Integrated hydrogen production process from cellulose by combining dark fermentation, microbial fuel cells, and a microbial electrolysis cell

Hydrogen gas production from cellulose was investigated using an integrated hydrogen production process consisting of a dark fermentation reactor and microbial fuel cells (MFCs) as power sources for a microbial electrolysis cell (MEC). Two MFCs (each 25 mL) connected in series to an MEC (72 mL) produced a maximum of 0.43 V using fermentation effluent as a feed, achieving a hydrogen production rate from the MEC of 0.48 m³ H₂/m³/d (based on the MEC volume), and a yield of 33.2 mmol H₂/g COD removed in the MEC. The overall hydrogen production for the integrated system (fermentation, MFC and MEC) was increased by 41% compared with fermentation alone to 14.3 mmol H₂/g cellulose, with a total hydrogen production rate of 0.24 m³ H₂/m³/d and an overall energy recovery efficiency of 23% (based on cellulose removed) without the need for any external electrical energy input.     

A comparison study on the high-rate co-digestion of sewage sludge and food waste using a temperature-phased anaerobic sequencing batch reactor system

Assessing contemporary anaerobic biotechnologies requires proofs on reliable performance in terms of renewable bioenergy recovery such as methane (CH₄) production rate, CH₄ yield while removing volatile solid (VS) effectively. This study, therefore, aims to evaluate temperature-phased anaerobic sequencing batch reactor (TPASBR) system that is a promising approach for the sustainable treatment of organic fraction of municipal solid wastes (OFMSW). TPASBR system is compared with a conventional system, mesophilic two-stage anaerobic sequencing batch reactor system, which differs in operating temperature of 1st-stage. Results demonstrate that TPASBR system can obtain 44% VS removal from co-substrate of sewage sludge and food waste while producing 1.2 m³CH₄/m³_system/d (0.2 m³CH₄/kgVS_added) at organic loading rate of 6.1 gVS/L/d through the synergy of sequencing-batch operation, co-digestion, and temperature-phasing. Consequently, the rapid and balanced anaerobic metabolism at thermophilic stage makes TPASBR system to afford high organic loading rate showing superior performance on OFMSW stabilization.     

Acidogenic fermentation of vegetable based market waste to harness biohydrogen with simultaneous stabilization

Vegetable based market waste was evaluated as a fermentable substrate for hydrogen (H₂) production with simultaneous stabilization by dark-fermentation process using selectively enriched acidogenic mixed consortia under acidophilic microenvironment. Experiments were performed at different substrate/organic loading conditions in concurrence with two types of feed compositions (with and without pulp). Study depicted the feasibility of H₂ production from vegetable waste stabilization process. H₂ production was found to be dependent on the concentration of the substrate and composition. Higher H₂ production and substrate degradation were observed in experiments performed without pulp (23.96 mmol/day (30.0 kg COD/m³); 13.96 mol/kg COD_R (4.8 kg COD/m³)) than with pulp (22.46 mmol/day (32.0 kg COD/m³); 12.24 mol/kg COD_R (4.4 kg COD/m³)). Generation of higher concentrations of acetic acid and butyric acid was observed in experiments performed without pulp. Data enveloping analysis (DEA) was employed to study the combined process efficiency of system by integrating H₂ production and substrate degradation.     

Autotrophic denitrification using hydrogen generated from metallic iron corrosion

Hydrogenotrophic denitrification was demonstrated using hydrogen generated from anoxic corrosion of metallic iron. For this purpose, a mixture of hydrogenated water and nitrate solution was used as reactor feed. A semi-batch reactor with nitrate loading of 2000 mg m⁻³ d⁻¹ and hydraulic retention time (HRT) of 50 days produced effluent with nitrate concentration of 0.27 mg N L⁻¹ (99% nitrate removal). A continuous flow reactor with nitrate loading of 28.9 mg m⁻³ d⁻¹ and HRT of 15.6 days produced effluent with nitrate concentration of ∼0.025 mg N L⁻¹ (95% nitrate removal). In both cases, the concentration of nitrate degradation by-products, viz., ammonia and nitrite, were below detection limits. The rate of denitrification in the reactors was controlled by hydrogen availability, and hence to operate such reactors at higher nitrate loading rates and/or lower HRT than reported in the present study, hydrogen concentration in the hydrogenated water must be significantly increased.     

