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.     

Biohydrogen production from autoclaved fruit and vegetable wastes by dry fermentation under thermophilic condition

Dark fermentative hydrogen production from organic waste is an attractive technique that simultaneously treats waste along with generation of renewable fuel. In this study, a relative new technology named dark dry fermentation was tested in a 55-L reactor to treat fruit and vegetable waste (FVW) along with simultaneous generation of biohydrogen. To understand the effect of autoclaving as a pretreatment method on FVW for subsequent biohydrogen production, two independent experiments were performed; one with autoclaved waste (experiment I) and another by using non autoclaved waste (experiment II). From the analyses, it was found that maximum hydrogen % obtained for experiment I was 41% (v/v%) whereas, for experiment II was 21%. In terms of total hydrogen produced, around 30% higher production was observed with experiment I compared to experiment II. The hydrogen yields for experiment I and experiment II were respectively, 27.19 and 20.81 NmL H₂/gVS (VS = volatile solid added), and the metabolites (VFAs) preferentially produced were acetic acid and iso-butyric acid.     

Determining the effect of trace elements on biohydrogen production from fruit and vegetable wastes

Trace elements are one of the important parameters for dark fermentative H₂ production because they work as co-factors in H₂ formation biochemistry. Lack or excess of trace element and its concentrations could be an important reason for the low yield of H₂ production. In this study, the effects of 11 different trace elements (Fe, Ni, Zn, Co, Cu, Mn, Al, B, Se, Mo and W) were tested at two levels in terms of biohydrogen production from Fruit and Vegetable Wastes (FVW) with Biochemical Hydrogen Potential (BHP) Tests using Plackett-Burman statistical design. 1.1–2.8 times enhancement of biohydrogen production was determined with its addition. The most effective trace elements were found as Zn and Ni. In order to reveal the resident microbial flora, Denaturing Gradient Gel Electrophoresis (DGGE) analysis was carried out on all BHP effluent samples. Results of DGGE analysis, four microbial sequences evaluated as Clostridium sp., Clostridium baratii, Uncultured bacterium, Uncultured Streptococcus sp., and their similarity rates were 99%, 100%, 89%, 98%, respectively.     

Ferric oxide/date seed activated carbon nanocomposites mediated dark fermentation of date fruit wastes for enriched biohydrogen production

Biohydrogen production from biomass waste, not only addresses the energy demand in a renewable manner but also resolves the safe disposal issues associated with these biowastes. Also, scalable and low-cost techniques to enhance biohydrogen production have gained more attraction and are highly explored. In this research work, date-palm fruit wastes have been studied for their biohydrogen production potential using Enterobacter aerogenes by dark fermentation. Hydrogen yield and productivity were improved through the addition of iron oxide nanoparticles (Fe₃O₄ NPs) and its date seed activated carbon nanocomposites (Fe₃O₄/DSAC) to the fermentation media. Studies on discrete inclusions of these NPs showed that the appropriate dosage of NPs promoted, while higher dosages repressed the hydrogen production performance. Optimal dosage and fermentation time was observed as 150 mg/L and 24 h for both the additives. Fe₃O₄/DSAC nanocomposites showed better hydrogen production enhancement than Fe₃O₄ NPs. Maximum hydrogen yield of 238.7 mL/g was obtained for the 150 mg/L nanocomposites, which was 65.7% higher than that of the standalone Fe₃O₄ NPs and three folds higher than the yield of the control run without any NPs inclusion (78.4 mL/g). Metabolites analysis showed that the hydrogen evolution followed the ethanol-acetate pathway. Formation levels of longer chain propionate and butyrate co-metabolites were significantly low in the presence of Fe₃O₄/DSAC than Fe₃O₄. The carbon support in the nanocomposites acted as an adsorbent-buffer, which favored the medium pH in-addition to the stimulatory effects of Fe₃O₄ NPs. Cell growth and specific hydrogenase activity analysis were also performed to supplement the hydrogen production results. Gompertz and modified Logistic kinetic models were employed for kinetic modeling of experimental hydrogen production values. The Fe₃O₄/DSAC nanocomposites exhibited significant application potential for the production of biohydrogen from date fruit wastes.     

