Biological hydrogen production in an anaerobic sequencing batch reactor: pH and cyclic duration effects

An anaerobic sequencing batch reactor (ASBR) was used to evaluate biological hydrogen production from carbohydrate-rich organic wastes. The goal of the proposed project was to investigate the effects of pH (4.9, 5.5, 6.1, and 6.7), and cyclic duration (4, 6, and 8 h) on hydrogen production. With the ASBR operated at 16-h HRT, 25 g COD/L, and 4-h cyclic duration, the results showed that the maximum hydrogen yield of 2.53 mol H₂/mol sucrose_consumed appeared at pH 4.9. The carbohydrate removal efficiency declined to 56% at pH 4.9, which indirectly resulted in the reduction of total volatile fatty acid production. Acetate fermentation was the dominant metabolic pathway at pH 4.9. The concentration of mixed liquor volatile suspended solid (MLVSS) also showed a decrease from nearly 15,000 mg/L between pHs 6.1 and 6.7 to 6000 mg/L at pH 4.9. Investigation of the effect of cyclic duration found that hydrogen yield reached the maximum of 1.86 mol H₂/mol sucrose_consumed at 4-h cyclic duration while ASBR was operating at 16-h HRT, 15 g COD/L, and pH 4.9. The experimental results showed that MLVSS concentration increased from 6200 mg/L at 4-h cyclic duration to 8500 mg/L at 8-h cyclic duration. However, there was no significant change in effluent volatile suspended solid concentration. The results of butyrate to acetate ratio showed that using this ratio to correlate the performance of hydrogen production is not appropriate due to the growth of homoacetogens. In ASBR, the operation is subject to four different phases of each cycle, and only the complete mix condition can be achieved at react phase. The pH and cyclic duration under the unique operations profoundly impact fermentative hydrogen production.     

Hydrogen production from alcohol wastewater with added fermentation residue by an anaerobic sequencing batch reactor (ASBR) under thermophilic operation

The objective of this study was to investigate the enhancement of hydrogen production from alcohol wastewater by adding fermentation residue using an anaerobic sequencing batch reactor (ASBR) under thermophillic operation (55 °C) and at a constant pH of 5.5. The digestibility of the added fermentation residue was also evaluated. For a first set of previous experiments, the ASBR system was operated to obtain an optimum COD loading rate of 50.6 kg/m³ d of alcohol wastewater without added fermentation residue and the produced gas contained 31% H₂ and 69% CO₂. In this experiment, the effect of added fermentation residue (100–1200 mg/l) on hydrogen production performance was investigated under a COD loading rate of 50.6 kg/m³ d of the alcohol wastewater. At a fermentation residue concentration of 1000 mg/l, the produced gas contained 40% H₂ and 60% CO₂ without methane and the system gave the highest hydrogen yield and specific hydrogen production rate of 128 ml/g COD removed and 2880 ml/l d, respectively. Under thermophilic operation with a high total COD loading rate (51.8 kg/m³ d) and a short HRT (21 h) at pH 5.5, the ASBR system could only break down cellulose (41.6%) and hemicellulose (21.8%), not decompose lignin.     

Optimization of biohydrogen production from sweet sorghum syrup using statistical methods

This study employed statistically based experimental designs to optimize fermentation conditions for hydrogen production from sweet sorghum syrup by anaerobic mixed cultures. Initial screening of important factors influencing hydrogen production, i.e., total sugar, initial pH, nutrient solution, iron (II) sulphate (FeSO₄), peptone and sodium bicarbonate was conducted by the Plackett–Burman method. Results indicated that only FeSO₄ had statistically significant (P ≤ 0.005) influences on specific hydrogen production (P_s) while total sugar and initial pH had an interdependent effect on P_s. Optimal conditions for the maximal P_s were 25 g/L total sugar, 4.75 initial pH and 1.45 g/L FeSO₄ in which P_s of 6897 mL H₂/L was estimated. Estimated optimum conditions revealed only 0.04% difference from the actual P_s of 6864 mL H₂/L which suggested that the optimal conditions obtained can be practically applied to produce hydrogen from sweet sorghum syrup with the least error.     

