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Methane production from solid-state anaerobic digestion (SS-AD) of empty fruit bunches (EFB) using recycling solid-anaerobic digested (S-AD) sludge and liquid-anaerobic digested (L-AD) sludge as inoculum was investigated under thermophilic (55 °C) and mesophilic conditions (40 °C). The methane production rate of L-AD sludge (2.98–3.27 L-CH4 L−1 reactor d −1) was 1–2 times higher than recycling S-AD sludge (1.36 L-CH4 L−1 reactor d −1) as inoculum. Methane production of L-AD and S-AD sludge inoculum was 53.5–54.1 and 31.2–39.0 m3 CH4 tonne-1 EFB, respectively. The SS-AD of EFB with L-AD and S-AD sludge under thermophilic condition had specific methane activity of 34–96 times higher than the mesophilic condition. Clostridium sp., Methanosaeta sp., and Methanoculleus sp. were responsible for the SS-AD of EFB with L-AD sludge inoculum. The L-AD sludge was suitable inoculum for SS-AD of EFB in terms of methane production rate, cellulose degradation efficiency, biogas production, and microbial activity.

The impact of the heavy metal contamination and acidity on the bacterial community was studied in samples collected from the Akhtala copper mine tailing using molecular approaches. The bacterial community structure analysis by PCR-DGGE fingerprinting revealed an abundance of Firmicutes, Acidobacteria, and Proteobacteria in different layers of the Akhtala tailing. 454 pyrotag sequence analyses revealed that a significant part of the sequences originated from Proteobacteria (49%) and Bacteroidetes (43%). Bacterial taxa are distributed also in phyla Saccharibacteria (2%), Verrucomicrobia (1.5%), Gammatimonadetes (1%), and some minor additional bacterial groups. The main primary producers in the Akhtala tailing appear to be obligate autotrophic Thiobacillus and Sulfuritalea species. Representatives of Lutibacter and Lysobacter genera are the most abundant acid-tolerant heterotrophs in the studied tailing. The presence of a large number of yet-uncultivated species emphasizes the importance of the future exploration of the tailing as an important source of novel bacteria.

The effect of trace metals supplementation into palm oil mill effluent on biohythane production and responsible microbial communities in thermophilic two-stage anaerobic fermentation was investigated. High biohythane yields were linked to Ni/Co/Fe supplementation (10, 6 and 20 mg L−1, respectively) with maximum H2 and CH4 yields of 139 mL H2 gVS−1 and 454 mL CH4 gVS−1, respectively. The Ni/Co/Fe supplementation resulted in higher numbers of Bacillus sp., Clostridium sp. and Thermoanaerobacterium sp. together with increasing hydrogenase expression level leading to increasing hydrogen yields of 90.4%. The numbers of Methanosarcina, Methanomassiliicoccus, and Methanoculleus were enhanced by Ni/Co/Fe addition, accompanied by 21.7% higher methane yields. No correlation between methyl coenzyme-M reductase expression level and methane yields was observed. The Ni/Co/Fe supplementation improved gas production in the two-stage biohythane process via enhancing a number of viable hydrogen-producing bacteria together with hydrogenase activity in H2 stage and enhancing number methanogens in the CH4 stage.

A sudden pH drops always inhibits the anaerobic digestion (AD) reactor for biogas production from palm oil mill effluent (POME). The pH adjustment of POME by oil palm ash addition and the biogas effluent recycling effect on the preventing of pH drop and change of the archaea community was investigated. The pH adjustment of POME to 7.5 increased the methane yield two times more than raw POME (pH 4.3). The optimal dose for pH adjustment by oil palm ash addition was 5% w/v with a methane yield of 440 mL-CH4/gVS. The optimal dose for pH adjustment by biogas effluent recycling was 20% v/v with a methane yield of 351 mL-CH4/gVS. Methane production from POME in a continuous reactor with pH adjustment by 5% w/v oil palm ash and 20% v/v biogas effluent recycling was 19.1 ± 0.25 and 13.8 ± 0.3 m3 CH4/m3-POME, respectively. The pH adjustment by oil palm ash enhanced methane production for the long-term operation with the stability of pH, alkalinity, and archaea community. Oil palm ash increased the number of Methanosarcina mazei and Methanothermobacter defluvii. Oil palm ash is a cost-effective alkali material as a source of buffer and trace metals for preventing the pH drop and the increased methanogen population in the AD process.