Effects of key operational parameters on biohydrogen production via anaerobic fermentation in a sequencing batch reactor

In this study, a series of tests were conducted in a 6 L anaerobic sequencing batch reactor (ASBR) to investigate the effect of pH, hydraulic retention time (HRT) and organic loading rate on biohydrogen production at 28 °C. Sucrose was used as the main substrate to mimic carbohydrate-rich wastewater and inoculum was prepared from anaerobic digested sludge without pretreatment. The reactor was operated initially with nitrogen sparging to form anaerobic condition. Results showed that methanogens were effectively suppressed. The optimum pH value would vary depending on the HRT. Maximum hydrogen production rate and yield of 3.04 L H₂/L reactor d and 2.16 mol H₂/mol hexose respectively were achieved at pH 4.5, HRT 30 h, and OLR 11.0 kg/m³ d. Two relationships involving the propionic acid/acetic acid ratio and ethanol/acetic acid ratio were derived from the analysis of the metabolites of fermentation. Ethanol/acetic acid ratio of 1.25 was found to be a threshold value for higher hydrogen production.     

Influence of carbohydrates and proteins concentration on fermentative hydrogen production using canteen based waste under acidophilic microenvironment

Functional role of biomolecules viz., carbohydrates and proteins on acidogenic biohydrogen (H₂) production was studied through the treatment of canteen based composite food waste. The performance was evaluated in an anaerobic sequencing batch reactor (AnSBR) at pH 6 with five variable organic loading conditions (OLR1, 0.854; OLR2, 1.69; OLR3, 3.38; OLR4, 6.54 and OLR5, 9.85 kg COD/m³-day). Experimental data depicted the feasibility of H₂ production from the stabilization of food waste and was found to depend on the substrate load. Among the five loading conditions studied, OLR4 documented maximum H₂ production (69.95 mmol), while higher substrate degradation (3.99 kg COD/m³-day) was observed with OLR5. Specific hydrogen yield (SHY) vary with the removal of different biomolecules and was found to decrease with increase in the OLR. Maximum SHY was observed with hexose removal at OLR1 (139.24 mol/kg Hexose_R at 24 h), followed by pentoses (OLR1, 108.26 mol/kg Pentose_R at 48 h), proteins (OLR1, 109.71 mol/kg Protein_R at 48 h) and total carbohydrates (OLR1, 58.31 mol/kg CHO_R at 24 h). Proteins present in wastewater helped to maintain the buffering capacity but also enhanced the H₂ production by supplying readily available organic nitrogen to the consortia. Along with carbohydrates and proteins, total solids also registered good removal.     

Hydrogen production by Rhodopseudomonas palustris WP 3-5 in a serial photobioreactor fed with hydrogen fermentation effluent

In this study, a lab-scale serial photobioreactor composed of three column reactors was constructed and continuously operated to investigate several parameters influencing photohydrogen production when using the synthetic wastewater and the anaerobic hydrogen fermentation effluents as the influents. The results indicated that better hydrogen production rate was obtained when the serial photobioreactor was operated under cellular recycling at a short HRT of 8 h. The serial photobioreactor maintained high hydrogen content ca. 80% in the produced gas and 0.4× dilution ratio was the suitable ratio for hydrogen production. When the photobioreactor fed with the real wastewater (Effluent 1) containing 100 mg/L NH₄Cl, Column 1 reactor successfully reduced ammonia concentration to about 60 mg/L for cell synthesis, resulting in a steady hydrogen production in the following two column reactors. The average hydrogen production rate was 205 mL-H₂/L/d.     

