Effects of pretreatment methods on cassava wastewater for biohydrogen production optimization

Batch production of biohydrogen from cassava wastewater pretreated with (i) sonication, (ii) OPTIMASH BG^® (enzyme), and (iii) α-amylase (enzyme) were investigated using anaerobic seed sludge subjected to heat pretreatment at 105 °C for 90 min. Hydrogen yield at pH 7.0 for cassava wastewater pretreated with sonication for 45 min using anaerobic seed sludge was 0.913 mol H₂/g COD. Results from pretreatment with OPTIMASH BG^® at 0.20% and pH 7 showed a hydrogen yield of 4.24 mol H₂/g COD. Superior results were obtained when the wastewater was pretreated with α-amylase at 0.20% at pH 7 with a hydrogen yield of 5.02 mol H₂/g COD. In all cases, no methane production was observed when using heat-treated sludge as seed inoculum. Percentage COD removal was found to be highest (60%) using α-amylase as pretreatment followed by OPTIMASH BG^® at 54% and sonication (40% reduction rate). Results further suggested that cassava wastewater is one of the potential sources of renewable biomass to produce hydrogen.     

Enhanced methane fermentation of municipal sewage sludge by microbial electrochemical systems integrated with anaerobic digestion

Sewage sludge from a municipal wastewater treatment plant was fed into a microbial electrochemical system, combined with an anaerobic digester (MES-AD), for enhanced methane production and sludge stabilization. The effect of thermally pretreating the sewage sludge on MES-AD performance was investigated. These results were compared to those obtained from control operations, in which the sludge was not pretreated or MES integration was absent. The soluble chemical oxygen demand (SCOD) in the raw sewage sludge after pretreatment was 31% higher than the SCOD in untreated sludge (5804.85 mg/L vs. 4441.46 mg/mL). The methane yield and proportion of methane in biogas generated by the MES-AD were higher than those of the control systems, regardless of the pretreatment process. The maximum methane yield (0.28 L CH₄/g COD) and methane production (1139 mL) were obtained with the MES inoculated with pretreated sewage sludge. Methane yield and production with this system using pretreated sewage were 47% and 56% higher, respectively, than those of the control (0.19 L CH₄/g COD, 730 mL). Additionally, the maximum SCOD removal (89%) and current generation were obtained with the MES inoculated with a pretreated substrate. These results suggested that sewage sludge could be efficiently stabilized with enhanced methane production by synergistic combination of MES-AD system with pretreatment process.     

Hydrogen production from the fermentation of corn stover biomass pretreated with a steam-explosion process

Lignocellulosic biomass contains 70–80% carbohydrates and could serve as the ideal feedstock for fermentative hydrogen production. We conducted the pretreatment of corn stover using a steam-explosion process and studied its fermentability for hydrogen production. Using natural inoculant obtained from the heated sludge of a local wastewater treatment plant, we demonstrated that the indigenous microbes were capable of efficiently fermenting the aqueous hydrolyzate derived from the hemicellulose fraction of the steam-pretreated corn stover with and without acid during pretreatment. Biogas contained equal amounts of hydrogen and carbon dioxide. The carbon mass balance is approximately 84%, with acetic and butyric acids as the major carbon byproducts along with carbon dioxide. Hydrogen molar yields of 2.84 and 3.0 were obtained using the mixed sugars present in the hydrolyzate derived from neutral and acidic steam explosion, respectively. These findings verify that hemicellulose from corn stover could be a suitable feedstock for hydrogen production via dark fermentation.     

Enhancement of biohydrogen producing using ultrasonication

Ultrasonication was evaluated as a pretreatment for biological hydrogen production from glucose in batch studies, in comparison with heat-shock pretreatment, acid pretreatment, and base pretreatment. The optimized sonication energy for hydrogen production using anaerobic digester sludge was 79 kJ/gTS. Sonication with temperature control (less than 30 °C) increased volumetric hydrogen production by 120% over the untreated sludge, and by 40% over the heat-shock and acid pretreated sludge, with a marginal (∼10%) increase in hydrogen production rate. Upon comparing the molar hydrogen yield in sonicated sludge with and without temperature control, the deleterious effect of heat on some hydrogen producers as reflected by a 30% decrease in yield to 1.03 mol H₂/mol glucose is evident. Sonication with temperature control affected a 45% increase in molar hydrogen yield to 1.55 mol H₂/mol glucose over heat-shock pretreatment at 70 °C for 30 min and acidification to pH 3.0 for 24 h at 4 °C. Sonication with temperature control produced a biomass yield of 0.13 g VSS/g COD, as compared to 0.24 g VSS/g COD for the untreated sludge. The hydrogen yield increased linearly with the molar acetate to butyrate ratio and decreased linearly with the biomass yield.     

