For the production of edible microbial protein (MP), ammonia generated by the Haber-Bosch process or reclaimed ammonia from waste streams is typically considered as the nitrogen source. These ...processes for ammonia production are highly energy intensive. In this study, the potential for using nitrogen gas (N2) as a direct nitrogen source for MP production by hydrogen-oxidizing bacteria (HOB) was evaluated. The use of N2 versus ammonium as nitrogen source during the enrichment process resulted in differentiation of the bacterial community composition of the enrichments. A few previously unknown potential N2-fixing HOB taxa (i.e., representatives of the genus Azonexus and the family Comamonadaceae) dominated the enrichments. The biomass yield of a N2-fixing HOB enrichment was 30–50% lower than that of the ammonium-based HOB enrichment from the same inoculum source. The dried biomass of N2-fixing HOB had a high protein content (62.0 ± 6.3%) and an essential amino acid profile comparable to MP from ammonium-based HOB. MP from N2-fixing HOB could potentially be produced in situ without entailing the emissions caused by ammonia production and transportation by conventional means. It could be a promising substitute for N2-fixing protein-rich soybean because it has 70% higher protein content and double energy conversion efficiency from solar energy to biomass.
Microbial fuel cells (MFCs) have the potential to combine wastewater treatment efficiency with energetic efficiency. One of the major impediments to MFC implementation is the operation of the cathode ...compartment, as it employs environmentally unfriendly catalysts such as platinum. As recently shown, bacteria can facilitate sustainable and cost-effective cathode catalysis for nitrate and also oxygen. Here we describe a carbon cathode open to the air, on which attached bacteria catalyzed oxygen reduction. The bacteria present were able to reduce oxygen as the ultimate electron acceptor using electrons provided by the solid-phase cathode. Current densities of up to 2.2 A m(-2) cathode projected surface were obtained (0.303+/-0.017 W m(-2), 15 W m(-3) total reactor volume). The cathodic microbial community was dominated by Sphingobacterium, Acinetobacter and Acidovorax sp., according to 16S rRNA gene clone library analysis. Isolates of Sphingobacterium sp. and Acinetobacter sp. were obtained using H(2)/O(2) mixtures. Some of the pure culture isolates obtained from the cathode showed an increase in the power output of up to three-fold compared to a non-inoculated control, that is, from 0.015+/-0.001 to 0.049+/-0.025 W m(-2) cathode projected surface. The strong decrease in activation losses indicates that bacteria function as true catalysts for oxygen reduction. Owing to the high overpotential for non-catalyzed reduction, oxygen is only to a limited extent competitive toward the electron donor, that is, the cathode. Further research to refine the operational parameters and increase the current density by modifying the electrode surface and elucidating the bacterial metabolism is warranted.
Typically, nitrification is a two-stage microbial process and is key in wastewater treatment and nutrient recovery from waste streams. Changes in salinity represent a major stress factor that can ...trigger response mechanisms, impacting the activity and the physiology of bacteria. Despite its pivotal biotechnological role, little information is available on the specific response of nitrifying bacteria to varying levels of salinity. In this study, synthetic communities of ammonia-oxidizing bacteria (AOB
and/or
) and nitrite-oxidizing bacteria (NOB
and/or
) were tested at 5, 10, and 30 mS cm
by adding sodium chloride to the mineral medium (0, 40, and 200 mM NaCl, respectively). Ammonia oxidation activity was less affected by salinity than nitrite oxidation. AOB, on their own or in combination with NOB, showed no significant difference in the ammonia oxidation rate among the three conditions. However,
improved the absolute ammonia oxidation rate of both
and
.
