Enhancing bio-hydrogen and bio-methane production of concentrated latex wastewater (CLW) by Co-digesting with palm oil mill effluent (POME): Batch and continuous performance test and ADM-1 modeling.

This study aimed at investigating the biohydrogen and biomethane potential of co-digestion from palm oil mill effluent (POME) and concentrated latex wastewater (CLW) in a two-stage anaerobic digestion (AD) process under thermophilic (55 ± 3 °C) and at an ambient temperature (30 ± 3 °C) conditions, respectively. The batch experiments of POME:CLW mixing ratios of 100:0, 70:30, 50:50, 30:70, and 0:100 was investigated with the initial loadings at 10 g-VS/L. The highest hydrogen yield of 115.57 mLH2/g-VS was obtained from the POME: CLW mixing ratio of 100:0 with 29.0 of C/N ratio. While, the highest subsequent methane production yield of 558.01 mLCH4/g-VS was achieved from hydrogen effluent from POME:CLW mixing ratio of 70:30 0 with 21.8 of C/N ratio. This mixing ratio revealed the highest synergisms of about 9.21% and received maximum total energy of 19.70 kJ/g-VS. Additionally, continuous hydrogen and methane production were subsequently performed in a series of continuous stirred tank reactor (CSTR) and up-flow anaerobic sludge blanket reactor (UASB) to treat the co-substate. The results indicated that the highest hydrogen yield of POME:CLW mixing ratio at 70:30 of 95.45 mL-H2/g-VS was generated at 7-day HRT, while methane production was obtained from HRT 15 days with a yield of 204.52 mL-CH4/g-VS. Thus, the study indicated that biogas production yield of CLW could be enhanced by co-digesting with POME. In addition, the two-stage AD model under anaerobic digestion model no. 1 (ADM-1) framework was established, 9.10% and 2.43% of error fitting of hydrogen and methane gas between model simulation data and experimental data were found. Hence, this research work presents a novel approach for optimization and feasibility for co-digestion of POME with CLW to generate mixed gaseous biofuel potentially.

Potential of butanol production from Thailand marine macroalgae using Clostridium beijerinckii ATCC 10132-based ABE fermentation.

The economical bio-butanol-based fermentation process is mainly limited by the high price of first-generation biomass, which is an intensive cost for the pretreatment of second-generation biomass. As third-generation biomass, marine macroalgae could be potentially advantageous for conversion to clean and renewable bio-butanol through acetone-butanol-ethanol (ABE) fermentation. In this study, butanol production from three macroalgae species (Gracilaria tenuistipitata, Ulva intestinalis, and Rhizoclonium sp.) by Clostridium beijerinckii ATCC 10132 was assessed comparatively. The enriched C beijerinckii ATCC 10132 inoculum produced a high butanol concentration of 14.07 g L-1 using 60 g L-1 of glucose. Among the three marine seaweed species, G. tenuistipitata exhibited the highest potential for butanol production (1.38 g L-1 ). Under the 16 conditions designed using the Taguchi method for low-temperature hydrothermal pretreatment (HTP) of G. tenuistipitata, the maximum reducing sugar yield rate of 57.6% and ABE yield of 19.87% were achieved at a solid to liquid (S/L) ratio of 120, temperature of 110°C, and holding time of 10 min (Severity factor, R0 1.29). In addition, pretreated G. tenuistipitata could be converted to 3.1 g L-1 of butanol using low-HTP at an S/L ratio of 50 g L-1 , temperature of 80°C (R0 0.11), and holding time of 5 min.

Effect of organic loading rate and effluent recirculation on biogas production of desulfated skim latex serum using up-flow anaerobic sludge blanket reactor.