Empty fruit bunches (EFB) have low biodegradability and restrict their commercial utilization in biogas plants. Integration of straw mushroom (Volvariella volvacea) cultivation as a function of bio-pretreatment on EFB to improve biodegradability and methane production by solid-state anaerobic digestion (SS-AD) was investigated. The mushroom yield was 47.3 kg·tonne−1 EFB with remaining weight in spent mushroom-EFB (S-mEFB) of 82%. The cellulose, hemicellulose, and lignin of EFB were degraded by 3.3%, 21.3%, and 17.6%, respectively, with an increased surface area of S-mEFB. The biodegradability of S-mEFB (62.7%) was 2 times higher than raw EFB (33.5%) with the highest methane yield and production of 281 mL CH4·g−1 VS and 50.6 m3·tonne−1 S-mEFB, respectively. The co-digestion of S-mEFB with 5% v/w POME had highest methane yield of 405 mL CH4·g−1 VS with biodegradability of 90.8%. Integrating straw mushroom cultivation with SS-AD is a promising strategy for achieving an environmentally friendly and economically feasible process.

This study aimed to evaluate the possibility of acidogenic phase as the pretreatment method of filter cake (FC) before being co-digested with biogas effluent (BE), and anaerobic sludge (AS) for methane production. The effects of separation of acidogenic and methanogenic phases on the improvement of the methane production process from a co-digestion of FC with BE, and AS were also investigated. FC was pretreated through acidogenic phase anaerobic digestion (AD) using a different substrate to inoculum ratios (SIRs) and substrate concentrations. The optimum conditions for acidogenic phase AD of FC were SIRs of 4:1 and substrate concentration of 15.0% total solid (TS). Resultantly, the maximum volatile fatty acids (VFA) yield of 0.17 g-VFAs/g-VS (volatile solids) was obtained. The major VFA components were butyric and acetic acid. The acidogenic phase of FC was semi-continuously conducted in a continuous stirred tank reactor (CSTR) at various hydraulic retention time (HRT). The highest concentration and yield of VFAs of 7.29 g/L and 0.39 g-VFAs/g-VS, respectively, were observed at HRT of 5 d. The major VFAs observed were butyric acid (48.8%), and acetic acid (36.8%). Subsequently, the proportions of pretreated FC, BE, and AS to produce methane were optimized in batch experiments. The highest methane yield (MY) of 195.9 mL-CH4/g-VS was achieved at the proportions of pretreated FC and BE of 37.75 and 22.25 g-VS/L, respectively, without the addition of AS. The optimum substrate proportion was further used to determine the MY in a 10 L CSTR with the various HRT. The maximum MY of 130.5 mL-CH4/g-VS was achieved at an HRT of 20 d which produced an EY of 4.7 kJ/g-VS. Analysis of microbial community by 16S rRNA amplicon sequencing on an illumination platform indicated that genera Rummeliibacillus, Lactobacillus, and Bacillus were the dominant bacteria found in the acidogenic phase. Methanoculleus and Methanosarcina were found to be the most dominant genera in methane fermentation. Results suggested that acidogenic phase digestion can be used as an alternative method for FC pretreatment to improve methane production. Furthermore, the separation of acidogenic and methanogenic phases improves the process stability.

Homoacetogens consortium was enriched from peatland soil, digested sludge, and rumen fluid for simultaneous carbon dioxide (CO2) consumption and acetic acid production in the biogas upgrading process. The homoacetogens consortium enriched from peatland soil (PL3) has a maximum CO2 consumption and acetic acid production of 95% and 120 mg/L, respectively. The methane content in upgraded biogas and acetic acid production of 98% and 543 mg/L was obtained at optimum hydrogen (H2) to CO2 at a ratio of 2:1, pH 8, and temperature 30 °C. The upgraded biogas (CH4 > 95%) can be used as vehicle fuel or injected into the natural gas grid. The homoacetogens consortium PL3 was dominated by Clostridium sp., Proteiniclasticum sp., and Petrimonas sp. Clostridium species are the main homoacetogens responsible for CO2 reduction using H2 as an energy source. The homoacetogens consortium PL3 could provide an opportunity for simultaneous biogas upgrading to biomethane and acetic acid production.

