Dark Fermentation Research Articles

Novel Insights into Alkyl Polyglucoside Biosurfactant Promoting Anaerobic Dark Fermentation for Hydrogen Production in Sludge.

Anaerobic fermentation of excess sludge (ES) for hydrogen production is a crucial strategy for resource utilization and environmentally friendly treatment. However, the low hydrolysis efficiency of ES and the depletion of produced hydrogen have become the limiting factors for low hydrogen yield. This study innovatively applied the bio-based surfactant alkyl polyglucoside (APG) to enhance the efficiency of dark fermentation for hydrogen production from ES. When the APG content was 100mg/g (calculated based on total suspended solids), the maximum hydrogen production reached 17.8 mL/g VSS, approximately 3.7 times that in the control group. Mechanistic analysis revealed that APG promoted the release of organic matter from ES. APG also facilitated the release of soluble protein and soluble polysaccharide, increasing the organic matter reduction rate to 34.8%, significantly higher than other groups. APG enhanced the accumulation of volatile fatty acids and promoted the proportion of small molecular carboxylic acids. Enzyme activity analysis revealed that APG promoted the activity of hydrolytic enzymes but inhibited the activity of hydrogen-consuming enzymes. The research results provide a green and environmentally friendly strategy for the efficient resource utilization of ES.

Biohydrogen production by co-digestion of food waste and corn industry wastewater

Food waste (FW) and corn industry wastewater (cornWW) are residues with a high potential for hydrogen production in the dark fermentation process. The study aimed to identify the co-digestion of FW and cornWW ratios for improving hydrogen production and evaluating the synergistic effect on biohydrogen production potential (BHP) in batch test. The inoculum consisted of a mixed culture plus the native microbiota of each substrate. The adjustment of the initial pH to 7.5 was compared to the co-digestion without control of the initial pH of the experiments. The highest hydrogen and metabolites production were obtained with a co-digestion FW:cornWW ratio. When no pH adjustment is applied, pH values above 10 completely inhibit the metabolic activity of bacteria in the process. The cornWW not only provided alkalinity to the system but also can be applied to adjust the organic matter content of FW, adjusting the initial concentration of organic matter. The growth of microbial communities was formed mainly by Clostridium sp. The co-digestion of FW:cornWW can be technically feasible to improve the production of H2 through the dark fermentation process.

Salt and heat stress enhances hydrogen production in cyanobacteria.

Cyanobacteria are among the most suitable organisms for the capture of excessive amounts of CO2 and can be grown in extreme environments. In our research we use the single-celled freshwater cyanobacteria Synechococcus elongatus PCC7942 PAMCOD strain and Synechocystis sp. PCC6714 for the production of carbohydrates and hydrogen. PAMCOD strain and Synechocystis sp. PCC6714 synthesize sucrose when exposed to salinity stress, as their main compatible osmolyte. We examined the cell proliferation rate and the sucrose accumulation in those two different strains of cyanobacteria under salt (0.4M NaCl) and heat stress (35 0C) conditions. The intracellular sucrose (mol sucrose content per Chl a) was found to increase by 50% and 108% in PAMCOD strain and Synechocystis sp. PCC6714 cells, respectively. As previously reported, PAMCOD strain has the ability to produce hydrogen through the process of dark anaerobic fermentation (Vayenos D, Romanos GE, Papageorgiou GC, Stamatakis K (2020) Photosynth Res 146, 235-245). In the present study, we demonstrate that Synechocystis sp. PCC6714 has also this ability. We further examined the optimal conditions during the dark fermentation of PAMCOD and Synechocystis sp. PCC6714 regarding H2 formation, increasing the PAMCOD H2 productivity from 2 nmol H2 h- 1 mol Chl a- 1 to 23 nmol H2 h- 1 mol Chl a- 1. Moreover, after the dark fermentation, the cells demonstrated proliferation in both double BG-11 and BG-11 medium enriched in NaNO3, thus showing the sustainability of the procedure.

