Denitrification Research Articles

Salt stimulates sulfide−driven autotrophic denitrification: Microbial network and metagenomics analyses

Sulfur autotrophic denitrification (SADN) is a promising biological wastewater treatment technology for nitrogen removal, and its performance highly relies on the collective activities of the microbial community. However, the effect of salt (a prevailing characteristic of some nitrogen−containing industrial wastewaters) on the microbial community of SADN is still unclear. In this study, the response of the sulfide−SADN process to different salinities (i.e., 1.5 % salinity, 0.5 % salinity, and without salinity) as well as the involved microbial mechanisms were investigated by molecular ecological network and metagenomics analyses. Results showed that the satisfactory nitrogen removal efficiency (>97 %) was achieved in the sulfide−SADN process (S/N molar ratio of 0.88) with 1.5 % salinity. In salinity scenarios, the genus Thiobacillus significantly proliferated and was detected as the dominant sulfur−oxidizing bacteria in the sulfide−SADN system, occupying a relative abundance of 29.4 %. Network analysis further elucidated that 1.5 % salinity had enabled the microbial community to form a more densely clustered network, which intensified the interactions between microorganisms and effectively improved the nitrogen removal performance of the sulfide−SADN. Metagenomics sequencing revealed that the abundance of functional genes encoding for key enzymes involved in SADN, dissimilatory nitrate reduction to ammonium, and nitrification was up−regulated in the 1.5 % salinity scenario compared to that without salinity, stimulating the occurrence of multiple nitrogen transformation pathways. These multi−paths contributed to a robust SADN process (i.e., nitrogen removal efficiency >97 %, effluent nitrogen <2.5 mg N/L). This study deepens our understanding of the effect of salt on the SADN system at the community and functional level, and favors to advance the application of this sustainable bioprocess in saline wastewater treatment.

Insights into the abundance, expression and diversity of key denitrification genes in an ecologically managed greenhouse agricultural soil

Understanding the bacteria associated with nitrification and denitrification is crucial for comprehending the processes that lead to nitrous oxide emissions in agricultural greenhouse soils. Therefore, it is important to determine their abundance and expression to gain insight into these processes.The aim of this study was to explore the bacterial communities associated with denitrification in a greenhouse agricultural soil amended with crop residues and manure for six years. For this purpose, we proceeded to detect and quantify the genes nirK and nirS and the gene nosZ through clone library construction, sequencing, phylogenetic analysis, and quantitative polymerase chain reaction (qPCR). Sequence analysis based on the clone library revealed that many of the nirS or nirK genes detected were not closely related to known denitrifier bacteria, but some of the nosZ sequences were related to the genera such as Pseudomonas, Halomonas, and Marinobacter. Furthermore, the qPCR revealed a high abundance of DNA copies in nirK, 6.08 × 109 ± 1.16 × 109, while nirS and nosZ showed lower values, 9.05 × 106 ± 1.65 × 106 and 8.71 × 106 ± 1.44 × 106, respectively. However, the highest expression rate was observed for nirS (mRNA/DNA ratio = 3.10 × 10− 3), while nirK and nosZ showed 10-fold lower expression rates (4.4 × 10− 4 and 3.5 × 10− 4, respectively). The results of this work provide a preliminary overview of the diversity, abundance and expression of key genes associated with the denitrification process in this type of soil and are a starting point for further studies to understand how this type of soil management can influence the denitrification process.

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.

Biodegradable film mulching increases soil microbial network complexity and decreases nitrogen-cycling gene abundance

Biodegradable plastic films have emerged as a substitute for conventional plastic films. Nevertheless, responses of plant-associated microbiomes to the application of biodegradable film mulching at field scale have received little attention. A field experiment was conducted to assess the influence of different film mulching treatments on various microbial attributes and nitrogen (N) cycling functional genes in bulk and rhizosphere soils. Biodegradable film mulching raised the bacterial Shannon index in bulk soils but not in rhizosphere soils. Biodegradable film mulching has led to an increase in the complexity and connectivity of microbial networks, as well as an enhancement in the positive association among microorganisms owing to raised soil nutrients and increased crop biomass. In biodegradable film-treated soils, both bacterial and fungal communities were primarily influenced by stochastic processes associated with dispersal limitation. Moreover, conventional plastic film mulching increased denitrification, anammox, N fixation, and dissimilatory nitrate-reduction (DNRA) gene abundance in bulk soils. In rhizosphere soils, biodegradable film mulching reduced nitrification, denitrification, anammox, N fixation, and DNRA gene abundance. Furthermore, keystone genera (e.g., Nitrosospira, Truepera, Adhaeribacter, Opitutus, and Fusarium) were affected by edaphic variables, contributing to decreased N-cycling gene abundance in biodegradable film-treated soils. Collectively, biodegradable film mulching transformed soil microbiome assembly and functional adaptation, and soil nutrient availability and plant biomass were the critical factors influencing the microbial community. These findings present a novel perspective on the diverse impacts of biodegradable and conventional film mulching on soil microbiome and N-cycling processes.

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.

