Dissimilatory Nitrate Reduction To Ammonium Research Articles

Contrasting effects of climate change on denitrification and nitrogen load reduction in the Po River (Northern Italy)

An increase in water temperature is one of the main factors that can potentially modify biogeochemical dynamics in lowland rivers, such as the removal and recycling of nitrogen (N). This effect of climate change on N processing deserves attention, as it may have unexpected impacts on eutrophication in the coastal zones. Intact sediment cores were collected seasonally at the closing section of the Po River, the largest Italian river and one of the main N inputs to the Mediterranean Sea. Benthic oxygen fluxes, denitrification, and dissimilatory nitrate reduction to ammonium (DNRA) rates were measured using laboratory dark incubations. Different temperature treatments were set up for each season based on historical data and future predictions. Higher water temperatures enhanced sediment oxygen demand and the extent of hypoxic conditions in the benthic compartment, favoring anaerobic metabolism. Indeed, warming water temperature stimulated nitrate (NO3−) reduction processes, although NO3− and organic matter availability were found to be the main controlling factors shaping the rates between seasons. Denitrification was the main process responsible for NO3− removal, mainly supported by NO3− diffusion from the water column into the sediments, and much more important than N recycling via DNRA. The predicted increase in the water temperature of the Po River due to climate change may exert an unexpected negative feedback on eutrophication by strongly controlling denitrification and contributing to partial buffering of N export in the lagoons and coastal areas, especially in spring.Graphical

Inhibition, Recovery and Stabilization of MBR-Anammox Process: Enhancing Nitrogen Removal by Biochar-supported Nano Zero-valent Iron Adoption

Anaerobic ammonium oxidation (Anammox) is recognized as a promising option to energy-neutral wastewater treatment, yet difficulties encountered in practice due to its poor stability. This work investigated the effects of biochar-supported nano zero-valent iron (nZVI-BC) on nitrogen removal performance of anammox process. Three membrane bioreactors (MBRs) suitable for slowly growing anammox bacteria was operated for 100 days, undergoing startup, dissolved oxygen (DO) inhibition, performance recovery and stabilization periods. The results showed that nZVI-BC adoption could improve the nitrogen removal efficiency (NRE) by 5.5–27.3 % and 3.1–20.7 % and nitrogen removal rate (NRR) by 5.4–28.6 % and 3.2–22.9 %, compared with control group and biochar adoption, respectively. Throughout the experimental periods, nZVI-BC was conducive to enrich anammox bacteria and increased the functional gene copies associated with anammox metabolism. Meanwhile, co-occurrence network analysis promulgated that nZVI-BC boosted the robustness and interaction of microbial community. Furthermore, the electrochemical analysis of the extracellular polymeric substance (EPS) revealed that nZVI-BC adoption augmented the extracellular electron transport (EET) process of anammox biomass. Metagenomic sequencing revealed that nZVI-BC reinforced the complete dissimilatory nitrate reduction to ammonium (DNRA) pathway in Candidatus Brocadia, the dominant anammox bacteria, potentially reducing nitrate production and enhancing nitrogen removal. The study manifested the possibility of nZVI-BC to improve the anammox robustness and nitrogen removal performance, and provided operational guidance for its engineering application.

Hypoxia exerts greater impacts on shallow groundwater nitrogen cycling than seawater mixture in coastal zone.

There is no doubt that hypoxia and seawater mixture are profoundly affecting the global nitrogen (N) cycle. However, their mechanisms for altering N cycling patterns in shallow coastal groundwater are largely unknown. Here, we examined shallow groundwater N transformation characteristics (dissolved inorganic N and related chemical properties) in the coastal area of east and west Shenzhen City. Results showed that common hypoxic conditions exist in this study area. Ions/Cl- ratios indicated varying levels of saltwater mixture and sulfide formation across this study area. Dissolved oxygen (DO) affects the N cycle process by controlling the conditions of nitrification and the formation of sulfides. Salinity affects nitrification and denitrification processes by physiological effects, while sulfide impacts nitrification, denitrification, and dissimilatory nitrate reduction to ammonium (DNRA) processes through its own toxicity mechanism and the provision of electron donors for DNRA organisms. Redundancy analysis (RDA) results indicate that the influence magnitude is in the following order: DO > sulfide > salinity. Seawater mixture weakened the nitrification and denitrification of groundwater by changing salinity, while hypoxia and its controlled sulfide formation not only weaken nitrification and denitrification but also stimulated the DNRA process and promotes N regeneration. In this study area, hypoxia is considered to exert greater impacts on N cycling in the coastal shallow groundwater than seawater mixture. These findings greatly improve our understanding of the consequences of hypoxia and seawater mixture on coastal groundwater N cycling.

