Effect of hydraulic retention time on the hydrogen production in a horizontal and vertical continuous stirred-tank reactor

Hydraulic retention time (HRT) is the main process parameter for biohydrogen production by anaerobic fermentation. This paper investigated the effect of the different HRT on the hydrogen production of the ethanol-type fermentation process in two kinds of CSTR reactors (horizontal continuous stirred-tank reactor and vertical continuous stirred-tank reactor) with molasses as a substrate. Two kinds of CSTR reactors operated with the organic loading rates (OLR) of 12kgCOD/m3•d under the initial HRT of the 8 h condition, and then OLR was adjusted as 6kgCOD/m3•d when the pH drops rapidly. The VCSTR and HCSTR have reached the stable ethanol-type fermentation process within 21 days and 24 days respectively. Among the five HRTs settled in the range of 2–8 h, the maximum hydrogen production rate of 3.7LH₂/Ld and 5.1LH₂/Ld were investigated respectively in the VCSTR and HCSTR. At that time the COD concentration and HRT were 8000 mg/L and 5 h for VCSTR, while 10000 mg/L and 4 h for HCSTR respectively.Through the analysis on the composition of the liquid fermentation product and biomass under the different HRT condition in the two kinds of CSTR, it can found that the ethanol-type fermentation process in the HCSTR is more stable than VCSTR due to enhancing biomass retention of HCSTR at the short HTR.     

Hydrogen production from xylose by moderate thermophilic mixed cultures using granules and biofilm up-flow anaerobic reactors

Continuous H₂ production from xylose by granules and biofilm up-flow anaerobic reactor using moderate thermophilic mixed cultures was investigated. The maximum H₂ yield of 251 mL H₂/g-xylose with H₂production rate of 15.1 L H₂/L⋅d was obtained from granules reactor operating at the organic loading rate (OLR) of 60 g-xylose/L⋅d and hydraulic retention time (HRT) of 4 h. Meanwhile the highest H₂ production rate of 13.3 L H₂/L⋅d with an H₂ yield of 221 mL H₂/g–xylose was achieved from the biofilm reactor. Both reactors were dominated by Thermoanaerobacterium species with acetate and butyrate as main fermentation products. The microbial community of the biofilm reactor was composed of Thermoanaerobacterium species, while granules reactor was composed of Clostridium sp., Thermoanaerobacterium sp. and Caloramator sp. The granular reactor was more microbial diversity and more balance between economic efficiency in term of the hydrogen production rate and technical efficiency in term of hydrogen yield.     

Adaptation of biohydrogen producing reactor to higher substrate load: Redox controlled process integration strategy to overcome limitations

Adaptation of acidogenic sequencing batch biofilm reactor (AcSBBR) to higher loading conditions of vegetable waste extract was studied during biohydrogen production at pH 6.0 under ambient conditions. H₂ production rate (HPR) and cumulative H₂ production (CHP) were found to improve with increase in organic load from 4.50 to 26.44 kg COD/m³ and later at 35.25 kg COD/m³ stabilization was observed. Acid metabolic intermediates production tends to lower the system pH which limits the substrate degradation and H₂ production at higher loading conditions. To overcome these limitations, redox controlled strategy (pH 7.0) was applied by integrating another AcSBBR. Upon redox controlled integration, CHP and substrate degradation were found to improve by 42.81% and 36.82% respectively. This approach helped to maintain the favorable redox microenvironment for fermentation at higher VFA concentrations. This process integration methodology will help to overcome some persistent limitation observed during biohydrogen production and make the process sustainable especially with high strength waste/wastewaters.     

Immobilized biofilm used as seeding source in batch biohydrogen fermentation

Anaerobic hydrogen fermentation converting carbohydrate-rich biomass to clean hydrogen energy is a renewable technology for hydrogen production. Applying immobilization technology on hydrogen fermentation could increase the cell concentration, hydraulic loading and organic loading. This study developed a novel technology to immobilize hydrogen producing bacteria in a thin film and attached on a carrier by polymer. The thin attached biofilm provides a stable hydrogen producing bacteria seeding source and makes the good hydrogen production performance. After 115 days, 9 generation transfer culture, the hydrogen yield could maintain a yield over 1.8 mol-H₂/mol-hexose. Moreover, immobilized biofilm decreased the lag phase for proximately 65–70%. The bacteria wrapped in the immobilized biofilm could grow through the biofilm and then cover the surface of biofilm. The immobilized biofilm provides seed bacteria to bulk solution continuously. Therefore, immobilized biofilm could be a promising immobilization technology and a good seeding source for biohydrogen fermentation.     