High sulfate contents in skim latex serum (SLS) can be reduced by rubber wood ash (RWA). Subsequently, the desulfated skim latex serum (DSLS) can be further anaerobically treated more effectively with the accompanying generated biomethane. In this study, DSLS was treated using an up-flow anaerobic sludge blanket (UASB) reactor operated at 10-day HRT and under mesophilic (37 °C) conditions. The effect of organic loading rates (OLR) at 0.89, 1.79 and 3.57 g-COD/L-reactor∙d on DSLS biodegradability was investigated in Phase I-IV using NaHCO3 as an external buffering agent. Maximum methane production yield of 226.35 mL-CH4/g-CODadded corresponding to 403.25 mL-CH4/L reactor·d was achieved at the suitable OLR of 1.79 g-COD/L-reactor∙d. UASB effluent recirculation which was then applied to replace the NaHCO3. It was found that with 53% effluent recirculation similar to an OLR of 2.01 g-COD/L-reactor∙d, an average of 185.70 mL-CH4/g-CODadded corresponding to 371.40 mL/L reactor·d of methane production was reached. The dominant bacteria in UASB reactor were members of Proteobacteria, Bacteroidota, Firmicutes, and Desulfobacterota phyla. Meanwhile, the archaeal community was majorly dominated by the genera Methanosaeta sp. and Methanomethylovorans sp. The study clearly indicates the capabilities of UASB reactor with effluent recirculation to treat DSLS anaerobically.

Deploying two-stage anaerobic process to co-digest greasy sludge and waste activated sludge for effective waste treatment and biogas recovery.

High-strength waste activated sludge (WAS) and greasy sludge (GS) were largely generated from canned tuna processing. This study reports the performance of the two-stage anaerobic process for co-digesting WAS and GS. Various WAS:GS mixing ratios of 0:100, 10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10, and 100:00 (volatile solids (VS) basis) were investigated in batch acidogenic stage at ambient (30 °C ± 3 °C), 55 °C, and 60 °C temperatures. Subsequently, the effluents from the first stage were used to produce methane in the second methanogenic stage at an ambient temperature. The highest methane yield of 609 mL CH4/g-VSadded was achieved using acidogenic effluents generated from a WAS:GS mixing ratio of 40:60 at an ambient temperature. The first-order kinetic constants (k) for the first (k1) and second (k2) stages were subsequently estimated to be 0.457 d-1 and 0.139 d-1, respectively. The obtained k constants were further used to predict the hydraulic retention time (HRT) for the two continuously stirred tank reactors (CSTR) in series. Consequently, the calculated 4-day HRT and 20-day HRT for 50-L CSTR1 and 250-L CSTR2, respectively, were used to operate the continuous two-stage process at an ambient temperature by feeding with a 40:60-WAS:GS mixing ratio. A satisfactory methane yield of 470-mL CH4/g-VS along with 75% chemical oxygen demand (COD) removal was generated. Furthermore, the predicted methane yield of 450-mL CH4/g-VS obtained from the simple kinetic CSTR model resembled the experimental yield with 96% accuracy. The obtained experimental results demonstrate that WAS and GS co-digestion could be successfully accomplished using a practical two-stage anaerobic process operated at an ambient temperature.

Performance of pilot scale two-stage anaerobic co-digestion of waste activated sludge and greasy sludge under uncontrolled mesophilic temperature.

Waste-activated sludge (WAS) and greasy sludge (GS) discharged from the canned tuna industry are considerably characterized as harsh organic wastes to be individually treated by using traditional anaerobic digestion. This study was attempted to anaerobically co-digest WAS and GS in continuous pilot scale two-stage process, comprising the first 50 L continuous stir tank reactor (CSTR1) and the second 250 L continuous stir tank reactor (CSTR2). The two-stage co-digesting operation of dewatered WAS:GS ratio of 0.4:1 (g-VS) at ambient temperature with the organic loading rate (OLR) of 12.6 ± 0.75 g-VS/L·d and 2.26 ± 0.13 g-VS/L·d, corresponding to 3-day and 17-day hydraulic retention time (HRT) for the first and second stage, respectively generated highest methane production rate of 957 ± 86 mL-CH4/L·d, corresponding to methane yield of 423.4 ± 36 mL-CH4/g-VS. Organic removal efficiency obtained was around 67.5% on COD basis. The microbial diversity was depended on the process's activity. Bacteria were mostly detected in the CSTR1, dominating with the phylum Firmicutes and Proteobacteria, whereas genus Methanosaeta archaea were found dominantly in the CSTR2. The economic analysis of process shows payback period (PBP), internal rate of return (IRR), and net present value (NPV) of 3 years, 30%, and 250,177 USD, respectively. This study demonstrated the potential approach to applying the two-stage anaerobic co-digestion process to stabilize both WAS and GS along with generating valuable bioenergy carriers.

Application of palm oil mill waste to enhance biogas upgrading and hornwort cultivation.

