Fermentative hydrogen production by diverse microflora

In this study, hydrogen production with activated sludge, a diverse bacterial source has been investigated and compared to microflora from anaerobic digester sludge, which is less diverse. Batch experiments were conducted at mesophilic (37 °C) and thermophilic (55 °C) temperatures. The hydrogen production yields with activated sludge at 37 °C and 55 °C were 0.56 and 1.32 mol H₂/mol glucose consumed, respectively. While with anaerobically digested sludge hydrogen yield was 2.18 mol H₂/mol glucose consumed at 37 °C and 1.25 mol H₂/mol glucose consumed at 55 °C. The results of repeated batch experiments for 615 h resulted in average yields of 1.21 ± 0.62 and 1.40 ± 0.16 mol H₂/mol glucose consumed for activated sludge and anaerobic sludge, respectively. The hydrogen production with activated sludge was not stable during the repeated batches and the fluctuation in hydrogen production was attributed to formation of lactic acid as the predominant metabolite in some batches. The presence of lactic acid bacteria in microflora was confirmed by PCR-DGGE.     

Isolation of bacteria capable of hydrogen production in dark fermentation and intensification of anaerobic granular sludge activity

In the anaerobic biological treatment of pulp and papermaking wastewater, the gradual deposition of CaCO₃ eventually leads to the inhibition of the activity of anaerobic granular sludge. In this study, a hydrogen production bacterial Raoultella DW01 was isolated from domesticated anaerobic granular sludge. The fermentation conditions were designed using central composite design, and the optimum conditions obtained by response surface analysis encompassed an initial pH 5.77, 4.13 g/L l-glutamic acid and an inoculation amount of 15%. The H₂ production yield represented a 29.5% increase over the unoptimized conditions. Finally, the effect of adding DW01 on the biogas production in anaerobic granular sludge with different sludge ages was investigated. The cumulative biogas yield and the max biogas production rate increased by 27.8% and 53.5% after adding DW01 to a sludge with an age of 335 days compared with the on-intensified sludge. This paper provides a way to alleviate the CaCO₃ deposition by intensifying the activity of H₂ and acid-producing bacteria via improving the activity of granular sludge.     

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%.     

Effect of organic loading on a novel hydrogen bioreactor

This study investigated the impact of six organic loading rates (OLR) ranging from 6.5 gCOD/L-d to 206 gCOD/L-d on the performance of a novel integrated biohydrogen reactor clarifier systems (IBRCSs) comprised a continuously stirred reactor (CSTR) for biological hydrogen production, followed by an uncovered gravity settler for decoupling of solids retention time (SRT) from hydraulic retention time (HRT). The system was able to maintain a high molar hydrogen yield of 2.8 mol H₂/mol glucose at OLR ranging from 6.5 to 103 gCOD/L-d, but dropped precipitously to approximately 1.2 and 1.1 mol H₂/mol glucose for the OLRs of 154 and 206 gCOD/L-d, respectively. The optimum OLR at HRT of 8 h for maximizing both hydrogen molar yield and volumetric hydrogen production was 103 gCOD/L-d. A positive statistical correlation was observed between the molar hydrogen production and the molar acetate-to-butyrate ratio. Biomass yield correlated negatively with hydrogen yield, although not linearly. Analyzing the food-to-microorganisms (F/M) data in this study and others revealed that, both molar hydrogen yields and biomass specific hydrogen rates peaked at 2.8 mol H₂/mol glucose and 2.3 L/gVSS-d at F/M ratios ranging from 4.4 to 6.4 gCOD/gVSS-d. Microbial community analysis for OLRs of 6.5 and 25.7 gCOD/L-d showed the predominance of hydrogen producers such as Clostridium acetobutyricum, Klebsiella pneumonia, Clostridium butyricum, Clostridium pasteurianum. While at extremely high OLRs of 154 and 206 gCOD/L-d, a microbial shift was clearly evident due to the coexistence of the non-hydrogen producers such as Lactococcus sp. and Pseudomonas sp.     

Combined effects of temperature and pH on biohydrogen production by anaerobic digested sludge

A full factorial design was conducted to investigate the combined effects of temperatures and initial pH on fermentative hydrogen production by mixed cultures in batch tests. The experimental results showed that the modified Logistic model can be used to describe the progress of cumulative hydrogen production in the batch tests of this study. The modified Ratkowsky model can be used to describe the combined effects of the temperatures and initial pH on the substrate degradation efficiency, hydrogen yield and average hydrogen production rate. The temperatures and initial pH had interactive impact on fermentative hydrogen production. The maximum substrate degradation efficiency, the maximum hydrogen yield and the maximum average hydrogen production rate was predicted at the temperature of 37.8 °C and the initial pH of 7.1, 37.4 °C and 6.9, and 38.2 °C and 7.2, respectively. In general, the optimal temperature for the fermentative hydrogen production was around 37.8 °C and the optimal initial pH for the fermentative hydrogen production was around 7.1.     