Enhancing biohydrogen production from xylose through natural FeS2 ore: Mechanistic insights

In this study, we investigated the advantages of utilizing natural FeS2 ore in the context of dark fermentative hydrogen production within a fermentation system employing heat-treated anaerobic granular sludge with xylose as the carbon source. The results demonstrated a significant improvement in both hydrogen production and the maximum rate, with increases of 2.58 and 4.2 times, respectively. Moreover, the presence of FeS2 ore led to a reduction in lag time by more than 2–3 h. The enhanced biohydrogen production performance was attributed to factors such as the intracellular NADH/NAD+ ratio, redox-active components of extracellular polymeric substances, secreted flavins, as well as the presence of hydrogenase and nitrogenase. Furthermore, the FeS2 ore served as a direct electron donor and acceptor during biohydrogen production. This study shed light on the underlying mechanisms contributing to the improved performance of biohydrogen production from xylose during dark fermentation through the supplementation of natural FeS2 ore.

A bibliometric analysis of the role of nanotechnology in dark fermentative biohydrogen production.

Recently, nanoparticles have drawn a lot of interest as catalysts to enhance the effectiveness and output of biohydrogen generation processes. This review article provides a comprehensive bibliometric analysis of the significance of nanotechnology in dark fermentative biohydrogen production. The study examines the scientific literature from the database of The Web of Science© while the bibliometric investigation utilized VOSviewer© and Bibliometrix software tools to conduct the analysis. The findings revealed that a total of 232 articles focused on studying dark fermentation for hydrogen production throughout the entire duration. The extracted data was used to analyze publication trends, authorship patterns, and geographic distribution along with types and effects of nanoparticles on the microbial community responsible for dark fermentative biohydrogen production. The findings of this bibliometric analysis provide valuable insights into the advancements and achievements in the utilization of nanoparticles in the dark fermentation process used to produce biohydrogen.

The Design of a Sustainable Industrial Wastewater Treatment System and The Generation of Biohydrogen from E. crassipes.

Water scarcity is a significant global issue caused by the prolonged disregard and unsustainable management of this essential resource by both public and private bodies. The dependence on fossil fuels further exacerbates society's bleak environmental conditions. Therefore, it is crucial to explore alternative solutions to preserve our nation's water resources properly and promote the production of biofuels. Research into the utilization of E. crassipes to remove heavy metals and generate biofuels is extensive. The combination of these two lines of inquiry presents an excellent opportunity to achieve sustainable development goals. This study aims to develop a sustainable wastewater treatment system and generate biohydrogen from dry, pulverized E. crassipes biomass. A treatment system was implemented to treat 1 L of industrial waste. The interconnected compartment system was built by utilizing recycled PET bottles to generate biohydrogen by reusing the feedstock for the treatment process. The production of biological hydrogen through dark fermentation, using biomass containing heavy metals as a biohydrogen source, was studied. Cr (VI) and Pb (II) levels had a low impact on hydrogen production. The uncontaminated biomass of E. crassipes displayed a significantly higher hydrogen yield (81.7 mL H2/g glucose). The presence of Cr (IV) in E. crassipes leads to a decrease in biohydrogen yield by 14%, and the presence of Pb (II) in E. crassipes leads to a decrease in biohydrogen yield of 26%. This work proposes a strategy that utilizes green technologies to recover and utilize contaminated water. Additionally, it enables the production of bioenergy with high efficiency, indirectly reducing greenhouse gases. This strategy aligns with international programs for the development of a circular economy.

Strategies to enhance biohydrogen production from microalgae: A comprehensive review

Microalgae represent a promising renewable feedstock for the sustainable production of biohydrogen. Their high growth rates and ability to fix carbon utilizing just sunlight, water, and nutrients make them well-suited for this application. Recent advancements have focused on improving microalgal hydrogen yields and cultivation methods. This review aims to summarize recent developments in microalgal cultivation techniques and genetic engineering strategies for enhanced biohydrogen production. Specific areas of focus include novel microalgal species selection, immobilization methods, integrated hybrid systems, and metabolic engineering. Studies related to microalgal strain selection, cultivation methods, metabolic engineering, and genetic manipulations were compiled and analyzed. Promising microalgal species with high hydrogen production capabilities such as Synechocystis sp., Anabaena variabilis, and Chlamydomonas reinhardtii have been identified. Immobilization techniques like encapsulation in alginate and integration with dark fermentation have led to improved hydrogen yields. Metabolic engineering through modulation of hydrogenase activity and photosynthetic pathways shows potential for enhanced biohydrogen productivity. Considerable progress has been made in developing microalgal systems for biohydrogen. However, challenges around process optimization and scale-up remain. Future work involving metabolic modeling, photobioreactor design, and genetic engineering of electron transfer pathways could help realize the full potential of this renewable technology.