The potential of oil palm ash (OPA) to enhance H2S and CO2 removal from biogas by scrubbing with maturation pond effluent (MPE), and further the treatment of biogas scrubber effluent (BSE) by Ceratophyllum demersum L. (hornwort) cultivation were investigated in this study. The results show that OPA + MPE solution with pH 9.3 and alkalinity 7525 mg CaCO3/L was obtained with 0.7 kg/L OPA loading. A pilot scale scrubber was used to study the effects of absorbent flow rates of 60-210 L/h on upgrading to 300 L/h field biogas stream. At 210 L/h, the CO2 removal efficiencies were 33% and 53% for MPE and OPA + MPE, respectively. To approach 100% H2S removal efficiency, the minimum flow rates were 120 L/h for MPE and 90 L/h for OPA + MPE. 50-150 g wet weight of hornwort in 30 L diluted POME were loaded to investigate appropriate initial hornwort loading level for hornwort cultivation. The highest specific growth rate of 0.045 day-1 with biomass production of 3.8 g/day were obtained with a 50 g initial loading. Among the wastewaters (MPE, OPA + MPE, and BSE) treatment using hornwort cultivation, the highest 0.035 day-1 specific growth rate and 2.6 g/day biomass production of hornwort were obtained in diluted BSE cultivation, and in 3 weeks of cultivation. COD, nitrate, phosphate, and alkalinity decreased by 76%, 76%, 55%, and 5%, respectively. The Eco-Efficiency concept for palm oil mill waste utilization proposed in this study has a high potential for enhanced biogas upgrading by using OPA + MPE, and hornwort is a good candidate for BSE post-treatment integrated with biomass production.

Strategies for recovery of imbalanced full-scale biogas reactor feeding with palm oil mill effluent.

BACKGROUND: Full-scale biogas production from palm oil mill effluent (POME) was inhibited by low pH and highly volatile fatty acid (VFA) accumulation. Three strategies were investigated for recovering the anaerobic digestion (AD) imbalance on biogas production, namely the dilution method (tap water vs. biogas effluent), pH adjustment method (NaOH, NaHCO3, Ca(OH)2, oil palm ash), and bioaugmentation (active methane-producing sludge) method. The highly economical and feasible method was selected and validated in a full-scale application. RESULTS: The inhibited sludge from a full-scale biogas reactor could be recovered within 30-36 days by employing various strategies. Dilution of the inhibited sludge with biogas effluent at a ratio of 8:2, pH adjustment with 0.14% w/v NaOH, and 8.0% w/v oil palm ash were considered to be more economically feasible than other strategies tested (dilution with tap water, or pH adjustment with 0.50% w/v Ca(OH)2, or 1.25% NaHCO3 and bioaugmentation) with a recovery time of 30-36 days. The recovered biogas reactor exhibited a 35-83% higher methane yield than self-recovery, with a significantly increased hydrolysis constant (kH) and specific methanogenic activity (SMA). The population of Clostridium sp., Bacillus sp., and Methanosarcina sp. increased in the recovered sludge. The imbalanced full-scale hybrid cover lagoon reactor was recovered within 15 days by dilution with biogas effluent at a ratio of 8:2 and a better result than the lab-scale test (36 days). CONCLUSION: Dilution of the inhibited sludge with biogas effluent could recover the imbalance of the full-scale POME-biogas reactor with economically feasible and high biogas production performance.

Biogas production from palm oil mill effluent and empty fruit bunches by coupled liquid and solid-state anaerobic digestion.

Biogas production of palm oil mill effluent (POME) and empty fruit bunches (EFB) was performed by coupled liquid (L-AD) and solid-state (SS-AD) anaerobic digestion processes. POME was fed to L-AD digester, while mixed of effluent from L-AD and EFB was fed to SS-AD digester. The maximum overall methane production of 60.9 m3-CH4·ton-1 waste was obtained at an optimal hydraulic retention time of 30 days and an organic loading rate of 1.66 gVS·L-1-reactor·d-1 for L-AD and 6.03 gVS·L-1-reactor·d-1 for SS-AD with L-AD effluent recycling rate of 16.7 mL·L-1-reactor·d-1. The bacterial community in the L-AD reactor was different from the SS-AD reactor, while the archaeal community was similar in both reactors. Synergistaceae, Caldicoprobacteraceae and Lachnospiraceae were increased in the SS-AD reactor. Coupling L-AD and SS-AD is able to increase energy production by 29% and 71% compared to the L-AD and SS-AD alone, respectively, with no outsource SS-AD inoculum required.