Denitrificimonas halotolerans sp. nov., a novel species isolated from UASB sludge treating landfill leachate.

A Gram-stain-negative, facultative anaerobe, rod-shaped strain JX-1T was isolated from UASB sludge treating landfill leachate in Wuhan, China. The isolate is capable of growing under conditions of pH 6.0-11.0 (optimum, pH 7.0-8.0), temperature 4-42℃ (optimum, 20-30℃), 0-8.0% (w/v) NaCl (optimum, 5.0%), and ammonia nitrogen concentration of 200-5000mg/L (optimum, 500mg/L) on LB plates. The microorganism can utilize malic acid, D-galactose, L-rhamnose, inosine, and L-glutamic acid as carbon sources, but does not reduce nitrates and nitrites. The major fatty acids are C18:1ω7c/C18:1ω6c, iso-C15:0, and anteiso-C15:0. The respiratory quinones are Q9 (91.92%) and Q8 (8.08%). Polar lipids include aminolipid, aminophospholipid, diphosphatidylglycerol, glycolipid, phosphatidylethanolamine, phosphatidylglycerol, and phospholipid. Compared with other strains, strain JX-1T and Denitrificimonas caeni HY-14T have the highest values in terms of 16S rRNA gene sequence similarity (96.79%), average nucleotide identity (ANI; 76.06%), and average amino acid identity (AAI; 78.89%). Its digital DNA-DNA hybridization (dDDH) result is 20.3%. The genome of strain JX-1T, with a size of 2.78Mb and 46.12mol% G + C content, lacks genes for denitrification and dissimilatory nitrate reduction to ammonium (DNRA), but contains genes for ectoine synthesis as a secondary metabolite. The results of this polyphasic study allow genotypic and phenotypic differentiation of the analysed strain from the closest related species and confirm that the strain represents a novel species within the genus Denitrificimonas, for which the name Denitrificimonas halotolerans sp. nov. is proposed with JX-1T (= MCCC 1K08958T = KCTC 8395T) as the type strain.

Spatiotemporal patterns and co-occurrence patterns of dissimilatory nitrate reduction to ammonium community in sediments of the Lancang River cascade reservoirs.

Dissimilatory nitrate reduction to ammonium (DNRA) is an important nitrate reduction pathway in freshwater sediments. Many studies have focused on the DNRA process in various natural habitats. However, the joint operation of cascade reservoirs will affect the physical and chemical properties of sediments, which may change the DNRA process and bacterial community pattern in the surface sediments of cascade reservoirs. Our study was the first to investigate the spatiotemporal distribution patterns of potential DNRA rate, nrfA gene abundances, and DNRA bacterial community diversity in surface sediments of the Lancang River cascade reservoirs. The results of slurry incubation experiments combined with the 15N isotope tracer experiment ascertained that the potential rates of DNRA were 0.01-0.15 nmol-N cm-3 h-1, and qPCR results indicated that the abundance range of nrfA was 1.08 × 105-2.51 × 106 copies g-1 dry weight. High throughput sequencing of the nrfA gene revealed that the relative abundance of Anaeromyxobacter (4.52% on average), Polyangium (4.09%), Archangium (1.86%), Geobacter (1.34%), and Lacunisphaera (1.32%) were high. Pearson and RDA correlation analysis exhibited that nrfA gene abundance was positively correlated with altitude, pH, OC, and sand concentration. Anaeromyxobacter was positively correlated with reservoir age and DNRA potential rate. The deterministic environmental selection process plays a crucial role in the formation of the DNRA bacterial community. Network analysis displayed that the dominant DNRA genus was the key population of the DNRA microbial community in the sediments of Lancang River cascade reservoirs. This study reveals that the variation of DNRA bacterial activity and community structure is largely driven by the construction of cascade reservoirs, and provides a new idea for further understanding the characteristics of the DNRA community in the cascade reservoir ecosystem.

