Microbial electrosynthesis of acetate from CO2 in three-chamber cells with gas diffusion biocathode under moderate saline conditions.

The industrial adoption of microbial electrosynthesis (MES) is hindered by high overpotentials deriving from low electrolyte conductivity and inefficient cell designs. In this study, a mixed microbial consortium originating from an anaerobic digester operated under saline conditions (∼13 g L-1 NaCl) was adapted for acetate production from bicarbonate in galvanostatic (0.25 mA cm-2) H-type cells at 5, 10, 15, or 20 g L-1 NaCl concentration. The acetogenic communities were successfully enriched only at 5 and 10 g L-1 NaCl, revealing an inhibitory threshold of about 6 g L-1 Na+. The enriched planktonic communities were then used as inoculum for 3D printed, three-chamber cells equipped with a gas diffusion biocathode. The cells were fed with CO2 gas and operated galvanostatically (0.25 or 1.00 mA cm-2). The highest production rate of 55.4 g m-2 d-1 (0.89 g L-1 d-1), with 82.4% Coulombic efficiency, was obtained at 5 g L-1 NaCl concentration and 1 mA cm-2 applied current, achieving an average acetate production of 44.7 kg MWh-1. Scanning electron microscopy and 16S rRNA sequencing analysis confirmed the formation of a cathodic biofilm dominated by Acetobacterium sp. Finally, three 3D printed cells were hydraulically connected in series to simulate an MES stack, achieving three-fold production rates than with the single cell at 0.25 mA cm-2. This confirms that three-chamber MES cells are an efficient and scalable technology for CO2 bio-electro recycling to acetate and that moderate saline conditions (5 g L-1 NaCl) can help reduce their power demand while preserving the activity of acetogens.

A meta-analysis of acetogenic and methanogenic microbiomes in microbial electrosynthesis.

A meta-analysis approach was used, to study the microbiomes of biofilms and planktonic communities underpinning microbial electrosynthesis (MES) cells. High-throughput DNA sequencing of 16S rRNA gene amplicons has been increasingly applied to understand MES systems. In this meta-analysis of 22 studies, we find that acetogenic and methanogenic MES cells share 80% of a cathodic core microbiome, and that different inoculum pre-treatments strongly affect community composition. Oxygen scavengers were more abundant in planktonic communities, and several key organisms were associated with operating parameters and good cell performance. We suggest Desulfovibrio sp. play a role in initiating early biofilm development and shaping microbial communities by catalysing H2 production, to sustain either Acetobacterium sp. or Methanobacterium sp. Microbial community assembly became more stochastic over time, causing diversification of the biofilm (cathodic) community in acetogenic cells and leading to re-establishment of methanogens, despite inoculum pre-treatments. This suggests that repeated interventions may be required to suppress methanogenesis.

Dark fermentative volatile fatty acids production from food waste: A review of the potential central role in waste biorefineries.

Volatile fatty acids (VFAs) are high-value chemicals that are increasingly demanded worldwide. Biological production via food waste (FW) dark fermentation (DF) is a promising option to achieve the sustainability and environmental benefits typical of biobased chemicals and concurrently manage large amounts of residues. DF has a great potential to play a central role in waste biorefineries due to its ability to hydrolyze and convert complex organic substrates into VFAs that can be used as building blocks for bioproducts, chemicals and fuels. Several challenges must be faced for full-scale implementation, including process optimization to achieve high and stable yields, the development of efficient techniques for selective recovery and the cost-effectiveness of the whole process. This review aims to critically discuss and statistically analyze the existing relationships between process performance and the main variables of concern. Moreover, opportunities, current challenges and perspectives of a FW-based and fermentation-centred biorefinery layout are discussed.

Membrane-based technologies for biohydrogen production: A review.

