Biohydrogen and methane production by co-digestion of cassava stillage and excess sludge under thermophilic condition

Thermophilic anaerobic hydrogen and methane production by co-digestion of cassava stillage (CS) and excess sludge (ES) was investigated in this study. The improved hydrogen and subsequent methane production were observed by co-digestion of CS with certain amount of ES in batch experiments. Compared with one phase anaerobic digestion, two phase anaerobic digestion offered an attractive alternative with more abundant biogas production and energy yield, e.g., the total energy yield in two phase obtained at VS_CS/VS_ES of 3:1 was 25% higher than the value of one phase. Results from continuous experiments further demonstrated that VS_CS/VS_ES of 3:1 was optimal for hydrogen production with the highest hydrogen yield of 74 mL/g total VS added, the balanced nutrient condition with C/N ratio of 1.5 g carbohydrate-COD/g protein-COD or 11.9 g C/g N might be the main reason for such enhancement. VS_CS/VS_ES of 3:1 was also optimal for continuous methane production considering the higher methane yield of 350 mL/g total VS added and the lower propionate concentration in the effluent.     

Enhancement of methane production from cassava residues by biological pretreatment using a constructed microbial consortium

In the study, a stable thermophilic microbial consortium with high cellulose-degradation ability was successfully constructed. That several species of microbes coexisted in this consortium was proved by DGGE (denaturing gradient gel electrophoresis) and sequence analysis. The cooperation and symbiosis of these microbes in this consortium enhanced their cellulose-degradation ability. The pretreatment of cassava residues mixing with distillery wastewater prior to anaerobic digestion was investigated by using this microbial consortium as inoculums in batch bioreactors at 55 °C. The experimental results showed that the maximum methane yield (259.46 mL/g-VS) of cassava residues was obtained through 12 h of pretreatment by this microbial consortium, which was 96.63% higher than the control (131.95 mL/g-VS). In addition, it was also found that the maximum methane yield is obtained when the highest filter paper cellulase (FPase), carboxymethyl cellulase (CMCase) and xylanase activity and soluble COD (sCOD) are produced.     

Potential for methane production from anaerobic co-digestion of swine manure with winery wastewater

This work examines the methane production potential for the anaerobic co-digestion of swine manure (SM) with winery wastewater (WW). Batch and semi-continuous experiments were carried out under mesophilic conditions. Batch experiments revealed that the highest specific methane yield was 348 mL CH₄ g⁻¹ COD added, obtained at 85.4% of WW and 0.7 g COD g⁻¹ VS. Specific methane yield from SM alone was 27 mL CH₄ g⁻¹ COD added d⁻¹. Furthermore, specific methane yields were 49, 87 and 107 mL CH₄ g⁻¹ COD added d⁻¹ for the reactors co-digesting mixtures with 10% WW, 25% WW and 40% WW, respectively. Co-digestion with 40% WW improved the removal efficiencies up to 52% (TCOD), 132% (SCOD) and 61% (VSS) compared to SM alone. These results suggest that methane can be produced very efficiently by the co-digestion of swine manure with winery wastewater.     

Enhanced methane and hydrogen production from municipal solid waste and agro-industrial by-products co-digested with crude glycerol

The effects of crude glycerol on the performance of single-stage anaerobic reactors treating different types of organic waste were examined. A reactor treating the organic fraction of municipal solid waste produced 1400 mL CH₄/d before the addition of glycerol and 2094 mL CH₄/d after the addition of glycerol. An enhanced methane production rate was also observed when a 1:4 mixture of olive mill wastewater and slaughterhouse wastewater was supplemented with crude glycerol. Specifically, by adding 1% v/v crude glycerol to the feed, the methane production rate increased from 479 mL/d to 1210 mL/d. The extra glycerol-COD added to the feed did not have a negative effect on the reactor performance in either case. Supplementation of the feed with crude glycerol also had a significant positive effect on anaerobic fermentation reactors. Hydrogen yield was 26 mmole H₂/g VS added and 15 mmole H₂/g VS added in a reactor treating the organic fraction of municipal solid waste and a 1:4 mixture of olive mill and slaughterhouse wastewater. The addition of crude glycerol to the feed enhanced hydrogen yield at 2.9 mmole H₂/g glycerol added and 0.7 mmole H₂/g glycerol added.     