Effect of percolation frequency on biohydrogen production from fruit and vegetable wastes by dry fermentation

Organic solid wastes are the most abundant sources for biohydrogen production. Dry fermentation system has many advantages over continuously fed reactor systems for treatment of organic solid wastes. In this study the effect of percolation frequency on yield of biohydrogen production from fruit and vegetable wastes using dry fermentation system was examined. For this purpose 2 times percolation per day, 1 time percolation per day and 1 time percolation per 2 days frequency were compared and the hydrogen yields were observed as; 57 mL H₂/gVS_removed, 53 mL H₂/gVS_removed and 68 mL H₂/gVS_removed respectively. The percolation frequency didn't affect the overall yield but significantly affected the biohydrogen producing reactor of the dry fermentation system. 80% of the hydrogen was produced in percolation tank during 1 time per 2 days feeding and almost all hydrogen production was conducted in dry fermenter during 2 times per day percolation. Therefore the percolation frequency is found to be very important for system operation characteristics.     

Effect of hydraulic retention time on a continuous biohydrogen production in a packed bed biofilm reactor with recirculation flow of the liquid phase

The present paper reports on results obtained from experiments carried out in a laboratory-scale anaerobic packed bed biofilm reactor (APBR), with recirculation of the liquid phase, for continuously biohydrogen production via dark fermentation. The reactor was filled with Kaldnes^® biofilm carrier and inoculated with an anaerobic mesophilic sludge from a urban wastewater treatment plant (WWTP). The APBR was operated at a temperature of 37 °C, without pH buffering. The effect of theoretical hydraulic retention time (HRT) from 1 to 5 h on hydrogen yield (HY), hydrogen production rate (HPR), substrate conversion and metabolic pathways was investigated. This study indicates the possibility of enhancing hydrogen production by using APBR with recirculation flow. Among respondents values of HRT the highest average values of HY (2.35 mol H₂/mol substrate) and HPR (0.085 L h^?1L^?1) have been obtained at HRT equal to 2 h.     

Dark fermentative biohydrogen production from confectionery wastewater in continuous-flow reactors

The production of biohydrogen from industrial wastewater through the dark fermentation (DF) process has attracted increased interest in recent years. To implement a DF process on a large scale, a thorough knowledge of laboratory scale process control is required. The operating parameters and design features of the reactors have a great influence on the efficiency of the process. In this work, the possibility of continuous production of biohydrogen from confectionery wastewater was evaluated. The DF process was carried out at 37 ± 1 °C in two different reactors: an upflow anaerobic filter (AF) and a fluidized bed reactor (AFB). Polyurethane foam (PU) was used to immobilize the biomass. The DF process was studied at four hydraulic retention times (HRT) (1.5, 2.5, 7.5 and 15 days) and the corresponding organic loading rates (OLR) (9.21, 6.12, 2.04 and 1.02 g COD_init/(L day)). The highest hydrogen yield (HY) (44.73 ml/g COD_init) and hydrogen production rate (HPR) (92.5 ml/(L day)) was observed in AFB at HRT of 7.5 days and 2.5 days, respectively. The highest concentration of hydrogen in biogas was 34% in AF and 36% in AFB at HRT of 7.5 days. In contrast to AF, the COD removal efficiency in AFB increased with increasing HRT. The pH of the effluent was low (3.95–4.38). However, due to the use of PU for biomass immobilization, it is possible that there were local zones in the reactor that were optimal for the functioning of not only acidogens, but also methanogens. This was evidenced by a rather high content of methane in biogas (2.5% in AF and 9.6% in AFB at HRT of 15 days). These results provide valuable data for optimizing the continuous DF of wastewater from confectionery and other food industries to produce biohydrogen or biohythane.     