's nitrite oxidation rate decreased to 42% residual activity upon exposure to 30 mS cm
, also showing a similar behavior when tested with
spp. The nitrite oxidation rate of
, as a single species, was not affected when adding sodium chloride up to 30 mS cm
, however, its activity was completely inhibited when combined with
spp. in the presence of ammonium/ammonia. The proteomic analysis of a co-culture of
and
revealed the production of osmolytes, regulation of cell permeability and an oxidative stress response in
and an oxidative stress response in
, as a result of increasing the salt concentration from 5 to 30 mS cm
. A specific metabolic response observed in
suggests the role of carbon metabolism in the production of reducing power, possibly to meet the energy demands of the stress response mechanisms, induced by high salinity. For the first time, metabolic modifications and response mechanisms caused by the exposure to salinity were described, serving as a tool toward controllability and predictability of nitrifying systems exposed to salt fluctuations.
Microbial fuel cells (MFCs) have applications other than electricity production, including the capacity to power desirable reactions in the cathode chamber. However, current knowledge of the ...microbial ecology and physiology of biocathodes is minimal, and as a result more research dedicated to understanding the microbial communities active in cathode biofilms is required. Here we characterize the microbiology of denitrifying bacterial communities stimulated by reducing equivalents generated from the anodic oxidation of acetate. We analyzed biofilms isolated from two types of cathodic denitrification systems: (1) a loop format where the effluent from the carbon oxidation step in the anode is subjected to a nitrifying reactor which is fed to the cathode chamber and (2) an alternative non-loop format where anodic and cathodic feed streams are separated. The results of our study indicate the superior performance of the loop reactor in terms of enhanced current production and nitrate removal rates. We hypothesized that phylogenetic or structural features of the microbial communities could explain the increased performance of the loop reactor. We used PhyloChip with 16S rRNA (cDNA) and fluorescent in situ hybridization to characterize the active bacterial communities. Our study results reveal a greater richness, as well as an increased phylogenetic diversity, active in denitrifying biofilms than was previously identified in cathodic systems. Specifically, we identified Proteobacteria, Firmicutes and Chloroflexi members that were dominant in denitrifying cathodes. In addition, our study results indicate that it is the structural component, in terms of bacterial richness and evenness, rather than the phylogenetic affiliation of dominant bacteria, that best corresponds to cathode performance.
Operation of microbial electrolysis cells (MECs) without an ion exchange membrane could help to lower the construction costs while lowering the ohmic cell resistance and improving MEC conversion ...rates by minimizing the pH gradient between anode and cathode. In this research, we demonstrate that membraneless MECs with plain graphite can be operated for methane production without pH adjustment and that the ohmic cell resistance could be lowered with approximately 50% by removing the cation exchange membrane. As a result, the current production increased from 66 ± 2 to 156 ± 1 A m⁻³ MEC by removing the membrane with an applied voltage of -0.8 V. Methane was the main energetic product despite continuous operation under carbonate-limited and slightly acidified conditions (pH 6.1-6.2). Our results suggest that continuous production of hydrogen in membraneless MECs will be challenging since methane production might not be avoided easily. The electrical energy invested was not always completely recovered under the form of an energy-rich biogas; however, our results indicate that membraneless MECs might be a viable polishing step for the treatment of the effluent of anaerobic digesters as methane was produced under low organic loading conditions and at room temperature.
Recovery of nutrients from source-separated urine can truncate our dependency on synthetic fertilizers, contributing to more sustainable food production. Urine-derived fertilizers have been ...successfully applied in soilless cultures. However, little is known about the adaptation of the plant to the nutrient environment. This study investigated the impact of urine-derived fertilizers on plant performance and the root-associated bacterial community of hydroponically grown lettuce (Lactuca sativa L.). Shoot biomass, chlorophyll, phenolic, antioxidant, and mineral content were associated with shifts in the root-associated bacterial community structures. K-struvite, a high-performing urine-derived fertilizer, supported root-associated bacterial communities that overlapped most strongly with control NPK fertilizer. Contrarily, lettuce performed poorly with electrodialysis (ED) concentrate and hydrolyzed urine and hosted distinct root-associated bacterial communities. Comparing the identified operational taxonomic units (OTU) across the fertilizer conditions revealed strong correlations between specific bacterial genera and the plant physiological characteristics, salinity, and NO3−/NH4+ ratio. The root-associated bacterial community networks of K-struvite and NPK control fertilized plants displayed fewer nodes and node edges, suggesting that good plant growth performance does not require highly complex ecological interactions in hydroponic growth conditions.