Effect of acid,heat and combined acid-heat pretreatments of anaerobic sludge on hydrogen production by anaerobic mixed cultures

Activated sludge (AS) from wastewater treatment plant of brewery industry was used as substrate for hydrogen production by anaerobic mixed cultures in batch fermentation process. The AS (10% TS) was pretreated by acid, heat and combined acid and heat. Combined acid- heat treatment (0.5% (w/v) HCl, 110 °C, 60 min) gave the highest soluble COD (sCOD) of 1785.6 ± 27.1 mg/L with the highest soluble protein and carbohydrate of 8.1 ± 0.1 and 38.5 ± 0.8 mg/L, respectively. After the pretreatment, the pretreated sludge was used to produce hydrogen by heat treated upflow anaerobic sludge blanket (UASB) granules. A maximum hydrogen production potential of 481 mL H₂/L was achieved from the AS pretreated with acid (0.5% (w/v) HCl) for 6 h.     

Hydrogen production from sludge of poultry slaughterhouse wastewater treatment plant pretreated with microwave

This study aims to produce hydrogen from sludge of poultry slaughterhouse wastewater treatment plant (5% total solid) by anaerobic batch fermentation with Enterobactor aerogenes or mixed cultures from hot spring sediment as the inoculums. Sludge was heated in microwave at 850 W for 3 min. Results indicated that a soluble chemical oxygen demand (sCOD) of pretreated sludge was higher than that of raw sludge. Pretreated sludge inoculated with E. aerogenes and supplemented with the Endo nutrient had a higher hydrogen yield (12.77 mL H₂/g tCOD) than the raw sludge (0.18 mL H₂/g tCOD). When considered the hydrogen yield, the optimum initial pH for hydrogen production from microwave pretreated sludge was 5.5 giving the maximum value of 12.77 mL H₂/g tCOD. However, when considered the hydrogen production rate (R_m), the optimum pH for hydrogen production would be 9.0 with the maximum R_m of 22.80 mL H₂/L sludge·h.     

Enriching hydrogen-producing bacteria from digested sludge by different pretreatment methods

The pretreatment of digested sludge by different methods, including ionizing irradiation, heat-shock, acid and base, was performed for enriching hydrogen-producing bacteria. These methods were evaluated and compared based on their suitability in the enrichment of hydrogen-producing bacteria in dark fermentation with glucose as a substrate in batch tests. The experimental results showed that the seed sludge pretreated by ionizing irradiation achieved the best hydrogen production among the different pretreatment methods, and the maximum hydrogen production potential, maximum hydrogen production rate, hydrogen yield and substrate degradation rate were 525.6 mL, 37.2 mL/h, 267.7 mL/g glucose (2.15 mol/mol glucose) and 98.9%, respectively. Ionizing irradiation can be a good optional pretreatment method for enriching hydrogen-producing bacteria from digested sludge. The effect of ionizing irradiation on the microbial community structure dynamics of the pretreated sludge deserves further study, which will help us to understand the mechanisms leading to the effect of high bio-hydrogen production.     

Catalytic gasification of textile wastewater treatment sludge for hydrogen production in supercritical water with K2CO3/H2O2: Reaction variables,mechanism and kinetics

As the main by-product, sludge was generated during the process of textile wastewater treatment. Due to containing organic compounds, textile sludge has great potential to be effectively reused for production of clean energy. In this work, gasification of textile sludge in supercritical water for hydrogen production was investigated. In order to improve hydrogen production, H₂O₂ and K₂CO₃ were used as catalysts. Effects of reaction variables (including temperature, retention time, oxidation coefficient and alkali catalyst dosage) on hydrogen yield were studied. Experimental results indicate that hydrogen yield increases with rise of temperature. When reaction temperature reaches 500 °C, the maximum value of hydrogen yield being 10.6 mol/kg was obtained. When excessive H₂O₂ was added, decrease of hydrogen yield was achieved. However, the addition of K₂CO₃ is favor to hydrogen yield, which is about 1.5 times as much as that of without catalyst. Meanwhile, reaction mechanism and kinetics of textile sludge gasification in supercritical water were explored. Reaction activation energy and Arrhenius constants were obtained in the established kinetic model.     