Methane free biohydrogen production from carbon monoxide using a continuously operated moving bed biofilm reactor

Biological water-gas shift (WGS) reaction is a green and sustainable alternative to thermochemical-catalytic WGS process for hydrogen production from carbon monoxide (CO). However, CO tolerant carboxydotrophic microbes for hydrogen production and scaling up the technology using a bioreactor system present challenges in successful application of this technology. This study demonstrated the capability of anaerobic microbial consortium for biohydrogen production from CO using a moving bed biofilm reactor (MBBR). The CO conversion pathway followed by the anaerobic biomass was first elucidated by inhibiting the methanogens present using 2-bromoethanesulfonate (BES) at an optimum concentration of 10 mmol/L. An increase in inlet CO concentration to the MBBR enhanced the H₂ production, but the CO conversion efficiency was low. More than 80% CO conversion efficiency was obtained only for a low inlet CO concentration. A maximum H₂ concentration of 19.5 mmol/L along with 2 mmol/L of acetate were obtained for 36 mmol/L of inlet CO concentration in the bioreactor. The carbon flux analysis showed that the CO was mainly utilized for methane free H₂ production, and only <10% of carbon flux was diverted towards acetate formation. Overall, this study demonstrated that MBBR system can be used for steady state biohydrogen production over a prolonged operation period.     

Approbation of an innovative method of pretreatment of dark fermentation feedstocks

According to the International Energy Agency, only a small part of the full potential of biomass energy is currently used in the world. The annual amount of agricultural waste in the Russian Federation is estimated at about 152 million tons, and the energy potential of animal waste is 201 PJ/year. Anaerobic digestion is an efficient method of converting organic waste into renewable energy sources. Previously, the positive effect of pretreatment of various organic feedstocks in vortex layer apparatus (VLA) on the characteristics of anaerobic digestion and energy efficiency was shown. Currently, there is a significant interest in the world in obtaining biohydrogen from organic waste using the dark fermentation (DF) process. During pretreatment in the VLA, the iron working bodies are abraded and iron particles are introduced into the feedstock of the DF reactor. This may have a positive effect on the production rate and yield of hydrogen, which has not been previously studied. This work is aimed at evaluating the possibility of using the VLA as a method for pretreatment of a dark fermentation feedstock for the intensification of biohydrogen production. To achieve this goal, an experimental setup was constructed. It consisted of a 45 L DF reactor, a VLA and a process control system to collect data on the DF process parameters every 5 min. At a hydraulic retention time in the DF reactor of 24 h and in the VLA of 30 s, the hydrogen content in the biogas increased from 51.1% to 52.2%. At the same time, the pH increased from 3.85 to 4.8–4.9, and the hydrogen production rate increased by 16% to 1.941 L/(L day). The hydrogen yield was 80.9 ml/g VS. Thus, pretreatment of the feedstock in VLA can be an effective way to intensify the DF process; however, further study of the VLA operating modes is required in order to optimize the concentrations of iron particles introduced into the feedstock for the most efficient continuous production of dark fermentative biohydrogen.     

Continuous biohydrogen production in immobilized biofilm system versus suspended cell culture

In this study, the performance of a new cell immobilization material, namely ceramic ball, was examined for continuous biohydrogen production in comparison to suspended cell culture system (CSTR). Production of biohydrogen in both systems was assessed under thermophilic conditions. Both systems were operated at varying hydraulic retention times (HRT) by shortening HRT values from 24 to 1.5 h at an influent sucrose concentration of 10 g/l. The immobilized bioreactor configuration outcompeted the CSTR bioreactor in terms of both volumetric hydrogen production (2.7 l H₂/l/day for immobilized system @ HRT = 3 h and 0.5 l H₂/l/day for CSTR @ HRT = 24 h) and resistance to cell-washout (CSTR reactor lost significant amount of biomass at short HRT values). It was concluded that immobilized bioreactor configuration is much more robust than CSTR against high organic loading rates and 5 fold more volumetric hydrogen production was achieved in 8 fold smaller immobilized bioreactor.     