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 15–20% 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 P 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. Normal 0 false false false ES-CL X-NONE X-NONE /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Tabla normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-parent:""; mso-padding-alt:0cm 5.4pt 0cm 5.4pt; mso-para-margin-top:0cm; mso-para-margin-right:0cm; mso-para-margin-bottom:8.0pt; mso-para-margin-left:0cm; line-height:107%; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri",sans-serif; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:"Times New Roman"; mso-bidi-theme-font:minor-bidi; mso-ansi-language:ES-CL;} Normal 0 false false false EN-US X-NONE TH

Valorization of biodiesel-derived crude glycerol and its hydrolysis product (ethanol) as a co-substrate supplemented to palm oil mill effluent (POME) to enhance biogas production efficiency under mesophilic anaerobic digestion was investigated. Based on our previous studies, the concentrations of each co-substrate were varied in the range of 1–5% v⋅v−1. The optimum concentrations of crude glycerol and ethanol were 1% and 5% v/v, respectively. They could increase both methane production (nearly 2 fold) and methane yield (1.98–2.09 fold based on volatile solids (VS) and 1.6–1.87 fold based on COD). However, they exhibited oppositely in the methane production rate; the 1% crude glycerol was 38% lower while 5% ethanol exhibited a 67% higher methane production rate than the control (POME). The results clearly indicated that the methane production rate from 5% ethanol was 2.7 times higher than that from the 1% crude glycerol. Microbial community analysis revealed that the dominant bacteria in both co-substrates biogas production process were Sandarakinotalea sp., Lachnobacterium sp., Blautia sp., Marinobacter sp., Anaerotruncus sp., Clostridium sp., Enterobacter sp., and Acinetobacter sp. The Blautia sp. was dominant in the presence of 1% crude glycerol than 5% ethanol. The archaea communities were dominated by hydrogenotrophic methanogen (Methanoregula sp. and Methanoculleus sp.) and acetoclastic methanogen (Methanosarcina sp. and Methanospirillum sp.). Therefore, ethanol was a better co-substrate than crude glycerol. Nevertheless, biogas production from POME with crude glycerol as co-substrate was anticipated to strongly increase by enhancing the growth (or bioaugmentation) of Blautia sp. and Methanosarcina sp.

Co-fermentation of palm oil mill effluent (POME) for hydrogen production by Thermoanaerobacterium thermosaccharolyticum PSU-2 under thermophilic condition was investigated. Hydrogen production from co-fermentation POME with 5 g/L decanter cake and without was 2.6 and 1.7 L-H 2 /L-POME. Co-fermentation of POME with decanter cake enhanced hydrogen production. Tbm. thermosaccharolyticum PSU-2 augmentation at 20% v/v was found to be an efficient start-up strategy in continuous hydrogen production from co-fermentation of POME with decanter cake. Tbm. thermosaccharolyticum PSU-2 augmentation could suppress normal flora bacteria in POME and decanter cake. Hydrogen production of 7.2 L-H 2 /L-POME and hydrogen production rate of 18.1 mmol-H 2 /L/h was achieved from continuous hydrogen in long term operation. The soluble metabolites were dominated by acetic acid and butyric acid. Tbm. thermosaccharolyticum PSU-2 augmentation could use for control undesired bacteria in the hydrogen production system with flavored Thermoanaerobacterium sp. as dominant species.

Thermoanaerobacterium sp. strain PSU-2 was isolated from thermophilic hydrogen producing reactor and subjected to draft genome sequencing on 454 pyrosequencing and annotated on RAST. The draft genome sequence of strain PSU-2 contains 2,552,497 bases with an estimated G + C content of 35.2%, 2555 CDS, 8 rRNAs and 57 tRNAs. The strain had a number of genes responsible for carbohydrates metabolic, amino acids and derivatives, and protein metabolism of 17.7%, 14.39% and 9.81%, respectively. Strain PSU-2 also had gene responsible for hydrogen biosynthesis as well as the genes related to Ni-Fe hydrogenase. Comparative genomic analysis indicates strain PSU-2 shares about 94% genome sequence similarity with Thermoanaerobacterium xylanolyticum LX-11. The nucleotide sequence of this draft genome was deposited into DDBJ/ENA/GenBank under the accession MSQD00000000.