Application of immobilized biological fillers for the hydrolytic acidification and denitrification of sulfide-containing tannery wastewater

This study investigated the application of immobilized biological fillers in the hydrolytic acidification (HA) and denitrification (DN) of actual sulfide-containing tannery wastewater (TWW) through pilot-scale systematic experiments, which provided technical insights for the engineering application of immobilized bio-filler in practical TWW treatment. HA fillers and DN fillers could effectively realize the enrichment and maintenance of functional flora even under long-term inhibition of high sulfide concentration. The HA fillers demonstrated efficient conversion of organic nitrogen. At a hydraulic retention time (HRT) in the range of 3–6 h, the HA reactor achieved an average organic nitrogen conversion concentration of about 100 mg/L, accounting for 56.2 %-68.5 % of the total organic nitrogen in the TWW. Under this condition, the highest average organic nitrogen conversion and mean chemical oxygen demand (COD) degradation efficiencies were achieved, at 32.95 mgNH4+-N/(L·h) and 179.42 mgCOD/(L·h), respectively. Free ammonia (FA) may contribute to pH stabilization in the HA reactor. Denitrification fillers could utilize the endogenous organic matter in the actual TWW hydrolyzed acidified solution, and realize stable biological nitrogen removal effect with no additional carbon source. The denitrification rate reached 76.3 mg NO3--N/(L·h) and the ΔC/ΔN of the denitrification ranged from 3 to 4.

Sulfate-reducing bacteria decreases fractional pressure of H2 to accelerate short-chain fatty acids production from waste activated sludge fermentation assisted with zero-valent iron activated sulfite pretreatment

The production of short-chain fatty acids (SCFAs) is constrained by substrate availability and the increased fractional pressure of H2 emitted by acidogenic/fermentative bacteria during anaerobic fermentation of waste activated sludge (WAS). This study introduced a novel approach employing zero-valent iron (ZVI)-activated sulfite pretreatment combined with H2-consuming sulfate-reducing bacteria (SRB) mediation to improve SCFAs, especially acetate production from WAS fermentation. Experimental results showed that the combined ZVI-activated sulfite and incomplete-oxidative SRB (io-SRB) process achieved a peak SCFAs production of 868.11 mg COD/L, with acetate accounting for 80.55 %, which was 7.90- and 2.18-fold higher than that obtained from raw WAS fermentation, respectively. This could be firstly attributed to the SO4− and OH generated by ZVI-activated sulfite, which significantly promoted WAS decomposition, e.g., soluble proteins and carbohydrates increased 14.3- and 10.8-fold, respectively, over those in raw WAS. The biodegradation of dissolved organic matter was subsequently enhanced by the synergistic interaction and H2 transfer between anaerobic fermentation bacteria (AFB) and io-SRB. The positive and negative correlations among AFB, nitrate-reducing bacteria (NRB) and the io-SRB consortia were revealed by molecular ecological network (MEN) and Mantel test. Moreover, the expression of functional genes was also improved, for instance, in relation to acetate formation, the relative abundances of phosphate acetyltransferase and acetate kinase was 0.002 % and 0.005 % higher than that in the control test, respectively. These findings emphasized the importance of sulfate radicals-based oxidation pretreatment and the collaborative relationships of multifunctional microbes on the value-added chemicals and energy recovery from sludge fermentation.

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.

Effects of sliver nanoparticles on nitrogen removal by the heterotrophic nitrification-aerobic denitrification bacteria Zobellella sp. B307 and their toxicity mechanisms

Due to the widespread use of sliver nanoparticles (AgNPs), a large amount of AgNPs has inevitably been released into the environment, and there is growing concern about the toxicity of AgNPs to nitrogen-functional bacteria. In addition to traditional anaerobic denitrifying bacteria, heterotrophic nitrification-aerobic denitrification (HNAD) bacteria are also important participants in the nitrogen cycle. However, the mechanisms by which AgNPs influence HNAD bacteria have yet to be explicitly demonstrated. In this study, the inhibitory effects of different concentrations of AgNPs on a HNAD bacteria Zobellella sp. B307 were investigated, and the underlying mechanism was explored by analyzing the antioxidant system and the activities of key denitrifying enzymes. Results showed that AgNPs could inhibit the growth and the HNAD ability of Zobellella sp. B307. AgNPs could accumulate on the surface of bacterial cells and significantly destroyed the cell membrane integrity. Further studies demonstrated that the presence of high concentration of AgNPs could result in the overproduction of reactive oxygen species (ROS) and related oxidative stress in the cells. Furthermore, the catalytic activities of key denitrifying enzymes (nitrate reductase (NAR), nitrite reductase (NIR), and nitrous oxide reductase (N2OR)) were significantly suppressed under exposure to a high concentration of AgNPs (20 mg·L−1), which might be responsible for the inhibited nitrogen removal performance of strain B307. This work could improve our understanding of the inhibitory effect and underlying mechanism of AgNPs on HNAD bacteria.