Overcoming the existing environmental issues and the gradual depletion of energy sources is a priority at global level, biohydrogen can provide a sustainable and reliable energy reserve. However, the process instability and low biohydrogen yields are still hindering the adoption of biohydrogen production plants at industrial scale. In this context, membrane-based biohydrogen production technologies, and in particular fermentative membrane bioreactors (MBRs) and microbial electrolysis cells (MECs), as well as downstream membrane-based technologies such as electrodialysis (ED), are suitable options to achieve high-rate biohydrogen production. We have shed the light on the research efforts towards the development of membrane-based technologies for biohydrogen production from organic waste, with special emphasis to the reactor design and materials. Besides, techno-economic analyses have been traced to ensure the suitability of such technologies in bio-H2 production. Operation parameters such as pH, temperature and organic loading rate affect the performance of MBRs. MEC and ED technologies also are highly affected by the chemistry of the membrane used and anode material as well as the operation parameters. The limitations and future directions for application of membrane-based biohydrogen production technologies have been individuated. At the end, this review helps in the critical understanding of deploying membrane-based technologies for biohydrogen production, thereby encouraging future outcomes for a sustainable biohydrogen economy.

Cathodic biofilms - A prerequisite for microbial electrosynthesis.

Cathodic biofilms have an important role in CO2 bio-reduction to carboxylic acids and biofuels in microbial electrosynthesis (MES) cells. However, robust and resilient electroactive biofilms for an efficient CO2 conversion are difficult to achieve. In this review, the fundamentals of cathodic biofilm formation, including energy conservation, electron transfer and development of catalytic biofilms, are presented. In addition, strategies for improving cathodic biofilm formation, such as the selection of electrode and carrier materials, cell design and operational conditions, are described. The knowledge gaps are individuated, and possible solutions are proposed to achieve stable and productive biofilms in MES cathodes.

Microbial electrosynthesis: Towards sustainable biorefineries for production of green chemicals from CO2 emissions.

Decarbonisation of the economy has become a priority at the global level, and the resulting legislative pressure is pushing the chemical and energy industries away from fossil fuels. Microbial electrosynthesis (MES) has emerged as a promising technology to promote this transition, which will further benefit from the decreasing cost of renewable energy. However, several technological challenges need to be addressed before the MES technology can reach its maturity. The aim of this review is to critically discuss the bottlenecks hampering the industrial adoption of MES, considering the whole production process (from the CO2 source to the marketable products), and indicate future directions. A flexible stack design, with flat or tubular MES modules and direct CO2 supply, is required for site-specific decentralised applications. The experience gained for scaling-up electrochemical cells (e.g. electrolysers) can serve as a guideline for realising pilot MES stacks to be technologically and economically evaluated in industrially relevant conditions. Maximising CO2 abatement rate by targeting high-rate production of acetate can promote adoption of MES technology in the short term. However, the development of a replicable and robust strategy for production and in-line extraction of higher-value products (e.g. caproic acid and hexanol) at the cathode, and meaningful exploitation of the currently overlooked anodic reactions, can further boost MES cost-effectiveness. Furthermore, the use of energy storage and smart electronics can alleviate the fluctuations of renewable energy supply. Despite the unresolved challenges, the flexible MES technology can be applied to decarbonise flue gas from different sources, to upgrade industrial and wastewater treatment plants, and to produce a wide array of green and sustainable chemicals. The combination of these benefits can support the industrial adoption of MES over competing technologies.

Carboxylic acids production and electrosynthetic microbial community evolution under different CO2 feeding regimens.

Microbial electrosynthesis (MES) is a potential technology for CO2 recycling, but insufficient information is available on the microbial interactions underpinning electrochemically-assisted reactions. In this study, a MES reactor was operated for 225 days alternately with bicarbonate or CO2 as carbon source, under batch or continuous feeding regimens, to evaluate the response of the microbial communities, and their productivity, to dynamic operating conditions. A stable acetic acid production rate of 9.68 g m-2 d-1, and coulombic efficiency up to 40%, was achieved with continuous CO2 sparging, higher than the rates obtained with bicarbonate (0.94 g m-2 d-1) and CO2 under fed-batch conditions (2.54 g m-2 d-1). However, the highest butyric acid production rate (0.39 g m-2 d-1) was achieved with intermittent CO2 sparging. The microbial community analyses focused on differential amplicon sequence variants (ASVs), allowing detection of ASVs significantly different across consecutive samples. This analysis, combined with co-occurence network analysis, and cyclic voltammetry, indicated that hydrogen-mediated acetogenesis was carried out by Clostridium, Eubacterium and Acetobacterium, whereas Oscillibacter and Caproiciproducens were involved in butyric acid production. The cathodic community was spatially inhomogeneous, with potential electrotrophs, such as Sulfurospirillum and Desulfovibrio, most prevalent near the current collector. The abundance of Sulfurospirillum positively correlated with that of Acetobacterium, supporting the syntrophic metabolism of both organisms.

