Anaerobic treatment of cassava stillage for hydrogen and methane production in continuously stirred tank reactor (CSTR) under high organic loading rate (OLR)

Anaerobic hydrogen and methane production from cassava stillage in continuously stirred tank reactor (CSTR) were investigated in this study. Results showed that the heat-pretreatment of inoculum did not enhance hydrogen yield compared to raw inoculum under mesophilic condition after continuous operation. However, the hydrogen yield increased from about 14 ml H₂/gVS under mesophilic condition to 69.6 ml H₂/gVS under thermophilic condition due to the decrease of propionate concentration and inhibition of homoacetogens. Therefore, temperature was demonstrated to be more important than pretreatment of inoculum to enhance the hydrogen production. Under high organic loading rate (OLR) (>10 gVS/(L·d)), the two-phase thermophilic CSTR for hydrogen and methane production was stable with hydrogen and methane yields of 56.6 mlH₂/gVS and 249 mlCH₄/gVS. The one-phase thermophilic CSTR for methane production failed due to the accumulation of both acetate and propionate, leading to the pH lower than 6. Instead of propionate alone, the accumulations of both acetate and propionate were found to be related to the breakdown of methane reactor.     

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.     

Hydrogen and methane production in extreme thermophilic conditions in two-stage (upflow anaerobic sludge bed) UASB reactor system

Two-stage hydrogen and methane production in extreme thermophilic (70 °C) conditions was demonstrated for the first time in UASB-reactor system. Inoculum used in hydrogen and methane reactors was granular sludge from mesophilic internal circulation reactor and was first acclimated for extreme thermophilic conditions. In hydrogen reactor, operated with hydraulic retention time (HRT) of 5 h and organic loading rate (OLR) of 25.1 kg COD/m³/d, hydrogen yield was 0.73 mol/mol glucose_added. Methane was produced in second stage from hydrogen reactor effluent. In methane reactor operated with HRT of 13 h and OLR of 7.8 kg COD/m³/d, methane yield was 117.5 ml/g COD_added. These results prove that hydrogen and methane can be produced in extreme thermophilic temperatures, but as batch experiments confirmed, for methane production lower temperature would be more efficient.     

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.     

Direct fermentation of Laminaria japonica for biohydrogen production by anaerobic mixed cultures

A few studies have been made on fermentative hydrogen production from marine algae, despite of their advantages compared with other biomass substrates. In this study, fermentative hydrogen production from Laminaria japonica (one brown algae species) was investigated under mesophilic condition (35 ± 1 °C) without any pretreatment method. A feasibility test was first conducted through a series of batch cultivations, and 0.92 mol H₂/mol hexose_added, or 71.4 ml H₂/g TS of hydrogen yield was achieved at a substrate concentration of 20 g COD/L (based on carbohydrate), initial pH of 7.5, and cultivation pH of 5.5. Continuous operation for a period of 80 days was then carried out using anaerobic sequencing batch reactor (ASBR) with a hydraulic retention time (HRT) of 6 days. After operation for approximately 30 days, a stable hydrogen yield of 0.79 ± 0.03 mol H₂/mol hexose_added was obtained. To optimize bioenergy recovery from L. japonica, an up-flow anaerobic sludge blanket reactor (UASBr) was applied to treat hydrogen fermentation effluent (HFE) for methane production. A maximum methane yield of 309 ± 12 ml CH₄/g COD was achieved during the 90 days operation period, where the organic loading rate (OLR) was 3.5 g COD/L/d.     

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.     

An online-monitored thermophilic hydrogen production UASB reactor for long-term stable operation

In this study, the H₂ production and chemical oxygen demand (COD) removal performances of a thermophilic upflow anaerobic sludge blanket (UASB) reactor with online monitoring system were investigated. The online monitoring system enabled a rapid monitoring and timely control of the process. As a consequence, high operating stability was achieved despite of the varied hydraulic retention time (HRT) during 310-day operation. The COD efficiency remained at above 98%, and the hydrogen yield fluctuated slightly within the range of 2.42–3.06 mol H₂ mol⁻¹ sucrose. Thermophilic H₂-producing granules were successfully cultivated in this reactor, which showed better physical and microbial properties than floc sludge and higher H₂ production rate than mesophilic granules. An analysis of the microbial growth kinetics further demonstrated a possibly higher synthesis and metabolism activity of microbes in the thermophilic granule state.     