Multiscale mixing analysis and modeling of biohydrogen production by dark fermentation

Hydrogen production by dark fermentation (DF) from wastewater, food waste, and agro-industrial waste combines the advantages to be renewable, sustainable and environmentally friendly. But this attractive process involves a three-phase gas-liquid-solid system highly sensitive to mixing conditions. However, mixing is usually disregarded in the conventional strategies for enhancing biohydrogen productivity, even though H₂ production can be doubled, e.g. versus of reactor design (0.6–1.5 mol H₂/mol hexose). The objective of this review paper is, therefore, to highlight the key effects of mixing on biohydrogen production among the abiotic parameters of DF. First, the pros and cons of the different modes of mixing in anaerobic digesters are described. Then, the influence of mixing on DF is discussed using recent data from the literature and theoretical analysis, focusing on the multiphase and multiscale aspects of DF. The methods and tools available to quantify experimentally the role of mixing both at the local and global scales are summarized. The 0-D to 3-D strategies able to implement mixing in fermentation modeling and scale-up procedures are examined. Finally, the perspectives in terms of process intensification and scale-up tools using mixing optimization are discussed with the issues that are still to be solved.     

Enhanced biohydrogen production from corn stover hydrolyzate by pretreatment of two typical seed sludges

Five individual pretreatment methods (heat, ultrasonic, ultraviolet, acid, and base) were performed on two typical seed sludges (river sediments and anaerobic granular sludge) to evaluate their effectiveness on enriching efficient hydrogen (H₂)-producing bacteria and enhancing H₂ production using corn stover hydrolyzate. Results indicated that pretreatment processes caused more remarkable improvements for river sediments than anaerobic granular sludge. Among the five protocols, heat pretreatment reached high H₂ yield for both river sediments (4.17 mmol H₂/g utilized sugar) and anaerobic granular sludge (2.84 mmol H₂/g utilized sugar). Ultraviolet and ultrasonic pretreatments were conditionally effective for river sediments and anaerobic granular sludge, respectively. In most cases, pretreatment processes altered soluble metabolites distribution towards more acetate and less ethanol production. Microbial community analysis indicated that heat and ultrasonic pretreatments can respectively lead to significant and indistinctive change on original microbial community. Besides frequently detected Escherichia spp., Serratia spp., and Klebsiella spp., some species of Clostridium spp. and Bacillus spp. might be efficient H₂ producer responsible for better H₂-producing performances.     

Kinetic and performance study of a batch two-phase anaerobic digestion of fruit and vegetable wastes

Anaerobic digestion of a simulated organic fraction of the waste of a central market selling fruit and vegetables was carried out in two-phase digesters in the mesophilic range of temperatures. Batch digestion was prolonged until no biogas was produced (33 days). With digested pig manure as inoculum, maximum biogas production was obtained around day 10, and within 3 weeks the digestion was almost complete. A kinetic analysis was carried out using first-order, Monod and Chen-Hashimoto models. The Chen-Hashimoto model represents the best fit, whereas a first-order model was not consistent with the experimental results.     

High efficient biohydrogen production from palm oil mill effluent by two-stage dark fermentation and microbial electrolysis under thermophilic condition

Biohydrogen production from palm oil mill effluent by two-stage dark fermentation and microbial electrolysis was investigated under thermophilic condition. The optimum chemical oxygen demand (COD) concentration and pH for dark fermentation were 66 g·L⁻¹ and 6.5 with a hydrogen yield of 73 mL-H₂·gCOD⁻¹. The dark fermentation effluent consisted of mainly acetate and butyrate. The optimum voltage for microbial electrolysis was 0.7 V with a hydrogen yield of 163 mL-H₂·gCOD⁻¹. The hydrogen yield of continuous two-stage dark fermentation and microbial electrolysis was 236 mL-H₂·gCOD⁻¹ with a hydrogen production rate of 7.81 L·L⁻¹·d⁻¹. The hydrogen yield was 3 times increased when compared with dark fermentation alone. Thermoanaerobacterium sp. was dominated in the dark fermentation stage while Geobacter sp. and Desulfovibrio sp. dominated in the microbial electrolysis cell stage. Two-stage dark fermentation and microbial electrolysis under thermophilic condition is a highly promising option to maximize the conversion of palm oil mill effluent into biohydrogen.     

