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This study was carried out to evaluate the forage quantity and quality of several halophyte species grown in arid-saline environments. After identifying 44 halophytic species in the region and considering the potential of quantitative and qualitative forage production, 13 species from four families, i.e. Amaranthaceae, Asteraceae, Leguminosae and Convolvulaceae, and eight genera were selected for further evaluation. These species differed significantly in terms of both forage quantity, measured in terms of fresh (FW) and dry weight (DW), and forage quality assessed in terms of tissue water content (TWC), ash, nitrogen content (N), crude protein (CP), acid detergent fiber (ADF), neutral detergent fiber (NDF), dry matter digestibility and metabolizable energy (ME). The highest fresh and dry weights were obtained from Suaeda ferticosa (1006.3 g and 306.3 g, respectively) and Noaea mucronata (909.3 g and 309 g, respectively). However, based on forage quality characteristics, Alhagi maurorum, Bassia scoparia, Noaea mucronata, Halostachys belangriana and Cressa cretica showed the best forage potential. Values of ash, CP, ADF, NDF and ME measured in the halophytes species ranged between 7.9% and 33.2%, 6.2% and 15.8%, 30.0% and 50.3%, 33.2% and 56.4%, 5.6 and 8.7 MJ kg−1, respectively. The forage quality of the evaluated halophytic plants was influenced by unfavorable environmental conditions such as high soil salinity and low rainfall, however, these species can be considered as new sources of forage. Nevertheless, further studies are needed to improve the quality of such halophytic species by reducing the ash content and increasing the ME.

Operations in copper sulfide flotation plants (CSFP) are complex and governed by several variables such as available technologies, reagents, and environmental conditions. However, few investigations are related to studying the microbial communities. These aspects provide a reason to compare the bacterial communities of two CSFP operated with freshwater (FwFlo) and seawater (SwFlo), and study whether indigenous bacteria could be used as pyrite bioreagents. Analyses were determined through next-generation sequencing by Illumina MiSeq System and conducted throughout the entire process: (i) minerals before and after grinding; (ii) final concentrate and concentrate thickener overflow; (iii) final tailings and tailings thickener overflow; and (iv) intake water. Bacterial strains from both plants were tested as potential bioreagents, given their tendency to adhere to pyrite after 5 min. In both CSFP, Proteobacteria (relative abundance from 45.48% to 79.22%), followed by Bacteroidetes (9.37%–44.7%), were the most abundant phyla. Regarding species, Algoriphagus olei (11.35%–43.52%) was present exclusively in FwFlo samples in contact with process water and absent in the mineral before grinding, where Cupriavidus metallidurans (16.05%) and Pseudomonas_uc (11.79%) predominated. In SwFlo samples, Marinobacter flavimaris (3.47%–41.1%), and GU061212-s (10.92%–27.63%), were the most abundant microorganisms. All of them were also detected in intake seawater. The strains with the highest adhesion rate (from 29.84% ± 0.14–100%) were phylogenetically identified as species of the genera Marinobacter, Pseudomonas, Idiomarina, Halomonas, Bacillus, Aerocuccus, and Peribacillus. Our results reveal that bacterial communities are critically dependent on process waters during mining activities, and our data depicted that indigenous bacteria could be used as potential pyrite bioreagents, evidenced by a high adhesion rate. It is thus possible to propose that different indigenous bacterial strains could be considered as new bioreagents to reduce the impact of conventional flotation reagents on health from an environment friendly perspective.

International audience; Cyanobacteria and microalgae are considered as interesting feedstocks for either the production of high value bio-based compounds and biofuels or wastewater treatment. Nevertheless, the high costs of production, mainly due to the harvesting process, hamper a wide commercialization of industrial cyanobacteria and microalgae based products. Recent studies have found in autoflocculation and bioflocculation promising spontaneous processes for a low-cost and environmentally sustainable cyanobacteria and microalgae biomass harvesting process. In the present work, bioflocculation process has been studied for three different inocula: filamentous cyanobacteria, microalgae and their mixture. Their cultivation has been conducted in batch mode using two different cultivation media: synthetic aqueous solution and urban wastewater. The removal of nutrients and flocculation process performance were monitored during the entire cultivation time. Results have proved that bioflocculation and sedimentation processes occur efficiently for filamentous cyanobacteria cultivated in synthetic aqueous solution, whereas such processes are less efficient in urban wastewater due to the specific characteristics of this medium that prevent bioflocculation to occur. Besides different efficiencies associated to cultivation media, this work highlighted that bioflocculation of sole microalgae is not as effective as when they are cultivated together with filamentous cyanobacteria.