Thermotolerant cellulolytic Clostridiaceae and Lachnospiraceae rich consortium enhanced biogas production from oil palm empty fruit bunches by solid-state anaerobic digestion.

Thermotolerant cellulolytic consortium for improvement biogas production from oil palm empty fruit bunches (EFB) by prehydrolysis and bioaugmentation strategies was investigated via solid-state anaerobic digestion (SS-AD). The prehydrolysis EFB with Clostridiaceae and Lachnospiraceae rich consortium have maximum methane yield of 252 and 349 ml CH4 g-1 VS with total EFB degradation efficiency of 62% and 86%, respectively. Clostridiaceae and Lachnospiraceae rich consortium augmentation in biogas reactor have maximum methane yield of 217 and 85.2 ml CH4 g-1 VS with degradation efficiency of 42% and 16%, respectively. The best improvement of biogas production was achieved by prehydrolysis EFB with Lachnospiraceae rich consortium with maximum methane production of 113 m3 CH4 tonne-1 EFB. While, Clostridiaceae rich consortium was suitable for augmentation in biogas reactor with maximum methane production of 70.6 m3 CH4 tonne-1 EFB. Application of thermotolerant cellulolytic consortium into the SS-AD systems could enhance biogas production of 3-11 times.

Effects of volatile fatty acids in biohydrogen effluent on biohythane production from palm oil mill effluent under thermophilic condition

Background: Biohydrogen effluent contains a high concentration of volatile fatty acid (VFA) mainly as butyric, acetic, lactic and propionic acids. The presence of various VFAs (mixture VFAs) and their cooperative effects on two-stage biohythane production need to be further studied. The effect of VFA concentrations in biohydrogen effluent of palm oil mill effluent (POME) on methane yield in methane stage of biohythane production was investigated. Results: The methane yield obtained in low VFA loading (0.9 and 1.8 g/L) was 15­20% times greater than that of high VFA loading (3.6 and 4.7 g/L). Butyric acid at high concentrations (8 g/L) has the individual significantly negative effect the methane production process (P b 0.05). Lactic, acetic and butyric acid mixed with propionic acid at a concentration higher than 0.5 g/L has an interaction significantly negative effect on the methanogenesis process (P b 0.05). Inhibition condition had a negative effect on both bacteria and archaea with inhibited on Geobacillus sp., Thermoanaerobacterium thermosaccharolyticum, Methanoculleus thermophilus and Methanothermobacter delfuvii resulting in low methane yield. Conclusion: Preventing the high concentration of butyric acid, and propionic acid in the hydrogenic effluent could enhance methane production in two-stage anaerobic digestion for biohythane production.

Effect of biogas sparging on the performance of bio-hydrogen reactor over a long-term operation.

This study aimed to enhance hydrogen production from sugarcane syrup by biogas sparging. Two-stage continuous stirred tank reactor (CSTR) and upflow anaerobic sludge blanket (UASB) reactor were used to produce hydrogen and methane, respectively. Biogas produced from the UASB was used to sparge into the CSTR. Results indicated that sparging with biogas increased the hydrogen production rate (HPR) by 35% (from 17.1 to 23.1 L/L.d) resulted from a reduction in the hydrogen partial pressure. A fluctuation of HPR was observed during a long term monitoring because CO2 in the sparging gas and carbon source in the feedstock were consumed by Enterobacter sp. to produce succinic acid without hydrogen production. Mixed gas released from the CSTR after the sparging can be considered as bio-hythane (H2+CH4). In addition, a continuous sparging biogas into CSTR release a partial pressure in the headspace of the methane reactor. In consequent, the methane production rate is increased.

Optimization and microbial community analysis for production of biogas from solid waste residues of palm oil mill industry by solid-state anaerobic digestion.

This study investigated the improvement of biogas production from solid-state anaerobic digestion (SS-AD) of oil palm biomass by optimizing of total solids (TS) contents, feedstock to inoculum (F:I) ratios and carbon to nitrogen (C:N) ratios. Highest methane yield from EFB, OPF and OPT of 358, 280 and 324m(3)CH4ton(-1)VS, respectively, was achieved at TS content of 16%, C:N ratio of 30:1 and F:I ratio of 2:1. The main contribution to methane from biomass was the degradation of cellulose and hemicellulose. The highest methane production of 72m(3)CH4ton(-1) biomass was achieved from EFB. Bacteria community structure in SS-AD process of oil palm biomass was dominated by Ruminococcus sp. and Clostridium sp., while archaea community was dominated by Methanoculleus sp. Oil palm biomass has great potential for methane production via SS-AD.

