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.     

Buffer requirements for enhanced hydrogen production in acidogenic digestion of food wastes

The requirements for pH buffer addition for hydrogen production and acidogenesis in batch acidogenic digestion of a food waste (FW) feedstock with limited alkalinity was studied at various initial pH conditions (6.0–8.0). The results showed that, without buffer addition, hydrogen production from this feedstock was insignificant regardless of the initial pH. With buffer addition, hydrogen production improved significantly if the initial pH was greater than 6.0. Substantial hydrogen production occurred when the pH at the end of the batch digestion was higher than 5.5. The maximum hydrogen production was found to be 120 mL/g VS added when the initial pH was 6.5 and buffer addition was in the range of 15–20 mmol/g VS. The effect of pH buffering on the formation of volatile fatty acids (acetic acid, propionic acid and butyric acid) was similar to its effect on hydrogen production. The results of this study clearly indicated shifts in the metabolic pathways with the pH of fermentation. The changes in metabolic pathways impacted upon the dosage of buffer that was required to achieve maximum hydrogen generation.     

Hydrogen production by the newly isolated Clostridium beijerinckii RZF-1108

A fermentative hydrogen-producing strain, RZF-1108, was isolated from a biohydrogen reactor, and identified as Clostridium beijerinckii on the basis of the 16S rRNA gene analysis and physiobiochemical characteristics. The effects of culture conditions on hydrogen production by C. beijerinckii RZF-1108 were investigated in batch cultures. The hydrogen production and growth of strain RZF-1108 were highly dependent on temperature, initial pH and substrate concentration. Yeast extract was a favorable nitrogen source for hydrogen production and growth of RZF-1108. Hydrogen production corresponded to cell biomass yield in different culture conditions. The maximum hydrogen evolution, yield and production rate of 2209 ml H₂/l medium, 1.97 mol H₂/mol glucose and 104.20 ml H₂/g CDW h⁻¹ were obtained at 9 g/l of glucose, initial pH of 7.0, inoculum volume of 8% and temperature of 35 °C, respectively. These results demonstrate that C. beijerinckii can efficiently produce H₂, and is another model microorganism for biohydrogen investigations.     

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.     

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.     

Eliminating methanogenic activity in hydrogen reactor to improve biogas production in a two-stage anaerobic digestion process co-digesting municipal food waste and sewage sludge

Laboratory scale two-stage anaerobic digestion process model was operated for 280 days to investigate the feasibility to produce both hydrogen and methane from a mixture feedstock (1:1 (v/v)) of municipal food waste and sewage sludge. The maximum hydrogen and methane yields obtained in the two stages were 0.93 and 9.5 mL/mL feedstock. To eliminate methanogenic activity and obtain substantial hydrogen production in the hydrogen reactor, both feedstock and mixed liquor required treatment. The heat treatment (100 °C, 10 min) for feedstock and a periodical treatment (every 2-5 weeks, either heating, removal of biomass particles or flushing with air) for mixed liquor were effective in different extent. The methane production in the second stage was significantly improved by the hydrogen production in the first stage. The maximum methane production obtained in the period of high hydrogen production was more than 2-fold of that observed in the low hydrogen production period.     

Co-digestion of intermediate landfill leachate and sewage sludge as a method of leachate utilization

This study examines the co-digestion of intermediate landfill leachate and sewage sludge from a municipal wastewater treatment plant. Application of leachate as a co-fermentation component increased the concentrations of soluble organic compounds (expressed as total organic carbon), ammonium nitrogen, and alkalinity in the digester influents.The biogas yield obtained from the co-fermentation of a 20:1 sewage sludge: intermediate leachate mixture was 1.30 m³ per kg of removed volatile solids (VS), while that from a 10:1 mixture was 1.24 m³ per kg of removed VS. These values exceeded the biogas yield for the sludge alone by 13% and 8%, respectively. The leachate addition influenced the proportion of methane to a minor extent. Increased methane yields of 16.9% and 6.2% per kg of removed VS were found for the two sewage sluge:intermediate leachate mixtures, respectively.     

