Anaerobic fluidized bed reactor with expanded clay as support for hydrogen production through dark fermentation of glucose

This study evaluated hydrogen production in an anaerobic fluidized bed reactor (AFBR) fed with glucose-based synthetic wastewater. Particles of expanded clay (2.8–3.35 mm) were used as a support material for biomass immobilization. The reactor was operated with hydraulic retention times (HRT) ranging from 8 to 1 h. The hydrogen yield production increased from 1.41 to 2.49 mol H₂ mol⁻¹ glucose as HRT decreased from 8 to 2 h. However, when HRT was 1 h, there was a slight decrease to 2.41 mol H₂ mol⁻¹ glucose. The biogas produced was composed of H₂ and CO₂, and the H₂ content increased from 8% to 35% as HRT decreased. The major soluble metabolites during H₂ fermentation were acetic acid (HAc) and butyric acid (HBu), accounting for 36.1–53.3% and 37.7–44.9% of total soluble metabolites, respectively. Overall, the results demonstrate the potential of using expanded clay as support material for hydrogen production in AFBRs.     

Performance and composition of bacterial communities in anaerobic fluidized bed reactors for hydrogen production: Effects of organic loading rate and alkalinity

This study evaluated the effects of the organic loading rate (OLR) and pH buffer addition on hydrogen production in two anaerobic fluidized bed reactors (AFBRs) operated simultaneously. The AFBRs were fed with glucose, and expanded clay was used as support material. The reactors were operated at a temperature of 30 °C, without the addition of a buffer (AFBR1) and with the addition of a pH buffer (AFBR2, sodium bicarbonate) for OLRs ranging from 19.0 to 140.6 kg COD m⁻³ d⁻¹ (COD: chemical oxygen demand). The maximum hydrogen yields for AFBR1 and AFBR2 were 2.45 and 1.90 mol H₂ mol⁻¹ glucose (OLR of 84.3 kg COD m⁻³ d⁻¹), respectively. The highest hydrogen production rates were 0.95 and 0.76 L h⁻¹ L⁻¹ for AFBR1 and AFBR2 (OLR of 140.6 kg COD m⁻³ d⁻¹), respectively. The operating conditions in AFBR1 favored the presence of such bacteria as Clostridium, while the bacteria in AFBR2 included Clostridium, Enterobacter, Klebsiella, Veillonellaceae, Chryseobacterium, Sporolactobacillus, and Burkholderiaceae.     

Continuous hydrogen production from cofermentation of sugarcane vinasse and cheese whey in a thermophilic anaerobic fluidized bed reactor

The co-fermentation of vinasse and cheese whey (CW) was evaluated in this study by using two thermophilic (55° C) anaerobic fluidized bed reactors (AFBRs). In AFBR using vinasse and CW (AFBR-V-CW), the CW was added in increasing proportions (2, 4, 6, 8, and 10 g COD.L^?1) to vinasse (10 g COD.L^?1) to assess the advantage of adding CW to vinasse. By decreasing the hydraulic retention time (HRT) from 8 h to 1 h in AFBR-V, maximum hydrogen yield (HY), production rate (HPR), and H₂ content (H₂%) of 1.01 ± 0.06 mmol H₂.g COD^?1, 2.54 ± 0.39 L H₂.d^?1.L^?1, and 47.3 ± 2.9%, respectively, were observed at an HRT of 6 h. The increase in CW concentration to values over 2 g COD.L^?1 in AFBR-V-CW decreased the HY, PVH, and H₂%, with observed maximum values of 0.82 ± 0.07 mmol H₂.g COD^?1, 1.41 ± 0.24 L H₂.d^?1.L^?1, and 55.5 ± 3.7%, respectively, at an HRT of 8 h. The comparison of AFBR-V-CW and AFBR-V showed that the co-fermentation of vinasse with 2 g COD.L^?1 of CW increased the HPR, H₂%, and HY by 117%, 68%, and 82%, respectively.     

