hypothesized to form multi‐layer along depth due to the upflow plug flow; use of a recirculation system and a rotating distributor and baffles aimed to assist treating high‐solids influent. The chemical oxygen demand (COD) removal efficiency and methane (CH₄) production rate were 89% and 2.10 L CH₄/L/d, respectively, at the peak influent COD concentration (110.4 g/L) and organic loading rate (7.5 g COD/L/d). The 454 pyrosequencing results clearly indicated heterogeneous distribution of bacterial communities at different vertical locations (upper, middle, and bottom) of the UMAR. Firmicutes was the dominant (>70%) phylum at the middle and bottom parts, while Deltaproteobacteria and Chloroflexi were only found in the upper part. Potential functions of the bacteria were discussed to speculate on their roles in the anaerobic performance of the UMAR system. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:1226–1234, 2017     

Production and oxidation of methane in a boreal mire after a decade of increased temperature and nitrogen and sulfur deposition

Natural wetlands are the single largest source of atmospheric methane (CH₄). Both a changed climate and deposition of anthropogenic nitrogen and sulfur can alter the production and oxidation of CH₄ respectively and thereby also CH₄ exchange. We used a long‐term (12 years) factorial field experiment in a boreal oligotrophic mire to evaluate the effects of greenhouse cover and addition of ammonium nitrate and sodium sulfate on the production and oxidation of CH₄ by applying laboratory incubations of samples from five depths in the mire. The rates of CH₄ production were measured without amendments and after the addition of either glucose or sulfate. Twelve years of increased nitrogen deposition has changed the mire from a Sphagnum‐dominated plant community to one dominated by sedges and dwarf shrubs. The deposition of nitrogen to the field plots caused increased production of CH₄ in incubations without amendments (34%), and also after amendments with glucose (40%) or sulfate (42%). This indicates increased substrate availability (without amendments) but also a greater abundance of methanogens (glucose amendment). The greenhouse cover caused a decrease in CH₄ production in incubations without amendments (34%), after glucose amendment (20%) and after sulfate amendment (31%). These responses indicate decreased substrate availability (without amendment) accompanied by the reduced abundance of methanogens (glucose amendment). The field application of sulfur had no effect on CH₄ production at the depth where maximal CH₄ production occurred. Closer to the mire surface, however, the rate of CH₄ production was significantly reduced by 32–45%. These results suggest that the deposition of sulfate has altered the vertical distribution of methanogens and sulfate‐reducing bacteria. The oxidation of CH₄ was not significantly affected by any of the long‐term field treatments.     

Integrated biogas upgrading and hydrogen utilization in an anaerobic reactor containing enriched hydrogenotrophic methanogenic culture

Biogas produced by anaerobic digestion, is mainly used in a gas motor for heat and electricity production. However, after removal of CO₂, biogas can be upgraded to natural gas quality, giving more utilization possibilities, such as utilization as autogas, or distant utilization by using the existing natural gas grid. The current study presents a new biological method for biogas upgrading in a separate biogas reactor, containing enriched hydrogenotrophic methanogens and fed with biogas and hydrogen. Both mesophilic‐ and thermophilic anaerobic cultures were enriched to convert CO₂ to CH₄ by addition of H₂. Enrichment at thermophilic temperature (55°C) resulted in CO₂ and H₂ bioconversion rate of 320 mL CH₄/(gVSS h), which was more than 60% higher than that under mesophilic temperature (37°C). Different dominant species were found at mesophilic‐ and thermophilic‐enriched cultures, as revealed by PCR–DGGE. Nonetheless, they all belonged to the order Methanobacteriales, which can mediate hydrogenotrophic methanogenesis. Biogas upgrading was then tested in a thermophilic anaerobic reactor under various operation conditions. By continuous addition of hydrogen in the biogas reactor, high degree of biogas upgrading was achieved. The produced biogas had a CH₄ content, around 95% at steady‐state, at gas (mixture of biogas and hydrogen) injection rate of 6 L/(L day). The increase of gas injection rate to 12 L/(L day) resulted in the decrease of CH₄ content to around 90%. Further study showed that by decreasing the gas–liquid mass transfer by increasing the stirring speed of the mixture the CH₄ content was increased to around 95%. Finally, the CH₄ content around 90% was achieved in this study with the gas injection rate as high as 24 L/(L day). Biotechnol. Bioeng. 2012; 109: 2729–2736. © 2012 Wiley Periodicals, Inc.     