Hydrogen production from alcohol distillery wastewater containing high potassium and sulfate using an anaerobic sequencing batch reactor

In this study, the feasibility of hydrogen production from alcohol distillery wastewater containing high potassium and sulfate was investigated using an anaerobic sequencing batch reactor (ASBR). The seed sludge taken from an anaerobic tank treating the distillery wastewater was boiled for 15 min before being fed to the ASBR. The ASBR system was operated under different feed chemical oxygen demand (COD) values and different COD loading rates at a mesophilic temperature of 37 °C, a controlled pH at 5.5, and a cycle time of 6 cycles per day. When the studied ASBR was operated under the best conditions (providing a maximum hydrogen production efficiency) of a feed COD of 40,000 mg/l, a COD loading rate of 60 kg/m³ d, and a hydraulic retention time of 16 h, the produced gas was found to contain 34.7% H₂ and 65.3% CO_2, without any methane being detected. Under these best conditions, the specific hydrogen production rate (SHPR) of 270 ml H₂/g MLVSS d (or 3310 ml H₂/l d), and hydrogen yield of 172 ml H₂/g COD removed, were obtained. When the feed COD exceeded 40,000 mg/l, the process performance in terms of hydrogen production decreased because of the potassium and sulfate toxicity.     

Effect of pH on continuous biohydrogen production from liquid swine manure with glucose supplement using an anaerobic sequencing batch reactor

pH is considered as one of the most important factors governing the hydrogen fermentation process. In this project, five pH levels, ranging from 4.4 to 5.6 at 0.3 increments, were tested to evaluate the pH effect on hydrogen production from swine manure supplemented with glucose in an anaerobic sequencing batch reactor system with 16 h of hydraulic retention time (HRT). The optimal hydrogen yield (1.50 mol H₂/mol glucose) was achieved at pH 5.0 when the maximum production rate of 2.25 L/d/L was obtained. Continuous hydrogen production was achieved for over 3 weeks for pH 5.0, 4.7, and 4.4, with no significant methane produced. However, as pH increased to 5.3 and 5.6, methane production was observed in the biogas with concurrent reductions in hydrogen production, indicating that methanogens could become increasingly activated for pH 5.3 or higher. Acetate, propionate, butyrate, valerate, and ethanol were the main aqueous products whose distribution was significantly affected by pH as well.     

Enhanced bio‐hydrogen production from sweet sorghum stalk with alkalization pretreatment by mixed anaerobic cultures

Bio‐hydrogen production from sweet sorghum stalk using mixed anaerobic sludge was reported in this paper. Batch tests were carried out to analyze influences of several environmental factors on yield of H₂ from sweet sorghum stalk under constant mesophillic temperature (36±1°C). The experimental results showed that, for the raw stalk, the cumulative hydrogen yield was 52.1 ml g⁻¹·TVS with utilization percentages of sugars, hemi‐cellulose and cellulose in the stalk being 89.12, 15.23 and 13.89% respectively; whereas for the stalk pretreated by 0.4% NaOH solution at room temperature for 24 h, the cumulative hydrogen yield was 127.26 ml g⁻¹·TVS with utilization percentages of sugars, hemi‐cellulose and cellulose being 99.17, 53.64, 41.56%, respectively. The hydrogen content in the biogas was about 53% while the methane content was less than 4% throughout the study. Besides hydrogen and methane, the main metabolic products detected were ethanol, propionate and butyrate. The experimental results suggested that the alkalization pretreatment of the substrate plays a crucial role in the conversion of the sweet sorghum stalk wastes into bio‐hydrogen by the mixed anaerobic sludge. Copyright © 2009 John Wiley & Sons, Ltd.     