Different ratios of carbon sources in the fermentation of cheese whey and glucose as substrates for hydrogen and ethanol production in continuous reactors

The fermentation of glucose, cheese whey and the mixture of glucose and cheese whey were evaluated in this study from two inocula sources (sludge from a UASB reactor for swine wastewater treatment and poultry slaughterhouse) for hydrogen production in continuous anaerobic fluidized bed reactors (AFBR). For all fermentations, a hydraulic retention time (HRT) of 6 h and a substrate concentration of 5 g COD L⁻¹ were used. In glucose fermentation, the maximum hydrogen yield (HY) was 1.37 mmol H₂ g⁻¹ COD. The co-fermentation of the cheese whey and glucose mixture was favorable for the concomitant production of hydrogen and ethanol, with yields of up to 1.7 mmol H₂ g⁻¹ COD and 3.45 mol EtOH g⁻¹ COD in AFBR2. The utilization of cheese whey as a sole substrate resulted in an HY of 1.9 mmol H₂ g⁻¹ COD. Throughout the study, ethanol fermentation was evident.     

Fermentative hydrogen production from starch using natural mixed cultures

Batch and continuous tests were conducted to evaluate fermentative hydrogen production from starch (at a concentration of chemical oxygen demand (COD) 20 g/L) at 35 °C by a natural mixed culture of paper mill wastewater treatment sludge. The optimal initial cultivation pH (tested range 5–7) and substrate concentration (tested range 5–60-gCOD/L) were evaluated by batch reactors while the effects of hydraulic retention time (HRT) on hydrogen production, as expressed by hydrogen yield (HY) and hydrogen production rate (HPR), were evaluated by continuous tests. The experimental results indicate that the initial cultivation pH markedly affected HY, maximum HPR, liquid fermentation product concentration and distribution, butyrate/acetate concentration ratio and metabolic pathway. The optimal initial cultivation pH was 5.5 with peak values of HY 1.1 mol-H₂/mol-hexose maximum HPR 10.4 mmol-H₂/L/h and butyrate concentration 7700 mg-COD/L. In continuous hydrogen fermentation, the optimal HRT was 4 h with peak HY of 1.5 mol-H₂/mol-hexose, peak HPR of 450 mmol-H₂/L/d and lowest butyrate concentration of 3000 mg-COD/L. The HPR obtained was 280% higher than reported values. A shift in dominant hydrogen-producing microbial population along with HRT variation was observed with Clostridium butyricum, C. pasteurianum, Klebshilla pneumoniae, Streptococcus sp., and Pseudomonas sp. being present at efficient hydrogen production at the HRTs of 4–6 h. Strategies based on the experimental results for optimal hydrogen production from starch are proposed.     

Hydrogen production from diluted and raw sugarcane vinasse under thermophilic anaerobic conditions

Two continuous anaerobic fluidized bed reactors (AFBRs) were operated under thermophilic (55 °C) temperature for 150 days to investigate the effect of dark H₂ fermentation of diluted and raw sugarcane vinasse on H₂ production using mixed seed sludge. Although effective H₂ production (52.8% of H₂; 0.80 L H₂ h⁻¹ L⁻¹; 0.79 mmol gCOD_added⁻¹) was observed using an elevated substrate concentration (30,000 mg COD L⁻¹), the optimal operational conditions were found for the AFBR₁ (10,000 mg COD L⁻¹) fed with diluted sugarcane vinasse (HRT 6 h; OLR 40 kg COD m⁻³ d⁻¹), achieving a H₂ yield of 2.86 mmol H₂ g COD_added⁻¹. H₂ production was inhibited by elevated volatile fatty acid (VFA) concentrations (butyric and acetic acids = 3.7 and 3.0 g L⁻¹, respectively) in the raw feedstock. Denaturing gradient gel electrophoresis (DGGE) analyses revealed changes in the bacterial population of the expanded clay biofilm as a function of the substrate concentration.     