Mining activities are among the main sources of potentially toxic elements (PTEs) in the environment which constitute a real concern worldwide, especially in developing countries. These activities have been carried out for more than a century in Chile, South America, where, as evidence of incorrect waste disposal practices, several abandoned mining waste deposits were left behind. This study aimed to understand multi-elements geochemistry, source patterns and mobility of PTEs in soils of the Taltal urban area (northern Chile). Topsoil samples (n = 125) were collected in the urban area of Taltal city (6km2) where physicochemical properties (redox potential, electric conductivity and pH) as well as chemical concentrations for 35 elements were determined by inductively coupled plasma optical emission spectrometer. Data were treated following a robust workflow, which included factor analysis (based on ilr-transformed data), a new robust compositional contamination index (RCCI), and fractal/multi-fractal interpolation in GIS environment. This approach allowed to generate significant elemental associations, identifying pool of elements related either to the geological background, pedogenic processes accompanying soil formation or to anthropogenic activities. In particular, the study eventually focused on a pool of 6 PTEs (As, Cd, Cr, Cu, Pb, and Zn), their spatial distribution in the Taltal city, and the potential sources and mechanisms controlling their concentrations. Results showed generally low baseline values of PTEs in most sites of the surveyed area. On a smaller number of sites, however, higher values concentrations of As, Cd, Cu, Zn and Pb were found. These corresponded to very high RCCI contamination level and were correlated to potential anthropogenic sources, such as the abandoned mining waste deposits in the north-eastern part of the Taltal city. This study highlighted new and significant insight on the contamination levels of Taltal city, and its links with anthropogenic activities. Further research is considered to be crucial to extend this assessment to the entire region. This would provide a comprehensive overview and vital information for the development of intervention limits and guide environmental legislation for these pollutants in Chilean soils.

Bioflocculation phenomena for filamentous cyanobacteria were studied and analysed in two different cultivation systems (i.e. based on air-bubbling and on shaking) and for different initial biomass concentrations. Floc formation and biomass settling were monitored during batch cultivation tests according to an innovative protocol. Results showed that the two cultivation systems enhanced two different flocculation behaviours: air bubbling led to the formation of small and dense flocs, while the shaking table resulted in larger (14 mm2 vs 4 mm2) but mechanically weaker flocs. Floc analysis evidenced that the different mixing systems also affected the speciation of biomass. A mathematical model was developed to simulate and predict the settling performance during the bioflocculation process of filamentous cyanobacteria. Natural settling was examined at different phases of biomass growth. Optimal conditions were obtained at the end of the exponential growth phase, when 70% of the total cultivated biomass could be recovered.

This paper proposes an innovative bioaugmentation approach for the remediation of polycyclic aromatic hydrocarbon (PAH) contaminated soils, based on a novel habitat-based strategy. This approach was tested using two inocula (i-24 and i-96) previously enriched through an anaerobic digestion process on wheat straw. It relies on the application of allochthonous microorganisms characterized by specific functional roles obtained by mimicking a natural hydrolytic environment such as the rumen. The inocula efficiency was tested in presence of naphthalene alone, benzo[a]pyrene alone, and a mix of both of them. In single-contamination tests, i-24 inoculum showed the highest biodegradation rates (84.7% for naphthalene and 51.7% for benzo[a]pyrene). These values were almost 1.2 times higher than those obtained for both contaminants with i-96 inoculum and in the control test in presence of naphthalene alone, while they were 3 times higher compared to the control test in presence of benzo[a]pyrene alone. In mixed-contamination tests, i-96 inoculum showed final biodegradation efficiencies for naphthalene and benzo[a]pyrene between 1.1 and 1.5 higher than i-24 inoculum or autochthonous biomass. Total microbial abundances increased in the bioaugmented tests in line with the PAH degradation. The microbial community structure showed the highest diversity at the end of the experiment in almost all cases. Values of the Firmicutes active fraction up to 7 times lower were observed in the i-24 bioaugmented tests compared to i-96 and control tests. This study highlights a successful bioaugmentation strategy with biological components that can be reused in multiple processes supporting an integrated and environmentally sustainable bioremediation system.