Bio-hydrogen and bio-methane potentials of skim latex serum in batch thermophilic two-stage anaerobic digestion.

Anaerobic digestion by two-stage process, containing hydrogen-producing (acidogenic) first stage and methanogenic second stage, has been proposed to degrade substrates which are difficult to be treated by single stage anaerobic digestion process. This research was aimed to evaluate the bio-hydrogen and the bio-methane potentials (BHP and BMP) of skim latex serum (SLS) by using sequential batch hydrogen and methane cultivations at thermophilic conditions (55°C) and with initial SLS concentrations of 37.5-75.0% (v/v). The maximal 1.57 L H2/L SLS for BHP and 12.2L CH4/L SLS for BMP were both achieved with 60% (v/v) SLS. The dominant hydrogen-producing bacteria in the H2 batch reactor were Thermoanaerobacterium sp. and Clostrdium sp. Meanwhile, the CH4 batch reactor was dominated by the methanogens Methanosarcina mazei and Methanothermobacter defluvii. The results demonstrate that SLS can be degraded by conversion to form hydrogen and methane, waste treatment and bioenergy production are thus combined.

Submersible microbial fuel cell for electricity production from sewage sludge.

A submersible microbial fuel cell (SMFC) was utilized to treat sewage sludge and simultaneously generate electricity. Stable power generation (145 +/- 5 mW/m2, 470 omega) was produced continuously from raw sewage sludge for 5.5 days. The maximum power density reached 190 +/- 5 mW/m2. The corresponding total chemical oxygen demand (TCOD) removal efficiency was 78.1 +/- 0.2% with initial TCOD of 49.7 g/L. The power generation of SMFC was depended on the sludge concentration, while dilution of the raw sludge resulted in higher power density. The maximum power density was saturated at sludge concentration of 17 g-TCOD/L, where 290 mw/m2 was achieved. When effluents from an anaerobic digester that was fed with raw sludge were used as substrate in the SMFC, a maximum power density of 318 mW/m2, and a final TCOD removal of 71.9 +/- 0.2% were achieved. These results have practical implications for development of an effective system to treat sewage sludge and simultaneously recover energy.

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-H(2)/g-VS (190 ml-H(2)/g-sugars) and 307 ml-CH(4)/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 H(2)-UASB reactor were Thermoanaerobacter wiegelii, Caldanaerobacter subteraneus, and Caloramator fervidus. Meanwhile, the CH(4)-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.

Extreme thermophilic biohydrogen production from wheat straw hydrolysate using mixed culture fermentation: effect of reactor configuration.

Hydrogen production from hemicellulose-rich wheat straw hydrolysate was investigated in continuously-stirred tank reactor (CSTR), up-flow anaerobic sludge bed (UASB) reactor, and anaerobic filter (AF) reactor. The CSTR was operated at an hydraulic retention time (HRT) of 3 days, and the UASB and AF reactors were operated at 1 day HRT, using mixed extreme thermophiles at 70 °C. The highest hydrogen production yield of 212.0±6.6 mL-H2/g-sugars, corresponding to a hydrogen production rate of 821.4±25.5 mL-H2/dL was achieved with the UASB reactor. Lowering the HRT to 2.5 days caused cell mass washout in the CSTR, while the UASB and AF reactors gave fluctuating and reducing hydrogen production at a 0.5-day HRT. The original rate and yield were recovered when the HRT was increased back to 1 day. These results demonstrate that reactor configuration is an important factor for enhancing and stabilizing H2 production.