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.     

Effects of key operational parameters on biohydrogen production via anaerobic fermentation in a sequencing batch reactor

In this study, a series of tests were conducted in a 6 L anaerobic sequencing batch reactor (ASBR) to investigate the effect of pH, hydraulic retention time (HRT) and organic loading rate on biohydrogen production at 28 °C. Sucrose was used as the main substrate to mimic carbohydrate-rich wastewater and inoculum was prepared from anaerobic digested sludge without pretreatment. The reactor was operated initially with nitrogen sparging to form anaerobic condition. Results showed that methanogens were effectively suppressed. The optimum pH value would vary depending on the HRT. Maximum hydrogen production rate and yield of 3.04 L H₂/L reactor d and 2.16 mol H₂/mol hexose respectively were achieved at pH 4.5, HRT 30 h, and OLR 11.0 kg/m³ d. Two relationships involving the propionic acid/acetic acid ratio and ethanol/acetic acid ratio were derived from the analysis of the metabolites of fermentation. Ethanol/acetic acid ratio of 1.25 was found to be a threshold value for higher hydrogen production.     

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.     

Hydrogen and methane production through two-stage mesophilic anaerobic digestion of olive pulp

The present study focused on the anaerobic biohydrogen production from olive pulp (two phase olive mill wastes, TPOMW) and the subsequent anaerobic treatment of the effluent for methane production under mesophilic conditions in a two-stage process. Biohydrogen production from water-diluted (1:4) olive pulp was investigated at hydraulic retention times (HRT) of 30 h, 14.5 h and 7.5 h while methane production from the effluent of hydrogenogenic reactor was studied at 20 d, 15 d, 10 d and 5 d HRT. In comparison with previous studies, it has been shown that the thermophilic hydrogen production process was more efficient than the mesophilic one in both hydrogen production rate and yield. The methanogenic reactor was successfully operated at 20, 15 and 10 days HRT while it failed when an HRT of 5 days was applied. Methane productivity reached the maximum value of 1.13 ± 0.08 L/L/d at 10 days HRT whereas the methane yield increased with the HRT. The Anaerobic Digestion Model no. 1 (ADM1) was applied to the obtained experimental data from the methanogenic reactor to simulate the digester response at all HRT tested. The ability of the model to predict the experimental results was evident even in the case of the process failure, thus implying that the ADM1 could be a valuable tool for process design even in the case of a complex feedstock. In general, the two-stage anaerobic digestion proved to be a stable, reliable and effective process for energy recovery and stabilization treatment of olive pulp.     

Biohydrogen from thermophilic co-fermentation of swine manure with fruit and vegetable waste: maximizing stable production without pH control

Hydrogen production by dark fermentation may suffer of inhibition or instability due to pH deviations from optimality. The co-fermentation of promptly degradable feedstock with alkali-rich materials, such as livestock wastes, may represent a feasible and easy to implement approach to avoid external adjustments of pH.Experiments were designed to investigate the effect of the mixing ratio of fruit-vegetable waste with swine manure with the aim of maximizing biohydrogen production while obtaining process stability through the endogenous alkalinity of manure.Fruit-vegetable/swine manure ratio of 35/65 and HRT of 2 d resulted to give the highest production rate of 3.27 ± 0.51 L_H2 L⁻¹ d⁻¹, with a corresponding hydrogen yield of 126 ± 22 mL_H2 g⁻¹_VS-added and H₂ content in the biogas of 42 ± 5%. At these operating conditions the process exhibited also one of the highest measured stability, with daily productions deviating for less than 14% from the average.     