Evaluation of bioenergy recovery processes treating organic residues from ethanol fermentation process

This study evaluates a two-stage bioprocess for recovering bioenergy in the forms of hydrogen and methane while treating organic residues of ethanol fermentation from tapioca starch. A maximum hydrogen production rate of 0.77 mmol H₂/g VSS/h can be achieved at volumetric loading rate (VLR) of 56 kg COD/m³/day. Batch results indicate that controlling conditions at S₀/X₀ = 12 with X₀ = 4000 mg VSS/L and pH 5.5-6 are important for efficient hydrogen production from fermentation residues. Hydrogen-producing bacteria enriched in the hydrogen bioreactor are likely utilizing lactate and acetate for biohydrogen production from ethanol-fermentation residues. Organic residues remained in the effluent of hydrogen bioreactor can be effectively converted to methane with a rate of 0.37 mmol CH₄/g VSS/h at VLR of 8 kg COD/m³/day. Approximately 90% of COD in ethanol-fermentation residues can be removed and among that 2% and 85.1% of COD can be recovered in the forms of hydrogen and methane, respectively.     

Performance and characterization of a newly developed self-agitated anaerobic reactor with biological desulfurization

The continuous operation of a newly developed methane fermentation reactor, which requires no electricity for the agitation of the fermentation liquid was investigated, and the extent of the biological desulfurization was monitored. Inside the reactor, the continual change in the liquid level and the self-agitation, occurring between 5 and 16 times every day, distributed the organic load near the inlet port of the reactor, as well as providing a nutrient supply to the hydrogen sulfide oxidizing bacteria. At different COD_Cr loading rates (5, 7, 10 kg m³ d⁻¹), the reactor achieved a biogas production yield of 0.72-0.82 m³ g⁻¹-TS, a COD_Cr reduction of 79.4-85.5% and an average of 99% hydrogen sulfide removal. This investigation demonstrated that the self-agitated reactor is comparable in digestion performance to the completely stirred tank reactor (CSTR) investigated in a previous study, and that the desulfurization performance was significantly enhanced compared to the CSTR.     

Enhanced methane and hydrogen production from municipal solid waste and agro-industrial by-products co-digested with crude glycerol

The effects of crude glycerol on the performance of single-stage anaerobic reactors treating different types of organic waste were examined. A reactor treating the organic fraction of municipal solid waste produced 1400 mL CH₄/d before the addition of glycerol and 2094 mL CH₄/d after the addition of glycerol. An enhanced methane production rate was also observed when a 1:4 mixture of olive mill wastewater and slaughterhouse wastewater was supplemented with crude glycerol. Specifically, by adding 1% v/v crude glycerol to the feed, the methane production rate increased from 479 mL/d to 1210 mL/d. The extra glycerol-COD added to the feed did not have a negative effect on the reactor performance in either case. Supplementation of the feed with crude glycerol also had a significant positive effect on anaerobic fermentation reactors. Hydrogen yield was 26 mmole H₂/g VS added and 15 mmole H₂/g VS added in a reactor treating the organic fraction of municipal solid waste and a 1:4 mixture of olive mill and slaughterhouse wastewater. The addition of crude glycerol to the feed enhanced hydrogen yield at 2.9 mmole H₂/g glycerol added and 0.7 mmole H₂/g glycerol added.     

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.     

Two stage anaerobic baffled reactors for bio-hydrogen production from municipal food waste

A two-step anaerobic baffled reactor (ABR-1 and ABR-2) for H₂ production from municipal food waste (MFW) was investigated at a temperature of 26 °C. In ABR-1, the average yield of H₂ at an HRT of 26 h and OLR of 58 kg COD/m³ d was 250 ml H₂/g VS removed. As unexpected; the H₂ production in the ABR-2 was further increased up to 370 ml H₂/gVS removed at a HRT of 26 h and OLR of 35 kgCOD/m³ d. The total H₂ yield in the two-step process was estimated to be 4.9 mol H₂/mol hexose. The major part of H₂ production in the ABR-1 was due to the conversion of COD_particulate (36%). In the ABR-2 the H₂ yield was mainly due to the conversion of COD in the soluble form (76%). Based on these results MFW could be ideal substrate for H₂ production in a two-step ABR processes.     