Optimization of fermentative biohydrogen production by response surface methodology using fresh leachate as nutrient supplement

Fresh compost leachate was used as a nutrients source to facilitate anaerobic fermentative hydrogen production from glucose inoculated with mixed culture. The optimum condition for hydrogen production was predicted by response surface methodology (RSM). The model showed the maximum cumulative hydrogen volume (469.74 mL) and molar hydrogen yield (1.60 mol H₂/mol glucose) could be achieved at 6174.93 mg/L glucose and 3383.20 mg COD/L leachate. According to the predicted optimal condition, four tests were carried out to validate the predicted values and evaluate the leachate’s effect on co-fermentation with juice wastewater. A maximum cumulative hydrogen volume of 587.05 ± 15.08 mL was obtained in co-fermentation test, and the molar hydrogen yield reached 2.06 ± 0.06 mol H₂/mol glucose. The co-fermentation of fresh leachate and glucose/juice wastewater was a combination of acetic acid and butyric acid type-fermentation. The results demonstrated that leachate can serve as a nutrients source for biohydrogen production.     

Methane recovery from the anaerobic codigestion of municipal sludge and FOG

The anaerobic biodegradability of a mix of municipal primary sludge (PS), thickened waste activated sludge (TWAS) and fat, oil, and grease (FOG) was assessed using semi-continuous feed, laboratory-scale anaerobic digesters operated at mesophilic (35 °C) and thermophilic (52 °C) temperature. Addition of a large FOG fraction (48% of the total VS load) to a PS + TWAS mix, resulted in 2.95 times larger methane yield, 152 vs. 449 mL methane @ STP/g VS added at 35 °C and 2.6 times larger methane yield, 197 vs. 512 mL methane @ STP/g VS added at 52 °C. The high FOG organic load fraction was not inhibitory to the process. The results of this study demonstrate the benefit of sludge and FOG codigestion.     

Evaluation of bioenergy recovery processes treating organic residues from ethanol fermentation process

This study evaluates a two-stage bioprocess for recovering bioenergy in the forms of hydrogen and methane while treating organic residues of ethanol fermentation from tapioca starch. A maximum hydrogen production rate of 0.77 mmol H₂/g VSS/h can be achieved at volumetric loading rate (VLR) of 56 kg COD/m³/day. Batch results indicate that controlling conditions at S₀/X₀ = 12 with X₀ = 4000 mg VSS/L and pH 5.5-6 are important for efficient hydrogen production from fermentation residues. Hydrogen-producing bacteria enriched in the hydrogen bioreactor are likely utilizing lactate and acetate for biohydrogen production from ethanol-fermentation residues. Organic residues remained in the effluent of hydrogen bioreactor can be effectively converted to methane with a rate of 0.37 mmol CH₄/g VSS/h at VLR of 8 kg COD/m³/day. Approximately 90% of COD in ethanol-fermentation residues can be removed and among that 2% and 85.1% of COD can be recovered in the forms of hydrogen and methane, respectively.     