Effects of rice husk particle size on biohydrogen production under solid state fermentation

As a renewable energy source bio-hydrogen production from lignocellulosic wastes is a promising approach which can produce clean fuel with no CO₂ emissions. Utilization of agro-industrial residues in solid state fermentation (SSF) is offering a solution to solid wastes disposal and providing an economical process of value-added products such as hydrogen.In this study three different particle size of rice husk (<2000 μm, <300 μm, <74 μm) was subjected to batch SSF with a Clostridium termitidis: Clostridium intestinale ratio of 5:1. C. termitidis is a cellulolytic microorganism that has the ability to hydrolyze cellulosic substances and C. intestinale is able to grow on glucose having a potential of enhancing hydrogen production when used in the co-culture. 5 g dw rice husk with 75% humidity was used as substrate in SSF under mesophilic conditions. The highest HF Volume (29.26 mL) and the highest yield (5.9 mL H₂ g⁻¹ substrate) were obtained with the smallest particle size (<74 μm). The main metabolites obtained from the fermentation media were acetic, butyric, propionic and lactic acids. The second best production yield (3.99 mL H₂ g⁻¹ substrate) was obtained with the middle particle size (<300 μm) rice husk with a HF of 19.71 mL.     

Maximizing biohydrogen production from water hyacinth by coupling dark fermentation and electrohydrogenesis

Biohydrogen production via dark fermentation has shown immense potential for simultaneous energy generation and waste remediation. However, the low substrate conversion rates limit its practical feasibility. Therefore, the present work attempts to develop a single chamber microbial electrolysis cell (MEC) as an additional means for biohydrogen production. Different organic substrates including simple sugars and volatile fatty acids were demonstrated as potential substrates for H₂ production in MEC. The use of water hyacinth as sole substrate for H₂ production was examined. Furthermore, the feasibility of using MEC for second stage energy recovery after dark fermentation was explored. The two-stage process exhibited improved performance as compared to single stage MEC process with overall hydrogen yield of 67.69 L H₂/kg COD_consumed, COD removal of 70.33% and energy recovery of 46%. These results suggest that coupled dark fermentation-MEC process can be a promising means for obtaining high yield biohydrogen from water hyacinth.     

Development of a method for biohydrogen production from wheat straw by dark fermentation

Lignocellulosic biomass contains approximately 70-80% carbohydrates. If properly hydrolyzed, these carbohydrates can serve as an ideal feedstock for fermentative hydrogen production. In this research, batch tests of biohydrogen production from acid-pretreated wheat straw were conducted to analyze the effects of various associated bioprocesses. The objective of the pretreatment phase was to investigate the effects of various sulfuric acid pretreatments on the conversion of wheat straw to biohydrogen. When sulfuric acid-pretreated solids at a concentration of 2% (w/v) were placed in an oven for 90 min at 120 °C, they degraded substantially to fermentative gas. Therefore, wheat straw that is pre-treated under the evaluated conditions is suitable for hydrolysis and fermentation in a batch test apparatus. Five different conditions were evaluated in the tests, which were conducted in accordance with standard batch test procedures (DIN 38414 S8): fresh straw, pre-treated straw, supernatants derived from acid hydrolyzation, Separate Hydrolysis and Fermentation (SHF) and Simultaneous Saccharification and Fermentation (SSF). The SSF method proved to be the most effective and economical way to convert wheat straw to biohydrogen. The hydrogen yield by this method was 1 mol H₂/mol glucose, which resulted from 5% carbon degradation (η_{C, gas}) or the equivalent of 64% of the hydrogen volume that was produced in the reference test (glucose equivalent test). This method also proved to have the shortest lag phase for gas production. The supernatants derived from acid hydrolysis were very promising substances for continuous tests and presented excellent characteristics for the mass production of biohydrogen. For example, a 1.19 mol H₂/mol glucose (76% glucose equivalent) yield was achieved along with a 52% carbon degradation.     