预处理温度对活性污泥发酵产氢特性的影响

为寻求适宜的种泥热处理方法,利用摇瓶发酵实验,考察了城市污水处理厂好氧活性污泥分别经65、80、95、110℃热处理30min后,其利用葡萄糖发酵产氢的特性。结果表明:在初始pH=7.0、葡萄糖浓度10g/L、接种量2gMLVSS/L条件下,35℃培养72h,经65℃和95℃处理的种泥表现出较好的发酵产氢性能,其葡萄糖的氢气转化率分别达到1.08和1.11mol/mol,污泥的比产氢率分别为8.36和9.05mmol/gMLVSS;经65℃预处理的种泥发酵体系,表现为丁酸型发酵,其葡萄糖降解率和最大产氢速率分别高达82%和11.29mL/h,而经95℃预处理的种泥发酵体系则呈现混合酸发酵特征,其葡萄糖转化率和最大产氢速率分别仅为76%和4.45mL/h。     

The effect of heat pretreatment temperature on fermentative hydrogen production using mixed cultures

The effect of heat treatment at different temperatures on two types of inocula, activated sludge and anaerobically digested sludge, was investigated in batch cultures. Heat treatments were conducted at 65, 80 and 95 °C for 30 min. The untreated inocula produced less amount of hydrogen than the pretreated inocula, with lactic acid as the main metabolite. The maximum yields of 2.3 and 1.6 mol H₂/mol glucose were achieved for the 65 °C pretreated anaerobically digested and activated sludges, respectively. Approximately a 15% decrease in yield was observed with increasing pretreatment temperature from 65 to 95 °C concomitant with an increase in butyrate/acetate ratio from 1.5 to 2.4 for anaerobically digested sludge. The increase of pretreatment temperature of activated sludge to 95 °C suppressed the hydrogen production by lactic acid fermentation. DNA analysis of the microbial community showed that the elevated pretreatment temperatures reduced the species diversity.     

Co-hydrothermal gasification of microbial sludge and algae Kappaphycus alvarezii for bio-hydrogen production: Study on aqueous phase reforming

In this study, wastewater obtained from a sewage treatment plant was treated successively by using microbial consortium and macroalgae Kappaphycus alvarezii to generate microbial sludge and algal biomass. The production of green fuel was carried out via co-gasification of microbial sludge and macroalgae Kappaphycus alvarezii for a duration of 60 min, feedstock to solvent ratio (5 to 20 g of feedstock in 200 mL), sludge to algae ratio (ranging from 1:1 to 3:1) and temperature (300–400 °C) respectively. Maximum bio-hydrogen yield was 36.1% and methane yield was 38.4% at a temperature of 360 °C at a feedstock to solvent ratio of 15:200 g/mL and sludge to algae ratio of 2:1 individually. The liquid by product of co-gasification process was later subjected to photocatalytic reforming, resulted in an enhanced hydrogen composition of 61.25%.     

Photocatalytic pretreatment for the redox conversion of waste activated sludge to enhance biohydrogen production

Photocatalytic pretreatment of waste activated sludge (WAS) using a flat photocatalytic reactor was undertaken. Photocatalytic pretreatment enhanced the release of soluble substances from WAS, in which the soluble protein and soluble carbohydrate concentration increased by about 50% and 80%, respectively. Significant removal of heavy metal ions from the liquid phase of WAS was also achieved after photocatalytic pretreatment. In addition, the highest hydrogen yield and the highest concentration of volatile fatty acids (VFAs) were achieved from the photocatalysis pretreated WAS by batch anaerobic digestion (55 °C). The cumulative hydrogen yield from photocatalysis pretreated WAS was 211.0 ml/l-sludge, much higher than those from UV pretreated WAS (111.0 ml/l-sludge) and from raw WAS (93.0 ml/l-sludge). The results indicate that photocatalysis is a promising WAS pretreatment method for the enhancement of biohydrogen production, probably due to the photo-oxidation of organics and simultaneous photo-reduction of heavy metal ions in WAS.     