Effect of feed to inoculum ratios on biogas yields of food and green wastes

Biogas and methane yields of food and green wastes and their mixture were determined using batch anaerobic digesters at mesophilic (35 ± 2 °C) and thermophilic (50 ± 2 °C) temperatures. The mixture was composed of 50% food waste and 50% green waste, based on the volatile solids (VS) initially added to the reactors. The thermophilic digestion tests were performed with four different feed to inoculum (F/I) ratios (i.e., 1.6, 3.1, 4.0 and 5.0) and the mesophilic digestion was conducted at one F/I (3.1). The results showed that the F/I significantly affected the biogas production rate. At four F/Is tested, after 25 days of thermophilic digestion, the biogas yield was determined to be 778, 742, 784 and 396 mL/g VS for food waste, respectively; 631, 529, 524 and 407 mL/g VS for green waste, respectively; and 716, 613, 671 and 555 mL/g VS for the mixture, respectively. About 80% of the biogas production was obtained during the first 10 days of digestion. At the F/I of 3.1, the biogas and methane yields from mesophilic digestion of food waste, green waste and their mixture were lower than the yields obtained at thermophilic temperature. The biogas yields were 430, 372 and 358 mL/g VS, respectively, and the methane yields were 245, 206, and 185 mL/g VS, respectively.     

Effect of enzyme additions on methane production and lignin degradation of landfilled sample of municipal solid waste

Operation of waste cells as landfill bioreactors with leachate recirculation is known to accelerate waste degradation and landfill gas generation. However, waste degradation rates in landfill bioreactors decrease with time, with the accumulation of difficult to degrade materials, such as lignin-rich waste. Although, potential exists to modify the leachate quality to promote further degradation of such waste, very little information is available in literature. The objective of this study was to determine the viability of augmenting leachate with enzymes to increase the rate of degradation of lignin-rich waste materials. Among the enzymes evaluated MnP enzyme showed the best performance in terms of methane yield and substrate (lignin) utilization. Methane production of 200 mL CH₄/g VS was observed for the MnP amended reactor as compared to 5.7 mL CH₄/g VS for the control reactor. The lignin reduction in the MnP amended reactor and control reactor was 68.4% and 6.2%, respectively.     

Effect of pig manure to grass silage ratio on methane production in batch anaerobic co-digestion of concentrated pig manure and grass silage

Anaerobic co-digestion of concentrated pig manure (PM) with grass silage (GS) at five different PM to GS volatile solid (VS) ratios of 1:0, 3:1, 1:1, 1:3 and 0:1 was evaluated by examining operation stability and methane (CH₄) production potentials. The highest specific CH₄ yields were 304.2 and 302.8 ml CH₄/g VS at PM to GS ratios of 3:1 and 1:1, respectively. The digestion systems failed at the ratio of 0:1. The lag phase lasted 29.5, 28.1, 24.6 and 21.3 days at the ratios of 1:0, 3:1, 1:1 and 1:3, respectively. The daily methane yield was linearly correlated with the acetic acid concentration, indicating methane production was probably associated with acetoclastic methanogenesis. The hydrolysis constant linearly decreased with increasing the fraction of GS in the feedstock. This study recommends applying the PM to GS ratio of 1:1 in practice due to a high specific methane yield and a short lag phase.     

Efficiency of pretreatments for optimal enzymatic saccharification of soybean fiber

The effectiveness of several pretreatments [high-power ultrasound, sulfuric acid (H₂SO₄), sodium hydroxide (NaOH), and ammonium hydroxide (NH₃OH)] to enhance glucose production from insoluble fractions recovered from enzyme-assisted aqueous extraction processing of extruded full-fat soybean flakes (FFSF) was investigated. Sonication of the insoluble fraction at 144 μm_{pp (peak-to-peak)} for 30 and 60 s did not improve the saccharification yield. The solid fractions recovered after pretreatment with H₂SO₄ [1% (w/w), 90 °C, 1.5 h], NaOH [15% (w/w), 65 °C, 17 h], and NH₃OH [15% (w/w), 65 °C, 17 h] showed significant lignin degradation, i.e., 81.9%, 71.2%, and 75.4%, respectively, when compared to the control (7.4%). NH₃OH pretreatment resulted in the highest saccharification yield (63%) after 48 h of enzymatic saccharification. A treatment combining the extraction and saccharification steps and applied directly to the extruded FFSF, where oil extraction yield and saccharification yield reached 98% and 43%, respectively, was identified.     

