Second-Generation Magnesium Phosphates as Water Extractant Agents in Forward Osmosis and Subsequent Use in Hydroponics.

The recovery of nutrients from wastewater streams for their later use in agricultural fertilization is an interesting approach. Wastewater recovered magnesium phosphate (MgP) salts were used in a forward osmosis (FO) system as draw solution in order to extract water and to produce a nutrient solution to be used in a hydroponic system with lettuces (Lactuca sativa, L.). Owing to the low solubility of the MgP salts (i.e., struvite, hazenite and cattiite) in water, acid dissolution was successfully tested using citric and nitric acids to reach pH 3.0. The dilution by FO of the dissolved salts reached levels close to those needed by a hydroponic culture. Ion migration through the membrane was medium to high, and although it did not limit the dilution potential of the system, it might decrease the overall feasibility of the FO process. Functional growth of the lettuces in the hydroponic system was achieved with the three MgP salts using the recovered water as nutrient solution, once properly supplemented with nutrients with the desired concentrations. This is an innovative approach for promoting water reuse in hydroponics that benefits from the use of precipitated MgP salts as a nutrient source.

Exploring the recovery of potassium-rich struvite after a nitrification-denitrification process in pig slurry treatment.

The integration of biological nitrogen (N) removal with struvite-type material recovery, which contained phosphorus (P) and potassium (K), was proved to be technically feasible in pig slurry treatment. Phosphate (PO4) salts were precipitated by raising the pH-value, using denitrified effluent and waste sludge purged from the bioreactor. When P was limiting, the unbalanced composition of the denitrified effluent resulted in low K-removal efficiency from the liquid phase; 10 % maximum when the initial pH-value was adjusted to 11.5 (93 % PO4-P recovery). By processing the waste sludge in two steps, using first ethylenediaminetetraacetic acid (EDTA) as an acidifier to release PO4 while preventing calcium interference, the K-removal efficiency reached 25 % (75 % PO4-P recovery). When K was limiting, the addition of newberyite particles resulted in the highest K-removal efficiency, up to 90 % (under online pH control to 10.5). Overall, new opportunities are envisaged for producing second-generation fertilizers potentially containing 0-1 % N, 11-17 % P and 6-8 % K.

Assessment of zooplankton-based eco-sustainable wastewater treatment at laboratory scale.

The combination of the filtration capacity of zooplankton (e.g. Daphnia) with the nutrient removal capacity of bacterial/algal biofilm in a zooplankton-containing reactor could provide a natural-based alternative for wastewater treatment. A laboratory-scale zooplankton-based reactor was tested at different HRTs resulting in a significant reduction in nutrient concentrations in wastewater when the system was operated at HRTs longer than 1.1 days (preferably of between 2 and 4 days). However, the presence of high concentrations of organic matter (>250 mg COD L-1) in the wastewater inhibited zooplankton activity, limiting its use to tertiary treatment. Therefore, in combination with other natural treatments that can perform primary and secondary treatments, zooplankton may provide a solution for wastewater clarification and nutrient polishing. The effect of a common metal such as copper on the filtration capacity of Daphnia was also evaluated. Daphnia, as well as the whole zooplankton-based reactor, adapted to copper concentrations of up to 70 µg Cu L-1 but an overload of 380 µg Cu L-1 for two-weeks severely affected the biological system.

Biogas upgrading, CO2 valorisation and economic revaluation of bioelectrochemical systems through anodic chlorine production in the framework of wastewater treatment plants.

Biogas production in wastewater treatment plants (WWTPs) plays a decisive role in the reduction of CO2 emissions and energy needs in the context of the water-energy nexus. The biogas obtained from sewage sludge digestion can be converted into biomethane by the use of biogas upgrading technologies. In this regard, an innovative water scrubbing based technology, known as ABAD Bioenergy® is presented and considered in this work. The effluents resulting from this system consist of biomethane and treated wastewater with a high CO2 concentration. Therefore, the study explores the feasibility of using this CO2-containing effluent in the cathode of a bioelectrochemical system (BES) for the transformation of CO2 into methane. Techno-economic assessment of the process is presented, including the valorisation of anode reactions through the production of chlorine compounds. Finally, the potential impacts of applying this technology in a WWTP operated by FCC Aqualia are (i) increasing biomethane production by 17.4%, (ii) decreasing CO2 content by 42.8% and (iii) producing over 60 ppm of chlorine compounds to disinfect all the treated wastewater of the plant.