Optimization of biohydrogen production of palm oil mill effluent by ozone pretreatment

The biohydrogen (H₂) production in batch experiments under varying concentrations of raw and ozonated palm oil mill effluent (POME) of 5000–30,000 mg COD.L⁻¹, at initial pH 6, under mesophilic (37 °C), thermophilic (55 °C) and extreme-thermophilic (70 °C) conditions. Effects of ozone pretreatment, substrate concentration and fermentation temperature on H₂ production using mesophilic seed sludge was undertaken. The results demonstrated that H₂ can be produced from both raw and ozonated POME, and the amounts of H₂ production were directly increased as the POME concentrations were increased. H₂ was successfully produced under the mesophilic fermentation of ozonated POME, with maximum H₂ yield, and specific H₂ production rate of 182 mL.g⁻¹ COD_removed (30,000 mg COD.L⁻¹) and 6.2 mL.h⁻¹.g⁻¹ TVS (25,000 mg COD.L⁻¹), respectively. Thus, indicating that the ozone pretreatment could elevate on the biodegradability of major constituents of the POME, which significantly enhanced yields and rates of the H₂ production. H₂ production was not achieved under the thermophilic and extreme-thermophilic fermentation. In both fermentation temperatures with ozonated POME, the maximum H₂ yield was 62 mL.g⁻¹ COD_removed (30,000 mg COD.L⁻¹) and 63 mL.g⁻¹ COD_removed (30,000 mg COD.L⁻¹), respectively. The highest efficiency of total and soluble COD removal was obtained at 44 and 37%, respectively following the mesophilic fermentation, of 24 and 25%, respectively under the thermophilic fermentation, of 32 and 20%, respectively under the extreme-thermophilic fermentation. The production of volatile fatty acids increased with an increased fermentation time and temperature in both raw and ozonated POME under all three fermentation temperatures. The accumulation of volatile fatty acids in the reactor content were mostly acetic and butyric acids. H₂ fermentation under the mesophilic condition of 37 °C was the better selection than that of the thermophilic and extreme-thermophilic fermentation.     

Effect of temperature on continuous hydrogen production of cellulose

The effect of temperature on the hydrogen fermentation of cellulose was evaluated by a continuous experiment using a mixed culture without pretreatment. The experiments were conducted at three different temperatures, which were mesophilic [37 ± 2 °C], thermophilic [55 ± 2 °C] and hyper-thermophilic [80 ± 2 °C], with an influent concentration of cellulose of 5 g/l and a hydraulic retention time [HRT] of 10 days. A stable hydrogen production was observed at each condition. At 37 ± 2 °C, the maximum hydrogen yield was 0.6 mmol H₂/g cellulose. However, at 55 ± 2 °C and 80 ± 2 °C, the maximum hydrogen yields were 15.2 and 19.02 mmol H₂/g cellulose, respectively. While 26% of the biogas was methane under the mesophilic temperature, no methane gas was detected under both the thermophilic and hyper-thermophilic temperatures. The results show that operational temperature is a key to sustainable bio-hydrogen production and that the thermophilic and hyper-thermophilic conditions produced better results than mesophilic condition.     

A bench scale study of fermentative hydrogen and methane production from food waste in integrated two-stage process

A two-stage fermentation process combining hydrogen and methane production for the treatment of food waste was investigated in this paper. In hydrogen fermentation reactor, the indigenous mixed microbial cultures contained in food waste were used for hydrogen production. No foreign inoculum was used in the hydrogen fermentation stage, the traditional heat treatment of inoculum was not applied either in this bench scale experiment. The effects of the stepwise increased organic loading rate (OLR) and solid retention time (SRT) on integrated two-stage process were investigated. At steady state, the optimal OLR and SRT for the integrated two-stage process were found to be 22.65 kg VS/m³ d (160 h) for hydrogen fermentation reactor and 4.61 (26.67 d) for methane fermentation reactor, respectively. Under the optimum conditions, the maximum yields of hydrogen (0.065 m³ H₂/kg VS) and methane (0.546 m³ CH₄/kg VS) were achieved with the hydrogen and methane contents ranging from 29.42 to 30.86%, 64.33 to 71.48%, respectively. Biodegradability analysis showed that 5.78% of the influent COD was converted to the hydrogen in H₂-SCRD and 82.18% of the influent COD was converted to the methane in CH₄-SCSTR under the optimum conditions.     