A tubular, single-chambered, continuous microbial fuel cell (MFC) that generates high power outputs using a granular graphite matrix as the anode and a ferricyanide solution as the cathode is ...described. The maximal power outputs obtained were 90 and 66 W m-3 net anodic compartment (NAC) (48 and 38 W m-3 total anodic compartment (TAC)) for feed streams based on acetate and glucose, respectively, and 59 and 48 W m-3 NAC for digester effluent and domestic wastewater, respectively. For acetate and glucose, the total Coulombic conversion efficiencies were 75 ± 5% and 59 ± 4%, respectively, at loading rates of 1.1 kg chemical oxygen demand m-3 NAC volume day-1. When wastewater was used, of the organic matter effectively removed (i.e., 22% at a loading of 2 kg organic matter m-3 NAC day-1), up to 96% was converted to electricity on a Coulombic basis. The lower overall efficiency of the wastewater-treating reactors is related to the presence of nonreadily biodegradable organics and the interference of alternative electron acceptors such as sulfate present in the wastewater. To further improve MFCs, focus has to be placed on the enhanced conversion of nonrapidly biodegradable material and the better directing of the anode flow toward the electrode instead of to alternative electron acceptors. Also the use of sustainable, open-air cathodes is a critical issue for practical implementation.
•85% COD & 100% urea removal from fresh urine with a microbial electrolysis cell.•Full urine nitrification in membrane-aerated reactor with DO<0.1 mg O2 L−1.•Bio-anodic pre-treatment avoids N loss in ...membrane-aerated nitrification.•The two reactors contained very distinct and diverse microbial communities.•MEC and MABR treatment enables highly resource-efficient urine stabilization.
Resource recovery from source-separated urine can shorten nutrient cycles on Earth and is essential in regenerative life support systems for deep-space exploration. In this study, a robust two-stage, energy-efficient, gravity-independent urine treatment system was developed to transform fresh real human urine into a stable nutrient solution. In the first stage, up to 85% of the COD was removed in a microbial electrolysis cell (MEC), converting part of the energy in organic compounds (27–46%) into hydrogen gas and enabling full nitrogen recovery by preventing nitrogen losses through denitrification in the second stage. Besides COD removal, all urea was hydrolysed in the MEC, resulting in a stream rich in ammoniacal nitrogen and alkalinity, and low in COD. This stream was fed into a membrane-aerated biofilm reactor (MABR) in order to convert the volatile and toxic ammoniacal nitrogen to non-volatile nitrate by nitrification. Bio-electrochemical pre-treatment allowed to recover all nitrogen as nitrate in the MABR at a bulk-phase dissolved oxygen level below 0.1 mg O2 L−1. In contrast, feeding the MABR directly with raw urine (omitting the first stage), at the same nitrogen loading rate, resulted in nitrogen loss (18%) due to denitrification. The MEC and MABR were characterised by very distinct and diverse microbial communities. While (strictly) anaerobic genera, such as Geobacter (electroactive bacteria), Thiopseudomonas, a Lentimicrobiaceae member, Alcaligenes and Proteiniphilum prevailed in the MEC, the MABR was dominated by aerobic genera, including Nitrosomonas (a known ammonium oxidiser), Moheibacter and Gordonia. The two-stage approach yielded a stable nitrate-rich, COD-low nutrient solution, suitable for plant and microalgae cultivation.