Continuous production of hydrogen from oat straw hydrolysate in a biotrickling filter

Hydrogen was produced in a biotrickling filter (BF) packed with perlite and fed with oat straw acid hydrolysate at 30 °C. Inlet chemical oxygen demand (COD) from 1.2 to 35 g/L and hydraulic retention time (HRT) between 24 h and 6 h were assayed. With increasing inlet COD or decreasing HRT, H₂ production rate (HPR) increased but H₂ production yield (HY) decreased. Maximum HPR of 81.4 mL H₂/L_reactor h (3.3 mmol H₂/L_reactor h) and HY of 2.9 mol H₂/mol_{hexose consumed} were found at an inlet COD of 0.05 g_COD/L h (HRT 24 h) and 2.9 g_COD/L h (HRT 12 h), respectively. Maximum hydrogen composition in gas was 45 ± 4% (v/v) with CO₂ as balance. Methane was not detected. Maximum HPR and inlet COD used in this work were higher than others reported for reactors with suspended or fixed biomass. However, implementation of strategies for biomass control to avoid reactor clogging is needed.     

Improving hydrogen production from cassava starch by combination of dark and photo fermentation

The combination of dark and photo fermentation was studied with cassava starch as the substrate to increase the hydrogen yield and alleviate the environmental pollution. The different raw cassava starch concentrations of 10–25 g/l give different hydrogen yields in the dark fermentation inoculated with the mixed hydrogen-producing bacteria derived from the preheated activated sludge. The maximum hydrogen yield (HY) of 240.4 ml H₂/g starch is obtained at the starch concentration of 10 g/l and the maximum hydrogen production rate (HPR) of 84.4 ml H₂/l/h is obtained at the starch concentration of 25 g/l. When the cassava starch, which is gelatinized by heating or hydrolyzed with α-amylase and glucoamylase, is used as the substrate to produce hydrogen, the maximum HY respectively increases to 258.5 and 276.1 ml H₂/g starch, and the maximum HPR respectively increases to 172 and 262.4 ml H₂/l/h. Meanwhile, the lag time (λ) for hydrogen production decreases from 11 h to 8 h and 5 h respectively, and the fermentation duration decreases from 75–110 h to 44–68 h. The metabolite byproducts in the dark fermentation, which are mainly acetate and butyrate, are reused as the substrates in the photo fermentation inoculated with the Rhodopseudomonas palustris bacteria. The maximum HY and HPR are respectively 131.9 ml H₂/g starch and 16.4 ml H₂/l/h in the photo fermentation, and the highest utilization ratios of acetate and butyrate are respectively 89.3% and 98.5%. The maximum HY dramatically increases from 240.4 ml H₂/g starch only in the dark fermentation to 402.3 ml H₂/g starch in the combined dark and photo fermentation, while the energy conversion efficiency increases from 17.5–18.6% to 26.4–27.1% if only the heat value of cassava starch is considered as the input energy. When the input light energy in the photo fermentation is also taken into account, the whole energy conversion efficiency is 4.46–6.04%.     

Non-sterile bio-hydrogen fermentation from food waste in a continuous stirred tank reactor (CSTR): Performance and population analysis

Bio-hydrogen production from food waste by anaerobic mixed cultures was conducted in a continuous stirred tank reactor (CSTR). The hydraulic retention time (HRT) was optimized in order to maximize hydrogen yield (HY) and hydrogen production rate (HPR). The maximum hydrogen content (38.6%), HPR (379 mL H₂/L. d) and HY (261 mL H₂/g-VS_added) were achieved at the optimum HRT of 60 h. The major soluble metabolite products were butyric and acetic acids which indicated a butyrate-acetate type fermentation. Operation of CSTR at HRT 60 h could select hydrogen producing bacteria and eliminate lactic acid bacteria and acetogenic bacteria. The microbial community analyzed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) revealed that the predominant hydrogen producer was Clostridium sp.     