This study investigated the effect of the salinity level, buffering agent and carbon source on the hydrogen (H2) and lactic acid synthesis under capnophilic (CO2-assisted) lactic fermentation (CLF) by Thermotoga neapolitana cf capnolactica (DSM 33003). Several series of batch fermentation experiments were performed either in 0.12 L serum bottles for selection of the best performing conditions or in a 3 L fermenter for the best possible combination of conditions. The serum bottle study revealed that change in the salinity level of the culture medium from 0 to 35 g L−1 NaCl increased lactic acid synthesis by 7.5 times without affecting the H2 yield. Use of different buffers (MOPS, TRIS or HEPES) did not affect the average H2 yield of 3.0 ± 0.24 mol H2 mol−1 of glucose and lactic acid synthesis of 13.7 ± 1.03mM when the cultures were sparged by CO2. Among the carbonsources investigated, glucose was found to be the best performing carbon source for the CLF fermentation with 35 g L−1 of NaCl and 0.01M of phosphate buffer. Hence, an up-scale experiment using a 3 L fermenter and the combination of the best performing conditions showed a 2.2 times more lactic acid synthesis compared to the 0.12 L serum bottle experiments. The study reveals the robustness and flexibility of the CLF-based technology using T. neapolitana cf capnolactica fermentation under various operating environmental conditions.

Food waste (FW) represents a source of high potential renewable energy if properly treated with anaerobic digestion (AD). Pretreating the substrates could yield a higher biomethane production in a shorter time. In this study, the effects of thermal (heating the FW in a separate chamber) and thermophilic (heating the full reactor content containing both FW and inoculum) pretreatments at 50, 60, 70 and 80. °C prior to mesophilic AD were studied through a series of batch experiments. Pretreatments at a lower temperature (50. °C) and a shorter time (55. °C) and longer operating times (>12. h) yielded higher soluble chemical oxygen demand (CODs), but had a negative effect on the methanogenic activity. The thermal pretreatments at the same conditions resulted in a lower solubilization of COD. Based on net energy calculations, the enhanced biomethane production is sufficient to heat up the FW for the thermal, but not for the thermophilic pretreatment.

The use of bacterial strains as agents in bioremediation processes could reduce the harmfulness of potential toxic elements (PTEs) from water and soil with low or even no impact on the natural ecosystems. In this study, two new metal resistant-bacterial strains (Q3 and Q5) of Bacillus sp. were isolated from a sulfurous spring and their potential (as pure cultures or mixed) to remove Pb(II) and Cd(II) from an aqueous matrix was evaluated and optimized using response surface methodology (RSM). The optimal conditions for Cd(II) removal from all tested strains combinations were observed at an initial pH 5, a temperature of 38 &deg, C, and an initial Cd(II) concentration of 50 mg L&minus, 1, while the performance of bacterial strains on Pb(II) removal was strongly correlated to initial pH and temperature conditions. Moreover, the efficiency of bacterial strains in removing both PTEs, Pb(II) and Cd(II), from an aqueous matrix was considerably higher when they were used as a mixed culture rather than pure. According to field emission SEM (FESEM) and EDS analysis, the two bacterial strains showed different mechanisms in removing Cd(II): Bacillus sp. Q5 bio-accumulated Cd(II) in its periplasmic space, whereas Bacillus sp. Q3 bio-accumulated Cd(II) on its cell surface. On the other hand, Pb(II) is removed by chemical precipitation (lead sulfide) induced by both Bacillus sp. Q3 and Q5. This study discloses new aspects of Pb(II) and Cd(II) bioremediation mechanisms in Bacillus species that can be extremely useful for designing and operating novel PTEs bioremediation processes.

The objective of this study was to enhance the hydrogen production rate of dark fermentation in batch operation. For the first time, the hyperthermophilic pure culture of Thermotoga neapolitana cf. Capnolactica was applied at elevated biomass concentrations. The increase of the initial biomass concentration from 0.46 to 1.74 g cell dry weight/L led to a general acceleration of the fermentation process, reducing the fermentation time of 5 g glucose/L down to 3 h with a lag phase of 0.4 h. The volumetric hydrogen production rate increased from 323 (+/- 11) to 654 (+/- 30) mL/L/h with a concomitant enhancement of the biomass growth and glucose consumption rate. The hydrogen yield of 2.45 (+/- 0.09) mol H-2/mol glucose, the hydrogen concentration of 68% in the produced gas and the composition of the end products in the digestate, i.e. 62.3 (+/- 2.5)% acetic acid, 23.5 (+/- 2.9)% lactic acid and 2.3 (+/- 0.1)% alanine, remained unaffected at increasing biomass concentrations. (C) 2018 Hydrogen Energy Publications LLC. Published by Elsevier Ltd. All rights reserved.