Mechanisms underlying the interactions and adaptability of nitrogen removal microorganisms in freshwater sediments

Microorganisms in eutrophic water play a vital role in nitrogen (N) removal, which contributes significantly to the nutrient cycling and sustainability of eutrophic ecosystems. However, the mechanisms underlying the interactions and adaptation strategies of the N removal microorganisms in eutrophic ecosystems remain unclear. We thus analyzed field sediments collected from a eutrophic freshwater ecosystem, enriched the N removal microorganisms, examined their function and adaptability through amplicon, metagenome and metatranscriptome sequencing. We found that the N removal activities could be affected through potential competition and inhibition among microbial metabolic pathways. High-diversity microbial communities generally increased the abundance and expression of N removal functional genes. Further enrichment experiments showed that the enrichment of N removal microorganisms led to a development of simplified but more stable microbial communities, characterized by similar evolutionary patterns among N removal microorganisms, tighter interactions, and increased adaptability. Notably, the sustained provision of NH4+ and NO2− during the enrichment could potentially strengthen the interconnections among denitrification, anaerobic ammonium oxidation (anammox) and dissimilatory nitrate reduction to ammonium (DNRA) processes. Moreover, the identification of shared metabolic traits among denitrification, anammox and DNRA implies important cooperative associations and adaptability of N removal microorganisms. Our findings highlight the microbial interactions affect the adaptive strategies of key microbial taxa involved in N removal.

Nitrate reduction to ammonium: a phylogenetic, physiological, and genetic aspects in Prokaryotes and eukaryotes.

The microbe-mediated conversion of nitrate (NO3-) to ammonium (NH4+) in the nitrogen cycle has strong implications for soil health and crop productivity. The role of prokaryotes, eukaryotes and their phylogeny, physiology, and genetic regulations are essential for understanding the ecological significance of this empirical process. Several prokaryotes (bacteria and archaea), and a few eukaryotes (fungi and algae) are reported as NO3- reducers under certain conditions. This process involves enzymatic reactions which has been catalysed by nitrate reductases, nitrite reductases, and NH4+-assimilating enzymes. Earlier reports emphasised that single-cell prokaryotic or eukaryotic organisms are responsible for this process, which portrayed a prominent gap. Therefore, this study revisits the similarities and uniqueness of mechanism behind NO3- -reduction to NH4+ in both prokaryotes and eukaryotes. Moreover, phylogenetic, physiological, and genetic regulation also shed light on the evolutionary connections between two systems which could help us to better explain the NO3--reduction mechanisms over time. Reports also revealed that certain transcription factors like NtrC/NtrB and Nit2 have shown a major role in coordinating the expression of NO3- assimilation genes in response to NO3- availability. Overall, this review provides a comprehensive information about the complex fermentative and respiratory dissimilatory nitrate reduction to ammonium (DNRA) processes. Uncovering the complexity of this process across various organisms may further give insight into sustainable nitrogen management practices and might contribute to addressing global environmental challenges.

Potential coupling of microbial methane, nitrogen, and sulphur cycling in the Okinawa Trough cold seep sediments.