Enhancing biohydrogen production from chemical wastewater treatment in anaerobic sequencing batch biofilm reactor (AnSBBR) by bioaugmenting with selectively enriched kanamycin resistant anaerobic mixed consortia

The basic aim of this study was to investigate the feasibility of bioaugmentation strategy in the process of enhancing biohydrogen (H₂) production from chemical wastewater treatment (organic loading rate (OLR)—6.3 kg COD/m³-day) in anaerobic sequencing batch biofilm reactor (AnSBBR) operated at room temperature (28±2$28\pm 2$ °C) under acidophilic microenvironment (pH 6) with a total cycle period of 24 h. Parent augmented inoculum (kanamycin resistant) was acquired from an operating upflow anaerobic sludge blanket (UASB) reactor treating chemical wastewater and subjected to selective enrichment by applying repetitive/cyclic pre-treatment methods [altering between heat-shock treatment (100 °C; 2 h) and acid treatment (pH 3; 24 h)] to eliminate non-spore forming bacteria and to inhibit the growth of methanogenic bacteria (MB). Experimental data revealed the positive influence of bioaugmentation strategy on the overall H₂ production. Specific H₂ production almost doubled after augmentation from 0.297 to 0.483 mol H₂/kg COD_R-day. Chemical wastewater acted as primary carbon source in the metabolic reactions involving molecular H₂ generation leading to substrate degradation. The augmented culture persisted in the system till the termination of the experiments. The survival and retention of the augmented inoculum and its positive effect on process enhancement may be attributed to the adopted reactor configuration and operating conditions. Scanning electron microscope (SEM) images documented the selective enrichment of morphologically similar group of bacteria capable of producing H₂ under acidophilic conditions in anaerobic microenvironment. This depicted work corroborated successful application of bioaugmentation strategy to improve H₂ production rate from anaerobic chemical wastewater treatment.     

Hydrogen production from alcohol wastewater by an anaerobic sequencing batch reactor under thermophilic operation: Nitrogen and phosphorous uptakes and transformation

The objective of this study was to investigate hydrogen production from alcohol wastewater using an anaerobic sequencing batch reactor (ASBR) under thermophilic operation and at a constant pH of 5.5. Under the optimum COD loading rate of 68 kg/m³d, the produced gas contained 43% H₂ without methane and the system provided a hydrogen yield and specific hydrogen production rate of 130 ml H₂/g COD removed and 2100 ml H₂/l d, respectively, which were much higher than those obtained under the mesophilic operation. Under thermophilic operation, both nitrogen and phosphate uptakes were minimal at the optimum COD loading rate for hydrogen production and most nitrogen uptake was derived from organic nitrogen. Under the thermophilic operation for hydrogen production, the nutrient requirement in terms of COD:N:P was found to be 100:6:0.5, which was much higher than that for the methenogenic step for methane production under both thermophilic and mesophilic operations and for the acidogenic step for hydrogen production under mesophilic operation.     

Hydrogen production from cassava wastewater using an anaerobic sequencing batch reactor: Effects of operational parameters,COD:N ratio,and organic acid composition

In this work, hydrogen production from cassava wastewater using anaerobic sequencing batch reactors (ASBR) was investigated to determine the optimum number of cycles per day, chemical oxygen demand (COD) loading rate, and COD:N ratio. The system operated at a COD loading rate of 30 kg/m³d and 6 cycles per day provided maximum hydrogen production performance in terms of specific hydrogen production rate (SHPR) (388 ml H₂/g VSS d or 3800 ml H₂/l d) and hydrogen yield (186 ml H₂/g COD removed). The effect of nitrogen supplementation was also studied by adding NH₄HCO₃ into the system at the COD:N ratios of 100:2.2, 100:3.3, and 100:4.4 under the COD loading rate of 30 kg/m³d and 6 cycles per day. The maximum SHPR and hydrogen yield of 524 ml H₂/g VSS d (5680 ml H₂/l d) and 438 ml H₂/g COD removed, respectively, were obtained at the stoichiometric COD:N ratio of 100:2.2. An excess nitrogen was found to promote the productions of higher organic acids and ethanol, resulting in lowering hydrogen production efficiency.     