Comparison of thermophilic and hyperthermophilic dark fermentation with subsequent mesophilic methanogenesis in expanded granular sludge bed reactors

Herein, dark fermentation (DF, V = 5.5 L) and subsequent mesophilic methanogenesis (V = 43.5 L) are run as expanded granular sludge bed reactors (EGSB) at thermophilic (υDF = 60 °C) and hyperthermophilic (υDF = 80 °C) temperatures. A synthetic glucose wastewater is run with a 22.5 g/L chemical oxygen demand (COD) and 48–9 h hydraulic retention times (HRTs), giving organic loading rates (OLRs) of 11–60 g COD/L/d for DF. The maximum hydrogen production rate (HPR) is HPR = 3.0 m³/m³/d for HRT = 9 h with a 50 L/kg COD hydrogen yield (HY) and 40 vol% H₂. Methane production rate (MPR) reaches MPR = 2.6 m³/m³/d with 70 vol% CH₄ at HRT = 2.8 d. The highest H₂ yields are HY = 180 L/kg COD with 53 vol% H₂ (thermophilic, HRT = 48 h). Hyperthermophilic temperatures led to lower HPRs (0.7 m³/m³/d) and MPRs (1.6 m³/m³/d). 53% of Thermoanaerobacterium thermosaccharolyticum as an H₂ producer are found. Discoloration of granular sludge from black to white and granule stability was observed in DF.     

Two-stage UASB reactor converting coffee drink manufacturing wastewater to hydrogen and methane

In this study, the feasibility of a continuous two-stage up-flow anaerobic sludge blanket (UASB) reactor system, consisted of thermophilic (55 °C) dark fermentative H₂ production and mesophilic (35 °C) CH₄ production from coffee drink manufacturing wastewater (CDMW), was tested. A recently proposed operational strategy was used to overcome a major drawback of the long start-up period of the UASB reactor. Firstly, a completely stirred tank reactor (CSTR) was operated for 8 days to prepare seeding. The seed was then directly transferred to the UASB reactor. Microbial aggregation took place in the initial period, and the floc size was gradually increased over time. In UASB reactor, the maximum H₂ yield of 2.57 mol H₂/mol hexose_added and a stable H₂ production rate of 4.24 L H₂/L/h were observed at a hydraulic retention time (HRT) of 6 h and substrate concentration of 20 g Carbo. COD/L. In this novel method using CDMW, thermophilic H₂-producing granules with an average particle size of 1.3 mm was successfully developed after 100 days. The more bioenergy recovery was attempted in a post-treatment process using a mesophilic UASB reactor for CH₄ production from the H₂ fermented effluent. The maximum CH₄ yield of 325 mL of CH₄/g COD was achieved with removing 93% of the COD at an organic loading rate of 3.5 g COD/L/d. The developed two-stage UASB reactor system achieved biogas conversion by 88.2% (H₂ 15.2% and CH₄ 73%) and COD removal by 98%.     

Effects of solid retention time on anaerobic digestion of dewatered-sewage sludge in mesophilic and thermophilic conditions

Anaerobic digestion of dewatered-sewage sludge using continuous stirred tank reactors (CSTRs) in duplicates was evaluated under thermophilic (50 °C) and mesophilic (37 °C) conditions over a range of nine solid retention times (SRTs). The 35- and 30-day SRTs were designed to simulate a full-scale plant operation while 25-, 20-, 15- and 12-day SRTs were planned to evaluate process performance at the various SRTs. The 9-, 5- and 3-day SRTs were performed to push the reactors to extend their degradation capacity and test the threshold for process imbalance. The corresponding organic loading rates (OLR) varied from 1.6 to 20.5 kg VS m^?3 day^?1. Biogas production rate could be tripled when the SRT was shortened from 30 to 12 days and more than doubled from 35- to 15-day SRT because of a concomitant increase in OLR. In general, higher biogas productivity was realized under thermophilic, but methane yields were comparable due to the higher methane content in the biogas under mesophilic digestion. The methane content in biogas fluctuated between 55 and 65% and the methane yield ranged from 0.314 to 0.348 Nm³ CH₄ kg VS_added^?1 day^?1 for both thermophilic and mesophilic digestion. The VS-reduction at 12- and 15-day SRT ranged from 45 to 52% and there was no accumulation of VFAs. Increasing concentrations of VFAs, decreasing concentration of partial alkalinity and decrease in pH were noted as signs of reactor instability. Process imbalance started at 9-day SRT, souring of the reactors, cell wash-out and foaming was noted as the principal causes of process failure under both thermophilic and mesophilic conditions. This study projected the possibility of using CSTRs in treating dewatered-sewage sludge at a shorter SRT to achieve reasonable biogas production and VS-reduction without encountering adverse operation conditions as foaming and wash-out of cells.     