[NiFe]-hydrogenases are constitutively expressed in an enriched Methanobacterium sp. population during electromethanogenesis.

Electromethanogenesis is the bioreduction of carbon dioxide (CO2) to methane (CH4) utilizing an electrode as electron donor. Some studies have reported the active participation of Methanobacterium sp. in electron capturing, although no conclusive results are available. In this study, we aimed at determining short-time changes in the expression levels of [NiFe]-hydrogenases (Eha, Ehb and Mvh), heterodisulfide reductase (Hdr), coenzyme F420-reducing [NiFe]-hydrogenase (Frh), and hydrogenase maturation protein (HypD), according to the electron flow in independently connected carbon cloth cathodes poised at- 800 mV vs. standard hydrogen electrode (SHE). Amplicon massive sequencing of cathode biofilm confirmed the presence of an enriched Methanobacterium sp. population (>70% of sequence reads), which remained in an active state (78% of cDNA reads), tagging this archaeon as the main methane producer in the system. Quantitative RT-PCR determinations of ehaB, ehbL, mvhA, hdrA, frhA, and hypD genes resulted in only slight (up to 1.5 fold) changes for four out of six genes analyzed when cells were exposed to open (disconnected) or closed (connected) electric circuit events. The presented results suggested that suspected mechanisms for electron capturing were not regulated at the transcriptional level in Methanobacterium sp. for short time exposures of the cells to connected-disconnected circuits. Additional tests are needed in order to confirm proteins that participate in electron capturing in Methanobacterium sp.

Specific detection of "Clostridium autoethanogenum", Clostridium ljungdahlii and Clostridium carboxidivorans in complex bioreactor samples.

The high genetic similarity between some carboxydotrophic bacteria does not allow for the use of common sequencing techniques targeting the 16S rRNA gene for species identification. 16S rRNA sequencing fails to discriminate among Clostridium ljungdahlii and 'Clostridium autoethanogenum', despite this two species exhibit significant differences in CO2 assimilation and alcohol production. In this work we designed PCR primers targeting for the DNA gyrase subunit A (gyrA) and a putative formate/nitrite transporter (fnt) to specifically detect the presence of 'C. autoethanogenum', C. ljungdahlii or Clostridium carboxidivorans. We could confirm the simultaneous presence of C. ljungdahlii and 'C. autoethanogenum' in different bioreactors, and a preference of the latter for high CO2 content.

Effect of ethanol and butanol on autotrophic growth of model homoacetogens.

Research efforts aimed at increasing ethanol and butanol productivity from syngas are currently gaining attention. For most model carboxydotrophic bacteria, production rates, yields and maximum product titres have been studied in detail, but little is known on alcohol toxicity in these bacteria. The aim of this work was to investigate the inhibitory effects of ethanol and butanol on the growth of Clostridium ljungdahlii PETC, C. carboxidivorans P7, and 'Butyribacterium methylotrophicum DSM3468'. Experiments to determine inhibitory effects due to product accumulation were carried out using a synthetic mixture of CO:CO2:H2 as a substrate. These conditions were chosen to mimic gaseous effluents of biomass and waste gasification plants. Inhibition effects were recorded as changes in growth parameters. No significant inhibition was observed for ethanol at concentrations below 15 g/L. The three species exhibited higher sensitivity to butanol. Half inhibition constants for butanol could be estimated for P7 (IC50 = 4.12 g/L), DSM3468 (IC50 = 1.79 g/L), and PETC (IC50 = 9.75 g/L). In conclusion, at least for the tested strains, alcohol toxicity is not an immediate handicap for increasing alcohol production of the tested homoacetogenic strains.

Opportunities for groundwater microbial electro-remediation.

Groundwater pollution is a serious worldwide concern. Aromatic compounds, chlorinated hydrocarbons, metals and nutrients among others can be widely found in different aquifers all over the world. However, there is a lack of sustainable technologies able to treat these kinds of compounds. Microbial electro-remediation, by the means of microbial electrochemical technologies (MET), can become a promising alternative in the near future. MET can be applied for groundwater treatment in situ or ex situ, as well as for monitoring the chemical state or the microbiological activity. This document reviews the current knowledge achieved on microbial electro-remediation of groundwater and its applications.

Microbial electricity driven anoxic ammonium removal.