Feasibility studies on continuous hydrogen production using photo-fermentative sequencing batch reactor

A novel photo-fermentative sequencing batch reactor (PFSBR) process assisted by activated carbon fibers (ACFs) was used to continuously produce hydrogen gas by Rhodopseudomonas faecalis RLD-53. Feasibility of continuous hydrogen production in PFSBR operation at different hydraulic retention times (HRTs) (48, 96, 144 and 192 h) and influent acetate concentrations (20, 40, 60 and 80 mmol/l) was investigated. The rate and yield of hydrogen production increased with HRTs from 48 to 144 h, and then decreased with the HRTs from 144 to 192 h. Regulation of the proper influent acetate concentration (60 mmol/l) not only increased hydrogen production by PFSBR, but also maintained quality of the effluent with high substrate removal efficiency (97.70%). Free R. faecalis RLD-53 was adsorbed on the surface of ACFs, initially isolated cells, then monolayer, and finally mature biofilm with three dimensional multilayers structures. The PFSBR reached a maximum hydrogen yield (3.12 mol H₂/mol acetate), and achieved a steady state when mature biofilm developed on ACFs. Therefore, photo-fermentative sequencing batch reactor was a promising process for continuous photo-fermentative hydrogen production.     

Biological hydrogen production in suspended and attached growth anaerobic reactor systems

Biological production of hydrogen gas has received increasing interest from the international community during the last decade. Most studies on biological fermentative hydrogen production from carbohydrates using mixed cultures have been conducted in conventional continuous stirred tank reactors (CSTR) under mesophilic conditions. Investigations on hydrogen production in reactor systems with attached microbial growth have recently come up as well as investigations on hydrogen production in the thermophilic temperature range. The present study examines and compares the biological fermentative production of hydrogen from glucose in a continuous stirred tank type bioreactor (CSTR) and an upflow anaerobic sludge blanket bioreactor (UASB) at various hydraulic retention times (2–12 h HRT) under mesophilic conditions (35 °C). Also the biohydrogen production from glucose in the CSTR at mesophilic and thermophilic (55 °C) temperature range was studied and compared. From the CSTR experiments it was found that thermophilic conditions combine high hydrogen production rate with low production of microbial mass, thus giving a specific hydrogen production rate as high as 104 mmole _H2/h/l/g${\mathrm{H}}_2/\mathrm{h}/\mathrm{l}/\mathrm{g}$ VSS at 6 h retention time compared to a specific hydrogen production rate of 12 mmole _H2/h/l/g${\mathrm{H}}_2/\mathrm{h}/\mathrm{l}/\mathrm{g}$ VSS under mesophilic conditions. On the other hand, the UASB reactor configuration is more stable than the CSTR regarding hydrogen production, pH, glucose consumption and microbial by-products (e.g. volatile fatty acids, alcohols etc.) at the HRTs tested. Moreover, the hydrogen production rate in the UASB reactor was significantly higher compared to that of the CSTR at low retention times (19.05 and 8.42 mmole _H2/h/l${\mathrm{H}}_2/\mathrm{h}/\mathrm{l}$, respectively at 2 h HRT) while hydrogen yield (mmole _H2/mmole${\mathrm{H}}_2/\mathrm{mmole}$ glucose consumed) was higher in the CSTR reactor at all HRT tested. This implies that there is a trade-off between technical efficiency (based on hydrogen yield) and economic efficiency (based on hydrogen production rate) when the attached (UASB) and suspended (CSTR) growth configurations are compared.     

Effect of substrate concentration on fermentative hydrogen production from sweet sorghum extract

The aim of the present study was to assess the influence of substrate concentration on the fermentative hydrogen production from sweet sorghum extract, in a continuous stirred tank bioreactor. The reactor was operated at a Hydraulic Retention Time (HRT) of 12 h and carbohydrate concentrations ranging from 9.89 to 20.99 g/L, in glucose equivalents. The maximum hydrogen production rate and yield were obtained at the concentration of 17.50 g carbohydrates/L and were 2.93 ± 0.09 L H₂/L reactor/d and 0.74 ± 0.02 mol H₂/mol glucose consumed, corresponding to 8.81 ± 0.02 L H₂/kg sweet sorghum, respectively. The main metabolic product at all steady states was butyric acid, while ethanol production was high at high substrate concentrations. The experiments showed that hydrogen productivity depends significantly on the initial carbohydrate concentration, which also influences the distribution of the metabolic products.     