Evaluation of process performance on biohydrogen production in continuous fixed bed reactor (C-FBR) using acid algae hydrolysate (AAH) as feedstock

In this study, a novel inoculation method to mitigate the inhibition of 5-hydroxymethylfurfural (5-HMF) is proposed. Acid algae hydrolysate containing 1.5 g 5-HMF/L and 15 g hexose/L hexose was fed to a continuous fixed bed reactor (C-FBR) partially packed with hybrid-immobilized beads. The inoculation method enabled a high rate of H₂ production, due to the reduction of 5-HMF inhibition and enhanced biofilm formation. Maximum hydrogen production was achieved at a hydraulic retention time of 6 h with a hydrogen production rate (HPR) of 20.0 ± 3.3 L H₂/L-d and a hydrogen yield (HY) of 2.3 ± 0.4 mol H₂/mol hexose _added. Butyrate and acetate were the major soluble metabolic products released during fermentation. Quantitative real-time polymerase chain reaction analysis revealed that Clostridium butyricum comprised 94.3% of the total bacteria, which was attributed to the high rate of biohydrogen production.     

Biohydrogen production from hydrolyzed waste wheat by dark fermentation in a continuously operated packed bed reactor: The effect of hydraulic retention time

The aim of the study is biohydrogen production from hydrolyzed waste wheat by dark fermentation in a continuously operated up-flow packed bed reactor. For this purpose, the effect of hydraulic retention time (HRT) on the rate (R_H2) and yield (Y_H2) of hydrogen gas formation were investigated. In order to determine the most suitable hydraulic retention time yielding the highest hydrogen formation, the reactor was operated between HRT = 1 h and 8 h. The substrate was the acid hydrolyzed wheat powder (AHWP). Waste wheat was sieved down to 70 μm size (less than 200 mesh) and acid hydrolyzed at pH = 2 and 90 °C in an autoclave for 15 min. The sugar solution obtained from hydrolysis of waste wheat was used as substrate at the constant concentration of 15 g/L after neutralization and nutrient addition for biohydrogen production by dark fermentation. The microbial growth support particle was aquarium biological sponge (ABS). Heat-treated anaerobic sludge was used as inoculum. Total gas volume and hydrogen percentage in total gas, hydrogen gas volume, total sugar and total volatile fatty acid concentrations in the feed and in the effluent of the system were monitored daily throughout the experiments. The highest yield and rate of productions were obtained as Y_H2 = 645.7 mL/g TS and R_H2 = 2.51 L H₂/L d at HRT = 3 h, respectively.     

Biohydrogen production using horizontal and vertical continuous stirred tank reactor- a numerical optimization

This paper illustrates the method to predict the production of biohydrogen and biogas from the horizontal and vertical continuous mixed tank reactor using a numerical approach. Utilization of computational fluid dynamics on the estimation of bioreactor performance is very crucial owing to the uncertainty in the numerical results. Since there has been little work on CFD to determine the influence of hydraulic retention time, impeller speeds, vortex growth, and pH on biohydrogen yield rate the effort had been made. A series of simulations are done with optimal boundary conditions to ensure maximum hydrogen and biogas production rate. Two types of reactors HCSTR and VCSTR are operated at different impeller speed from 40 rpm to 120 rpm. Further, the rate of HRT and organic loading are varied. As the rpm of the impeller increase the rate of hydrogen and biomass production increases not later than 80 rpm. Meanwhile, the optimal range of HRT and pH are 4–8 h and 6.0–8.0. Running the impeller at optimized rate with derived conditions leads to high hydrogen and biogas production of 6 LH₂/Ld and 30 L/d. The obtained results are validated with the experimental findings to compare the uncertainty formed due to the numerical simulations. The optimum boundary condition obtained from the study is expected to provide the essential knowledge in establishing the full-scaled reactors.     

Discontinuous biomass recycling as a successful strategy to enhance continuous hydrogen production at high organic loading rates

A novel strategy to discontinuously increase the biomass concentration in a continuous stirred-tank reactor was evaluated to enhance the performance of dark fermentation. Different concentrations of biomass were evaluated at organic loading rates (OLR) ranging from 90 to 160 g lactose/L-d with a hydraulic retention time (HRT) of 6 h. The study revealed that the discontinuous increase of biomass enhanced the hydrogen (H₂) production rates and carboxylic acids concentrations by 19–25% and 8–23%, respectively. In particular, a maximum H₂ production rate of 30.8 L H₂/L-d with carboxylic acids concentration of 20 g/L was reached at an OLR of 138 g lactose/L-d with a biomass concentration of 15 g volatile suspended solids/L. The analysis of microbial communities showed the co-dominance of Clostridium and lactic acid bacteria. Overall, the discontinuous increase of biomass was an effective strategy to improve the performance of suspended-biomass reactors operated at high OLR and low HRT.     