The Okinawa Trough (OT) is a back-arc basin with a wide distribution of active cold seep systems. However, our understanding of the metabolic function of microbial communities in the cold seep sediments of the OT remains limited. In this study, we investigated the vertical profiles of functional genes involved in methane, nitrogen, and sulphur cycling in the cold seep sediments of the OT. Furthermore, we explored the possible coupling mechanisms between these biogeochemical cycles. The study revealed that the majority of genes associated with the nitrogen and sulphur cycles were most abundant in the surface sediment layers. However, only the key genes responsible for sulphur disproportionation (sor), nitrogen fixation (nifDKH), and methane metabolism (mcrABG) were more prevalent within sulfate-methane transition zone (SMTZ). Significant positive correlations (P < 0.05) were observed between functional genes involved in sulphur oxidation, thiosulphate disproportionation with denitrification, and dissimilatory nitrate reduction to ammonium (DNRA), as well as between AOM/methanogenesis and nitrogen fixation, and between sulphur disproportionation and AOM. A genome of Filomicrobium (class Alphaproteobacteria) has demonstrated potential in chemoautotrophic activities, particularly in coupling DNRA and denitrification with sulphur oxidation. Additionally, the characterized sulfate reducers such as Syntrophobacterales have been found to be capable of utilizing nitrate as an electron acceptor. The predominant methanogenic/methanotrophic groups in the OT sediments were identified as H2-dependent methylotrophic methanogens (Methanomassiliicoccales and Methanofastidiosales) and ANME-1a. This study offered a thorough understanding of microbial ecosystems in the OT cold seep sediments, emphasizing their contribution to nutrient cycling.IMPORTANCEThe Okinawa Trough (OT) is a back-arc basin formed by extension within the continental lithosphere behind the Ryukyu Trench arc system. Cold seeps are widespread in the OT. While some studies have explored microbial communities in OT cold seep sediments, their metabolic potential remains largely unknown. In this study, we used metagenomic analysis to enhance comprehension of the microbial community's role in nutrient cycling and proposed hypotheses on the coupling process and mechanisms involved in biogeochemical cycles. It was revealed that multiple metabolic pathways can be performed by a single organism or microbes that interact with each other to carry out various biogeochemical cycling. This data set provided a genomic road map on microbial nutrient cycling in OT sediment microbial communities.

Seawater intrusion effects on nitrogen cycling in the regulated Nakdong River Estuary, South Korea

Research on the impact of seawater intrusion on nitrogen (N) cycling in coastal estuarine ecosystems is crucial; however, there is still a lack of relevant research conducted under in-situ field conditions. The effects of elevated salinity on N cycling processes and microbiomes were examined in situ seawater intrusion experiments conducted from 2019 to 2021 in the Nakdong River Estuary (South Korea), where an estuarine dam regulates tidal hydrodynamics. After the opening of the Nakdong Estuary Dam (seawater intrusion event), the density difference between seawater and freshwater resulted in varying degrees of seawater trapping at topographically deep stations. Bottom-water oxygen conditions had been altered in normoxia, hypoxia, and weak hypoxia due to the different degrees of seawater trapping in 2019, 2020, and 2021, respectively. Denitrification mostly dominated the nitrate (NO3-) reduction process, except in 2020 after seawater intrusion. However, denitrification rates decreased because of reduced coupled nitrification after seawater intrusion due to the dissolved oxygen limitation in 2020. Dissimilatory nitrate reduction to ammonium (DNRA) rates immediately increased after seawater intrusion in 2020, replacing denitrification as the dominant pathway in the NO3- reduction process. The enhanced DNRA rate was mainly due to the abundant organic matter associated with seawater invasion and more reducing environment (maybe sulfide enhancement effects) under high seawater-trapping conditions. Denitrification increased in 2021 after seawater intrusion during weak hypoxia; however, DNRA did not change. Small seawater intrusion in 2019 caused no seawater trapping and overall normoxic condition, though a slight shift from denitrification to DNRA was observed. Metagenomic analysis revealed a decrease in overall denitrification-associated genes in response to seawater intrusion in 2019 and 2020, while DNRA-associated gene abundance increased. In 2021 after seawater intrusion, microbial gene abundance associated with denitrification increased, while that of DNRA did not change significantly. These changes in gene abundance align mostly with alterations in nitrogen transformation rates. In summary, ecological change effects in N cycling after the dam opening (N retention or release, that is, eutrophication deterioration or mitigation) depend on the degree of seawater intrusion and the underlying freshwater conditions, which constitute the extent of seawater-trapping.

Does invasive submerged macrophyte diversity affect dissimilatory nitrate reduction processes in sediments with varying microplastics?