Evolution of methane and hydrogen from ethanol wastewater with maximization of energy yield by three-stage anaerobic sequencing batch reactor system

A new anaerobic sequencing batch reactor (ASBR) unit with three stage concept was proposed and tested for spontaneous generation of hydrogen (H₂) and methane (CH₄) from ethanol wastewater with maximum energy yield. The system was controlled at 37 °C, a fixed pH of 5.5 in the first bioreactor and a recycle ratio of unity from the third bioreactor to the first bioreactor to lower the amount of sodium hydroxide for pH regulation in the first bioreactor and to increase total alkalinity and process stability. Under the optimum organic loading rate of 15 kg COD/m³d (based on the total liquid holding volume), it exhibited the highest yields and specific generation rates of both gases, resulting in the uppermost overall energy yield which was much higher than those of all other anaerobic systems. The lactic acid was found to be about 4–8 times more toxic than acetic acid to methanogens. The higher the solution pH in any anaerobic system, the greater the reduction of micronutrients.     

An anaerobic sequential batch reactor for enhanced continuous hydrogen production from fungal pretreated cornstalk hydrolysate

Anaerobic sequencing batch reactor (ASBR) process offers great potential for H₂ production from wastewaters. In this study, an ASBR was used at first time for enhanced continuous H₂ production from fungal pretreated cornstalk hydrolysate by Thermoanaerobacterium thermosaccharolyticum W16. The reactor was operated at different hydraulic retention times (HRTs) of 6, 12, 18, and 24 h by keeping the influent hydrolysate constant at 65 mmol sugars L⁻¹. Results showed that increasing the HRT from 6 to 12 h led to the H₂ production rate increased from 6.7 to the maximum of 9.6 mmol H₂ L⁻¹ h⁻¹ and the substrate conversion reached 90.3%, although the H₂ yield remained at the same level of 1.7 mol H₂ mol⁻¹ substrate. Taking into account both H₂ production and substrate utilization efficiencies, the optimum HRT for continuous H₂ production via an ASBR was determined at 12 h. Compared with other continuous H₂ production processes, ASBR yield higher H₂ production at relatively lower HRT. ASBR is shown to be another promising process for continuous fermentative H₂ production from lignocellulosic biomass.     

Effect of hydraulic retention time on hydrogen production and chemical oxygen demand removal from tapioca wastewater using anaerobic mixed cultures in anaerobic baffled reactor (ABR)

This study evaluated hydrogen production and chemical oxygen demand removal (COD removal) from tapioca wastewater using anaerobic mixed cultures in anaerobic baffled reactor (ABR). The ABR was conducted based on the optimum condition obtained from the batch experiment, i.e. 2.25 g/L of FeSO₄ and initial pH of 9.0. The effects of the varying hydraulic retention times (HRT: 24, 18, 12, 6 and 3 h) on hydrogen production and COD removal in a continuous ABR were operated at room temperature (32.3 ± 1.5 °C). Hydrogen production rate (HPR) increased with a reduction in HRT i.e. from 164.45 ± 4.14 mL H₂/L.d (24 h HRT) to 883.19 ± 7.89 mL H₂/L.d (6 h HRT) then decreased to 748.54 ± 13.84 mL H₂/L.d (3 h HRT). COD removal increased with reduction in HRT i.e. from 14.02 ± 0.58% (24 h HRT) to 29.30 ± 0.84% (6 h HRT) then decreased to 21.97 ± 0.94% (3 h HRT). HRT of 6 h was the optimum condition for ABR operation as indicated.     

Microbial community analysis of mixed anaerobic microflora in suspended sludge of ASBR producing hydrogen from palm oil mill effluent

This study investigated the microbial community of an anaerobic sequencing batch reactor (ASBR) operating at mesophilic temperature under varying hydraulic retention times (HRTs) for evaluating optimal hydrogen production conditions, using palm oil mill effluent (POME) as substrate. POME sludge enriched by heat treatment with hydrogen-producing bacteria was used as inoculum and acclimated with the POME. The microbial community was determined by first isolating cultivable bacteria at each operating HRT and then using polymerase chain reaction (PCR). The PCR products were sequenced and sequence identification was performed using the BLAST algorithm and Genbank database. The findings revealed that about 50% of the isolates present were members of the genus Streptococcus, about 30% were Lactobacillus species and around 20% were identified as species of genus Clostridium. Scanning electron microscopy (SEM) analysis also confirmed the presence of spherical and rod-shaped microbial morphologies in the sludge samples of bioreactor during prolonged cultivation.     