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.     

Two-stage anaerobic digestion of tomato, cucumber, common reed and grass silage in leach-bed reactors and upflow anaerobic sludge blanket reactors

Anaerobic digestion of tomato, cucumber, common reed and grass silage was studied in four separate two-stage reactor configuration consisting of leach bed reactor (LBR) and upflow anaerobic sludge blanket reactor (UASB). LBR studies showed that COD solubilization for cucumber and grass silage was higher (50%) than tomato (35%) and common reed (15%). Results also showed that 31-39% of initial TKN present in tomato and cucumber was solubilized in the leachates and 47-54% of the solubilized TKN was converted to NH₄-N. The corresponding values for common reed and grass silage were 38-50% and 18-36%, respectively. Biomethanation of the leachates in UASB reactors resulted in methane yields of 0.03-0.14 m³ CH₄ kg⁻¹VS_fed for the studied crop materials. Thus, high COD solubilization, high nitrogen mineralization and solubilization rates were feasible during anaerobic digestion of lignocellulosic materials in a two-stage LBR-UASB reactor system.     

Effect of starch addition on the biological conversion and microbial community in a methanol-fed UASB reactor during long-term continuous operation

The effect of starch addition on the microbial composition and the biological conversion was investigated using two upflow anaerobic sludge bracket (UASB) reactors treating methanolic wastewater: one reactor was operated with starch addition, and another reactor was operated without starch addition. Approximately 300 days of operation were performed at 30 kg COD/m³/d, and then, the organic load of the reactors was gradually increased to 120 kg COD/m³/d. Successful operation was achieved at 30 kg COD/m³/d in both reactors; however, the methanol-fed reactor did not perform well at 120 kg COD/m³/d while the methanol-starch-fed reactor did. The granule analysis revealed the granule developed further only in the methanol-starch-fed reactor. The results of the microbial community analysis revealed more Methanosaeta cells were present in the methanol-starch-fed reactor, suggesting the degradation of starch produced acetate as an intermediate, which stimulated the growth of Methanosaeta cells responsible for the extension of granules.     

Modelling mono-digestion of grass silage in a 2-stage CSTR anaerobic digester using ADM1

This paper examines 174 days of experimental data and modelling of mono-digestion of grass silage in a two stage wet process with recirculation of liquor; the two vessels have an effective volume of 312 L each. The organic loading rate is initiated at 0.5 kg VS m⁻³ d⁻¹ (first 74 days) and subsequently increased to 1 kg VS m⁻³ d⁻¹. The experimental data was used to generate a mathematical model (ADM1) which was calibrated over the first 74 days of operation. Good accuracy with experimental data was found for the subsequent 100 days. Results of the model would suggest starting the process without recirculation and thus building up the solids content of the liquor. As the level of VFA increases, recirculation should be employed to control VFA. Recirculation also controls solids content and pH. Methane production was estimated at 88% of maximum theoretical production.     

Hydrogen production by the newly isolated Clostridium beijerinckii RZF-1108

A fermentative hydrogen-producing strain, RZF-1108, was isolated from a biohydrogen reactor, and identified as Clostridium beijerinckii on the basis of the 16S rRNA gene analysis and physiobiochemical characteristics. The effects of culture conditions on hydrogen production by C. beijerinckii RZF-1108 were investigated in batch cultures. The hydrogen production and growth of strain RZF-1108 were highly dependent on temperature, initial pH and substrate concentration. Yeast extract was a favorable nitrogen source for hydrogen production and growth of RZF-1108. Hydrogen production corresponded to cell biomass yield in different culture conditions. The maximum hydrogen evolution, yield and production rate of 2209 ml H₂/l medium, 1.97 mol H₂/mol glucose and 104.20 ml H₂/g CDW h⁻¹ were obtained at 9 g/l of glucose, initial pH of 7.0, inoculum volume of 8% and temperature of 35 °C, respectively. These results demonstrate that C. beijerinckii can efficiently produce H₂, and is another model microorganism for biohydrogen investigations.     