The dairy biorefinery: Integrating treatment processes for cheese whey valorisation.

With an estimated worldwide production of 190 billion kg per year, and due to its high organic load, cheese whey represents a huge opportunity for bioenergy and biochemicals production. Several physical, chemical and biological processes have been proposed to valorise cheese whey by producing biofuels (methane, hydrogen, and ethanol), electric energy, and/or chemical commodities (carboxylic acids, proteins, and biopolymers). A biorefinery concept, in which several value-added products are obtained from cheese whey through a cascade of biotechnological processes, is an opportunity for increasing the product spectrum of dairy industries while allowing for sustainable management of the residual streams and reducing disposal costs for the final residues. This review critically analyses the different treatment options available for energy and materials recovery from cheese whey, their combinations and perspectives for implementation. Thus, instead of focusing on a specific valorisation platform, in the present review the most relevant aspects of each strategy are analysed to support the integration of different routes, in order to identify the most appropriate treatment train.

Organic waste biorefineries: Looking towards implementation.

The concept of biorefinery expands the possibilities to extract value from organic matter in form of either bespoke crops or organic waste. The viability of biorefinery schemes depends on the recovery of higher-value chemicals with potential for a wide distribution and an untapped marketability. The feasibility of biorefining organic waste is enhanced by the fact that the biorefinery will typically receive a waste management fee for accepting organic waste. The development and implementation of waste biorefinery concepts can open up a wide array of possibilities to shift waste management towards higher sustainability. However, barriers encompassing environmental, technical, economic, logistic, social and legislative aspects need to be overcome. For instance, waste biorefineries are likely to be complex systems due to the variability, heterogeneity and low purity of waste materials as opposed to dedicated biomasses. This article discusses the drivers that can make the biorefinery concept applicable to waste management and the possibilities for its development to full scale. Technological, strategic and market constraints affect the successful implementations of these systems. Fluctuations in waste characteristics, the level of contamination in the organic waste fraction, the proximity of the organic waste resource, the markets for the biorefinery products, the potential for integration with other industrial processes and disposal of final residues are all critical aspects requiring detailed analysis. Furthermore, interventions from policy makers are necessary to foster sustainable bio-based solutions for waste management.

OpenTCC: An open source low-cost temperature-control chamber.

Microbial electrochemical technologies (MET) are emerging systems for environmental applications such as renewable energy production or pollution remediation. MET research often requires stable temperatures and low levels of electromagnetic interference. Due to the presence of electrical wires and sensors, heating MET using water jacket recirculation can raise safety issues, whereas heating coils may affect the results of electrochemical analyses. The proposed open-source temperature-control chamber (OpenTCC) aims to provide a low-cost solution for controlling temperature (in the range 20-55 °C) while simultaneously reducing the electromagnetic interferences caused by switching mode power supplies. OpenTCC consists of a light and cheap structure, incorporating eight heating pads and two Peltier-cooling modules powered by open-source electronic circuits. Its hardware is controlled by an Arduino microcontroller and a Python interface which provides data-logging and serve as a basis for programable temperature cycles. The system has a modular design to allow stacking several independent modules. OpenTCC provides a reliable and tunable temperature control at lower costs than currently available commercial temperature controllers and provides a platform for field-specific upgrades. Though optimized for MET, Open-TCC can be adapted to other laboratory applications due to its flexible design.

Propionate Production by Bioelectrochemically-Assisted Lactate Fermentation and Simultaneous CO2 Recycling.