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.     

Screening of biohydrogen production based on dark fermentation in the presence of nano-sized Fe2O3 doped metal oxide additives

Increasing the biohydrogen production efficiency through the dark fermentation process has become the biggest challenge in recent years. We aim to enhance biohydrogen production yield by adding nano-sized iron oxide doped metal oxides prepared by the wet impregnation method. Biohydrogen production in the presence of additives was evaluated by the screening of (i) type (Fe₂O₃@Al₂O₃, Fe₂O₃@ZrO₂, and Fe₂O₃@TiO₂) and (ii) concentration (0–200 mg/L) of additives. During the screening of additive type, approximately 14 wt% nano-sized (6 nm) hematite (Fe₂O₃) doped Al₂O₃ showed the highest improvement for biohydrogen production yield among all additives. Batch experiments conducted through dark fermentation demonstrated that 50 mg/L Fe₂O₃@Al₂O₃ addition caused an acceleration in biohydrogen production by 34% and an increment in yield by 15%, despite even low concentrations Al₂O₃ addition inhibited the process.     

Lactate wastewater dark fermentation: The effect of temperature and initial pH on biohydrogen production and microbial community

Biohydrogen production using dark fermentation (hydrolysis and acidogenesis) is one of the ways to recover energy from lactate wastewater from the food-processing industry, which has high organic matter. Dark fermentation can be affected by the temperature, pH and the microbial community structure. This study investigated the effects of temperature and initial pH on the biohydrogen production and the microbial community from a lactate wastewater using dark fermentation. Biohydrogen production was successful only at lower temperature levels (35 and 45 °C) and initial pH 6.5, 7.5 and 8.5. The highest hydrogen yield (0.85 mol H₂/mol lactate consumed) was achieved at 45 °C and initial pH 8.5. The COD reduction achieved by fermenting the lactate wastewater at 35 °C ranged between 21 and 30% with the maximum COD reduction at pH 8.5, and at 45 °C, the COD reduction ranged between 12 and 21%, with the maximum at pH 7.5. At 35 °C, the lactate degradation ranged between 54 and 95%, while at 45 °C, it ranged between 77 and 99.8%. 16S rRNA sequencing revealed that at 35 °C, bacteria from the Clostridium genera were the most abundant at the end of the fermentation in the reactors that produced hydrogen, while at 45 °C Sporanaerobacter, Clostridium and Pseudomonas were the most abundant.     

Optimization of particle number,substrate concentration and temperature of batch immobilized reactor system for biohydrogen production by dark fermentation

Immobilized cell bioreactor was operated in batch mode for biohydrogen generation by dark fermentation from acid hydrolyzed waste wheat powder. It was aimed to optimize the fermentation conditions with the purpose of obtaining the highest hydrogen yield (Y_H2) and production rate (HPR) by applying Box–Wilson statistical experimental design method. Particle number (PN = 120–240; X₁), initial total sugar concentration (TS₀ = 10–30 g/l; X₂) and fermentation temperature (T = 35–55 °C; X₃) were selected as independent variables. Polyester fibers with particle diameter “Dp” = 0.5 cm were used as support material to immobilize microorganisms with heat-pretreated sludge. Quadratic equations for production yield and rate were developed by using experimental results. The maximum Y_H2 (3.21 mol H₂/mol glucose) and HPR (73.3 ml H₂/h) were predicted at the optimum conditions of PN = 240, TS₀ = 10 g/l and T = 44.9 °C. Also, analysis of variance, as well as sum of ranking difference test results demonstrated that fitting models were statistically significant.     