Bio-hydrogen production from cheese whey powder (CWP) solution: Comparison of thermophilic and mesophilic dark fermentations

Cheese whey powder (CWP) solution was used as the raw material for hydrogen gas production by mesophilic (35 °C) and thermophilic (55 °C) dark fermentations at constant initial total sugar and bacteria concentrations. Thermophilic fermentation yielded higher cumulative hydrogen formation (CHF = 171 mL), higher hydrogen yield (111 mL H₂ g⁻¹ total sugar), and higher hydrogen formation rate (3.46 mL H₂ L⁻¹ h⁻¹) as compared to mesophilic fermentation. CHF in both cases were correlated with the Gompertz equation and the constants were determined. Despite the longer lag phase, thermophilic fermentation yielded higher specific H₂ formation rate (2.10 mL H₂ g⁻¹cells h⁻¹). Favorable results obtained in thermophilic fermentation were probably due to elimination of H₂ consuming bacteria at high temperatures and selection of fast hydrogen gas producers.     

Biological saccharification by Clostridium thermocellum and two-stage hydrogen and methane production from hydrogen peroxide-acetic acid pretreated sugarcane bagasse

The present work aimed at establishing an efficient degradation and energy recovery system form sugarcane bagasse (SCB) through hydrogen peroxide-acetic acid (HPAC) pretreatment, thermophilic hydrogen production and mesophilic methane production. The degradation ratio of HPAC pretreated SCB (HPAC-SCB, 2%, w/v) exceeded 90% under the biological hydrolysis of C. thermocellum without enzyme addition. The hydrogen yield in the co-culture fermentation of T. thermosaccharolyticum and C. thermocellum from HPAC-SCB (2%, w/v) reached 226 mL/g substrate. The long-term hydrogen fermentation was successfully established with 1.59 L/(L·d), 0.159 L/g substrate for average hydrogen productivity and yield, respectively. Methane production of 0.341 L/g COD (chemical oxygen demand)_added was recovered by semi-continuous methane fermentation from hydrogen-producing effluent at 12 days of hydraulic retention time (HRT). Average energy recovery of 8.79 MJ/kg SCB was obtained under the optimal conditions. The present work indicated the promising application of the established process in valorization of lignocellulosic bio-waste.     

Thermophilic hydrogen and methane production from sugarcane stillage in two-stage anaerobic fluidized bed reactors

This study evaluated the feasibility of H₂ and CH₄ production in two-stage thermophilic (55 °C) anaerobic digestion of sugarcane stillage (5,000 to 10,000 mg COD.L⁻¹) using an acidogenic anaerobic fluidized bed reactor (AFBR-A) with a hydraulic retention time (HRT) of 4 h and a methanogenic AFBR (AFBR-S) with HRTs of 24 h–10 h. To compare two-stage digestion with single-stage digestion, a third methanogenic reactor (AFBR-M) with a HRT of 24 h was fed with increasing stillage concentrations (5,000 to 10,000 mg COD.L⁻¹). The AFBR-M produced a methane content of 68.4 ± 7.2%, a maximum yield of 0.30 ± 0.04 L CH₄.g COD⁻¹, a production rate of 3.78 ± 0.40 L CH₄.day⁻¹.L⁻¹ and a COD removal of 73.2 ± 5.0% at an organic loading rate (OLR) of 7.5 kg COD.m⁻³.day⁻¹. In contrast, the two-stage AFBR-A system produced a hydrogen content of 23.9 ± 5.6%, a production rate of 1.30 ± 0.16 L H₂.day⁻¹.L⁻¹ and a yield of 0.34 ± 0.08 mmol H₂.g CODap⁻¹. Additionally, the decrease in the HRT from 18 h to 10 h in the AFBR-S favored a higher methane production, improving the maximum methane content (74.5 ± 6.0%), production rate (5.57 ± 0.38 L CH₄.day⁻¹.L⁻¹) and yield (0.26 ± 0.06 L CH₄.g COD⁻¹) at an OLR of 21.6 kg COD.m⁻³.day⁻¹ (HRT of 10 h) with a total COD removal of 70.1 ± 7.1%. Under the applied COD of 10,000 mg L⁻¹, the two-stage system showed a 52.8% higher energy yield than the single-stage anaerobic digestion system. These results show that, relative to a single-stage system, two-stage anaerobic digestion systems produce more hydrogen and methane while achieving similar treatment efficiencies.     