Oxygen effects on methane production and oxidation in humid tropical forest soils

We investigated the effects of oxygen (O₂) concentration on methane (CH₄) production and oxidation in two humid tropical forests that differ in long‐term, time‐averaged soil O₂ concentrations. We identified sources and sinks of CH₄ through the analysis of soil gas concentrations, surface emissions, and carbon isotope measurements. Isotope mass balance models were used to calculate the fraction of CH₄ oxidized in situ. Complementary laboratory experiments were conducted to determine the effects of O₂ concentration on gross and net rates of methanogenesis. Field and laboratory experiments indicated that high levels of CH₄ production occurred in soils that contained between 9±1.1% and 19±0.2% O₂. For example, we observed CH₄ concentrations in excess of 3% in soils with 9±1.1% O₂. CH₄ emissions from the lower O₂ sites were high (22–101 nmol CH₄ m^?2 s^?1), and were equal in magnitude to CH₄ emissions from natural wetlands. During peak periods of CH₄ efflux, carbon dioxide (CO₂) emissions became enriched in ¹³C because of high methanogenic activity. Gross CH₄ production was probably greater than flux measurements indicated, as isotope mass balance calculations suggested that 48–78% of the CH₄ produced was oxidized prior to atmospheric egress. O₂ availability influenced CH₄ oxidation more strongly than methanogenesis. Gross CH₄ production was relatively insensitive to O₂ concentrations in laboratory experiments. In contrast, methanotrophic bacteria oxidized a greater fraction of total CH₄ production with increasing O₂ concentration, shifting the δ¹³C composition of CH₄ to values that were more positive. Isotopic measurements suggested that CO₂ was an important source of carbon for methanogenesis in humid forests. The δ¹³C value of methanogenesis was between ?84‰ and ?98‰, which is well within the range of CH₄ produced from CO₂ reduction, and considerably more depleted in ¹³C than CH₄ formed from acetate.     

Genotoxicity assessment of ammonia in cultured Oreochromis niloticus using RAPD assay

Limited efforts have been made to study the genotoxic effects of ammonia in cultured OOreochromi niloticus. Therefore, the present study was planned to assess the genotoxic effect of ammonia in cultured O. niloticus using random amplified polymorphic DNA (RAPD) assay. Fish was categorized into four groups. The 1st group exposed to 2.5 mg/L of total ammonia nitrogen (TAN) (0.16 NH₃ mg/L), the 2nd exposed to 5.0 mg/L of total ammonia nitrogen (TAN) (0.32 NH₃ mg/L) and the 3rd exposed to 10.0 mg/L of total ammonia nitrogen (TAN) (0.65 NH₃ mg/L), in addition to control group for the treatment period of 6 days. The results revealed that some genes in O. niloticus are susceptible to DNA disturbances/mutation as a result of exposure to high concentration of ammonia in water, this clearly indicated using RAPD screening assay.     

Biochemical methane potential (BMP) test for thickened sludge using anaerobic granular sludge at different inoculum/substrate ratios

The biochemical methane potential (BMP) test for thickened sludge was evaluated at three different inoculum/substrate (I:S) ratios. The cumulative methane yield was 51.4 mL CH₄/g VS_added at an I:S ratio of 1:1, 76.3 mL CH₄/g VS_added at an I:S ratio of 1:3, and 21.9 mL CH₄/g VS_added at an I:S ratio of 1:8. The greatest ultimate methane yield and methane production rate constant were achieved at an I:S ratio of 1:3, whereas the least was obtained at an I:S ratio of 1:8. The maximum methane production rate constant was 0.38/day and the minimum methane production rate constant was 0.0016/day. For the case of a lower I:S ratio, the biomass activity may be affected due to the low substrate concentration. On the other hand, for the case of higher I:S ratios, anaerobic digestion of thickened sludge was inhibited by higher concentrations of volatile fatty acids and lower pH.     