Evaluation of different pretreatment methods for preparing hydrogen-producing seed inocula from waste activated sludge

Waste activated sludge (WAS) is the most favorable inoculum for dark fermentative hydrogen-producing processes, because it can be collected economically. In order to accelerate the start-up process and develop the efficiency and stability of a hydrogen production system, pretreatment of the seed sludge has been examined to enrich hydrogen-producing bacteria. Six pretreatment methods including acid, base, heat-shock, aeration, chloroform and 2-bromoethanesulfonate (BES) were performed on WAS in batch cultures utilizing glucose as the substrate. The results showed that, at 35 °C and initial pH of 7.0, hydrogen yields of the pretreated sludge (except for BES) were higher than the control test. The pretreatment methods resulted in different distributions of soluble metabolites. Acid pretreatment at pH of 3 was the best among all six pretreatment methods, and the maximal hydrogen yield of 1.51 mol/mol-glucose-consumed and the maximal specific hydrogen production of 22.81 mmolH₂/g VSS were observed. The hydrogen yield of the acid treated sludge increased to 1.82 mol/mol-glucose-consumed after five repeated-batch cultivations. It was concluded that acid pretreatment is a simple, economic and effective method for enriching hydrogen-producing bacteria from WAS.     

Thermophilic hydrogen fermentation from Korean rice straw by Thermotoga neapolitana

Rice straw, a low-cost lignocellulosic biomass was used as feedstock for thermophilic hydrogen fermentation by Thermotoga neapolitana. Hydrogen production, the growth and cellulose digestibility of the hyperthermophile in batch mode from untreated as well as chemically pretreated (ammonia and dilute sulfuric acid) Korean rice straws were investigated. Pretreatment method using combination of 10% ammonia and 1.0% dilute sulfuric acid was developed to increase the digestibility of rice straw for the hyperthermophilic H₂ fermentation and to decrease the time consumption. In a typical fermentation using raw rice straw, 29% of the substrate was digested and 2.3 mmol H₂/g straw of hydrogen yield was consistently obtained. Compared with the pretreatments using only ammonia or dilute sulfuric acid, the combined pretreatment method using both chemical agents significantly increases the digestibility of rice straw with 85.4% of substrate consumption. H₂ production on rice straw from this combined pretreatment showed the highest yield (2.7 mmol H₂/g straw) and the highest sugar conversions (72.9% of glucose and 95.7% of xylose).     

Effects of different pretreatment methods on fermentation types and dominant bacteria for hydrogen production

In order to enrich hydrogen producing bacteria and to establish high-efficient communities of the mixed microbial cultures, inoculum needs to be pretreated before the cultivation. Four pretreatment methods including heat-shock pretreatment, acid pretreatment, alkaline pretreatment and repeated-aeration pretreatment were performed on the seed sludge which was collected from a secondary settling tank of a municipal wastewater treatment plant. In contrast to the control test without any pretreatment, the heat-shock pretreatment, acid pretreatment and repeated-aeration pretreatment completely suppressed the methanogenic activity of the seed sludge, but the alkaline pretreatment did not. Employing different pretreatment methods resulted in the change in fermentation types as butyric-acid type fermentation was achieved by the heat-shock and alkaline pretreatments, mixed-acid type fermentation was achieved by acid pretreatment and the control, and ethanol-type fermentation was observed by repeated-aeration pretreatment. Denaturing gradient gel electrophoresis (DGGE) profiles revealed that pretreatment method substantially affected the species composition of microbial communities. The highest hydrogen yield of 1.96 mol/mol-glucose was observed with the repeated-aeration pretreatment method, while the lowest was obtained as the seed sludge was acidified. It is concluded that the pretreatment methods led to the difference in the initial microbial communities which might be directly responsible for different fermentation types and hydrogen yields.     

Evaluation of ultrasonication as a treatment strategy for enhancement of biohydrogen production from complex distillery wastewater and process optimization

Dark fermentation of distillery wastewater (DWW) gives a lower hydrogen yield (HY) and hydrogen production rate (R), owing to the complexity and a recalcitrant nature of effluent. Therefore, an effective pretreatment of DWW becomes imperative for the improvement of biohydrogen production. In the present study, the efficacy of ultrasonic pretreatment for enhancement of biohydrogen production from DWW was evaluated in batch test. Several variables, such as COD input, ultrasonic density (UD), and ultrasonication time (UT) were studied for optimization using response surface methodology integrated with desirability function. The highest HY, 10.95 mmol/g COD, and R, 6.67 mmol/L h, were obtained for batch test of ultrasonically pretreated DWW under optimal conditions for COD, UD and UT at 56 g/L, 0.12 W/mL, and 17 min, respectively. The significant relative enhancement of HY, 101%, and R, 103%, implies that ultrasonically treated DWW is about 1.2–1.4 times more effective for enhanced biohydrogen production from complex DWW compared to unsonicated DWW.     