Long-term stability of hydrogen and organic acids production in an anaerobic fluidized-bed reactor using heat treated anaerobic sludge inoculum

This study evaluates the stability of hydrogen and organic acids production in an anaerobic fluidized-bed reactor (AFBR) that contains expanded clay (2.8–3.35 mm in diameter) as a support medium and is operated on a long-term basis. The reactor was inoculated with thermally pre-treated anaerobic sludge and operated with decreasing hydraulic retention time (HRT), from 8 h to 1 h, at a controlled temperature of 30 °C and a pH of about 3.8. Glucose (2000 mg L^?1) was used as the substrate, generating conversion rates of 92–98%. Decreasing the HRT from 8 h to 1 h led to an increase in average hydrogen-production rates, with a maximum value of 1.28 L h^?1 L^?1 for an HRT of 1 h. In general, hydrogen yield production increased as HRT decreased, reaching 2.29 mol of H₂/mol glucose at an HRT of 2 h and yielding a maximum hydrogen content of 37% in the biogas. No methane was detected in the biogas throughout the period of operation. The main soluble metabolites (SMP) were acetic acid (46.94–53.84% of SMP) and butyric acid (34.51–42.16% of SMP), with less than 15.49% ethanol. The steady performance of the AFBR may be attributed to adequate thermal treatment of the inoculum, the selection of a suitable support medium for microbial adhesion, and the choice of satisfactory environmental conditions imposed on the system. The results show that stable hydrogen production and organic acids production were maintained in the AFBR over a period of 178 days.     

Effect of hydraulic retention time on a continuous biohydrogen production in a packed bed biofilm reactor with recirculation flow of the liquid phase

The present paper reports on results obtained from experiments carried out in a laboratory-scale anaerobic packed bed biofilm reactor (APBR), with recirculation of the liquid phase, for continuously biohydrogen production via dark fermentation. The reactor was filled with Kaldnes^® biofilm carrier and inoculated with an anaerobic mesophilic sludge from a urban wastewater treatment plant (WWTP). The APBR was operated at a temperature of 37 °C, without pH buffering. The effect of theoretical hydraulic retention time (HRT) from 1 to 5 h on hydrogen yield (HY), hydrogen production rate (HPR), substrate conversion and metabolic pathways was investigated. This study indicates the possibility of enhancing hydrogen production by using APBR with recirculation flow. Among respondents values of HRT the highest average values of HY (2.35 mol H₂/mol substrate) and HPR (0.085 L h^?1L^?1) have been obtained at HRT equal to 2 h.     

Optimizing hydrogen production from a switchgrass steam exploded liquor using a mixed anaerobic culture in an upflow anaerobic sludge blanket reactor

In this study, the operation of an upflow anaerobic sludge blanket reactor (UASBR) producing hydrogen (H₂) from a steam-exploded switchgrass (SWG) liquor was statistically optimized. The factors consider included pH, hydraulic retention time (HRT) and linoleic acid (LA) concentration. Under optimal operational conditions (pH 5.0, 10 h HRT and 1.75 g L⁻¹ LA), which were close to the predicted conditions using the D-optimality index, the maximum H₂ and methane yield observed were 99.86 ± 5.6 mL g⁻¹ TVS and 0.5 ± 0.1 mL g⁻¹ TVS, respectively. Under maximum H₂-producing conditions, high levels of acetate plus butyrate were observed with low levels of ethanol and lactate. A principal component analysis revealed that clustering of the samples was based on the operating conditions and fermentation metabolites. The microbial profiles revealed that by lowering the HRT from 16 to 8 h or decreasing the pH from 7.0 to 5.0 in the controls caused a 50% reduction in the relative abundance of the terminal restriction fragments belonging to the methanogenic population (Methanobacteria, Methanomicrobia, Methanococci). With LA treatment, H₂ producers (Ruminococcaceae and Clostridiaceae) were dominant and methanogens were inhibited and/or washed-out from the UASBR.     

Continuous thermophilic hydrogen production and microbial community analysis from anaerobic digestion of diluted sugar cane stillage

The aim of this study was to promote biohydrogen production in an thermophilic anaerobic fluidized bed reactor (AFBR) at 55 °C using a mixture of sugar cane stillage and glucose at approximately 5000–5300 mg COD L⁻¹. During a reduction in the hydraulic retention time (HRT) from 8, 6, 4, 2 and 1 h, H₂ yields of 5.73 mmol g COD_added⁻¹ were achieved (at HRT of 4 h, with organic loading rate of 52.7 kg COD m⁻³ d⁻¹). The maximum volumetric H₂ production of 0.78 L H₂ h⁻¹ L⁻¹ was achieved using stillage as carbon source. In all operational phases, the H₂ average content in the biogas was between 31.4 and 52.0%. Butyric fermentation was the predominant metabolic pathway. The microbial community in accordance with the DGGE bands profile was found similarity coefficient between 91 and 95% without significant changes in bacterial populations after co-substrate removal. Bacteria like Thermoanaerobacterium sp. and Clostridium sp. were identified.     