Use of sulfate reducing cell suspension bioreactors for the treatment of SO2 rich flue gases

This paper describes a novel bioscrubber concept for biological flue gas desulfurization, based on the recycling of a cell suspension of sulfite/sulfate reducing bacteria between a scrubber and a sulfite/sulfate reducing hydrogen fed bioreactor. Hydrogen metabolism in sulfite/sulfate reducing cell suspensions was investigated using batch activity tests and by operating a completely stirred tank reactor (CSTR). The maximum specific hydrogenotrophic sulfite/sulfate reduction rate increased with 10% and 300%, respectively, by crushing granular inoculum sludge and by cultivation of this sludge as cell suspension in a CSTR. Operation of a sulfite fed CSTR (hydraulic retention time 4 days; pH 7.0; sulfite loading rate 0.5–1.5 g SO ₃ ^2- l^-1 d^-1) with hydrogen as electron donor showed that high (up to 1.6 g l^-1) H₂S concentrations can be obtained within 10 days of operation. H₂S inhibition, however, limited the sulfite reducing capacity of the CSTR. Methane production by the cell suspension disappeared within 20 days reactor operation. The outcompetition of methanogens in excess of H₂ can be attributed to CO₂ limitation and/or to sulfite or sulfide toxicity. The use of cell suspensions opens perspectives for monolith or packed bed reactor configurations, which have a much lower pressure drop compared to air lift reactors, to supply H₂ to sulfite/sulfate reducing bioreactors.     

Long term operation of pilot-scale biological nutrient removal process in treating municipal wastewater

The performance of a pilot-scale biological nutrient removal process has been evaluated for 336 days, receiving the real municipal wastewater with a flowrate of 6.8 m³/d. The process incorporated an intermittent aeration reactor for enhancing the effluent quality, and a nitrification reactor packed with the porous polyurethane foam media for supporting the attached-growth of microorganism responsible for nitrification. The observation shows that the process enabled a relatively stable and high performance in both organics and nutrient removals. When the SRT was maintained at 12 days, COD, nitrogen, and phosphorus removals averaged as high as 89% at a loading rate of 0.42–3.95 kg COD/m³ d (corresponding to average influent concentration of 304 mg COD/L), 76% at the loading rate of 0.03–0.27 kg N/m³ d (with 37.1 mg TN/L on average), and 95% at the loading rate of 0.01–0.07 kg TP/m³ d (with 5.4 mg TP/L on average), respectively.     

Process of inorganic nitrogen transformation and design of kinetics model in the biological aerated filter reactor

As one of the plug-flow reactors, biological aerated filter (BAF) reactor was divided into four sampling sectors to understand the characteristics of elemental nitrogen transformation during the reaction process, and then the different characteristics of elemental nitrogen transformation caused by different NH₃-N loadings, biological quantities and activities in each section were obtained. The results showed that the total transformation ratio in the nitrifying reactor was more than 90% in the absence of any organic carbon resource, at the same time, more than 65% NH₃-N in the influent were nitrified at the filter height of 70 cm below under the conditions of the influent runoff 9-19 L/h, the gas-water ratio 4-5:1, the dissolved oxygen 3.0-5.8 mg/L and the NH₃-N load 0.28-0.48 kg NH₃-N/m³ d. On the base of the Eckenfelder mode, the kinetics equation of the NH₃-N transformation along the reactor was S_e = S₀ exp(−0.0134D/L^1.2612).