Copper stressed anaerobic fermentation: biogas properties,process stability,biodegradation and enzyme responses

The effect of copper (added as CuCl₂) on the anaerobic co-digestion of Phragmites straw and cow dung was studied in pilot experiments by investigating the biogas properties, process stability, substrate degradation and enzyme activities at different stages of mesophilic fermentation. The results showed that 30 and 100 mg/L Cu²⁺ addition increased the cumulative biogas yields by up to 43.62 and 20.77% respectively, and brought forward the daily biogas yield peak, while 500 mg/L Cu²⁺ addition inhibited biogas production. Meanwhile, the CH₄ content in the 30 and 100 mg/L Cu²⁺-added groups was higher than that in the control group. Higher pH values (close to pH 7) and lower oxidation–reduction potential (ORP) values in the Cu²⁺-added groups after the 8th day indicated better process stability compared to the control group. In the presence of Cu²⁺, the degradation of volatile fatty acids (VFAs) and other organic molecules (represented by chemical oxygen demand, COD) generated from hydrolysis was enhanced, and the ammonia nitrogen (NH₄ ⁺-N) concentrations were more stable than in the control group. The contents of lignin and hemicellulose in the substrate declined in the Cu²⁺-added groups while the cellulose contents did not. Neither the cellulase nor the coenzyme F₄₂₀ activities could determine the biogas producing efficiency. Taking the whole fermentation process into account, the promoting effect of Cu²⁺ addition on biogas yields was mainly attributable to better process stability, the enhanced degradation of lignin and hemicellulose, the transformation of intermediates into VFA, and the generation of CH₄ from VFA.     

Effect of the solid content on anaerobic digestion of meat and bone meal

The effect of the solid content on anaerobic digestion of meat and bone meal (MBM) was investigated in batch reactors at MBM solid contents of 1%, 2%, 5% and 10%. There was no significant difference in the specific methane (CH₄) production potential with respect to the total volatile MBM solids (TVS) applied at these solid contents, which ranged from 351 to 381 ml CH₄/g TVS. However, the highest CH₄ yield with respect to the removed volatile MBM solids (RVS) was 482 ml CH₄/g RVS at the MBM solid content of 5%; the CH₄ yields were 384–448 ml CH₄/g RVS at the other MBM solid contents. The lag time of CH₄ production rose with the increase in the solid content. The longer lag time at MBM solid contents of 5% and 10% was due to inhibition caused by high concentrations of volatile fatty acids (VFAs) and free ammonia in the reactors, but the inhibition was reversible. The production of VFAs during the digestion varied with solid contents: at the solid content of 1%, only acetic acid was detected; at 2%, both acetic and propionic acids were detected; and at 5% and 10%, acetic, propionic, butyric and valeric acids were detected. After 93-day digestion, the volatile MBM solid reduction was 92%, 91%, 79% and 80% at MBM solid contents of 1%, 2%, 5% and 10%, respectively.     

Cultivation, nitrogen fertilization, and set-aside effects on methane uptake in a drained marsh soil in Northeast China