Evaluation of biohydrogen production from glucose and protein at neutral initial pH

Organic wastes are considered as potential substances for economical biohydrogen production, because the carbohydrate and protein are main components. Previous investigations indicate that an optimum hydrogen production appear in acidic conditions to carbohydrates, or in alkali condition to protein. However, in practice, the treatment of these organic wastes by anaerobic fermentation usually carries out at neutral pH condition, in which biohydrogen production is only a middle process. So, the purpose of this paper is to evaluate the biohydrogen production at neutral pH condition from carbohydrates or protein. Batch tests were conducted to investigate the differences in biohydrogen production by anaerobic fermentation at neutral initial pH using carbohydrate and protein (glucose and peptone) as the sole carbon source. The experimental results showed that the maximal hydrogen yields of two substrates were about 0.14 ml H₂/mg glucose and 0.077 ml H₂/mg protein, respectively, at neutral initial pH. Although the hydrogen yields of glucose is far greater than that of protein at neutral pH, they were lower than previous results of hydrogen production in acidic condition to carbohydrate or in alkali condition to protein. This result shows that the neutral pH is not an optimal condition for biohydrogen production. In this experiment of biohydrogen production, a phenomenon has been observed that the hydrogen production and hydrogen consumption occurred simultaneously in the fermentation of protein, whereas the hydrogen production occurred only in the fermentation of glucose. Furthermore, the different evaluation of the main components of the organic liquid by-products produced by fermentation of each substrate implied that the biohydrogen production pathways of these two substrates were different. Molecular analysis indicated that the dominant microorganisms during the anaerobic fermentation of these two substrates are greatly different.     

Start-up study of biohydrogen production from palm oil mill effluent in a lab-scale up-flow anaerobic sludge blanket fixed-film reactor

A start-up study of lab-scale up-flow anaerobic sludge blanket fixed-film reactor (UASFF) was conducted to produce biohydrogen from palm oil mill effluent (POME). The reactor was fed with POME at different hydraulic retention time (HRT) and organic loading rate (OLR) to obtain the optimum fermentation time for maximum hydrogen yield (HY). The results showed the HY, volumetric hydrogen production rate (VHPR), and COD removal of 0.5–1.1 L H₂/g COD_consumed, 1.98–4.1 L H₂ L^?1 day^?1, and 33.4–38.5%, respectively. The characteristic study on POME particles was analyzed by particle size distribution (PSD), Scanning electron microscopy (SEM), and Energy-dispersive X-ray spectroscopy (EDX). The microbial Shannon and Simpson diversity indices and Principal Component Analysis assessed the alpha and beta diversity, respectively. The results indicated the change of bacterial community diversity over the operation, in which Clostridium sensu stricto 1 and Lactobacillus species were contributed to hydrogen fermentation.     

Biohydrogen from molasses with ethanol-type fermentation: Effect of hydraulic retention time

Though ethanol-type fermentation has many advantages for improving hydrogen production rate (HPR) in continuously mode hydrogen producing system, information on this fermentation is very deficient. The effect of hydraulic retention time (HRT) on biohydrogen production and operational stability of ethanol-type fermentation was investigated in a continuous stirred tank reactor (CSTR) using molasses as substrate. Five HRTs were examined, ranging from 4 to 10 h. At HRT 5 h, the highest HPR of 12.27 mmol L⁻¹ h⁻¹ was obtained from ethanol-type fermentation in the pH range of 4.3–4.4. During the whole operation process, ethanol, butyrate and acetate were the predominant metabolites. A total COD concentration of ethanol and acetate accounted for above 73.3% of total soluble microbial products. Linear regression showed that HPR and ethanol production rate were proportionately correlated at all HRTs which could be expressed as y = 0.9821x − 3.5151 (r² = 0.9498). It is meaningful that the proposed recovery of both hydrogen and ethanol from fermentation process can improve energy production rate and economic profit. Results demonstrated that the best energy production rate was 15.50 kJ L⁻¹ h⁻¹, occurred at HRT = 5 h.     