Removal of nitrogen, mainly in form of ammonium (NH4+), in wastewater treatment plants (WWTPs) is a highly energy demanding process, mainly due to aeration. It causes costs of about half a million Euros per year in an average European WWTP. Alternative, more economical technologies for the removal of nitrogen compounds from wastewater are required. This study proves the complete anoxic conversion of ammonium (NH4+) to dinitrogen gas (N2) in continuously operated bioelectrochemical systems at the litre-scale. The removal rate is comparable to conventional WWTPs with 35 ± 10 g N m-3 d-1 with low accumulation of NO2-, NO3-, N2O. In contrast to classical aerobic nitrification, the energy consumption is considerable lower (1.16 ± 0.21 kWh kg-1 N, being more than 35 times less than for the conventional wastewater treatment). Biotic and abiotic control experiments confirmed that the anoxic nitrification was an electrochemical biological process mainly performed by Nitrosomonas with hydroxylamine as the main substrate (mid-point potential, Eox = +0.67 ± 0.08 V vs. SHE). This article proves the technical feasibility and reduction of costs for ammonium removal from wastewater, investigates the underlying mechanisms and discusses future engineering needs.

Employing Microbial Electrochemical Technology-driven electro-Fenton oxidation for the removal of recalcitrant organics from sanitary landfill leachate.

The feasibility of employing Microbial Electrochemical Technology (MET)-driven electro-Fenton oxidation was evaluated as a post-treatment of an anammox system treating sanitary landfill leachate. Two different MET configuration systems were operated using effluent from partial nitrification-anammox reactor treating mature leachate. In spite of the low organic matter biodegradability of the anammox's effluent (2401±562mgCODL-1; 237±57mgBOD5L-1), the technology was capable to reach COD removal rates of 1077-1244mgL-1d-1 with concomitant renewable electricity production (43.5±2.1Am-3NCC). The operation in continuous mode versus batch mode reinforced the removal capacity of the technology. The recirculation of acidic catholyte into anode chamber hindered the anodic efficiency due to pH stress on anodic electricigens. The obtained results demonstrated that the integrated system is a potentially applicable process to deal with bio-recalcitrant compounds present in mature landfill leachate.

Microbial electrosynthesis of butyrate from carbon dioxide: Production and extraction.

To date acetate is the main product of microbial electrosynthesis (MES) from carbon dioxide (CO2). In this work a tubular bioelectrochemical system was used to carry out MES and enhance butyrate production over the other organic products. Batch tests were performed at a fixed cathode potential of -0.8V vs SHE. The reproducibility of the results according to previous experiments was validated in a preliminary test. According to the literature butyrate production could take place by chain elongation reactions at low pH and high hydrogen partial pressure (pH2). During the experiment, CO2 supply was limited to build up pH2 and trigger the production of compounds with a higher degree of reduction. In test 1 butyrate became the predominant end-product, with a concentration of 59.7mMC versus 20.3mMC of acetate, but limitation on CO2 supply resulted in low product titers. CO2 limitation was relaxed in test 2 to increase the bioelectrochemical activity but increase pH2 and promote the production of butyrate, what resulted in the production of 87.5mMC of butyrate and 34.7mMC of acetate. The consumption of ethanol, and the presence of other products in the biocathode (i.e. caproate) suggested that butyrate production took place through chain elongation reactions, likely driven by Megasphaera sueciensis (>39% relative abundance). Extraction and concentration of butyrate was performed by liquid membrane extraction. A concentration phase with 252.4mMC of butyrate was obtained, increasing also butyrate/acetate ratio to 16.4. The results are promising for further research on expanding the product portfolio of MES.

On the Edge of Research and Technological Application: A Critical Review of Electromethanogenesis.

The conversion of electrical current into methane (electromethanogenesis) by microbes represents one of the most promising applications of bioelectrochemical systems (BES). Electromethanogenesis provides a novel approach to waste treatment, carbon dioxide fixation and renewable energy storage into a chemically stable compound, such as methane. This has become an important area of research since it was first described, attracting different research groups worldwide. Basics of the process such as microorganisms involved and main reactions are now much better understood, and recent advances in BES configuration and electrode materials in lab-scale enhance the interest in this technology. However, there are still some gaps that need to be filled to move towards its application. Side reactions or scaling-up issues are clearly among the main challenges that need to be overcome to its further development. This review summarizes the recent advances made in the field of electromethanogenesis to address the main future challenges and opportunities of this novel process. In addition, the present fundamental knowledge is critically reviewed and some insights are provided to identify potential niche applications and help researchers to overcome current technological boundaries.