To evaluate the effect of cultivation, nitrogen fertilizer, and set aside on CH₄ uptake after drained marshland was converted into agricultural fields, CH₄ fluxes and CH₄ concentrations in soil gas were in situ measured in a drained marsh soil, a set‐aside cultivated soil, and cultivated soils in Sanjiang Plain of Northeast China in August 2001. Over the measuring period, the highest CH₄ uptake rate was 120.7±6.2 μg CH₄ m^?2 h^?1 in the drained marsh soil and the lowest was 29.5±4.9 μg CH₄ m^?2 h^?1 in the set‐aside cultivated soil, showing that there was no significant recovery of CH₄ uptake ability 5 years after cultivation activity was stopped. CH₄ uptake rates were significantly less in the cultivated soils than in the drained marsh soil by 30.1–74.6%, which resulted mainly from cultivation and partly from nitrogen addition. A significantly negative correlation between CH₄ flux and bulk density in the cultivated soils tilled by machine suggests that cultivation reduced CH₄ uptake through compaction, because of the enhanced diffusion resistance for CH₄ and O₂. Nitrogen fertilization slowly reduced but persistently affected CH₄ uptake even after long‐term application of nitrogen.     

Influence of inoculum to substrate ratios (ISRs) on the performance of anaerobic digestion of algal residues

With increasing concerns of microalgal-biodiesel, algal residues after lipid extraction are raising great attention for energy production. A batch test of 15 days under mesophilic condition was conducted to evaluate the effects of inoculum to substrate ratios (ISRs) on the methane production by anaerobic digestion of Chlorella sp. residue. The stability and progress of the reaction from algal residue to methane were monitored by measuring the pH, volatile fatty acids (VFAs), total ammoniacal nitrogen (TAN), methane volume on a daily basis. The results indicated that the values obtained were 26.6, 191.6, 195.6 and 210.6 ml CH₄/g volatile solid (VS) for ISRs of 1:2, 1:1, 2:1 and 3:1. The methane production was significantly decreased as the ISR was lower than 1:1, which was resulting from the poor methanogenesis inhibited by NH₄ ⁺-N. It would be of great importance that determination of ISRs might provide useful information on how to initialize a batch digester with algal residue as material.     

Methane emissions from wetlands and their relationship with vascular plants: an Arctic example

This paper investigates the relationship between vascular plant production and CH₄ emissions from an arctic wet tundra ecosystem in north‐east Greenland. Light intensity was manipulated by shading during three consecutive growing seasons (1998–2000). The shading treatment resulted in lower carbon cycling in the ecosystem as mean seasonal net ecosystem exchange (NEE) decreased from ?336 to ?196 mg CO₂ m^?2 h^?1 and from ?476 to ?212 mg CO₂ m^?2 h^?1 in 1999 and 2000, respectively, and total ecosystem respiration decreased from 125 to 94 mg CO₂ m^?2 h^?1 in 1999 and from 409 to 306 mg CO₂ m^?2 h^?1 in 2000. Seasonal mean CH₄ emissions in controls and shaded plots were, respectively, 6.5 and 4.5 mg CH₄ m^?2 h^?1 in 1999 and 8.3 and 6.2 mg CH₄ m^?2 h^?1 in 2000. We found that CH₄ emission was sensitive to NEE and carbon turnover, and it is reasonable to assume that the correlation was due to a combined effect of vegetative CH₄ transport and substrate quality coupled to vascular plant production. Total above‐ground biomass was correlated to mean seasonal CH₄ emission, but separation into species showed that plant‐mediated CH₄ transport was highly species dependent. Potential CH₄ production peaked at the same depth as maximum root density (5–15 cm) and treatment differences further suggest that substrate quality was negatively affected by decreased NEE in the shaded plots. The concentration of dissolved CH₄ decreased in the control plots as the growing season progressed while it was relatively stable in the shaded plots. This suggests that a progressively better developed root system in the controls increased the capacity to transport CH₄ from the soil to the atmosphere. In conclusion, vascular plant photosynthetic rate and subsequent allocation of recently fixed carbon to below‐ground structures seemed to influence both vegetative CH₄ transport and substrate quality.     