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Electroactive biofilms (EABs) have recently attracted considerable research interest for their possible use as amperometric biosensors in environmental or bioprocess monitoring, for example for in ...situ detection of toxic compounds. Almost exclusively, corresponding research has focused on heterotrophic, anodic EABs. These biofilms require sufficiently high organic loads and anoxic conditions to deliver a stable baseline current. Conversely, electroautotrophic O2-reducing EABs have recently been proposed to monitor toxic shocks in oxic solutions that are poor or devoid of organic substrate. This was done in optimal media and only assessed for formaldehyde as a model toxic compound. Here we show that O2-reducing EABs can grow in unamended tap water on carbon electrodes at + 0.2 V vs. Ag/AgCl. They retained substantial electroactivity for at least eight months without adding exogenous compounds. The most represented operational taxonomic units were assigned to the phylum Gammaproteobacteria (25 ± 15%, n = 5 electrodes). Cyclic voltammograms showed a reproducible nernstian behavior for O2 reduction with a mid-wave potential at + 0.27 V and variable plateau current densities ranging from – 1 to – 22 µA cm−2 (n = 10 electrodes). The biocatalytic current was substantially impacted by the addition of either of three tested heavy metals (Hg(II), Cr(VI) or Pb(II)) or by organic pollutants (formaldehyde, 2,4-dichlorophenol, benzalkonium chloride), with limits of detection ranging from 0.5 to 10 mg L−1 (2.5–61 µmol L−1). Response times were typically around 1 min. Comparison with previous reports suggests that O2-reducing microbial cathodes may be more sensitive to toxic shocks than anodic, heterotrophic EABs.
•Electroactive biofilms electroreducing O2 at high potential could grow in tap water.•Microbially-induced current generated for more than 8 months in unamended tap water.•Toxic shocks monitored amperometrically for 6 heavy metals and organic pollutants.•Results suggest that cathodic biofilms are more sensitive than anodic to toxicants.
Electrochemical water softening was proposed as a sustainable alternative for ion exchange softening, avoiding the input of salt to drinking water and the production of a concentrated brine. Here we ...demonstrated two novel modes of operation combining an electrochemical cell with a fluidized bed crystallizer. The first approach relied on an electrochemical cell consisting of an anode and cathode separated by a cation or anion exchange membrane. The feed water was first directed into a crystallizer where it was blended with alkaline cathode effluent. The effluent of the crystallizer, softened water, was in part recirculated to the cathode to generate alkalinity, in part to the anode compartment, where the pH was again decreased. Average removal efficiencies for calcium and magnesium of 75–86% and 7–21% respectively, could be sustainably reached, at a specific energy consumption of 7.0–10.1 kWh kg−1 CaCO3 (0.86–1.39 kWh m−3 water). This configuration allowed reagent-free water softening, albeit with an effluent with a pH between 3.0 and 3.6. In a second mode of operation, the process influent to soften was also directed to the crystallizer and recirculated over the cathode, which was separated from the anode using an anion exchange membrane. In this mode of operation, the cathode effluent was sent through the crystallizing unit, and the anode compartment was operated in closed-loop. Average calcium and magnesium removal efficiencies of 73–78% and 40–44% were obtained at specific energy consumptions of 5.8–7.5 kWh kg−1 CaCO3 (0.77–0.88 kWh m−3 water). Although the softened water had an elevated pH (∼9.4), the advantage of this configuration is concomitant removal of anions and the formation of acids/disinfectant in the anode compartment. Both methods of operation thus showed reagent-free water softening at a relatively low specific energy consumption. These novel methods of softening could be used in remote locations where access to chemicals or discharge of ion exchange brines proves to be difficult, or in case addition of chemicals for softening is unwanted. Further research is needed to further decrease the specific energy consumption during long-term operation.
•Electrochemical softening enables removal of hardness from potable water without reagents.•Energy investment is 5.8–7.5 kWh kg−1 CaCO3 removed.•Applications range from softening in remote locations to addressing concerns of reagent addition.
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