Nitrogen removal is essential for restoring eutrophic lakes. Microorganisms and aquatic plants in lakes are both crucial for removing excess nitrogen. However, microplastic (MP) pollution and the invasion of exotic aquatic plants have become increasingly serious in lake ecosystems due to human activity and plant-dominant traits. This field mesocosm study explored how the diversity of invasive submerged macrophytes affects denitrification (DNF), anammox (ANA), and dissimilatory nitrate reduction to ammonium (DNRA) in lake sediments with varying MPs. Results showed that invasive macrophytes suppressed DNF rates, but DNRA and ANA were less sensitive than DNF to the diversity of invasive species. Sediment MPs increased the biomass of invasive species more than native species, but did not affect microbial processes. The effects of MPs on nitrate dissimilatory reduction were process-specific. MPs increased DNF rates and the competitive advantage of DNF over DNRA by changing the sediment environment. The decoupling of DNF and ANA was also observed, with increased DNF rates and decreased ANA rates. The study findings suggested new insights into how the invasion of exotic submerged macrophytes affects the sediment nitrogen cycle complex environments.

Characterizing the impact of reservoir storage and discharge on nitrogen dynamics in an upstream wetland using a δ15N and δ18O dual-isotope approach

The identification of nitrate sources in reservoir water is important for watershed-scale surface pollution management. Significant fluctuations in river water levels arising from reservoir storage and discharge influence nitrate sources and transport processes. The Sanmenxia Reservoir, in the middle reaches of the Yellow River in China, undergoes significant water level changes (290–316 m), altering the composition of the nitrogen sources. This study employed a δ15N and δ18O dual-isotope method and MixSIAR modeling to quantify the contributions of nitrate sources. This reveals the impact of reservoir water impoundment and discharge on nitrogen dynamics in the upstream region of the wetland and the model sensitivity for each nitrate source. The results showed that the average concentrations of nitrate‑nitrogen (NO- 3-N) were elevated during the impoundment period compared to the discharge period. Nitrogen sources exhibited varying proportions in surface water, groundwater, and soil water during both the impoundment and discharge periods. The predominant sources include manure and sewage (MS), with a maximum proportion of 57.4 % in surface water. Soil nitrogen (SN) accounted for 25.8 % of groundwater nitrogen and 32.1 % of soil water nitrogen during the impoundment period, whereas, during the discharge period, soil nitrogen made up 41.4 % of surface water nitrogen, manure and sewage contributed 44.8 % of groundwater nitrogen, and manure and sewage dominated with 56.7 % of soil water nitrogen. Sensitivity analysis of the MixSIAR model revealed that the isotopic composition of the manure and sewage primary source most significantly influenced the apportionment results of the riverine nitrate source. Reservoir discharge facilitates the dissimilatory nitrate reduction to ammonium (DNRA). The migration of NO- 3 from surface water to soil water and groundwater occurred from the impoundment period to the discharge period.

Overlooked in-situ sulfur disproportionation fuels dissimilatory nitrate reduction to ammonium in sulfur-based system: Novel insight of nitrogen recovery

Sulfur-based denitrification is a promising technology in treatments of nitrate-contaminated wastewaters. However, due to weak bioavailability and electron-donating capability of elemental sulfur, its sulfur-to-nitrate ratio has long been low, limiting the support for dissimilatory nitrate reduction to ammonium (DNRA) process. Using a long-term sulfur-packed reactor, we demonstrate here for the first time that DNRA in sulfur-based system is not negligible, but rather contributes a remarkable 40.5 %–61.1 % of the total nitrate biotransformation for ammonium production. Through combination of kinetic experiments, electron flow analysis, 16S rRNA amplicon, and microbial network succession, we unveil a cryptic in-situ sulfur disproportionation (SDP) process which significantly facilitates DNRA via enhancing mass transfer and multiplying 86.7–210.9 % of bioavailable electrons. Metagenome assembly and single-copy gene phylogenetic analysis elucidate the abundant genomes, including uc_VadinHA17, PHOS-HE36, JALNZU01, Thiobacillus, and Rubrivivax, harboring complete genes for ammonification. Notably, a unique group of self-SDP-coupled DNRA microorganism was identified. This study unravels a previously concealed fate of DNRA, which highlights the tremendous potential for ammonium recovery and greenhouse gas mitigation. Discovery of a new coupling between nitrogen and sulfur cycles underscores great revision needs of sulfur-driven denitrification technology.