Steady-state and dynamic modeling of biohydrogen production in an integrated biohydrogen reactor clarifier system

Steady-state operational data from the integrated biohydrogen reactor clarifier system (IBRCS) during anaerobic treatment of glucose-based synthetic wastewater at HRT of 8 h and SRT ranging from 26 to 50 h and organic loading rates of 6.5–206 gCOD/L-d were used to calibrate and verify a process model of the system developed using BioWin. The model accurately predicted biomass concentrations in both the bioreactor and the clarifier supernatant with average percentage errors (APEs) of 4.6% and 10%, respectively. Hydrogen production rates and hydrogen yields predicted by the model were in close agreement with the observed experimental results as reflected by an APE of less than 4%, while the hydrogen content was well correlated with an APE of 10%. The successful modeling culminated in the accurate prediction of soluble metabolites, i.e. volatile fatty acids in the reactor with an APE of 14%. The calibrated model confirmed the advantages of decoupling of the solids retention time (SRT) from the hydraulic retention time (HRT) in biohydrogen production, with the average hydrogen yield decreasing from 3.0 mol H₂/mol glucose to 0.8 mol H₂/mol glucose upon elimination of the clarifier. Dynamic modeling showed that the system responds favorably to short-term hydraulic and organic surges, recovering back to the original condition. Furthermore, the dynamic simulation revealed that with a prolonged startup periods of 10 and 30 days, the IBRCS can be operated at an HRT of 4 h and OLR as high as 206 gCOD/L-d without inhibition and/or marked performance deterioration.     

Strategies to improve the biohydrogen production from cassava wastewater in fixed-bed reactors

The biological production of hydrogen from cassava starch wastewater (CSW) was evaluated in an anaerobic fixed-bed reactor (UAFBR). The assays were carried out to evaluate the effects of organic loading rate (OLR) increase and strategies of inoculation (AS – anaerobic sludge thermally treated and NF – naturally fermented cassava starch wastewater) on UAFBR performance. The OLR increase (10–20 g L⁻¹ d⁻¹) associated with hydraulic retention time (HRT) decrease (4–2 h) improved the volumetric hydrogen production rate (VHPR, from 229 to 550 mLH₂.L⁻¹.d⁻¹), molar hydrogen flow rate (MHFR, from 1.0 to 2.5 mmolH₂.h⁻¹) and hydrogen yield (HY, from 0.2 to 0.3 molH₂.mol⁻¹Carb) from CSW due to increase in substrate availability. Both inoculation alternatives (AS and NF) were effective for the selection of acidogenic microorganisms, which demonstrates that NF could be considered a simple and economic alternative for the acquisition of inoculum for continuous acidogenic reactors. Hydrogen production decreased after 10 days of operation when the specific organic loading rate (SOLR) reached reduced values (<1 gCarb.g⁻¹VSS.d⁻¹), which impairs hydrogen production. For all assays, methane was present in the biogas after the 20th day of operation mainly due to biomass accumulation, which alters the biota of the reactor. Although many factors could influence the process performance in UAFBR for the production of biohydrogen, the accumulation of biomass have been pointed as the main factor in the determination of the production time, thus demanding the implementation of systematic practices to remove the excess of biomass to maintain the SOLR in levels adequate for hydrogen production.     

Bioproduction of hydrogen from food waste by pilot-scale combined hydrogen/methane fermentation

A pilot-scale two-phase hydrogen/methane fermentation system generated 3.9 L biogas per unit time and reactor volume from food waste, of which the fraction of H₂ was approximately 60% at a hydraulic retention time (HRT) of 21 h. As substrate, 90% of the carbohydrates in the organic compounds were consumed, based on COD removal efficiency, and the hydrogen yield was approximately 1.82 (H₂-mol/glucose-mol). The maximum decomposition rate coefficient of hydrogen fermentation was observed at an HRT of 21 h, indicating that reducing HRTs improves hydrogen production. Over 80% of the methane was produced in the methane fermentation tank and the predominant fraction of organic acids after methane fermentation comprised acetic acid. Based on our economic evaluation, two-phase hydrogen/methane fermentation has greater potential for recovering energy than methane-only fermentation.