Production of volatile fatty acids (VFAs), fundamental building blocks for the chemical industry, depends on fossil fuels but organic waste is an emerging alternative substrate. Lactate produced from sugar-containing waste streams can be further processed to VFAs. In this study, electrofermentation (EF) in a two-chamber cell is proposed to enhance propionate production via lactate fermentation. At an initial pH of 5, an applied potential of -1 V vs. Ag/AgCl favored propionate production over butyrate from 20 mM lactate (with respect to non-electrochemical control incubations), due to the pH buffering effect of the cathode electrode, with production rates up to 5.9 mM d-1 (0.44 g L-1 d-1). Microbial community analysis confirmed the enrichment of propionate-producing microorganisms, such as Tyzzerella sp. and Propionibacterium sp. Organisms commonly found in microbial electrosynthesis reactors, such as Desulfovibrio sp. and Acetobacterium sp., were also abundant at the cathode, indicating their involvement in recycling CO2 produced by lactate fermentation into acetate, as confirmed by stoichiometric calculations. Propionate was the main product of lactate fermentation at substrate concentrations up to 150 mM, with a highest production rate of 12.9 mM d-1 (0.96 g L-1 d-1) and a yield of 0.48 mol mol-1 lactate consumed. Furthermore, as high as 81% of the lactate consumed (in terms of carbon) was recovered as soluble product, highlighting the potential for EF application with high-carbon waste streams, such as cheese whey or other food wastes. In summary, EF can be applied to control lactate fermentation toward propionate production and to recycle the resulting CO2 into acetate, increasing the VFA yield and avoiding carbon emissions and addition of chemicals for pH control.

Microbial electrochemical technologies: Electronic circuitry and characterization tools.

Microbial electrochemistry merges microbiology, electrochemistry and electronics to provide a set of technologies for environmental engineering applications. Understanding the electronic concepts is crucial for effectively adopting these systems, but the importance of electronic circuitry is often overlooked by microbial electrochemistry researchers. This review provides the background on the electronics and electrochemical concepts involved in the study of microorganisms interacting with electricity, and their applications in microbial electrochemical technology (MET). The potentiostat circuitry is described along with its working principles. Electrochemical analyses are presented together with the rational and parameters employed to study MET devices and electroactive microorganisms. Finally, future directions are delineated towards the adoption of MET, and the related electronics, in environmental engineering applications.

Power production and microbial community composition in thermophilic acetate-fed up-flow and flow-through microbial fuel cells.

The microbial communities developed from a mixed-species culture in up-flow and flow-through configurations of thermophilic (55 °C) microbial fuel cells (MFCs), and their power production from acetate, were investigated. The up-flow MFC was operated for 202 days, obtaining an average power density of 0.13 W/m3, and Tepidiphilus was the dominant transcriptionally-active microorganisms. The planktonic community developed in the up-flow MFC was used to inoculate a flow-through MFC resulting in the proliferation of Ureibacillus, whose relative abundance increased from 1 to 61% after 45 days. Despite the differences between the up-flow and flow-through MFCs, including the anode electrode, hydrodynamic conditions, and the predominant microorganism, similar (p = 0.05) volumetric power (0.11-0.13 W/m3), coulombic efficiency (16-18%) and acetate consumption rates (55-69 mg/L/d) were obtained from both. This suggests that though MFC design can shape the active component of the thermophilic microbial community, the consortia are resilient and can maintain similar performance in different MFC configurations.

Composition and role of the attached and planktonic microbial communities in mesophilic and thermophilic xylose-fed microbial fuel cells.

A mesophilic (37 °C) and a thermophilic (55 °C) two-chamber microbial fuel cell (MFC) were studied and compared for their power production from xylose and the microbial communities involved. The anode-attached, membrane-attached, and planktonic microbial communities, and their respective active subpopulations, were determined by next generation sequencing (Illumina MiSeq), based on the presence and expression of the 16S rRNA gene. Geobacteraceae accounted for 65% of the anode-attached active microbial community in the mesophilic MFC, and were associated to electricity generation likely through direct electron transfer, resulting in the highest power production of 1.1 W m-3. A lower maximum power was generated in the thermophilic MFC (0.2 W m-3), likely due to limited acetate oxidation and the competition for electrons by hydrogen oxidizing bacteria and hydrogenotrophic methanogenic archaea. Aerobic microorganisms, detected among the membrane-attached active community in both the mesophilic and thermophilic MFC, likely acted as a barrier for oxygen flowing from the cathodic chamber through the membrane, favoring the strictly anaerobic exoelectrogenic microorganisms, but competing with them for xylose and its degradation products. This study provides novel information on the active microbial communities populating the anodic chamber of mesophilic and thermophilic xylose-fed MFCs, which may help in developing strategies to favor exoelectrogenic microorganisms at the expenses of competing microorganisms.