Hydrological and biogeochemical processes controlling riparian groundwater quantity and quality during riverbank filtration

Groundwater exploitation in a riparian zone causes water infiltration from the river into the aquifer. Owing to adsorption and redox reactions along the flow path, the quality of water flowing from the river to groundwater wells is variably altered. The riverbed composition often involves spatiotemporal differences due to frequent changes in hydrological conditions. These changes create uncertainties in the transport and removal of solutes in the river water. In this study, the hydrodynamic field associated with riparian groundwater, changes in the structure of riverbed sediments caused by erosion and deposition, fluctuations in surface water and groundwater levels, and the removal efficiency of pollutants from groundwater through pumping were investigated. This involved in situ monitoring and sample testing of the composition of the river water, riverbed sediments, riverbed pore water, and groundwater during dry and wet seasons. Implementation of field in situ column experiments and molecular biology evidences were conducive to identifying the main biogeochemical processes occurring in the riverbed. The findings indicated that riparian groundwater exploitation alters the natural groundwater flow field, while fine sand deposition and microbial adsorption can reduce river recharge to aquifers by diminishing riverbed hydraulic conductivity. Shallow sediments within 1 m depth mainly involve NO3− reduction and E. coli adsorption. Reductive dissolution of Mn dominates in the deeper sediments. Additionally, reductive dissolution of Fe and dissimilatory nitrate reduction to ammonium (DNRA) drive high Fe2+ and NH4+ concentrations in groundwater. The findings can improve the management of riparian groundwater and aid in the optimization of a plan for its exploitation.

Competitive partitioning of denitrification pathways during arrested methanogenesis: Implications in ammonium recovery, N2O emission, and volatile fatty acid production

The complex interaction between nitrate (NO3−) reduction and fermentation is poorly understood when high levels of NO3− are introduced into anaerobic systems. This study investigated the competitive distribution between conventional denitrification (DEN) and dissimilatory nitrate reduction to ammonium (DNRA) during simultaneous denitrification and fermentation in arrested methanogenesis. Up to 62% of initial NO3− (200 mg-N/L) was retained as ammonium through DNRA at a chemical oxygen demand (COD)/N ratio of 25. Significant N2O emission occurred (1.7 – 8.0% of the initial NO3−) with limited carbon supply (≤1600 mg COD/L) and sludge concentration (≤3000 mg COD/L). VFA composition shifted predominantly towards acetic acid (>50%) in the presence of nitrate. A novel kinetic model was developed to predict DNRA vs. DEN partitioning and NO2− accumulation. Overall, NO3− input, organic loading, and carbon source characteristics independently and collectively controlled competitive DNRA vs. DEN partitioning.

Nitrogen and sulfur cycling and their coupling mechanisms in eutrophic lake sediment microbiomes

Microorganisms play important roles in the biogeochemical cycles of lake sediment. However, the integrated metabolic mechanisms governing nitrogen (N) and sulfur (S) cycling in eutrophic lakes remain poorly understood. Here, metagenomic analysis of field and bioreactor enriched sediment samples from a typical eutrophic lake were applied to elucidate the metabolic coupling of N and S cycling. Our results showed significant diverse genes involved in the pathways of dissimilatory sulfur metabolism, denitrification and dissimilatory nitrate reduction to ammonium (DNRA). The N and S associated functional genes and microbial groups generally showed significant correlation with the concentrations of NH4+, NO2− and SO42, while with relatively low effects from other environmental factors. The gene-based co-occurrence network indicated clear cooperative interactions between N and S cycling in the sediment. Additionally, our analysis identified key metabolic processes, including the coupled dissimilatory sulfur oxidation (DSO) and DNRA as well as the association of thiosulfate oxidation complex (SOX systems) with denitrification pathway. However, the enriched N removal microorganisms in the bioreactor ecosystem demonstrated an additional electron donor, incorporating both the SOX systems and DSO processes. Metagenome-assembled genomes-based ecological model indicated that carbohydrate metabolism is the key linking factor for the coupling of N and S cycling. Our findings uncover the coupling mechanisms of microbial N and S metabolism, involving both inorganic and organic respiration pathways in lake sediment. This study will enhance our understanding of coupled biogeochemical cycles in lake ecosystems.