Buswell’s Equation for Quantifying Biohydrogen

Biohydrogen is widely generated by dark fermentation and two-stage anaerobic digestion. Anaerobic digestion can be chemically represented by the Buswell’s equation, which is based on elemental composition of organic matter. When compared to Buswell’s equation for biomethane, its equation for biohydrogen has been given less attention. In this article, the nature of Buswell’s equation for biohydrogen is introduced by using the H-atom method. The mean oxidation number of organic carbons is employed as a metric for counting theoretical quantity of biohydrogen and parameters of Buswell’s equation for biohydrogen. Based on an empirical formula and mean oxidation number of organic carbons, the general Buswell’s equation for biohydrogen is developed. The mathematical relationships among mean oxidation number of organic carbons, empirical formula, quantity of biohydrogen, theoretical biohydrogen potential, and biohydrogen percent yield are also established. Biowastes and bio-substrates are chosen to demonstrate this notion.

Biohydrogen Production from Waste Black Cumin (Nigella Sativa) Extract Liquid.

Hydrogen creates water during combustion. Therefore, it is expected to be the most promising environmentally friendly energy alternative in the coming years. This study used extract liquid obtained from the waste nigella sativa generated by the black cumin oil industry. The performance of biological hydrogen manufacturing via dark fermentation was investigated in the fluidized bed reactor (FBR) and completely stirred tank reactor (CSTR) under the operation conditions of pH 5.0, 4.0, and 6.0 and a hydraulic retention time (HRT) of 36 and 24 h. The performance of hydrogen manufacturing was determined to be good under an organic loading ratio (OLR) of 6.66 g.nigella sativa extract/L and pH 4.0. According to these conditions, the maximum amount of hydrogen in CSTR and FBR was found to be 20.8 and 7.6 mL H2/day, respectively. The operating process of the reactors displayed that a reduction in HRT augmented biohydrogen manufacturing. The work that used mixed culture found that the dominant microbial population at pH 4.0 involved Hydrogenimonas thermophila, Sulfurospirillum carboxydovorans, Sulfurospirillum cavolei, Sulfurospirillum alkalitolerans, and Thiofractor thiocaminus. No research on waste black cumin extract was found in biohydrogen studies, and it was determined that this substrate source is applicable for biological hydrogen manufacturing.

Development of a unique integrated bioreactor for simultaneous desalination and bioenergy and biohydrogen production

In the wastewater treatment challenge, it is really essential to develop integrated systems in reducing greenhouse gases, producing green energy and achieving sustainable development. In this regard, an integrated electro-biomembrane reactor was developed and performed in this study for simultaneous biohydrogen (bioH2) production from energetic poplar leaves using dark fermentation (DF) process, conventional H2 production, bioenergy production in the DF process, and saline water desalination in a single system. The results of this study showed that pH was the main controlling parameter in bioH2 production, and the superior production of 40.2 mL/g-biomass was obtained at a pH of 5.5. The maximum current and power density values were 2861.7 mW/m2 and 2819.4 mA/m2 at pH 5.5 under improved conditions. Furthermore, the maximum conventional H2 production was found to be 1341.6 mL using 2 M of KOH solution. Overall, the results further proved that the proposed integrated system can be a sustainable and promising process for industrial applications, considering its high desalination, energy production, and conventional and biological H2 production efficiencies.

Inhibition of norfloxacin on fermentative hydrogen production: Performance evaluation and metagenomic analysis

Toxic chemicals can inhibit the dark fermentation process for renewable biohydrogen production. Norfloxacin (NOR), a typical antibiotic, is present in various feedstocks of dark fermentation, which has excellent antibacterial properties. It is hypothesized that NOR could cause adverse effects on fermentative hydrogen production, while current understanding of this subject is scare. This study investigated the potential inhibitory effect of NOR on dark hydrogen fermentation, and explored the underlying mechanisms from the perspectives of bacterial community structure and functional gene abundance. The results showed that NOR significantly reduced the hydrogen-producing capacity upon exposure to concentrations exceeding 10 mg/L. The hydrogen yield decreased from 1.56 to 0.27 H2/mol-glucose at NOR concentrations of 50 mg/L. NOR also prolonged the hydrogen-producing lag period. Bacterial community analysis revealed that NOR reduced the abundance of high-yielding H2-generating bacteria, such as Clostridium sp., while increasing the abundance of H2-consuming bacteria (e.g. Enterococcus sp.). Metagenomic analysis further indicated that crucial genes involved in glycolysis (e.g. HK, tal-pgi, pfk and PK) and hydrogen formation (e.g. por and E1.12.7.2) were remarkably downregulated under NOR exposure, essentially causing the inhibition of hydrogen production. This study contributes to a better understanding of how antibiotics inhibit dark hydrogen fermentation and provides a theoretical basis for reducing potential inhibitory effects.