Biohydrogen production from xylose by fresh and digested activated sludge at 37, 55 and 70 °C.

Two heat-treated inocula, fresh and digested activated sludge from the same municipal wastewater treatment plant, were compared for their H2 production via dark fermentation at mesophilic (37 °C), thermophilic (55 °C) and hyperthermophilic (70 °C) conditions using xylose as the substrate. At both 37 and 55 °C, the fresh activated sludge yielded more H2 than the digested sludge, whereas at 70 °C, neither of the inocula produced H2 effectively. A maximum yield of 1.85 mol H2 per mol of xylose consumed was obtained at 55 °C. H2 production was linked to acetate and butyrate production, and there was a linear correlation (R2 = 0.96) between the butyrate and H2 yield for the fresh activated sludge inoculum at 55 °C. Approximately 2.4 mol H2 per mol of butyrate produced were obtained against a theoretical maximum of 2.0, suggesting that H2 was produced via the acetate pathway prior to switching to the butyrate pathway due to the increased H2 partial pressure. Clostridia sp. were the prevalent species at both 37 and 55 °C, irrespectively of the inoculum type. Although the two inocula originated from the same plant, different thermophilic microorganisms were detected at 55 °C. Thermoanaerobacter sp., detected only in the fresh activated sludge cultures, may have contributed to the high H2 yield obtained with such an inoculum.

Effect of temperature on selenium removal from wastewater by UASB reactors.

The effect of temperature on selenium (Se) removal by upflow anaerobic sludge blanket (UASB) reactors treating selenate and nitrate containing wastewater was investigated by comparing the performance of a thermophilic (55 °C) versus a mesophilic (30 °C) UASB reactor. When only selenate (50 µM) was fed to the UASB reactors (pH 7.3; hydraulic retention time 8 h) with excess electron donor (lactate at 1.38 mM corresponding to an organic loading rate of 0.5 g COD L(-1) d(-1)), the thermophilic UASB reactor achieved a higher total Se removal efficiency (94.4 ± 2.4%) than the mesophilic UASB reactor (82.0 ± 3.8%). When 5000 µM nitrate was further added to the influent, total Se removal was again better under thermophilic (70.1 ± 6.6%) when compared to mesophilic (43.6 ± 8.8%) conditions. The higher total effluent Se concentration in the mesophilic UASB reactor was due to the higher concentrations of biogenic elemental Se nanoparticles (BioSeNPs). The shape of the BioSeNPs observed in both UASB reactors was different: nanospheres and nanorods, respectively, in the mesophilic and thermophilic UASB reactors. Microbial community analysis showed the presence of selenate respirers as well as denitrifying microorganisms.

Entrapped elemental selenium nanoparticles affect physicochemical properties of selenium fed activated sludge.

Selenite containing wastewaters can be treated in activated sludge systems, where the total selenium is removed from the wastewater by the formation of elemental selenium nanoparticles, which are trapped in the biomass. No studies have been carried out so far on the characterization of selenium fed activated sludge flocs, which is important for the development of this novel selenium removal process. This study showed that more than 94% of the trapped selenium in activated sludge flocs is in the form of elemental selenium, both as amorphous/monoclinic selenium nanospheres and trigonal selenium nanorods. The entrapment of the elemental selenium nanoparticles in the selenium fed activated sludge flocs leads to faster settling rates, higher hydrophilicity and poorer dewaterability compared to the control activated sludge (i.e., not fed with selenite). The selenium fed activated sludge showed a less negative surface charge density as compared to the control activated sludge. The presence of trapped elemental selenium nanoparticles further affected the spatial distribution of Al and Mg in the activated sludge flocs. This study demonstrated that the formation and subsequent trapping of elemental selenium nanoparticles in the activated sludge flocs affects their physicochemical properties.