Treatment of food waste by anaerobic digestion can lead to an energy production coupled to a reduction of the volume and greenhouse gas emissions from this waste type. According to EU Regulation EC1774/2002, food waste should be pasteurized/sterilized before or after anaerobic digestion. With respect to this regulation and also considering the slow kinetics of the anaerobic digestion process, thermal and chemical pretreatments of food waste prior to mesophilic anaerobic digestion were studied. A series of batch experiments to determine the biomethane potential of untreated as well as pretreated food waste was carried out. All tested conditions of both thermal and ozonation pretreatments resulted in an enhanced biomethane production. The kinetics of the anaerobic digestion process were, however, accelerated by thermal pretreatment at lower temperatures (

Microalgae cultivation systems fed with wastewater as source of nutrients represents the principal sustainable condition to produce microalgal biomass to be converted conveniently to biofuels. In order to optimize microalgae growth and their lipid content, the effect of light intensity and nutrients load in real wastewater was investigated through batch microalgal cultivation tests. A microalgal polyculture was used as inoculum and grown for 10 days in batch at different conditions of light intensity (i.e. 20, 50 and 100 μmol s−1m−2) and nutrients concentration in wastewater. Experimental results showed that biomass productivity decreased for rich nutrients conditions and increased for high light intensities. The highest lipid mass content (29%) was found for high light intensity condition (100 μmol s−1m−2). Furthermore, microalgae settleability tests, conducted at the end of the cultivation time, resulted in the highest biomass recovery efficiency (72%) for low light intensity and nutrients supply conditions.

Dairy wastes can be conveniently processed and valorized in a biorefinery value chain since they are abundant, zero-cost and all year round available. For a comprehensive knowledge of the microbial species involved in producing biofuels and valuable intermediates from dairy wastes, the changes in bacterial and archaeal population were evaluated when H2, CH4 and chemical intermediates were produced. Batch anaerobic tests were conducted with a mixture of mozzarella cheese whey and buttermilk as organic substrate, inoculated with 1% and 3% w/v industrial animal manure pellets. The archaeal methanogens concentration increased in the test inoculated at 3% (w/v) when H2 and CH4 production occurred, being 1 log higher than that achieved in the test inoculated at 1% (w/v). Many archaeal species, mostly involved in the production of CH4, were identified by sequencing denaturing gradient gel electrophoresis (DGGE) bands. Methanoculleus, Methanocorpusculum and Methanobrevibacter genera were dominant archaea involved in the anaerobic process for bioenergy production from mozzarella cheese whey and buttermilk mixture.

Solid-liquid extraction (adsorption or ion exchange) is a promising approach for the in situ separation of organic acids from fermentation broths. In this study, a diluted concentration of lactic acid (< 10 g/L) separation from a model fermentation broth by granular activated carbon (GAC) as well as weak (Reillex (R) 425 or RLX425) and strong (Amberlite (R) IRA-400 or AMB400) base anion exchange resins under various operating conditions was experimentally investigated. Thermodynamic analysis showed that the best lactic acid adsorption performances were obtained at a pH below the pKa value of lactic acid (i. e., 3.86) for GAC and RLX425 by physical adsorption mechanism and above the pKa value for the AMB400 resin by an ion exchange mechanism, respectively. The adsorption capacity for GAC (38.2 mg/g) was the highest, followed by AMB400 (31.2 mg/g) and RLX425 (17.2 mg/g). As per the thermodynamic analysis, the lactic acid adsorbed onto GAC and RLX425 through a physical adsorption mechanism, whereas the lactic acid adsorbed onto AMB400 with an ion exchange mechanism. The Langmuir adsorption isotherm model (R-2 > 0.96) and the pseudo-second order kinetic model (R-2 similar to 1) fitted better to the experimental data than the other models tested. Postulating the conditions for the real fermentation broth (pH: 5.0-6.5 and temperature: 30-80 degrees C), the resin AMB400 represents an ideal candidate for the extraction of lactic acid during fermentation.