Magnetite accelerates syntrophic acetate oxidation in methanogenic systems with high ammonia concentrations

Ammonia accumulation is a major inhibitory substance causing anaerobic digestion upset and failure in CH₄ production. At high ammonia levels, CH₄ production through syntrophic acetate oxidization (SAO) pathways is more tolerant to ammonia toxicity than the acetoclastic methanogenesis pathway, but the low CH₄ production rate through SAO constitutes the main reason for the low efficiency of energy recovery in anaerobic digesters treating ammonia‐rich substrates. In this study, we showed that acetate fermentation to CH₄ and CO₂ occurred through SAO pathway in the anaerobic reactors containing a high ammonia concentration (5.0 g l^?1 NH₄⁺–N), and the magnetite nanoparticles supplementation increased the CH₄ production rates from acetate by 36–58%, compared with the anaerobic reactors without magnetite under the same ammonia level. The mechanism of facilitated methanogenesis was proposed to be the establishment of direct interspecies electron transfer (DIET) for SAO, in which magnetite facilitated DIET between syntrophic acetate oxidizing bacteria and methanogens. High‐throughput 16S rRNA gene sequencing analysis revealed that the bacterial Geobacteraceae and the archaeal Methanosarcinaceae and Methanobacteriaceae might be involved in magnetite‐mediated DIET for SAO and CH₄ production. This study demonstrated that magnetite supplementation might provide an effective approach to accelerate CH₄ production rates in the anaerobic reactors treating wastewater containing high ammonia.     

Biohydrogen production from wheat straw hydrolysate by dark fermentation using extreme thermophilic mixed culture

Hydrolysate was tested as substrate for hydrogen production by extreme thermophilic mixed culture (70°C) in both batch and continuously fed reactors. Hydrogen was produced at hydrolysate concentrations up to 25% (v/v), while no hydrogen was produced at hydrolysate concentration of 30% (v/v), indicating that hydrolysate at high concentrations was inhibiting the hydrogen fermentation process. In addition, the lag phase for hydrogen production was strongly influenced by the hydrolysate concentration, and was prolonged from approximately 11 h at the hydrolysate concentrations below 20% (v/v) to 38 h at the hydrolysate concentration of 25% (v/v). The maximum hydrogen yield as determined in batch assays was 318.4 ± 5.2 mL‐H₂/g‐sugars (14.2 ± 0.2 mmol‐H₂/g‐sugars) at the hydrolysate concentration of 5% (v/v). Continuously fed, and the continuously stirred tank reactor (CSTR), operating at 3 day hydraulic retention time (HRT) and fed with 20% (v/v) hydrolysate could successfully produce hydrogen. The hydrogen yield and production rate were 178.0 ± 10.1 mL‐H₂/g‐sugars (7.9 ± 0.4 mmol H₂/g‐sugars) and 184.0 ± 10.7 mL‐H₂/day L_reactor (8.2 ± 0.5 mmol‐H₂/day L_reactor), respectively, corresponding to 12% of the chemical oxygen demand (COD) from sugars. Additionally, it was found that toxic compounds, furfural and hydroxymethylfurfural (HMF), contained in the hydrolysate were effectively degraded in the CSTR, and their concentrations were reduced from 50 and 28 mg/L, respectively, to undetectable concentrations in the effluent. Phylogenetic analysis of the mixed culture revealed that members involved hydrogen producers in both batch and CSTR reactors were phylogenetically related to the Caldanaerobacter subteraneus, Thermoanaerobacter subteraneus, and Thermoanaerobacterium thermosaccharolyticum. Biotechnol. Bioeng. 2010;105: 899–908. © 2009 Wiley Periodicals, Inc.     