The ManureEcoMine pilot installation: advanced integration of technologies for the management of organics and nutrients in livestock waste.

Manure represents an exquisite mining opportunity for nutrient recovery (nitrogen and phosphorus), and for their reuse as renewable fertilisers. The ManureEcoMine proposes an integrated approach of technologies, operated in a pilot-scale installation treating swine manure (83.7%) and Ecofrit® (16.3%), a mix of vegetable residues. Thermophilic anaerobic digestion was performed for 150 days, the final organic loading rate was 4.6 kgCOD m-3 d-1, with a biogas production rate of 1.4 Nm3 m-3 d-1. The digester was coupled to an ammonia side-stream stripping column and a scrubbing unit for free ammonia inhibition reduction in the digester, and nitrogen recovery as ammonium sulphate. The stripped digestate was recirculated daily in the digester for 15 days (68% of the digester volume), increasing the gas production rate by 27%. Following a decanter centrifuge, the digestate liquid fraction was treated with an ultrafiltration membrane. The filtrate was fed into a struvite reactor, with a phosphorus recovery efficiency of 83% (as orthophosphate). Acidification of digestate could increment the soluble orthophosphate concentration up to four times, enhancing phosphorus enrichment in the liquid fraction and its recovery via struvite. A synergistic combination of manure processing steps was demonstrated to be technologically feasible to upgrade livestock waste into refined, concentrated fertilisers.

Tracking bio-hydrogen-mediated production of commodity chemicals from carbon dioxide and renewable electricity.

This study reveals that reduction of carbon dioxide (CO2) to commodity chemicals can be functionally compartmentalized in bioelectrochemical systems. In the present example, a syntrophic consortium composed by H2-producers (Rhodobacter sp.) in the biofilm is combined with carboxidotrophic Clostridium species, mainly found in the bulk liquid. The performance of these H2-mediated electricity-driven systems could be tracked by the activity of a biological H2 sensory protein identified at cathode potentials between -0.2V and -0.3V vs SHE. This seems to point out that such signal is not strain specific, but could be detected in any organism containing hydrogenases. Thus, the findings of this work open the door to the development of a biosensor application or soft sensors for monitoring such systems.

External Resistances Applied to MFC Affect Core Microbiome and Swine Manure Treatment Efficiencies.

Microbial fuel cells (MFCs) can be designed to combine water treatment with concomitant electricity production. Animal manure treatment has been poorly explored using MFCs, and its implementation at full-scale primarily relies on the bacterial distribution and activity within the treatment cell. This study reports the bacterial community changes at four positions within the anode of two almost identically operated MFCs fed swine manure. Changes in the microbiome structure are described according to the MFC fluid dynamics and the application of a maximum power point tracking system (MPPT) compared to a fixed resistance system (Ref-MFC). Both external resistance and cell hydrodynamics are thought to heavily influence MFC performance. The microbiome was characterised both quantitatively (qPCR) and qualitatively (454-pyrosequencing) by targeting bacterial 16S rRNA genes. The diversity of the microbial community in the MFC biofilm was reduced and differed from the influent swine manure. The adopted electric condition (MPPT vs fixed resistance) was more relevant than the fluid dynamics in shaping the MFC microbiome. MPPT control positively affected bacterial abundance and promoted the selection of putatively exoelectrogenic bacteria in the MFC core microbiome (Sedimentibacter sp. and gammaproteobacteria). These differences in the microbiome may be responsible for the two-fold increase in power production achieved by the MPPT-MFC compared to the Ref-MFC.

Low Fermentation pH Is a Trigger to Alcohol Production, but a Killer to Chain Elongation.