Hydrogen production by anaerobic co-digestion of rice straw and sewage sludge

In this study, the rich carbon content of rice straw and the high nitrogen content of sewage sludge make the straw a good potential substrate and the sludge a viable inoculum for biohydrogen production. Two treatment conditions for the sewage sludge (raw and heat-treated) were used in the present experiments. Batch test using a mixture of rice straw and sewage sludge were carried out to investigate the optimum carbon to nitrogen (C/N) ratio for effective biohydrogen production. The experimental results indicate that untreated sludge could be used as the inoculum for efficient hydrogen production when mixed with the appropriate proportion of rice straw. According to our results, biogas and hydrogen production in all raw sludge cases ramped up more quickly and also exhibited longer and higher hydrogen production in comparison with heat-treated cases. At the C/N ratio of 25 in untreated sludge, hydrogen production was 33% higher than heat-treated one. Additionally, under the same conditions, high and stable hydrogen content (58%) and the maximal hydrogen yield (0.74 mmol H₂/g-VS added straw) were obtained.     

热处理对污泥厌氧发酵产氢的影响

通过对污泥进行热处理来提高污泥厌氧发酵产氢的能力.结果表明:热处理是一种有效的污泥融胞方法,热处理对糖和蛋白质的水解效果好,热处理后污泥中可溶蛋白质浓度为原污泥的6.4～8.9倍,可溶糖浓度为原污泥的1.6～7.9倍.75℃热处理10 min效果最好,最大累积产氢量可达20.3 ml,较原污泥提高了19倍;VS最大比产氢率为152.2 ml·(kg·h)-1.并用SGompertz方程对实验数据进行拟合,定量说明在不同的热处理温度和时间下,厌氧发酵的累积产氢量(y)和时间(x)的关系.污泥厌氧发酵产氢前后各指标都发生明显变化,NH4 -N和总挥发性脂肪酸(TVFA)的浓度都增加了,而可溶糖和可溶蛋白质的浓度都降低了.热处理后的污泥在厌氧发酵产氢过程中,主要降解的有机物为蛋白质,发酵后蛋白质可降解20%～41%.     

替硝唑片对污泥厌氧发酵产氢作用的研究

研究表明，在污泥中添加替硝唑片可使污泥中的有机物质含量明显增加，特别是可溶的有机物质；替硝唑片对提高污泥产氢效果显著，替硝唑片添加量为1．40g时，污泥的产氢效果最佳，其累积产氢量和产氢潜能分别为580．64mL和98．41mL／gVS；厌氧发酵属于典型的乙醇型发酵；发酵代谢过程主要降解的有机物质为糖类物质，总糖降解率高达70．32％，蛋白质降解率只有26.25％。     

Biohydrogen production from cornstalk wastes by anaerobic fermentation with activated sludge

An anaerobic fermentation process to produce hydrogen from cornstalk wastes was systematically investigated in this work. Batch experiments numbered series I, II and III were designed to investigate the effects of acid pretreatment, enzymatic hydrolysis (enzymatic temperature, enzymatic time and enzymatic pH) on hydrogen production by using the natural sludge as inoculant. A maximum cumulative H₂ yield of 126.22 ml g⁻¹-CS (Cornstalk, or 146.94 ml g⁻¹-TS, Total Solid) and an average H₂ production rate of 9.58 ml g⁻¹-CS h⁻¹ were obtained from fermentation cornstalk with a concentration of 20 g/L and an initial pH of 7.0 at 36 °C through an optimal pretreatment process. The optimal process was that the substrate was soaked with an HCl concentration of 0.6 wt% at 90 °C for 2 h, and subsequently enzymatic hydrolysis for 72 h at 50 °C and pH 4.8 before fermentation. The biogas consisted of only H₂ and CO₂. In addition, the fermentation system was the typical ethanol-type fermentation according to ethanol and acetate as the main liquid by-products.     

Biological hydrogen production from sweet sorghum syrup by mixed cultures using an anaerobic sequencing batch reactor (ASBR)