A two-stage thermophilic fermentation and mesophilic methanogenic process for biohythane production from palm oil mill effluent (POME) was investigated. The hydrogen and methane potential from POME were 170–200 L H 2 kgCOD −1 and 210–292 L CH 4 kgCOD −1 , respectively. A continuous two-stage process with hydraulic retention time (HRT) of 2 d for hydrogen reactor and 15 d for methane reactor, obtained 34% higher energy yield than single stage methane production. The hydrogen and methane yields from two-stage were 210 L H 2 kgCOD −1 and 315 L CH 4 kgCOD −1 , respectively with total energy yield of 15.34 MJ kgCOD −1 . Mixed hydrogen and methane (biohythane) production rate was 4.4 L biogas L −1 d −1 with containing of 51% CH 4 , 14% H 2 and 35% CO 2 . Hydrogen reactor was dominated with hydrogen producing bacteria of Thermoanaerobacterium thermosaccharolyticum , while acetoclastic Methanoculleus sp. was the dominant methanogen in methane reactor. Two-stage process for biohythane production could efficiently for energy recovery from POME.

The microbial community structure of thermophilic mixed culture sludge used for biohydrogen production from palm oil mill effluent was analyzed by fluorescence in situ hybridization (FISH) and 16S rRNA gene clone library techniques. The hydrogen-producing bacteria were isolated and their ability to produce hydrogen was confirmed. The microbial community was dominated by Thermoanaerobacterium species (∼66%). The remaining microorganisms belonged to Clostridium and Desulfotomaculum spp. (∼28% and ∼6%, respectively). Three hydrogen-producing strains, namely HPB-1, HPB-2, and HPB-3, were isolated. 16S rRNA gene sequence analysis of HPB-1 and HPB-2 revealed a high similarity to Thermoanaerobacterium thermosaccharolyticum (98.6% and 99.0%, respectively). The Thermoanaerobacterium HPB-2 strain was a promising candidate for thermophilic fermentative hydrogen production with a hydrogen yield of 2.53 mol H2 mol−1hexose from organic waste and wastewater containing a mixture of hexose and pentose sugars. Thermoanaerobacterium species play a major role in thermophilic hydrogen production as confirmed both by molecular and cultivation-based analyses.

Thermoanaerobacterium-rich sludge was used for hydrogen production and phenol removal from palm oil mill effluent (POME) in the presence of phenol concentration of 100–1000 mg/L. Thermoanaerobacterium-rich sludge yielded the most hydrogen of 4.2 L H2/L-POME with 65% phenol removal efficiency at 400 mg/L phenol. Butyric acid and acetic acid were the main metabolites. The effects of oil palm ash, NH4NO3 and iron concentration (Fe2+) on hydrogen production and phenol removal efficiency from POME by Thermoanaerobacterium-rich sludge was investigated using response surface methodology (RSM). The RSM results indicated that the presence of 0.2 g Fe2+/L, 0.3 g/L NH4NO3 and 20 g/L oil palm ash in POME could improved phenol removal efficiency, with predicted hydrogen production and phenol removal efficiency of 3.45 L H2/L-POME and 93%, respectively. In a confirmation experiment under optimized conditions highly reproducible results were obtained, with hydrogen production and phenol removal efficiency of 3.43 ± 0.12 L H2/L-POME and 92 ± 1.5%, respectively. Simultaneous hydrogen production and phenol removal efficiency in continuous stirred tank reactor at hydraulic retention time (HRT) of 1 and 2 days were 4.0 L H2/L-POME with 85% and 4.2 L H2/L-POME with 92%, respectively. Phenol degrading Thermoanaerobacterium-rich sludge comprised of Thermoanaerobacterium thermosaccharolyticum, Thermoanaerobacterium aciditolerans, Desulfotomaculum sp., Bacillus coagulans and Clostridium uzonii. Phenol degrading Thermoanaerobacterium-rich sludge has great potential to harvest hydrogen from phenol-containing wastewater.