Impact of bioelectricity on DNRA process and microbial community composition within cathodic biofilms in dual-chambered bioelectrode microbial fuel cell (MFC)

The synchronous bioelectricity generation and dissimilatory nitrate reduction to ammonium (DNRA) pathway in Klebsiella variicola C1 was investigated. The presence of bioelectricity facilitated cell growth on the anodic biofilms, consequently enhancing the nitrate removal efficiency decreasing total nitrogen levels and causing a negligible accumulation of NO2– in the supernatant. Genomic analysis revealed that K. variicola C1 possessed a complete DNRA pathway and largely annotated electron shuttles. The up-regulated expression of genes narG and nirB, encoding nitrite oxidoreductase and nitrite reductase respectively, was closely associated with increased extracellular electron transfer (EET). High-throughput sequencing analysis was employed to investigate the impact of bioelectricity on microbial community composition within cathodic biofilms. Results indicated that Halomonas, Marinobacter and Prolixibacteraceae were enriched at the cathode electrodes. In conclusion, the integration of a DNRA strain with MFC facilitated the efficient removal of wastewater containing high concentrations of NO3– and enabled the environmentally friendly recovery of NH4+.

Resting cells of Skeletonema marinoi assimilate organic compounds and respire by dissimilatory nitrate reduction to ammonium in dark, anoxic conditions.

Diatoms can survive long periods in dark, anoxic sediments by forming resting spores or resting cells. These have been considered dormant until recently when resting cells of Skeletonema marinoi were shown to assimilate nitrate and ammonium from the ambient environment in dark, anoxic conditions. Here, we show that resting cells of S. marinoi can also perform dissimilatory nitrate reduction to ammonium (DNRA), in dark, anoxic conditions. Transmission electron microscope analyses showed that chloroplasts were compacted, and few large mitochondria had visible cristae within resting cells. Using secondary ion mass spectrometry and isotope ratio mass spectrometry combined with stable isotopic tracers, we measured assimilatory and dissimilatory processes carried out by resting cells of S. marinoi under dark, anoxic conditions. Nitrate was both respired by DNRA and assimilated into biomass by resting cells. Cells assimilated nitrogen from urea and carbon from acetate, both of which are sources of dissolved organic matter produced in sediments. Carbon and nitrogen assimilation rates corresponded to turnover rates of cellular carbon and nitrogen content ranging between 469 and 10,000 years. Hence, diatom resting cells can sustain their cells in dark, anoxic sediments by slowly assimilating and respiring substrates from the ambient environment.

Interspecies cooperation-driven photogenerated electron transfer processes and efficient multi-pathway nitrogen removal in the g-C3N4-anammox consortia biohybrid system

Photocatalytic materials-microbial biohybrid systems pave the way for solar-driven wastewater nitrogen removal. In this study, interspecies cooperation in photogenerated electron transfer and efficient nitrogen removal mechanism in the g-C3N4-anammox consortia biohybrid system were first deciphered. The results indicated that the essential extracellular electron carriers (cytochrome c and flavin) for anammox genomes were provided by associated bacteria (BACT3 and CHLO2). This cooperation, regulated by the ArcAB system and electron transfer flavoprotein, made anammox bacteria the primary photogenerated electron sink. Furthermore, an efficient photogenerated electron harness was used to construct a reductive glycine pathway (rGlyP) in anammox bacteria inventively, which coexisted with the Wood–Ljungdahl pathway (WLP), constituting a dual-pathway carbon fixation model, rGlyP-WLP. Carbon fixation products efficiently contributed to the tricarboxylic acid cycle, while inhibiting electron diversion in anabolism. Photogenerated electrons were targeted channeled into nitrogen metabolism-available electron carriers, enhancing anammox and dissimilatory nitrate reduction to ammonium (DNRA) processes. Moreover, ammonia assimilation by the glycine cleavage system in rGlyP established an alternative ammonia removal route. Ultimately, multi-pathway nitrogen removal involving anammox, DNRA, and rGlyP achieved 100 % ammonia removal and 94.25 % total nitrogen removal efficiency. This study has expanded understanding of anammox metabolic diversity, enhancing its potential application in carbon-neutral wastewater treatment.