Continuous biological hydrogen production from sweet sorghum syrup by mixed cultures was investigated by using anaerobic sequencing batch reactor (ASBR). The ASBR was conducted based on the optimum condition obtained from batch experiment i.e. 25 g/L of total sugar concentration, 1.45 g/L of FeSO₄ and pH of 5.0. Feasibility of continuous hydrogen fermentation in ASBR operation at room temperature (30 ± 3 °C) with different hydraulic retention time (HRT) of 96, 48, 24 and 12 hr and cycle periods consisting of filling (20 min), settling (20 min), and decanting (20 min) phases was analyzed. Results showed that hydrogen content decreased with a reduction in HRT i.e. from 42.93% (96 hr HRT) to 21.06% (12 hr HRT). Decrease in HRT resulted in a decrease of solvents produced which was from 10.77 to 2.67 mg/L for acetone and 78.25 mg/L to zero for butanol at HRT of 96 hr-12 hr, respectively. HRT of 24 hr was the optimum condition for ASBR operation indicated by the maximum hydrogen yield of 0.68 mol H₂/mol hexose. The microbial determination in DGGE analysis indicated that the well-known hydrogen producers Clostridia species were dominant in the reacting step. The presence of Sporolactobacillus sp. which could excrete the bacteriocins causing the adverse effect on hydrogen-producing bacteria might responsible for the low hydrogen content obtained.     

Bioelectrochemical enhancement of hydrogen and methane production from the anaerobic digestion of sewage sludge in single-chamber membrane-free microbial electrolysis cells

Batch tests were carried out to investigate the bioelectrochemical enhancement of hydrogen and methane production from the anaerobic digestion of sewage sludge in single-chamber membrane-free microbial electrolysis cells (MEC) and non-MECs. Hydrogen and methane were produced from the anaerobic digestion of sewage sludge in all reactors. Compared with controls, hydrogen production was enhanced 1.7–5.2-fold, and methane production 11.4–13.6-fold with Ti/Ru electrodes at applied voltages of 1.4 and 1.8 V, respectively. Most of hydrogen was produced in the first 5 days of digestion and most of methane was generated after 5 days. No oxygen was detected in the biogas and no hydrogen production was detected in the control test with water. The applied voltages can enhance the removal of suspended and volatile suspended solids, increase the transformation of soluble chemical oxygen demand, accelerate the conversion of volatile fatty acids and maintain an optimal pH range for methanogen growth.     

Biogas production from brown grease using a pilot-scale high-rate anaerobic digester

Food wastes are typically disposed of in landfills for convenience and economic reasons. However, landfilling food wastes increases the organic content of leachate and the risk of soil contamination. A sound alternative for managing food wastes is anaerobic digestion, which reduces organic pollution and produces biogas for energy recovery. In this study, anaerobic digestion of a common food waste, brown grease, was investigated using a pilot-scale, high-rate, completely-mixed digester (5.8 m³). The digestibility, biogas production and the impact of blending of liquid waste streams from a nearby pulp and paper mill were assessed. The 343-day evaluation was divided into 5 intensive evaluation stages. The organic removal efficiency was found to be 58 ± 9% in terms of COD and 55 ± 8% in terms of VS at a hydraulic retention time (HRT) of 11.6 ± 3.8 days. The removal was comparable to those found in organic solid digesters (45–60%), but at a much shorter HRT. Methane yield was estimated to be 0.40–0.77 m³-CH_4@STP kg-VS_removed⁻¹, higher than the typical range of other food wastes (0.11–0.42 m³-CH_4@STP kg-VS_removed⁻¹), with a mean methane content of 75% and <200 ppm of hydrogen sulfide in the biogas. The blending of selected liquid wastes from a paper mill at 10 vol% of brown grease slurry did not cause significant reduction in digester performance. Using a pseudo-first-order rate law, the observed degradation constant was estimated to be 0.10–0.19 d⁻¹ compared to 0.03–0.40 d⁻¹ for other organic solids. These results demonstrate that brown grease is a readily digestible substrate that has excellent potential for energy recovery through anaerobic digestion.     

Fermentative hydrogen production from cassava stillage by mixed anaerobic microflora: Effects of temperature and pH

Fermentative hydrogen production from cassava stillage was conducted to investigate the influences of temperature (37 °C, 60 °C, 70 °C) and initial pH (4–10) in batch experiments. Although the seed sludge was mesophilic anaerobic sludge, maximum hydrogen yield (53.8 ml H₂/gVS) was obtained under thermophilic condition (60 °C), 53.5% and 198% higher than the values under mesophilic (37 °C) and extreme-thermophilic (70 °C) conditions respectively. The difference was mainly due to the different VFA and ethanol distributions. Higher hydrogen production corresponded with higher ratios of butyrate/acetate and butyrate/propionate. Similar hydrogen yields of 66.3 and 67.8 ml H₂/gVS were obtained at initial pH 5 and 6 respectively under thermophilic condition. The total amount of VFA and ethanol increased from 3536 to 7899 mg/l with the increase of initial pH from 4 to 10. Initial pH 6 was considered as the optimal pH due to its 19% higher total VFA and ethanol concentration than that of pH 5. Homoacetogenesis and methonogenesis were very dependent on the initial pH and temperature even when the inoculum was heat-pretreated. Moreover, a difference between measured and theoretical hydrogen was observed in this study, which could be attributed to homoacetogenesis, methanogenesis and the degradation of protein.     