Biohydrogen production from crude glycerol by immobilized Klebsiella sp. TR17 in a UASB reactor and bacterial quantification under non-sterile conditions

Biohydrogen production from crude glycerol by immobilized Klebsiella sp. TR17 was investigated in an up-flow anaerobic sludge blanket (UASB) reactor. The reactor was operated under non-sterile conditions at 40^○C and initial pH 8.0 at different hydraulic retention times (HRTs) (2–12 h) and glycerol concentrations (10–30 g/L). Decreasing the HRT led to an increase in hydrogen production rate (HPR) and hydrogen yield (HY). The highest HPR of 242.15 mmol H₂/L/d and HY of 44.27 mmol H₂/g glycerol consumed were achieved at 4 h HRT and glycerol concentrations of 30 and 10 g/L, respectively. The main soluble metabolite was 1,3-propanediol, which implies that Klebsiella sp. was dominant among other microorganisms. Fluorescence in situ hybridization (FISH) revealed that the microbial community was dominated by Klebsiella sp. with 56.96, 59.45, and 63.47% of total DAPI binding cells, at glycerol concentrations of 10, 20, and 30 g/L, respectively.     

Bio-hydrogen production by photo-fermentation of dark fermentation effluent with intermittent feeding and effluent removal

Pure culture of Rhodobacter sphaeroides (NRRL- B1727) was used for continuous photo-fermentation of volatile fatty acids (VFA) present in the dark fermentation effluent of ground wheat starch. The feed contained 1950 ± 50 mg L⁻¹ total VFA with some nutrient supplementation. Hydraulic residence time (HRT) was varied between 24 and 120 hours. The highest steady-state daily hydrogen production (55 ml d⁻¹) and hydrogen yield (185 ml H₂ g⁻¹ VFA) were obtained at HRT = 72 hours (3 days). Biomass concentration increased with increasing HRT. Volumetric and specific hydrogen formation rates were also maximum at HRT = 72 h. High extent of TVFA fermentation at HRT = 72 h resulted in high hydrogen gas production.     

Seed inocula for biohydrogen production from biodiesel solid residues

This study aimed to optimize the hydrogen production from various seed sludges (two kinds of sewage sludges (S1, S2), cow dung (S3), granular sludge (S4) and effluent from condensed soluble molasses H₂ fermenter (S5)) and enhancement of hydrogen production via heat treatment for substrate and seed sludge by using the solid residues of biodiesel production (BDSR). Two batch assay tests were operated at a biodiesel solid residue concentration of 10 g/L, temperature of 55 °C and an initial cultivation pH of 8. The results showed that the peak hydrogen yield (HY) of 94.6 mL H₂/g volatile solid (VS) (4.1 mmolH₂/g VS) was obtained from S1 when substrate and seed sludge were both heat treated at 100 °C for 1 h. However, the peak hydrogen production rate (HPR) and specific hydrogen production rate (SHPR) of 1.48 L H₂/L-d and 0.30 L H₂/g VSS-d were obtained from S2 without any treatment. The heat treatment was found to increase the HY in both the cases of sewage sludges S1 and S2.The HY of 89.5 mL H₂/g VS (without treatment) was increased to 94.6 mL H₂/g VS and 82.6 mL H₂/g VS (without treatment) was increased to 85.7 mL H₂/g VS for S1 and S2. The soluble metabolic product (SMP) analysis showed that the fermentation followed mainly acetate–butyrate pathway with considerable production of ethanol. The total bioenergy production was calculated as 2.8 and 2.9 kJ/g VS for favorable hydrogen and ethanol production, respectively. The BDSR could be used as feedstock for dark fermentative hydrogen production.     