Comparative performance of enzymatic and combined alkaline-enzymatic pretreatments on methane production from ensiled sorghum forage

This study investigated the effect of enzymatic and combined alkaline-enzymatic pretreatments on chemical composition and methane production from ensiled sorghum forage. Four commercial enzymatic preparations were tested and the two yielding the highest sugars release were added to evaluate any hydrolytic effect on both untreated and alkaline pretreated samples. In the combined alkaline-enzymatic pretreatment trials, the highest sugar release was found with Primafast and BGL preparations (added at a final concentration 0.12 and 0.20 mL/g TS, respectively), with a total monomeric content of 12 and 6.5 g/L. Fibre composition analysis confirmed that the combined alkaline-enzymatic pretreatment led to cellulose (up to 32 %) and hemicelluloses (up to 56 %) solubilisation, compared to the enzymatic pretreatment alone. BMP tests were performed on both untreated and pretreated samples, and time courses of methane production were fitted. Both enzymatic and combined alkaline-enzymatic pretreatment led to a methane production increase (304 and 362 mL CH₄/g VS), compared to that of untreated sorghum (265 mL CH₄/g VS), as  +15 and  +37 %, respectively. Moreover, higher specific methane production rates, compared to that of untreated sorghum (20.31 mL CH₄/g VS/d), were obtained by applying the enzymatic and combined alkaline-enzymatic pretreatment (33.94 and 31.65 mL CH₄/g VS/d), respectively.     

Cryopreservation of zebrafish (Danio rerio) primordial germ cells by vitrification of yolk-intact and yolk-depleted embryos using various cryoprotectant solutions

The aim of this study was to examine the effects of partial removal of yolk and cryoprotectant mixtures on the viability of cryopreserved primordial germ cells (PGCs) and elucidated the differentiation ability of cryopreserved PGCs in zebrafish. First, dechorionated yolk-intact and yolk-depleted (partially yolk removed) embryos, PGCs of which were labeled with green fluorescence protein (GFP), were vitrified after serial exposures to pretreatment solution (PS) and vitrification solution (VS) that contained ethylene glycol (EG), dimethyl sulfoxide (Me₂SO) or propylene glycol at 3 and 5 M, respectively. Although partial removal of yolk improved the viability of cryopreserved PGCs, numbers of PGCs with pseudopodial movement were limited (0–2.6 cells/embryo). Next, yolk-depleted embryos were cryopreserved using mixtures of two types of cryoprotectants. The maximum survival rate of PGCs (81%; 9.6 cells/embryo) was obtained from the yolk-depleted embryos vitrified using PS containing 2 M EG + 1 M Me₂SO and VS containing 3 M EG + 2 M Me₂SO and 56% (5.3 cells/embryo) of PGCs showed pseudopodial movement. Finally, PGCs recovered from yolk-depleted embryos (wild-type) that were vitrified under the optimum condition were transplanted individually into 236 sterilized recipient blastulae (recessive light-colored). Seven recipients matured and generated progeny with characteristics inherited from the PGC donor. In conclusion, the authors confirmed the beneficial effects of partial removal of yolk on the viability of cryopreserved PGCs and that the viability of the PGCs was improved by using PS and VS that contained two types of cryoprotectants, especially PS containing 2 M EG + 1 M Me₂SO and VS containing 3 M EG + 2 M Me₂SO, and that recovered PGCs retained ability to differentiate into functional gametes.     

Multi stage high rate biomethanation of poultry litter with self mixed anaerobic digester

A multi stage high rate biomethanation process with novel self mixed anaerobic digester (SMAD) was developed in the present study to reduce the hydraulic residence time (HRT), increase the volatile solids (VS) loading rate, improve the VS destruction efficiency and enhance the methane yield. Specific design features of SMAD were useful in mixing the digester contents without consuming power and de-alienated the problem of scum formation. In the first phase, poultry litter having 10% total solids (TS) was subjected to high rate biomethanation in multi stage configuration (SMAD-I and II in series with UASB reactor). It was observed that gross VS reduction of 58%, gross methane yield of 0.16 m³ kg⁻¹ (VS reduced) and VS loading rate of 3.5 kg VS m⁻³ day⁻¹ at HRT of 13 days was obtained. In the second phase SMAD-II was bypassed from the process scheme keeping the other parameters same as in the first phase. The results obtained were not as encouraging as in the first phase. The study showed that multi stage configuration with SMAD design improved the anaerobic digestion process efficiency of poultry litter.     