Gasification of organic wastes coupled to syngas fermentation allows the recovery of carbon in the form of commodity chemicals, such as carboxylates and biofuels. Acetogenic bacteria ferment syngas to mainly two-carbon compounds, although a few strains can also synthesize four-, and six-carbon molecules. In general, longer carbon chain products have a higher biotechnological (and commercial) value due to their higher energy content and their lower water solubility. However, de-novo synthesis of medium-chain products from syngas is quite uncommon in acetogenic bacteria. An alternative to de-novo synthesis is bioproduction of short-chain products (C2 and C4), and their subsequent elongation to C4, C6, or C8 through reversed ß-oxidation metabolism. This two-step synergistic approach has been successfully applied for the production of up to C8 compounds, although the accumulation of alcohols in these mixed cultures remained below detection limits. The present work investigates the production of higher alcohols from syngas by open mixed cultures (OMC). A syngas-fermenting community was enriched from sludge of an anaerobic digester for a period of 109 days in a lab-scale reactor. At the end of this period, stable production of ethanol and butanol was obtained. C6 compounds were only transiently produced at the beginning of the enrichment phase, during which Clostridium kluyveri, a bacterium able to carry out carbon chain elongation, was detected in the community. Further experiments showed pH as a critical parameter to maintain chain elongation activity in the co-culture. Production of C6 compounds was recovered by preventing fermentation pH to decrease below pH 4.5-5. Finally, experiments showed maximal production of C6 compounds (0.8 g/L) and alcohols (1.7 g/L of ethanol, 1.1 g/L of butanol, and 0.6 g/L of hexanol) at pH 4.8. In conclusion, low fermentation pH is critical for the production of alcohols, although detrimental to C. kluyveri. Fine control of fermentation pH to final values around 4.8 could allow sustained production of higher alcohols.

Bidirectional microbial electron transfer: Switching an acetate oxidizing biofilm to nitrate reducing conditions.

Up to date a few electroactive bacteria embedded in biofilms are described to catalyze both anodic and cathodic reactions in bioelectrochemical systems (i.e. bidirectional electron transfer). How these bacteria transfer electrons to or from the electrode is still uncertain. In this study the extracellular electron transfer mechanism of bacteria within an electroactive biofilm was investigated by using cyclic voltammetry (CV) and differential pulse voltammetry (DPV). First, a mature anodic electroactive biofilm was developed from an activated sludge sample (inoculum), acetate as electron donor and a poised electrode (+397mV vs. SHE). Later, this biofilm was "switched" to biocathodic conditions by feeding it with a medium containing nitrates and poising the electrode at -303mV vs. SHE. The electrochemical characterization indicated that both, acetate oxidation and nitrate reduction took place at a similar formal potential of -175±05 and -175±34mV vs. SHE, respectively. The biofilm was predominantly composed by Geobacter sp. at both experimental conditions. Taken together, the results indicated that both processes could be catalyzed by using the same electron conduit, and most likely by the same bacterial consortium. Hence, this study suggests that electroactive bacteria within biofilms could use the same electron transfer conduit for catalyzing anodic and cathodic reactions.

Conversion of sewage sludge to commodity chemicals via syngas fermentation.

Gasification of sewage sludge allows the recovery of energy, and produces a mix of CO, CO2and H2called synthesis gas (or syngas), which can be fermented by acetogenic bacteria to added-value products. This work presents the conversion of syngas to organic acids and alcohols using both pure and mixed cultures. Pure culture kinetic experiments with Clostridium carboxidivorans P7 resulted in the production of high concentrations of acetate (454 mgC/L) and ethanol (167 mgC/L). The pH was the main factor driving solventogenesis, with about 50% of the products in the form of alcohols at pH 5. Conversely, laboratory-scale experiments using a carboxydotrophic mixed culture of the genus Clostridium enriched from anaerobic digester sludge of a municipal wastewater treatment plant was capable of producing mainly butyrate, with maximum concentration of 1,184 mgC/L.

Incubation at 25 °C prevents acid crash and enhances alcohol production in Clostridium carboxidivorans P7.

Incubation of carboxydotrophs at 37 °C provides optimal conditions for their growth. However, a fast accumulation of organic acids, specifically acetate, during the exponential growth phase may result in low alcohol production and substrate consumption due to a phenomenon known as "acid crash". The present work investigates growth and productivity of Clostridium carboxidivorans P7 at two incubation temperatures. At 37 °C the culture was not able to override the "acid crash", resulting in low ethanol titers (1.56 mM). On the other hand, lower metabolic rates at 25 °C enhanced ethanol and butanol production (32.1 and 14.5 mM, respectively). Moreover, at low temperatures, hexanol and caproic acid were also produced at significant concentrations, 8.21 and 9.02 mM respectively, among the highest values reported for P7. Our results demonstrate that production of biofuels with longer carbon chains molecules may be enhanced incubating syngas-fermenting acetogenic bacteria at sub-optimal temperatures.