Extreme-thermophilic biohydrogen production by an anaerobic heat treated digested sewage sludge culture

Biohydrogen production process from glucose using extreme-thermophilic H₂-producing bacteria enriched from digested sewage sludge was investigated for five cycles of repeated batch experiment at 70 °C. Heat shock pretreatment was used for preparation of hydrogen-producing bacteria comparing to an untreated anaerobic digested sludge for their hydrogen production performance and responsible microbial community structures. The results showed that the heat shock pretreatment completely repressed methanogenic activity and gave the maximum hydrogen production yield of 355-488 ml H₂/g COD in the second cycle of repeated batch cultivation with more stable gas production during the cultivation when compared with control. Hydrogen production was accompanied by production of acetic acid. The average specific hydrogen in five cycles experiment ranged from 150 to 200 ml H₂/g VSS. PCR-DGGE profiling showed that the extreme-thermophilic culture predominant species were closely affiliated to Thermoanaerobacter pseudethanolicus.     

Biohydrogen production from paper industry wastes by SSF: A study of the influence of temperature/enzyme loading

The present work focused in assessing the hydrogen production from pretreated wastes of the paper industry (PIW) by simultaneous saccharification and fermentation (SSF) using anaerobic biofilms developed in natural fibers (ixtle) at different conditions. Anaerobic sludge from brewery wastewater treatment was used for biofilms and they were developed in plastic spheres covered with fiber cord from ixtle. The solid wastes of paper industry were previously pretreated with H₂SO₄ at 2.5% (v/v) at 120 °C by 30 min. The solids pretreated were hydrolyzed and fermented in batch reactors. All reactors were kept at an initial pH of 5.0 and three levels of enzyme loadings (10, 40 and 70 FPU/mL) and temperatures (35, 45, 55 °C) were assessed. The maximum hydrogen obtained (60.75 mmol/h*g volatile solids) was at 45 °C and 70 FPU/mL, moreover, no methane was detected in all cases.     

Biohydrogen production from dual digestion pretreatment of poultry slaughterhouse sludge by anaerobic self-fermentation

Poultry slaughterhouse sludge from chicken processing wastewater treatment plant was tested for their suitability as a substrate and inoculum source for fermentation hydrogen production. Dual digestion of poultry slaughterhouse sludge was employed to produce hydrogen by batch anaerobic self-fermentation without any extra-seeds. The sludge (5% TS) was dual digested by aerobic thermophilic digestion at 55 °C with the varying retention time before using as substrate in anaerobic self-fermentation. The best digestion time for enriching hydrogen-producing seeds was 48 h as it completely repressed methanogenic activity and gave the maximum hydrogen yield of 136.9 mL H₂/g TS with a hydrogen production rate of 2.56 mL H₂/L/h. The hydrogen production of treated sludge at 48 h (136.9 mL H₂/g TS) was 15 times higher than that of the raw sludge (8.83 mL H₂/g TS). With this fermentation process, tCOD value in the activated sludge could be reduced up to 30%.     

Ultrasonic pretreatment of palm oil mill effluent: Impact on biohydrogen production,bioelectricity generation,and underlying microbial communities

Substrate bioavailabity is one of the critical factors that determine the relative biohydrogen (bioH₂) yield in fermentative hydrogen production and bioelectricity output in a microbial fuel cell (MFC). In the present undertaking, batch bioH₂ production and MFC-based biolectricity generation from ultrasonically pretreated palm oil mill effluent (POME) were investigated using heat-pretreated anaerobic sludge as seed inoculum. Maximum bioH₂ production (0.7 mmol H₂/g COD) and COD removal (65%) was achieved at pH 7, for POME which was ultrasonically pretreated at a dose of 195 J/mL. Maximum value for bioH₂ productivity and COD removal at this sonication dose was higher by 38% and 20%, respectively, than unsonicated treatments. In batch MFC experiments, the same ultrasound dose led to reduced lag-time in bioelectricity generation with concomitant 25% increase in bioelectricity output (18.3 W/m³) and an increase of COD removal from 30% to 54%, as compared to controls. Quantitative polymerase chain reaction (qPCR) tests on sludge samples from batch bioH₂ production reflected an abundance of gene fragments coding for both clostridial and thermoanaerobacterial [FeFe]-hydrogenase. Fluorescence in situ hybridization (FISH) tests on sludge from MFC experiments showed Clostridium spp. and Thermoanaerobacterium spp. as the dominant microflora. Results suggest the potential of ultrasonicated POME as sustainable feedstock for dark fermentation-based bioH₂ production and MFC-based bioelectricity generation.