Feasibility of biohydrogen production by anaerobic co-digestion of food waste and sewage sludge

Anaerobic co-digestion of food waste and sewage sludge for hydrogen production was performed in serum bottles under various volatile solids (VS) concentrations (0.5–5.0%) and mixing ratios of two substrates (0:100–100:0, VS basis). Through response surface methodology, empirical equations for hydrogen evolution were obtained. The specific hydrogen production potential of food waste was higher than that of sewage sludge. However, hydrogen production potential increased as sewage sludge composition increased up to 13–19% at all the VS concentrations. The maximum specific hydrogen production potential of 122.9 ml/g carbohydrate-COD was found at the waste composition of 87:13 (food waste:sewage sludge) and the VS concentration of 3.0%. The relationship between carbohydrate concentration, protein concentration, and hydrogen production potential indicated that enriched protein by adding sewage sludge might enhance hydrogen production potential. The maximum specific hydrogen production rate was 111.2 ml H₂/g VSS/h. Food waste and sewage sludge were, therefore, considered as a suitable main substrate and a useful auxiliary substrate, respectively, for hydrogen production. The metabolic results indicated that the fermentation of organic matters was successfully achieved and the characteristics of the heat-treated seed sludge were similar to those of anaerobic spore-forming bacteria, Clostridium sp.     

Effects of pH value and substrate concentration on hydrogen production from the anaerobic fermentation of glucose

A series of batch experiments were conducted to investigate the effects of pH and glucose concentrations on biological hydrogen production by using the natural sludge obtained from the bed of a local river as inoculant. Batch experiments numbered series I and II were designed at an initial and constant pH of 5.0–7.0 with 1.0 increment and four different glucose concentrations (5.0, 7.5, 10 and 20 g glucose/L). The results showed that the optimal condition for anaerobic fermentative hydrogen production is 7.5 g glucose/L and constant pH 6.0 with a maximum H₂ production rate of 0.22 mol H₂ mol⁻¹ glucose h⁻¹, a cumulative H₂ yield of 1.83 mol H₂ mol⁻¹ glucose and a H₂ percentage of 63 in biogas.     

Production of hydrogen and methane from wastewater sludge using anaerobic fermentation

Hydrogen and methane were produced from wastewater sludge using a clostridium strain. The original sludge and pre-treated (acidified, basified and freeze/thawed) sludges were the testing samples. Some pre-treatments enhanced hydrogen yield, whereas other treatments enhanced methane yield. Hydrogen fermentation can be used as a pre-stage to improve subsequent methane production from wastewater sludge.     

A comparison of treatment techniques to enhance fermentative hydrogen production from piggery anaerobic digested residues

To accelerate the start-up process and enhance the efficiency of a hydrogen production system, piggery anaerobic digested residues (PADRs) were subjected to several different treatment methods to enrich the hydrogen-producing bacteria. Eight treatment methods were performed on the PADRs, including acid, alkali, heat, drying, ultrasound, aeration, sodium 2-bromoethanesulfonate (BES), and chloroform. The best method was found to be drying at 60 °C for 48 h, which maximised the total biogas production and the hydrogen fraction without causing any methane production. The volatile fatty acids (VFAs) found after the drying treatment were acetate and butyrate, which together accounted for 91.9% of all VFAs, indicating that butyric acid fermentation was established. Due to the drying treatment, the metabolites produced from the biodegradable DOM were utilised more rapidly, more completely, and with the least amount of hard-degradation organic matter content obtained, according to EEM fluorescence spectra. This drying treatment offers a promising method to¹ improve bio-hydrogen production.     

Hydrogen production performance from food waste using piggery anaerobic digested residues inoculum in long-term systems

The objective of this study was to investigate the hydrogen production performance from food waste using piggery anaerobic digested residues (PADRs) inoculum. Multiple parameters were evaluated such as organic load rate (OLR), pH, and hydraulic retention time (HRT), over a wide range of values in long-term dark fermentation systems. Results showed that a value of 126.50 mL/gVS·d hydrogen yield was achieved at OLR 6 g VS/L·d under thermophilic condition. A relatively stable structural composition dominated by Thermoanaerobacterium was maintained even suffering from OLR and acid shock. On the contrary, mesophilic fermentation performed acetic acids accumulation and an average hydrogen yield of less than 80 mL/gVS·d. High OLR and low pH (range of 5.0–5.5) led to the establishment of Lactobacillus. Beyond this range, the relative abundance of Olsenella, Streptococcus, and other bacteria showed a significant difference under different operating conditions, which caused weak resistance to external shocks during mesophilic fermentation. It showed that PADRs was capable of obtaining optimal hydrogen production performance under thermophilic condition from food waste with a stable microbial community structure.