Characteristics of biohydrogen fermentation from various substrates

The characteristics of biohydrogen production from sucrose, slurry-type piggery waste and food waste under the effects of the reactor configurations and operational pHs (6 and 9) were examined by using heat-treated anaerobic sludge as a seed biomass. When sucrose was used in the batch test, the maximum hydrogen yield was 0.12–0.13 g COD (as H₂)/g COD (1.41–1.43 mol/mol hexose) at pH 6. In contrast, 0.10–0.11 g COD (as H₂)/g COD (1.12–1.21 mol/mol hexose) hydrogen yield was achieved from the reactor at pH 9. On the other hand, hydrogen production was not observed in the continuous sequencing batch mode fermenters fed with sucrose. Profile analysis at each cycle revealed hydrogen production at the initial operation periods but eventually only methane at 36 days. When slurry-type piggery waste was used as the substrate, the upflow elutriation-type fermenters produced methane but not hydrogen after 30 days operation. The fermentation intermediate profile showed that the hydrogen produced might have been consumed by homoacetogenic or propionate producing reactions, and eventually converted into methane by acetoclastic methanogens. The downflow leaching bed fermenters using food waste produced 0.013 L H₂/g volatile solids (VS) (0.0061 g COD (as H₂)/g COD) at pH 6 with 54% VS reduction whereas 0.0041 L H₂/g VS (0.0020 g COD (as H₂)/g COD) was produced at pH 9 with 86% VS reduction. The results show that the hydrogen produced should be released rapidly from the reactor before it can be consumed in other biochemical reactions, and substrates with high pH level (>9.0) can be used directly to produce hydrogen without needing to adjust the pH.     

Biohydrogen production from palm oil mill effluent (POME) by two stage anaerobic sequencing batch reactor (ASBR) system for better utilization of carbon sources in POME

The production of biohydrogen through dark fermentation of palm oil mill effluent (POME) was evaluated in two-stages of biohydrogen in an anaerobic sequencing batch reactor (ASBR) system using enriched mixed culture for the first time. This study attempts to examine the effect of HRT and its interaction behavior with the solid retention time (SRT), and the sugar consumption. The effluent after discharged from the thermophilic reactor contained 7.61 g/L TC and 22.87 g/L TSS was fed to the secondary mesophilic reactor system. Results indicated that the overall sugar consumption reached 88.62% at the optimum HRT of 12 h with the SRT set to 20 h. The optimum hydrogen yield and HPR in the thermophilic stage were 2.99 mol H₂/mol-sugar and 8.54 mmol H₂/L·h respectively, while for the mesophilic stage were 1.19 mol H₂/mol-sugar and 1.47 mmolH₂/L·h respectively. The overall HPR showed an improvement and increase from 8.54 mmol H₂/L·h to 10.34 mmol H₂/L.h. Microbial community analysis of mixed culture in the two-stage thermophilic (55.0 °C) and mesophilic (37.0 °C) ASBR reactor was dominated by Thermoanaerobacterium sp. based on the PCR-DGGE technique.     

Hydrogen-methane production from pulp & paper sludge and food waste by mesophilic–thermophilic anaerobic co-digestion

The present study focused on the mesophilic anaerobic bio-hydrogen production from PPS (pulp & paper sludge) and FW (food waste), and the subsequent anaerobic digestion of the effluent for the methane production under thermophilic conditions by a two-stage process. The maximum hydrogen yield of 64.48 mL g⁻¹ VS_fed and methane yield of 432.3 mL g⁻¹ VS_fed were obtained when PPS and FW were applied with 1: 1 VS ratio as the feedstock. No VFA were cumulated in the reactor during the period of hydrogen - methane fermentation, as well as no NH₃–N and Na⁺ inhibition were found in the process. 71%–87% removal efficiencies of SCOD were attained for hydrogen and methane co-production. pH 4.8–6.4 and alkalinity 794–3316 mg CaCO₃ L⁻¹ for H₂ fermentation, as well as pH 6.5–8.8 and alkalinity 4165–4679 mg CaCO₃ L⁻¹ for CH₄ fermentation, were achieved without any adjustment. This work showed that anaerobic co-digestion of PPS and FW for hydrogen-methane co-production was a stable, reliable and effective way for energy recovery and bio-solid waste stabilization by the two-stage mesophilic–thermophilic process.     

Effects of temperature and substrate concentration on biological hydrogen production from starch

This study investigates the effects of temperature and substrate concentration on biological hydrogen production from starch using mixed cultures. In this work, although hydrogen was successfully produced under the thermophilic condition, stable hydrogen production was not observed under the mesophilic condition. In the thermophilic reactor, the maximum hydrogen yield was 2.8 mol H₂/mol glucose at 20 g/l-starch; however, hydrogen yield decreased drastically with the change of by-product distribution when substrate concentration was over 30 g/l-starch. A negative correlation was observed between the hydrogen yield and the total concentration of undissociated acids.