Recently, issues concerning the sustainable and harmless disposal of organic solid waste have generated interest in microbial biotechnologies aimed at converting waste materials into bioenergy and biomaterials, thus contributing to a reduction in economic dependence on fossil fuels. To valorize biomass, waste materials derived from agriculture, food processing factories, and municipal organic waste can be used to produce biopolymers, such as biohydrogen and biogas, through different microbial processes. In fact, different bacterial strains can synthesize biopolymers to convert waste materials into valuable intracellular (e.g., polyhydroxyalkanoates) and extracellular (e.g., exopolysaccharides) bioproducts, which are useful for biochemical production. In particular, large numbers of bacteria, including Alcaligenes eutrophus, Alcaligenes latus, Azotobacter vinelandii, Azotobacter chroococcum, Azotobacter beijerincki, methylotrophs, Pseudomonas spp., Bacillus spp., Rhizobium spp., Nocardia spp., and recombinant Escherichia coli, have been successfully used to produce polyhydroxyalkanoates on an industrial scale from different types of organic by-products. Therefore, the development of high-performance microbial strains and the use of by-products and waste as substrates could reasonably make the production costs of biodegradable polymers comparable to those required by petrochemical-derived plastics and promote their use. Many studies have reported use of the same organic substrates as alternative energy sources to produce biogas and biohydrogen through anaerobic digestion as well as dark and photofermentation processes under anaerobic conditions. Therefore, concurrently obtaining bioenergy and biopolymers at a reasonable cost through an integrated system is becoming feasible using by-products and waste as organic carbon sources. An overview of the suitable substrates and microbial strains used in low-cost polyhydroxyalkanoates for biohydrogen and biogas production is given. The possibility of creating a unique integrated system is discussed because it represents a new approach for simultaneously producing energy and biopolymers for the plastic industry using by-products and waste as organic carbon sources.

The removal of polycyclic aromatic hydrocarbons (PAHs) from a spiked OECD (Organisation for Economic Co-operation and Development) artificial soil was investigated. Laboratory-scale thermally insulated bioremediation reactors were used to implement biostimulation strategy of composting. The selected PAHs included anthracene, chrysene, benzo(k)fluoranthene, and benzo(a)pyrene with an initial concentration of 658 mg of USEPA 16 PAHs kg−1 soil (d/w). The contaminants’ removal was improved by amending the contaminated soil with four different types of fresh organic waste. After 140 days of incubation, the removal of three-ring and four-ring PAHs in all reactors was higher than five-ring PAHs. The reactor displaying a mesophilic phase during bioremediation ended with a removal of 89 and 59 % for three-ring and four-ring PAHs, respectively. In contrast the reactor displaying a thermophilic phase ended with 71 and 41 % removal for three-ring and four-ring PAHs, respectively. The highest five-ring PAH removal was obtained for reactors with buffalo manure and sewage sludge amendments (40 and 33 %, respectively), while food and kitchen waste and fruit and vegetable waste amendments showed less efficiency (26 and 8 %, respectively). Microtox® test data indicated lower toxicity in reactor amended with sewage sludge considering that this setup reached the highest PAH removal and DHA (dehydrogenase activity) compared to others.

A kinetic model of fermentative hydrogen (H-2) production was developed for the hyper-thermophilic marine bacterium Thermotoga neapolitana DSMZ 4359(T) using glucose as the substrate. The model was built based on anaerobic digestion model no.1 (ADM1) by IWA using Monod-like kinetic equations. Several series of pH controlled batch fermentation tests were performed under pure CO2 or N-2 atmosphere in serum bottles to evaluate the growth kinetic parameters. The kinetic parameters were estimated by applying the nonlinear least square method to Monod and Michaelis Menten kinetic equations. The experimentally estimated value of k, k(s), Y and k(d) were 0.839 h(-1), 1.42 g/L, 0.1204 and 0.0043 h(-1), respectively. The kinetic parameters were analyzed for sensitivities and subsequently calibrated and validated by a separate set of experimental results. The model was particularly effective in estimating the biomass growth, substrate consumption, and H-2 production except for acetate and lactate production, due to capnophilic condition (under CO2 atmosphere). Copyright (C) 2016, Hydrogen Energy Publications, LLC. Published by Elsevier Ltd. All rights reserved.