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

Hydrolysate was tested as substrate for hydrogen production by extreme thermophilic mixed culture (70 degrees C) in both batch and continuously fed reactors. Hydrogen was produced at hydrolysate concentrations up to 25% (v/v), while no hydrogen was produced at hydrolysate concentration of 30% (v/v), indicating that hydrolysate at high concentrations was inhibiting the hydrogen fermentation process. In addition, the lag phase for hydrogen production was strongly influenced by the hydrolysate concentration, and was prolonged from approximately 11 h at the hydrolysate concentrations below 20% (v/v) to 38 h at the hydrolysate concentration of 25% (v/v). The maximum hydrogen yield as determined in batch assays was 318.4 +/- 5.2 mL-H(2)/g-sugars (14.2 +/- 0.2 mmol-H(2)/g-sugars) at the hydrolysate concentration of 5% (v/v). Continuously fed, and the continuously stirred tank reactor (CSTR), operating at 3 day hydraulic retention time (HRT) and fed with 20% (v/v) hydrolysate could successfully produce hydrogen. The hydrogen yield and production rate were 178.0 +/- 10.1 mL-H(2)/g-sugars (7.9 +/- 0.4 mmol H(2)/g-sugars) and 184.0 +/- 10.7 mL-H(2)/day L(reactor) (8.2 +/- 0.5 mmol-H(2)/day L(reactor)), respectively, corresponding to 12% of the chemical oxygen demand (COD) from sugars. Additionally, it was found that toxic compounds, furfural and hydroxymethylfurfural (HMF), contained in the hydrolysate were effectively degraded in the CSTR, and their concentrations were reduced from 50 and 28 mg/L, respectively, to undetectable concentrations in the effluent. Phylogenetic analysis of the mixed culture revealed that members involved hydrogen producers in both batch and CSTR reactors were phylogenetically related to the Caldanaerobacter subteraneus, Thermoanaerobacter subteraneus, and Thermoanaerobacterium thermosaccharolyticum.

Bioethanol, biohydrogen and biogas production from wheat straw in a biorefinery concept.

The production of bioethanol, biohydrogen and biogas from wheat straw was investigated within a biorefinery framework. Initially, wheat straw was hydrothermally liberated to a cellulose rich fiber fraction and a hemicellulose rich liquid fraction (hydrolysate). Enzymatic hydrolysis and subsequent fermentation of cellulose yielded 0.41 g-ethanol/g-glucose, while dark fermentation of hydrolysate produced 178.0 ml-H(2)/g-sugars. The effluents from both bioethanol and biohydrogen processes were further used to produce methane with the yields of 0.324 and 0.381 m(3)/kg volatile solids (VS)(added), respectively. Additionally, evaluation of six different wheat straw-to-biofuel production scenaria showed that either use of wheat straw for biogas production or multi-fuel production were the energetically most efficient processes compared to production of mono-fuel such as bioethanol when fermenting C6 sugars alone. Thus, multiple biofuels production from wheat straw can increase the efficiency for material and energy and can presumably be more economical process for biomass utilization.

Biohydrogen production from xylose at extreme thermophilic temperatures (70 degrees C) by mixed culture fermentation.

Biohydrogen production from xylose at extreme thermophilic temperatures (70 degrees C) was investigated in batch and continuous-mode operation. Biohydrogen was successfully produced from xylose by repeated batch cultivations with mixed culture received from a biohydrogen reactor treating household solid wastes at 70 degrees C. The highest hydrogen yield of 1.62+/-0.02 mol-H2/mol-xylose(consumed) was obtained at initial xylose concentration of 0.5 g/L with synthetic medium amended with 1g/L of yeast extract. Lower hydrogen yield was achieved at initial xylose concentration higher than 2g/L. Addition of yeast extract in the cultivation medium resulted in significant improvement of hydrogen yield. The main metabolic products during xylose fermentation were acetate, ethanol, and lactate. The specific growth rates were able to fit the experimental points relatively well with Haldane equation assuming substrate inhibition, and the following kinetic parameters were obtained: the maximum specific growth rate (mu(max)) was 0.17 h(-1), the half-saturation constant (K(s)) was 0.75g/L, and inhibition constant (K(i)) was 3.72 g/L of xylose. Intermittent N2 sparging could enhance hydrogen production when high hydrogen partial pressure (> 0.14 atm) was present in the headspace of the batch reactors. Biohydrogen could be successfully produced in continuously stirred reactor (CSTR) operated at 72-h hydraulic retention time (HRT) with 1g/L of xylose as substrate at 70 degrees C. The hydrogen production yield achieved in the CSTR was 1.36+/-0.03 mol-H2/mol-xylose(sonsumed), and the production rate was 62+/-2 ml/d x L(reactor). The hydrogen content in the methane-free mixed gas was approximately 31+/-1%, and the rest was carbon dioxide. The main intermediate by-products from the effluent were acetate, formate, and ethanol at 4.25+/-0.10, 3.01+/-0.11, and 2.59+/-0.16 mM, respectively.