The influence of inoculum source and pretreatment on the biohydrogen production in the dark fermentation process

The production of biohydrogen from food waste (FW) by dark fermentation (DF) is a promising technology for commercialisation, as it is both a clean fuel and a suitable means of sustainable waste management. The described experiments compared the biohydrogen production yields obtained after the use of inoculum from two different sources: digested sludge from the wastewater treatment plant (WWTP) in Lodz and sludge from the anaerobic treatment of dairy industry wastewater (DIW) (unconcentrated and double-concentrated). In addition, the effect of different temperatures (70, 90 and 121°C) of inoculum pretreatment on the biohydrogen production in DF was tested. The process was carried out batchwise at 37°C. The highest yield of hydrogen production was obtained after the inoculum pretreatment at 70°C. In addition, a higher amount of hydrogen could be obtained by using sludge from the WWTP as the inoculum (96 cm3 H2/gTVSFW) than unthickened sludge from the DIW (85 cm 3 H 2/g TVSFW). However, after thickening the sludge from the dairy industry, and at the same time balancing the dry matter of both sludges, the hydrogen production potential was comparable for bothsludges (for the WWTP sludge – 96 and for the DIW sludge – 93 cm 3 H 2/g TVSFW). The kinetics of hydrogen production was described by modified Gompertz equation, which showed a good fit (determination coefficient R2 between 0.909 and 0.999) to the experimental data.

Advances and Perspectives in Biohydrogen Production from Palm Oil Mill Effluent

Palm oil, the main vegetable oil produced globally, serves diverse purposes, ranging from cooking to the production of processed foods, cosmetics, and biodiesel. Despite contributing significantly to the economies of major producing nations, the escalating production of palm oil raises serious environmental concerns, including deforestation, biodiversity loss, and various forms of pollution. Palm oil mill effluent (POME), a byproduct of palm oil extraction, poses a severe environmental threat when left untreated. As an eco-friendly alternative, anaerobic digestion in controlled bioreactors has emerged, offering simultaneous POME treatment and biofuel generation, particularly hydrogen, with high energy efficiency. This review explores the challenges and opportunities associated with biohydrogen production from POME. Key considerations involve optimizing parameters through pretreatments, nanoparticle incorporation, defining optimal bioreactor conditions, determining hydraulic retention times, and integrating multi-stage processes like dark fermentation followed by photofermentation. This review also emphasizes the significance of sustainable practices and economic analyses in shaping the future of hydrogen production from POME, positioning it as a pivotal player in the palm oil industry’s circular economy and the global energy transition.

Impacts of micro- to nano-sized carbon supplements on mixed and archaea-free halophilic cultures when used for bioenergy recovery from saline wastewater

Due to the growing interest in enhancing dark fermentation and anaerobic digestion via carbon-based additives to exploit different lucrative synergies, process-based optimizations and mechanistic analyses are crucial. Herein, we studied the impacts of carbon supplements on halophilic microbial cultures as a process intensification way towards bioenergy recovery from saline wastewater. Three additives, i.e., powdered activated carbon, multiwall carbon nanotubes and graphene, were examined. Two sets of parallel batch experiments were conducted using archaea-free (for dark fermentation) and mixed (for anaerobic digestion) halophilic cultures at different supplement doses. Adding 30 mg/L of graphene and 100 mg/L of powdered activated carbon exhibited the maximum improvements in dark fermentation (H2-production) and anaerobic digestion (biogas production), i.e., ∼28% and ∼27% over the control, respectively. This underscored the dose-dependent impact of different-sized carbon amendments. Both the electron transport system activity and extracellular polymeric substances were improved with all amendments, indicating a stimulated electron transfer flux. However, distinct bacterial compositions were identified when using mixed and archaea-free cultures along with indicators for direct interspecies electron transfer establishment in the presence of archaea. Beyond this, such findings also supported the expanded role of conductive additives on H2-producing bacteria, including Clostridium. According to a techno-economic analysis, powdered activated carbon emerged as the best for optimizing bioenergy recovery (in terms of H2 or biogas) from saline wastewater, providing a solid basis for relevant applications.