Biological hydrogen production by the algal biomass Chlorella vulgaris MSU 01 strain isolated from pond sediment

Chlorella vulgaris MSU 01 strain isolated from the sediment of the pond is able to produce molecular hydrogen in a clean way. To relate the dynamic coupling between the cultural conditions and biological responses, an original lab scale set up has been developed for hydrogen production. Different sources like mannitol, glucose, alanine, citric acid, aspartic acid, l-alanine, l-cysteine, sodium succinate and sodium pyruvate were used for algal media optimization. Corn stalk, from 1 to 5 g/L was tested for the effective algal growth and hydrogen production. The cell concentration of 1.6-19 g/L dry cell weight (DCW) was found at the 10th day. The kinetic parameters involved in the hydrogen production at 4 g/L corn stalk using the algal inoculum (50 mL) in the bioreactor volume (500 mL) was found to be with the hydrogen production potential (P_s) of 7.784 mL and production yield of (P_r) 5.534 mL respectively. The growth profile of the algal biomass at the above mentioned condition expressed the logistic model with R² 0.9988. The final pH of the broth was increased from 7.0 to 8.5-8.7. The anaerobic fermentation by C. vulgaris MSU 01 strain involved in the conversion process of complex carbon source has increased the H₂ evolution rate and higher butyrate concentration in the fermentate.     

Enhancement of photo-hydrogen production in a biofilm photobioreactor using optical fiber with additional rough surface

In this study, a biofilm photobioreactor with optical fibers that have additional rough surface (OFBP-R) was developed and it was shown that additional rough surface greatly enhanced the biofilm formation and thus increased the cell concentration, leading to an improvement in the hydrogen production performance. The effects of operational conditions, including the influent substrate concentration, flow rate, temperature and influent medium pH, on the performance of OFBP-R were also investigated. The experimental results showed that the optimum operational conditions for hydrogen production were: the influent substrate concentration 60 mM, flow rate 30 mL/h, temperature 30 °C and influent medium pH 7. Under the optimal operation conditions discovered in this work, the OFBP-R yielded fairly good and stable long-term performance with hydrogen production rate of 1.75 mmol/L/h, light conversion efficiency of 9.3% and substrate degradation efficiency of 75%.     

Harvesting biohydrogen from cellobiose from sulfide or nitrite-containing wastewaters using Clostridium sp. R1

Harvesting biohydrogen from inhibiting wastewaters is of practical interest since the toxicity of compounds in a wastewater stream commonly prevents the bioenergy content being recovered. The isolated Clostridium sp. R1 is utilized to degrade cellobiose in sulfide or nitrite-containing medium for biohydrogen production. The strain can effectively degrade cellobiose free of severe inhibitory effects at up to 200 mg l⁻¹ sulfide or to 5 mg l⁻¹ nitrite, yielding hydrogen at >2.0 mol H₂ mol⁻¹ cellobiose. Principal metabolites of cellobiose fermentation are acetate and butyrate, with the concentration of the former increases with increasing sulfide and nitrite concentrations. The isolated strain can yield hydrogen from cellobiose in sulfide-laden wastewaters. However, the present of nitrite significantly limit the efficiency of the biohydrogen harvesting process.     

Bioconversion of wheat stalk to hydrogen by dark fermentation: effect of different mixed microflora on hydrogen yield and cellulose solubilisation

This study determined hydrogen production, volatile fatty acids (VFAs) generation and cellulose solubilisation from anaerobic dark fermentation of wheat stalk and showed the effect of different mixed microflora. The cumulative hydrogen yields of anaerobic digested activated sludge (AS)-inoculated and anaerobic digested dairy manure (DM)-inoculated system were 23.3 and 37.0 mL/g VS at 204 h, respectively. A modified Gompertz equation was able to adequately describe the production of hydrogen from the batch fermentation by both mixed microflora. During the process, acetate and butyrate accounted for more than 76.1% of total VFAs for both fermentations. The extent of cellulose solubilisation approached 46.6% and 75.2% for AS- and DM-inoculated fermentation, respectively. The X-ray diffraction (XRD) showed that the crystallinities of both fermented stalks were partly disrupted by the mixed microflora, and DM-inoculated fermentation had more disruption than AS-inoculated one.     