Influence of the initial proportion of carbohydrates,proteins, and lipids on biohydrogen production by dark fermentation: A multi-response optimization approach

The present study investigated the effect of the initial proportions of carbohydrates, proteins and lipids within the substrate on the resulting biohydrogen productivity by dark fermentation. Organic matter removal and the related metabolic by-products generated during the process were also assessed. The results obtained showed that initial substrate composition in terms of carbohydrates, proteins and lipids has a significant effect on maximal potential hydrogen production (H_max), hydrogen production rate (R_max), hydrogen yield (Y_H2) and metabolites distribution. Tests with proteins and lipids as unique substrate did not produce H₂. A simplex-centroid design (SCD) and compositional data analysis of the substrate was used to determine the best condition to convert the substrate into H₂. H_max, R_max and Y_H2 were significantly increased using an initial proportion of 56% carbohydrates (15 g/L), 22% proteins (6 g/L), and 22% lipids (6 g/L), which was concomitant with the generation of acetic and butyric acids. Protein and lipid proportions higher than 29% and lower than 12% led to decreased H_max, R_max and Y_H2 values with a consequent accumulation of propionic acid.     

Effects of various pretreatment methods of anaerobic mixed microflora on biohydrogen production and the fermentation pathway of glucose

The influence of different pretreatment methods on anaerobic mixed inoculum was evaluated for selectively enriching the hydrogen (H₂) producing mixed culture using glucose as the substrate. The efficiency of H₂ yield and the glucose fermentation pathway were found to be dependent on the type of pretreatment procedure adopted on the parent inoculum. The H₂ yield could be increased by appropriate pretreatment methods including the use of heat, alkaline or acidic conditions. Heat pretreatment of the inoculum for 30 min at 80 °C increased the H₂ yield to 53.20% more than the control.When the inoculum was heat-pretreated at 80 °C and 90 °C, the glucose degraded via ethanol (HEt) and butric acid (HBu) fermentation pathways. The degradation pathways shifted to HEt and propionate (HPr) types as the heat pretreatment temperature increased to 100 °C. When the inoculum was alkali- or acid-pretreated, the fermentation pathway shifted from glucose to a combination of the HPr and HBu types. This trend became obvious as the acidity increased. As the fermentation pathway shift from the HEt type to the HPr and HBu types, the H₂ yield decreased.     

The production of biohydrogen by a novel strain Clostridium sp. YM1 in dark fermentation process

In view of increasing attempts for the production of renewable energy, the production of biohydrogen energy by a new mesophilic bacterium Clostridium sp. YM1 was performed for the first time in the dark fermentation. Experimental results showed that the fermentative hydrogen was successfully produced by Clostridium sp. YM1 with the highest cumulative hydrogen volume of 3821 ml/L with a hydrogen yield of 1.7 mol H₂/mol glucose consumed. Similar results revealed that optimum incubation temperature and pH value of culture medium were 37 °C and 6.5, respectively. The study of hydrogen production from glucose and xylose revealed that this strain was able to generate higher hydrogen from glucose compared to that from xylose. The profile of volatile fatty acids produced showed that hydrogen generation by Clostridium sp. YM1 was butyrate-type fermentation. Moreover, the findings of this study indicated that an increase in head space of fermentation culture positively enhanced hydrogen production.     

Bio-hydrogen production from cheese whey powder (CWP) solution: Comparison of thermophilic and mesophilic dark fermentations

Cheese whey powder (CWP) solution was used as the raw material for hydrogen gas production by mesophilic (35 °C) and thermophilic (55 °C) dark fermentations at constant initial total sugar and bacteria concentrations. Thermophilic fermentation yielded higher cumulative hydrogen formation (CHF = 171 mL), higher hydrogen yield (111 mL H₂ g⁻¹ total sugar), and higher hydrogen formation rate (3.46 mL H₂ L⁻¹ h⁻¹) as compared to mesophilic fermentation. CHF in both cases were correlated with the Gompertz equation and the constants were determined. Despite the longer lag phase, thermophilic fermentation yielded higher specific H₂ formation rate (2.10 mL H₂ g⁻¹cells h⁻¹). Favorable results obtained in thermophilic fermentation were probably due to elimination of H₂ consuming bacteria at high temperatures and selection of fast hydrogen gas producers.