Comparison of Seven Chemical Pretreatments of Corn Straw for Improving Methane Yield by Anaerobic Digestion

Agriculture straw is considered a renewable resource that has the potential to contribute greatly to bioenergy supplies. Chemical pretreatment prior to anaerobic digestion can increase the anaerobic digestibility of agriculture straw. The present study investigated the effects of seven chemical pretreatments on the composition and methane yield of corn straw to assess their effectiveness of digestibility. Four acid reagents (H₂SO₄, HCl, H₂O₂, and CH₃COOH) at concentrations of 1%, 2%, 3%, and 4% (w/w) and three alkaline reagents (NaOH, Ca(OH)₂, and NH₃·H₂O) at concentrations of 4%, 6%, 8%, and 10% (w/w) were used for the pretreatments. All pretreatments were effective in the biodegradation of the lignocellulosic straw structure. The straw, pretreated with 3% H₂O₂ and 8% Ca(OH)₂, acquired the highest methane yield of 216.7 and 206.6 mL CH₄ g VS ⁻¹ in the acid and alkaline pretreatments, which are 115.4% and 105.3% greater than the untreated straw. H₂O₂ and Ca(OH)₂ can be considered as the most favorable pretreatment methods for improving the methane yield of straw because of their effectiveness and low cost.     

Synthesis of γ-cyclodextrin/chitosan composites for the efficient removal of Cd(II) from aqueous solution

The synthesis of chitosan-graft-γ-cyclodextrin (Ch-g-γ-CD) using persulfate/ascorbic acid redox system was done and characterized by FTIR, XRD, TGA and SEM/EDX. The optimum yield of the copolymer was obtained using 16 × 10⁻³ M γ-cyclodextrins (γ-CD), 2.8 × 10⁻² M ascorbic acid (AA), 1.8 × 10⁻² M K₂S₂O₈ and 0.1 g chitosan in 25 mL of 2% aqueous formic acid at 45 ± 0.2 °C. The highest percent grafting samples were evaluated for cadmium metal ion (Cd(II)) removal from the aqueous solutions where the sorption capacities were found proportional to the grafting extent. The sorption was pH and concentration dependent where, pH = 8.5 was found to be the optimum value. The adsorption data were modeled using Langmuir and Freundlich isotherms. The equilibrium data followed the Langmuir isotherm model with maximum sorption capacity of 833.33 mg/g. The influence of electrolytes, sodium chloride (NaCl) and sodium sulphate (Na₂SO₄) on Cd(II) uptake was also studied. Desorption of the cadmium loaded Ch-g-γ-CD was accomplished with 0.01 N H₂SO₄. The adsorbent exhibited high reusability and could be successfully recycled for nine cycles where in the ninth cycle 27% adsorption was feasible.     

Optimization of microwave-assisted FeCl3 pretreatment conditions of rice straw and utilization of Trichoderma viride and Bacillus pumilus for production of reducing sugars

In this study, Box-Behnken design (BBD) and response surface methodology (RSM) were used to optimize microwave-assisted FeCl₃ pretreatment conditions of rice straw with respect to FeCl₃ concentration, microwave intensity, irradiation time and substrate concentration. When rice straw was pretreated at the optimal conditions of FeCl₃ concentration, 0.14 mol/L; microwave intensity, 160 °C; irradiation time, 19 min; substrate concentration, 109 g/L; and inoculated with Trichoderma viride and Bacillus pumilus, the production of reducing sugars was 6.62 g/L. This yield was 2.9 times higher than that obtained with untreated rice straw. The microorganisms degraded 37.8% of pretreated rice straw after 72 h. The structural characteristic analyses suggest that microwave-assisted FeCl₃ pretreatment damaged the silicified waxy surface of rice straw, disrupted almost all the ether linkages between lignin and carbohydrates, and removed lignin.     