The aim of the present study was to develop a kinetic model for a recently proposed unique and novel metabolic process called capnophilic (CO2-requiring) lactic fermentation (CLF) pathway in Thermotoga neapolitana. The model was based on Monod kinetics and the mathematical expressions were developed to enable the simulation of biomass growth, substrate consumption and product formation. The calibrated kinetic parameters such as maximum specific uptake rate (k), semi-saturation constant (k(S)), biomass yield coefficient (Y) and endogenous decay rate (k(d)) were 1.30 h(-1), 1.42 g/L, 0.1195 and 0.0205 h(-1), respectively. A high correlation (>0.98) was obtained between the experimental data and model predictions for both model validation and cross validation processes. An increase of the lactate production in the range of 40-80% was obtained through CLF pathway compared to the classic dark fermentation model. The proposed kinetic model is the first mechanistically based model for the CLF pathway. This model provides useful information to improve the knowledge about how acetate and CO2 are recycled back by Thermotoga neapolitana to produce lactate without compromising the overall hydrogen yield. (C) 2016 Elsevier Ltd. All rights reserved.

This paper presents the results of an experimental study aimed at investigating the effect of operative parameters on the efficiency of a soil flushing process, conducted on real contaminated soil containing high amounts of Cu and Zn. Soil flushing tests were carried out with Ethylenediamine-N,N′ -disuccinic acid (EDDS) as a flushing agent due to its high biodegradability and environmentally friendly characteristics. Process parameters such as Empty-Bed Contact Time (EBCT) and EDDS solution molarity were varied from 21–33 h and from 0.36–3.6 mM, respectively. Effects on the mobility of cations such as Fe and Mn were also investigated. Results showed that very high performances can be obtained at [EDDS] = 3.6 mM and EBCT = 33 h. In these conditions, in fact, the amount of removed Cu was 53%, and the amount of removed Zn was 46%. Metal distribution at different depths from the top surface revealed that Cu has higher mobility than Zn. The process results were strongly dependent on the exchange of metals due to the different stability constants of the EDDS complexes. Finally, results from a comparative study showed that soil washing treatment reached the same removal efficiency of the flushing process in a shorter time but required a larger amount of the EDDS solution.

In an Anaerobic Digestion (AD) process treating particulate substrates, the size of solids is expected to negatively affect the rate of hydrolysis step and consequently influence the performance of the whole process. To avoid any disadvantage due to size of solids, expensive pre-treatments aimed at disintegrating and solubilizing substrates are commonly conducted prior to AD. This practice is doubtlessly successful, but not always necessary, since some organic substrates, although particulate, once immersed in water, tend to solubilize immediately. This aspect, if properly considered, could result in saving money and time in the AD process, as well as refining the development and calibration of AD mathematical models. The present study is actually aimed at demonstrating, through experiments and mathematical simulations, different results deriving from the AD process performed, under the same operating conditions, on two different substrates, i.e. homemade pasta and carrot batons, having the same particle size, but different chemical composition and texture. Experimental outcomes highlighted the effect of particles size on bio-methane production only from the bio-methanation potential tests (BMP) conducted on carrot batons. Similar results were obtained by mathematical model calibration, i.e. , different kinetic constants for differently-sized carrot batons and same kinetic constant for differently-sized homemade pasta solids.

A new procedure, recently proposed for on-line monitoring of copper released from metal pipes in household plumbing system for drinking water distribution during the development of corrosion processes, is tested experimentally. Experiments were carried out in laboratory controlled conditions, using synthetic water and varying the water alkalinity. The possibility of using the corrosion potential as a surrogate measure of copper concentration in stagnating water is shown, verifying, in the meantime, the effect of alkalinity on the development of passivation phenomena, which tend to protect the pipe from corrosion processes. Experimental data are discussed, highlighting the potentiality of the procedure, and recognizing its limitations. © IWA Publishing 2008.

This study focused on the supersaturation of hydrogen in the liquid phase (H2aq) and its inhibitory effect on dark fermentation by Thermotoga neapolitana cf. capnolactica by increasing the agitation (from 100 to 500 rpm) and recirculating H2-rich biogas (GaR). At low cell concentrations, both 500 rpm and GaR reduced the H2aq from 30.1 (±4.4) mL/L to the lowest values of 7.4 (±0.7) mL/L and 7.2 (±1.2) mL/L, respectively. However, at high cell concentrations (0.79 g CDW/L), the addition of GaR at 300 rpm was more efficient and increased the hydrogen production rate by 271%, compared to a 136% increase when raising the agitation to 500 rpm instead. While H2aq primarily affected the dark fermentation rate, GaR concomitantly increased the hydrogen yield up to 3.5 mol H2/mol glucose. Hence, H2aq supersaturation highly depends on the systems gas-liquid mass transfer and strongly inhibits dark fermentation.