Effect of hydraulic retention time on biohydrogen production from palm oil mill effluent in anaerobic sequencing batch reactor

The feasibility of hydrogen generation from palm oil mill effluent (POME), a high strength wastewater with high solid content, was evaluated in an anaerobic sequencing batch reactor (ASBR) using enriched mixed microflora, under mesophilic digestion process at 37 °C. Four different hydraulic retention times (HRT), ranging from 96 h to 36 h at constant cycle length of 24 h and various organic loading rate (OLR) concentrations were tested to evaluate hydrogen productivity and operational stability of ASBR. The results showed higher system efficiency was achieved at HRT of 72 h with maximum hydrogen production rate of 6.7 LH₂/L/d and hydrogen yield of 0.34 LH₂/g COD_feeding, while in longer and shorter HRTs, hydrogen productivity decreased. Organic matter removal efficiency was affected by HRT; accordingly, total and soluble COD removal reached more than 37% and 50%, respectively. Solid retention time (SRT) of 4-19 days was achieved at these wide ranges of HRTs. Butyrate was found to be the dominant metabolite in all HRTs. Low concentration of volatile fatty acid (VFA) confirmed the state of stability and efficiency of sequential batch mode operation was achieved in ASBR. Results also suggest that ASBR has the potential to offer high digestion rate and good stability of operation for POME treatment.     

Modeling of fermentative hydrogen production from sweet sorghum extract based on modified ADM1

The Anaerobic digestion model 1 (ADM1) framework can be used to predict fermentative hydrogen production, since the latter is directly related to the acidogenic stage of the anaerobic digestion process. In this study, the ADM1 model framework was used to simulate and predict the process of fermentative hydrogen production from the extractable sugars of sweet sorghum biomass. Kinetic parameters for sugars’ consumption and yield coefficients of acetic, propionic and butyric acid production were estimated using the experimental data obtained from the steady states of a CSTR. Batch experiments were used for kinetic parameter validation. Since the ADM1 does not account for metabolic products such as lactic acid and ethanol that are crucial during the fermentative hydrogen production process, the structure of the model was modified to include lactate and ethanol among the metabolites and to improve the predictions. The modified ADM1 simulated satisfactorily batch experiments although further modifications could be made in order to further improve the predictions for the hydrogenogenic process.     

Using a statistical approach to model hydrogen production from a steam exploded corn stalk hydrolysate fed to mixed anaerobic cultures in an ASBR

In this study, hydrogen (H₂) production from the fermentation of steam exploded corn stalk (CS) liquor was statistically optimized using a fractional factorial design approach. The factors under consideration included temperature, pH and hydraulic retention time (HRT). Under optimal conditions at 53 °C, a pH at 4.5 and a 9.5 h HRT, the observed maximum H₂ yield of 98 ± 2 mL g⁻¹ TVS together with negligible CH₄ were similar to the model predicted responses. A flux analysis revealed negligible homoacetogenic activity in cultures at 53 °C and low pH (≤5.5). Both homoacetogenic (R17) and methanogenic (R28 and R29) fluxes accounted for more than 68–90% of H₂ consumption in cultures at low temperatures. The low H₂ yields observed in cultures maintained at 21 °C and 37 °C was associated with high lactate and solvent levels. High H₂ yields in cultures at 53 °C were associated with a higher abundance of Clostrdium sp. and CH₄ production at low temperatures was due to the presence of hydrogenotrophic methanogens (Methanothermobacter marburgensis and Methanobrevibacter ruminatum) and aceticlastic methanogens (Methanosaeta sp. and Methanosarcina sp.). The results obtained from this study (within the factor ranges investigated) indicated that steam exploded CS liquor could be a potential substrate for H₂ production using mixed microbial cultures.