Batch anaerobic co-digestion of cow manure and waste milk in two-stage process for hydrogen and methane productions

Anaerobic co-digestion of cow manure (CM) and waste milk (WM), produced by sick cows during treatment with antibiotics, was evaluated in two-stage process under thermophilic condition (55 °C) to determine the effect of WM addition on hydrogen (H₂) and methane (CH₄) production potentials, volatile solids (VS) removal, and energy recovery. Six CM to WM VS ratios of 100:0, 90:10, 70:30, 50:50, 30:70, and 10:90 were examined using 1-L batch digesters. The WM VS ratio of 30 % was found to be the minimum limit for significant increases in specific H₂ and CH₄ yields, and VS removal as compared to digestion of manure alone (P < 0.05). The highest specific H₂ and CH₄ yields, VS removal and energy yield were 38.2 mL/g VS, 627.6 mL/g VS, 78.4 % and 25,459.8 kJ/kg VS, respectively, in CM:WM 30:70. Lag phases to H₂ and CH₄ productions were observed in CM–WM mixtures, increased with increasing the amount of WM in the feedstock and were greater than 72 h in CM:WM 50:50 and 30:70. The digestion system failed in CM:WM 10:90. The results suggest that CM:WM 30:70 was optimum, however, due to limited amount of WM usually generated and long lag phase at this ratio which may make the process uneconomical, CM:WM 70:30 is recommended in practice.     

3-Phenyllactic acid production by substrate feeding and pH-control in fed-batch fermentation of Lactobacillus sp. SK007

3-Phenyllactic acid (PLA), which is produced by some strains of lactic acid bacteria (LAB), is a known antimicrobial agent with a broad spectrum. Batch and fed-batch fermentation by the strain Lactobacillus sp. SK007 for PLA production have been reported. With batch fermentation without pH-control, PLA production yield was 2.42 g L⁻¹. When fed-batch fermentation by Lactobacillus sp. SK007 was conducted in 3 L initial volume with pH-control at 6.0 and intermittent feeding, which was developed after fermentation for 12 h and every 2 h with 120 mL 100 g L⁻¹ PPA phenylpyruvic acid (PPA) and 50 mL 500 g L⁻¹ glucose each time, PLA production yield reached 17.38 g L⁻¹. The final conversion ratio of PPA to PLA was 51.1%, and the PLA production rate was 0.241 g L⁻¹ h⁻¹. This indicated that PPA was the ideal substrate for PLA fermentation production, and fed-batch fermentation with intermittent PPA feeding and pH-control was an effective approach to improve PLA production yield.     

Feasibility of biogas production from anaerobic co-digestion of herbal-extraction residues with swine manure

The objective of this work was to examine the feasibility of biogas production from the anaerobic co-digestion of herbal-extraction residues with swine manure. Batch and semi-continuous experiments were carried out under mesophilic anaerobic conditions. Batch experiments revealed that the highest specific biogas yield was 294 mL CH₄ g⁻¹ volatile solids added, obtained at 50% of herbal-extraction residues and 3.50 g volatile solids g⁻¹ mixed liquor suspended solids. Specific methane yield from swine manure alone was 207 mL CH₄ g⁻¹ volatile solid added d⁻¹ at 3.50 g volatile solids g⁻¹ mixed liquor suspended solids. Furthermore, specific methane yields were 162, 180 and 220 mL CH₄ g⁻¹ volatile solids added d⁻¹ for the reactors co-digesting mixtures with 10%, 25% and 50% herbal-extraction residues, respectively. These results suggested that biogas production could be enhanced efficiently by the anaerobic co-digestion of herbal-extraction residues with swine manure.