Driving mechanism of different nutrient conditions on microbial mediated nitrate reduction in magnetite-present river infiltration zone

Significant research is focused on the ability of riparian zones to reduce groundwater nitrate contamination. Owing to the extremely high redox activity of nitrate, naturally existing electron donors, such as organic matter and iron minerals, are crucial in facilitating nitrate reduction in the riparian zone. Here, we examined the coexistence of magnetite, an iron mineral, and nitrate, a frequently observed coexisting system in sediments, to investigate nitrate reduction features at various C/N ratios and evaluate the response of microbial communities to these settings. Additionally, we aimed to use this information as a foundation for examining the effect of nutritional conditions on the nitrate reduction process in magnetite-present environments. These results emphasise the significance of organic matter in enabling dissimilatory nitrate reduction to ammonium (DNRA) and enhancing the connection between nitrate reduction and iron in sedimentary environments. In the later phases of nitrate reduction, nitrogen fixation was the prevailing process in low-carbon environments, whereas high-carbon environments tended to facilitate the breakdown of organic nitrogen. High-throughput sequencing analysis revealed a robust association between C/N ratios and alterations in microbial community composition, providing insights into notable modifications in essential functioning microorganisms. The nitrogen-fixing bacterium Ralstonia is more abundant in ecosystems with scarce organic matter. In contrast, in settings rich in organic matter, microorganisms, such as Acinetobacter and Clostridia, which may produce ammonia, play crucial roles. Moreover, the population of iron bacteria grows in such an environment. Hence, this study proposes that C/N ratios can influence Fe(II)/Fe(III) conversions and simultaneously affect the process of nitrate reduction by shaping the composition of specific microbial communities.

Uncovering the role of oxygen on organic carbon cycling: insights from a continuous culture study with a facultative anaerobic bacterioplankton species (Shewanella baltica)

Deoxygenation is tied to organic carbon (Corg) supply and utilization in marine systems. Under oxygen-depletion, bacteria maintain Corg respiration using alternative electron acceptors such as nitrate. Since anaerobic respiration’s energy yield is lower, Corg remineralization may be reduced and its residence time increased. We investigated the influence of oxygen and alternative electron acceptors’ availability on Corg cycling by heterotrophic bacteria during a continuous culture experiment with Shewanella baltica, a facultative anaerobic γ-Proteobacteria in the Baltic Sea. We tested six different oxygen levels, from suboxic (&amp;lt;5 µmol L-1) to fully oxic conditions, using a brackish (salinity=14 g L-1) media supplied with high (HighN) or low (LowN) inorganic nitrogen concentrations relative to glucose as labile Corg source. Our results show that suboxia limited DOC (glucose) uptake and cell growth only under LowN, while higher availability of alternative electron acceptors seemingly compensated oxygen limitation under HighN. N-loss was observed under suboxia in both nitrogen treatments. Under HighN, N-loss was highest and a C:N loss ratio of ~2.0 indicated that Corg was remineralized via denitrification. Under LowN, the C:N loss ratio under suboxia was higher (~5.5), suggesting the dominance of other anaerobic respiration pathways, such as dissimilatory nitrate reduction to ammonium (DNRA). Bacterial growth efficiency was independent of oxygen concentration but higher under LowN (34 ± 3.0%) than HighN (26 ± 1.6%). Oxygen concentration also affected dissolved organic matter (DOM) cycling. Under oxic conditions, the release of dissolved combined carbohydrates was enhanced, and the amino acid-based degradation index (DI) pointed to more diagenetically altered DOM. Our results suggest bacterial Corg uptake in low-oxygen systems dominated by S. baltica can be limited by oxygen but compensated by high nitrate availability. Hence, suboxia diminishes Corg remineralisation only when alternative electron acceptors are lacking. Under high nitrate:Corg supply, denitrification leads to a higher N:C loss ratio, potentially counteracting eutrophication in the long run. Low nitrate:Corg supply may favour other anaerobic respiration pathways like DNRA, which sustains labile nitrogen in the system, potentially intensifying the cycle of eutrophication. Going forward, it will be crucial to establish the validity of our findings for S. baltica in natural systems with diverse organic substrates and microbial consortia.