One-stage and two-stage anaerobic digestion of lipid-extracted algae

Waste-grown microalgae are a potentially important biomass for wastewater treatment. The lipid accumulated in microalgae could be utilized as feedstocks for biodiesel production. The algal residues, as major by-products derived from lipid extraction, mainly consist of carbohydrate and protein, making anaerobic digestion an efficient way to recover energy. The conversion of lipid-extracted algal residues into methane plays dual role in renewable energy production and sustainable development of microalgal biodiesel industry. Therefore, an anaerobic fermentation process for investigation of the methane production potential of algal residues was conducted in this paper. The effect of inoculum to substrate ratios (ISRs) on the methane production by anaerobic digestion of Chlorella sp. residue in a single stage was evaluated. The maximum methane yield of 195.6 ml CH₄/g volatile solid (VS) was obtained at an ISR of 1:1. The stability and progress of the reaction from algal residues to methane were monitored by measuring the pH, volatile fatty acids (VFAs), total ammoniacal nitrogen (TAN), and methane volume. Based on the results of one-stage experiments, two-stage technology was proposed and was found to be more suitable for high organic load. The optimum conditions for acidogenesis and methanogenesis are indicated in this paper.     

Optimization of culture media for large‐scale lutein production by heterotrophic Chlorella vulgaris

Lutein is a carotenoid with a purported role in protecting eyes from oxidative stress, particularly the high‐energy photons of blue light. Statistical optimization was performed to growth media that supports a higher production of lutein by heterotrophically cultivated Chlorella vulgaris. The effect of media composition of C. vulgaris on lutein was examined using fractional factorial design (FFD) and central composite design (CCD). The results indicated that the presence of magnesium sulfate, EDTA‐2Na, and trace metal solution significantly affected lutein production. The optimum concentrations for lutein production were found to be 0.34 g/L, 0.06 g/L, and 0.4 mL/L for MgSO₄·7H₂O, EDTA‐2Na, and trace metal solution, respectively. These values were validated using a 5‐L jar fermenter. Lutein concentration was increased by almost 80% (139.64 ± 12.88 mg/L to 252.75 ± 12.92 mg/L) after 4 days. Moreover, the lutein concentration was not reduced as the cultivation was scaled up to 25,000 L (260.55 ± 3.23 mg/L) and 240,000 L (263.13 ± 2.72 mg/L). These observations suggest C. vulgaris as a potential lutein source. © 2014 American Institute of Chemical Engineers Biotechnol. Prog., 30:736–743, 2014     

Energy recovery from lipid extracted,transesterified and glycerol codigested microalgae biomass

The production of methane (CH₄) via the anaerobic digestion of microalgae biomass residues from the biodiesel production process has the potential to meet some of the energy requirements of the primary biomass to fuel conversion process. This paper investigates the practical CH₄ yields achievable from the anaerobic conversion of the microalgae residues (as well as codigestion with glycerol) after biodiesel production using both the conventional and in situ transesterification methods. Results demonstrate that the type of lipid extraction solvent utilized in the conventional transesterification process could inhibit subsequent CH₄ production. On the basis of actual CH₄ production, a recoverable energy of 8.7–10.5 MJ kg^?1 of dry microalgae biomass residue was obtained using the lipid extracted and transesterified microalgae samples. On codigesting the microalgae residues with glycerol, a 4–7% increase in CH₄ production was observed.     

Hydrogen and methane production from desugared molasses using a two‐stage thermophilic anaerobic process

Hydrogen and methane production from desugared molasses by a two‐stage thermophilic anaerobic process was investigated in a series of two up‐flow anaerobic sludge blanket (UASB) reactors. The first reactor that was dominated with hydrogen‐producing bacteria of Thermoanaerobacterium thermosaccharolyticum and Thermoanaerobacterium aciditolerans could generate a high hydrogen production rate of 5600 mL H₂/day/L, corresponding to a yield of 132 mL H₂/g volatile solid (VS). The effluent from the hydrogen reactor was further converted to methane in the second reactor with the optimal production rate of 3380 mL CH₄/day/L, corresponding to a yield of 239 mL CH₄/g VS. Aceticlastic Methanosarcina mazei was the dominant methanogen in the methanogenesis stage. This work demonstrates that biohydrogen production can be very efficiently coupled with a subsequent step of methane production using desugared molasses. Furthermore, the mixed gas with a volumetric content of 16.5% H_2, 38.7% CO₂, and 44.8% CH₄, containing approximately 15% energy by hydrogen is viable to be bio‐hythane.