Assessment of a Low-Cost Hydrogen Sensor for Detection and Monitoring of Biohydrogen Production during Sugarcane Straw/Vinasse Co-Digestion

In this work, hydrogen production from the co-digestion of sugarcane straw and sugarcane vinasse in the dark fermentation (DF) process was monitored using a cost-effective hydrogen detection system. This system included a sensor of the MQ-8 series, an Arduino Leonardo board, and a computer. For the DF, different concentrations of sugarcane vinasse and volumetric ratios of vinasse/hemicellulose hydrolysate were used together with a thermally pretreated inoculum, while the hydrogen detection system stored the hydrogen concentration data during the fermentation time. The results showed that a higher concentration of vinasse led to higher inhibitors for the DF, resulting in a longer lag phase. Additionally, the hydrogen detection system proved to be a useful tool in monitoring the DF, showcasing a rapid response time, and providing reliable information about the period of adaptation of the inoculum to the substrate. The measurement system was assessed using the error metrics SE, RMSE, and MBE, whose values ranged 0.6 and 5.0% as minimum and maximum values. The CV (1.0–8.0%) and SD (0.79–5.62 ppm) confirmed the sensor’s robustness, while the ANOVA at the 5% significance level affirmed the repeatability of measurements with this instrument. The RMSE values supported the accuracy of the sensor for online measurements (6.08–14.78 ppm). The adoption of this straightforward and affordable method sped up the analysis of hydrogen in secluded regions without incurring the expenses associated with traditional measuring instruments while offering a promising solution for biomass valorization, contributing to the advancement of rural green energy initiatives in remote areas.

Investigating the efficiency of a two-stage anaerobic-aerobic process for the treatment of confectionery industry wastewaters with simultaneous production of biohydrogen and polyhydroxyalkanoates.

The scope of the current study was to investigate the efficiency of a two-stage anaerobic-aerobic process for the simultaneous treatment and valorization of selective wastewater streams from a confectionary industry. The specific wastewater (confectionary industry wastewater, CIW) was a mixture of the rinsing eluting during washing of the cauldrons in which jellies and syrups were produced, and contained mainly readily fermentable sugars, being thus of high organic load. The first stage of the process was the dark fermentation (DF) of the CIW in continuous, attached-biomass systems, in which the effect on hydrogen yields and distribution of metabolites were studied for different packing materials (ceramic or plastic), hydraulic retention times, HRTs (12 h–30 h) and feed substrate concentration (20 g COD/L- 50 g COD/L). In the second stage, the effectiveness of the aerobic treatment of the DF effluents was evaluated in terms of the reduction of the organic load and the production of polyhydroxyalkanoates (PHAs) through an enriched mixed microbial culture (MMC). The MMC was developed in a continuous draw and fill system, in which the accumulation potential of PHAs was studied. It was shown that the hydrogen production rates decreased for increasing substrate concentration and HRTs, with a maximum of 12.70 ± 0.35 m3 H2/m3 initial CIW achieved for the lowest HRT and feed concentration and using ceramic beads as packing material. Butyrate, acetate and lactate were the main metabolites generated in all cases, in different ratios. The distribution of metabolites during DF was shown to highly affect the efficiency of the second process in terms of both the reduction of organic load and the PHAs yields. The highest removal of organic load achieved after 48 h of aerobic treatment was 84.0 ± 0.9 %, whereas the maximum PHAs yield was 21.46 ± 0.13 kg PHAs/m3 initial CIW.

Biomethane and biohydrogen production from an anaerobic sludge used in the treatment of rice parboiling effluent: Specific methanogenic and hydrogenic activity