Feasibility of hydrogen production from ripened fruits by a combined two-stage (dark/dark) fermentation system

Anaerobic fermentation for hydrogen (H₂) production was studied in a two-stage fermentation system fed with different ripened fruit feedstocks (apple, pear, and grape). Among the feedstocks, ripened apple was the most efficient substrate for cumulative H₂ production (4463.7 mL-H₂ L⁻¹-culture) with a maximum H₂ yield (2.2 mol H₂ mol⁻¹ glucose) in the first stage at a hydraulic retention time (HRT) of 18 h. The additional cumulative biohydrogen (3337.4 mL-H₂ L⁻¹-culture) was produced in the second stage with the reused residual substrate from the first stage. The major byproducts in this study were butyrate, acetate, and ethanol, and butyrate was dominant among them in all test runs. During the two-stage system, the energy efficiency (H₂ conversion) obtained from mixed ripened fruits (RF) increased from 4.6% (in the first stage) to 15.5% (in the second stage), which indicated the energy efficiency can be improved by combined hydrogen production process. The RF could be used as substrates for biohydrogen fermentation in a two-stage (dark/dark) fermentation system.     

Evaluation of dairy cattle manure as a supplement to improve net energy gain in fermentative hydrogen production from sucrose

This study evaluated fermentative biohydrogen production from sucrose supplemented with dairy cattle manure at different sucrose:manure ratios. Hydrogen yields found in this study (2.9-5.3 M hydrogen/M sucrose) at ambient temperature are higher than literature results obtained at mesophilic temperatures. This study demonstrated that dairy cattle manure could serve as a buffering agent to maintain recommended pH levels; as a nutrient source to provide the required nutrients for hydrogen production; as a seed to produce hydrogen from sucrose; and as a co-substrate to improve the hydrogen yield. Based on an analysis of the net energy gain, it is concluded that positive net energy gains can be realized with non-thermal pretreatment and/or by combining dark fermentation with anaerobic digestion or microbial fuel cells to extract additional energy from the aqueous products of dark fermentation.     

Hydrogen and methane production from desugared molasses using a two‐stage thermophilic anaerobic process

Hydrogen and methane production from desugared molasses by a two‐stage thermophilic anaerobic process was investigated in a series of two up‐flow anaerobic sludge blanket (UASB) reactors. The first reactor that was dominated with hydrogen‐producing bacteria of Thermoanaerobacterium thermosaccharolyticum and Thermoanaerobacterium aciditolerans could generate a high hydrogen production rate of 5600 mL H₂/day/L, corresponding to a yield of 132 mL H₂/g volatile solid (VS). The effluent from the hydrogen reactor was further converted to methane in the second reactor with the optimal production rate of 3380 mL CH₄/day/L, corresponding to a yield of 239 mL CH₄/g VS. Aceticlastic Methanosarcina mazei was the dominant methanogen in the methanogenesis stage. This work demonstrates that biohydrogen production can be very efficiently coupled with a subsequent step of methane production using desugared molasses. Furthermore, the mixed gas with a volumetric content of 16.5% H_2, 38.7% CO₂, and 44.8% CH₄, containing approximately 15% energy by hydrogen is viable to be bio‐hythane.     

玉米芯发酵法生物制氢

在批式培养试验中, 以牛粪堆肥为天然产氢菌源, 玉米芯为底物, 通过厌氧发酵生产氢气。系统考察了底物预处理条件、初始pH值和底物浓度对玉米芯产氢能力的影响。在初始pH 8.0, 1.0%盐酸预处理底物30 min, 底物浓度10 g/L的最佳产氢条件下, 玉米芯最大产氢能力〔每克TVS（总挥发性固体物）产氢量〕和最大产氢速率（每克TVS每小时产氢量）分别为107.9 mL /g、4.20 mL/g·h-1。玉米芯经酸预处理后半纤维素含量由42.2%下降至3.0%, 而酸预处理的玉米芯产氢前后纤维素、半纤维素和木质素含量只有少量变化。产氢菌主要用酸预处理产生的可溶性糖产氢, 故底物的酸预处理对玉米芯的发酵产氢非常重要。用傅里叶变换红外光谱(FTIR)分析显示酸预处理和产氢过程中玉米芯的特征峰发生变化, 酸预处理过程降解了底物纤维素的无定形区和半纤维素, 产氢微生物对纤维素的结晶区有破坏作用。