Biohydrogen production from renewable agri-waste blend: Optimization using mixer design

Biohydrogen from untreated mixed renewable agri-waste using buffalo dung compost is reported. Corn husk (CH) supported 25% higher hydrogen (H₂) production and showed the maximum value (62.38%) with p value (1.2 × 10⁻⁶) revealing its significance at individual and interactive level, respectively, compared to ground nut shell (GNS) and rice husk (RH). Augmented-simplex-lattice design experimentation revealed that a partial supplementation of RH or GNS to CH improves H₂ yield. Multiple-linear-regression analysis indicated that a quadratic model (low p = 0.0023, high F value = 35.99 and R²_quadratic = 0.99) was more significant compared to other (linear, cubic and special cubic) models. Acetate and butyrate were accounted >80% of the volatile fatty acids (VFAs). A maximum accumulation of 65.78 ml H₂ g⁻¹ TVS was produced using agri-wastes in the ratio of CH:RH:GNS = 70:16:12.     

Production of hydrogen from sewage biosolids in a continuously fed bioreactor: Effect of hydraulic retention time and sparging

Hydrogen was produced from primary sewage biosolids via mesophilic anaerobic fermentation in a continuously fed bioreactor. Prior to fermentation the sewage biosolids were heated to 70 °C for 1 h to inactivate methanogens and during fermentation a cellulose degrading enzyme was added to improve substrate availability. Hydraulic retention times (HRT) of 18, 24, 36 and 48 h were evaluated for the duration of hydrogen production. Without sparging a hydraulic retention time of 24 h resulted in the longest period of hydrogen production (3 days), during which a hydrogen yield of 21.9 L H₂ kg⁻¹ VS added to the bioreactor was achieved. Methods of preventing the decline of hydrogen production during continuous fermentation were evaluated. Of the techniques evaluated using nitrogen gas to sparge the bioreactor contents proved to be more effective than flushing just the headspace of the bioreactor. Sparging at 0.06 L L min⁻¹ successfully prevented a decline in hydrogen production and resulted in a yield of 27.0  L H₂ kg⁻¹ VS added, over a period of greater than 12 days or 12 HRT. The use of sparging also delayed the build up of acetic acid in the bioreactor, suggesting that it serves to inhibit homoacetogenesis and thus maintain hydrogen production.     

HRT control as a strategy to enhance continuous hydrogen production from sugarcane juice under mesophilic and thermophilic conditions in AFBRs

The objective of this study was to evaluate the effects of hydraulic retention time (HRT) (8–1 h) on H₂ production from sugarcane juice (5000 mg COD L⁻¹) in mesophilic (30 °C, AFBR-30) and thermophilic (55 °C, AFBR-55) anaerobic fluidized bed reactors (AFBRs). At HRTs of 8 and 1 h in AFBR-30, the H₂ production rates were 60 and 116 mL H₂ h⁻¹ L⁻¹, the hydrogen yields were 0.60 and 0.10 mol H₂ mol⁻¹ hexose, and the highest bacterial diversities were 2.47 and 2.34, respectively. In AFBR-55, the decrease in the HRT from 8 to 1 h increased the hydrogen production rate to 501 mL H₂ h⁻¹ L⁻¹ at the HRT of 1 h. The maximum hydrogen yield of 1.52 mol H₂ mol⁻¹ hexose was observed at the HRT of 2 h and was associated with the lowest bacterial diversity (0.92) and highest bacterial dominance (0.52).     

COX-free H2 production via catalytic decomposition of CH4 over Ni supported on zeolite catalysts

Catalytic decomposition of methane produces CO_X-free hydrogen, which is necessary for PEM fuel-cell applications. In this paper, hydrogen production by catalytic decomposition of methane at 550 °C over Ni on HY, USY, SiO₂ and SBA-15 supports is examined at atmospheric pressure. The catalytic activities and the life times of the catalysts are evaluated and discussed. The relationships between catalyst performance and characterization of the fresh and used catalysts are discussed with the results obtained from SEM, XRD, TPR, solid acidity and the measured carbon contents generated of the used samples along with their H₂ production rates. Among all the catalysts tested, Ni supported on HY zeolite showed a higher activity of 955 mol H₂ (mol Ni)⁻¹ and a longevity of 720 min at 550 °C.