This study evaluated the biomethane and biohydrogen production capacity of an anaerobic sludge collected from a rice parboiling industry. Three main parameters were assess from an experimental batch with varying substrate-inoculum ratios (S/I): the final mass production, the maximum generation rate, and the reactors’ biomethane and biohydrogen yield. From the analysis of the final production and the maximum generation rate, the best S/I ratios found for the methanogenic and hydrogenic reactors were, respectively, 1.4 and 3.8 gCOD/gVSS. These proportions produced 102.65 mgCH4/gVSS and 27.96 mgH2/gVSS, reaching the following maximum generation rates: 32.91 mgCOD CH4/gVSS.h and 12.63 mgCOD H2/gVSS.h. In contrast to the methanogenic microorganisms of the studied sludge, the acetogenics allow the use of higher organic loads in the systems. However, the lowest S/I ratio (0.1 gCOD/gVSS) resulted in both the best biomethane and biohydrogen production yield, around 2.33 mol CH4/mol glucose and 2.46 mol H2/mol glucose. Compared to the maximum theoretical yields, whereas the sludge produced biohydrogen more effectively under high S/I ratios, biomethane was generated more efficiently in low S/I ratios. Finally, the anaerobic sludge can be employed as a satisfactory inoculum in anaerobic digestion and dark fermentation processes designed for biomethane and biohydrogen production.

Sustainable Management Strategies for Fruit Processing Byproducts for Biorefineries: A Review

The fruit processing industry generates enormous amounts of byproducts, which are primarily removed through landfill or incineration. However, these processes cause carbon dioxide and methane emissions and release dioxin into the environment. The management of fruit processing byproducts is important for reducing the amount of food waste that is sent to landfills and for developing strategies through the reuse of these products for valorization and economic added value. Fruit processing byproducts are rich sources of bioactive compounds and fermentable and nonfermentable sugars. Therefore, these materials are very attractive feedstocks for developing integrated multifeed biorefineries that coproduce a wide range of natural products and bioenergy. The studies presented here have shown sustainable strategies for managing fruit processing byproducts via a biorefinery approach to achieve full valorization via a circular economy. The full valorization project proposed five main phases, namely, pretreatment, extraction, dark or aerobic fermentation, anaerobic digestion, and post-treatment, as well as two additional pathways to generate additional bioelectricity. When choosing the appropriate directions for the presented concept, a technoeconomic analysis should be carried out, considering the type of biomass and its availability at the site of the biorefinery and throughout the year of production. Applying the proposed concept of biorefineries in closed-loop technology is a promising way to enhance economic efficiency and decrease environmental influence in accordance with sustainable development.

Influence of Inoculum Thermal Pretreatment Time on Hydrogen Production in Dark Fermentation

Dark fermentation (DF) of kitchen waste (KW) is a promising technology for the production of renewable biohydrogen. It can be both a method of obtaining clean energy and a sustainable waste management. Despite its potential, this process requires further research to improve efficiency. The aim of the research was to test the effect of thermal pretreatment of the inoculum on H2 and volatile fatty acids (VFAs) production in the DF process. The process was carried out at 37 °C, in batch mode. The digested sludge from the Group Wastewater Treatment Plant in Lodz was used as inoculum. KW from households was used as substrate. The inoculum was pre-treated at 70 °C for 15 and 30 min. Two control reference experiments were also used. The first without the inoculum, and the second without heating the inoculum. The thermal pretreatment inhibited methane production and increased hydrogen production. After the thermal pretreatment, the amount of CO2 produced during the process decreased compared to the bioreactor without inoculum pretreatment. Additionally, the main VFAs in the samples with pretreated inoculum were acetic and butyric acids, which are associated with hydrogen production in the biochemical pathways of the DF process. However, the time of thermal pretreatment had no significant effect on H2 production.

Valorization of organic carbon in primary sludge via semi-continuous dark fermentation: First step to establish a wastewater biorefinery

In this study, lab-scale, bench-scale, and pilot-scale experiments were carried out to optimize short-chain fatty acids production from primary sludge. Batch tests showed the requirement of short retention times and semi-continuous operation mode showed a plateau of maximum daily productivity at 36-hours hydraulic retention time with minimal methanation. Optimization from pH 5 to pH 10 at 36 h-hydraulic retention time under long-term semi-continuous operating mode revealed that production of short-chain fatty acids was pH dependent and highest yields could be achieved at pH 7 by establishing optimum redox conditions for fermentation. Pilot-scale experiments at 32 °C showed that daily productivity (3.1 g∙Lreactor-1∙dHRT-1) and yields (150 mg∙gVS-1; OLR = 21 gVS∙Lreactor-1∙dHRT-1; pH 7) of short-chain fatty acids could be significantly improved, specifically for acetic and propionic acids. From these results, a robust dark fermentation step for recovery of valuable products from the solids treatment step in a biorefinery can be achieved.