In situ electron generation through Fe/C supported sludge coupled with a counter-diffusion biofilm for electron-deficient wastewater treatment: Binding properties and catalytic competition mechanism of nitrate reductase

A membrane-aerated biofilm-coupled Fe/C supported sludge system (MABR-Fe/C) was constructed to achieve in situ electron production for NO3−-N reduction enhancement in different Fe/C loadings (10 g and 200 g). The anoxic environment formed in the MABR-Fe/C promoted a continual Fe2+release of Fe/C in 120 d operation (average Fe2+concentrations is 1.18 and 2.95 mg/L in MABR-Fe/C10 and MABR-Fe/C200, respectively). Metagenomics results suggested that the electrons generated from ongoing Fe2+ oxidation were transferred via the Quinone pool to EC 1.7.5.1 rather than EC 1.9.6.1 to complete the process of NO3−-N reduction to NO2−-N in Acidovorax, Ottowia, and Polaromonas. In the absence of organic matter, the NO3−-N removal in MABR-Fe/C10 and MABR-Fe/C200 increased by 11.99 and 12.52 mg/L, respectively, compared to that in MABR. In the further NO2−-N reduction, even if the minimum binding free energy (MBFE) was low, NO2−-N in Acidovorax and Dechloromonas preferentially bind the Gln-residues for dissimilatory nitrate reduction (DNR) in the presence of Fe/C. Increasing Fe/C loading (MABR-Fe/C200) caused the formation of different residue binding sites, further enhancing the already dominant DNR. When DNR in MABR-Fe/C200 intensified, the TN in the effluent increased by 3.75 mg/L although the effluent NO3−-N concentration was lower than that in MABR-Fe/C10. This study demonstrated a new MABR-Fe/C system for in situ electron generation to enhance biological nitrogen removal and analyzed the NO3−-N reduction pathway and metabolic mechanism, thus providing new ideas for nitrogen removal in electron-deficient wastewater.

Rapid conversion of heterotrophic denitrification to autotrophic denitrification in mariculture wastewater treatment: Denitrification performance and microbial communities under antibiotic stress

Mariculture wastewater, characterized by a low COD/Nitrogen (C/N) ratio and the occurrence of antibiotics, is commonly treated by a heterotrophic denitrification (HD) process with supplying substantial carbon sources. Compared to HD, sulfur-based autotrophic denitrification (AD) requires less external carbon sources and is more favorable for low-carbon wastewater treatment. However, the AD process often consumes more startup time. Thus, the present study was designed to explore a rapid startup strategy for the AD process by switching from HD to AD, treating mariculture wastewater. A rapid transformation from HD to AD was achieved by inoculating mixotrophic denitrifying sludge, reducing carbon sources and adding sulfur sources under the antibiotic sulfamethoxazole (SMX). From the functional microbial community point of view, our results showed that autotrophic bacteria exhibited greater adaptability to sulfamethoxazole than heterotrophic bacteria. During the transition from HD to AD, nitrate removal improved from 53.13 % to 68.57 %, and the secretion of tightly bound extracellular polymeric substances (TB-EPS) increased by 29.94 mg/g VSS. Meanwhile, in the heterotrophic phase, the presence of sulfate in mariculture wastewater maintained a high abundance of sulfate-reducing bacteria such as Desulfuromusa (0.8 %) and sulfur-based autotrophic denitrifying bacteria such as Sedimenticola (36.7 %) and Thiogranum (15.2 %). This increases the activity of sulfur metabolism during the heterotrophic phase and ensures a rapid transition from HD to AD. The rapid shift from HD to AD in seawater under antibiotic stress provides theoretical support for the improvement of mariculture wastewater treatment processes.

Promising membrane photobioreactor for continuous flow treatment of Low C/N wastewater: Treating process, community structure, and cooperation mechanism

Low carbon/nitrogen (C/N) wastewater treatment is an emerging issue for conventional treatment technologies. To achieve this goal, a novel membrane photobioreactor (MPBR), integrating microalgae consortium (MC) and activated sludge (AS) phases, was investigated. The separation of the MC and AS phases could improve the removal of pollutants in low C/N wastewater, which could achieve the rapid denitrogenation through NH4+-N assimilation in MC phase, and effectively reduce organic matter concentration in AS phase, resulting in better nutrient recovery, lower costs, and stable pH. The results in phase I showed that the pollutants, such as chemical oxygen demand (COD), ammonium (NH4+-N), and total phosphorus (TP), met the Environmental Quality Standards for Surface Water (GB3838–2002) by MPBR. The AS phase with low DO, which could enrich heterotrophic bacteria (e.g., Bacteroidetes vadinHA17, Spirochaeta and Ahniella) and nitrogen cycling bacteria (e.g., Mycobacterium, Bacillus and Ellin6067), could promote the removal of COD and NO3--N in the continuous effluent. Based on the co-occurrence pattern analysis, the main influencing factor of MC and AS phases was the reduction of COD, which also illustrated the synergistic removal of organic matter by the cooperative interaction of microalgae and heterotrophic bacteria. From a genetic perspective, the internal circulation mode also enhanced microbial nitrogen metabolism in the MPBRs, with higher abundances of nitrification- and denitrification-related functional genes in the MC phase, and higher abundances of nitrification, dissimilatory nitrate reduction, and denitrification genes in the AS phase. This study provided an experimental and theoretical basis for MPBR in low-C/N wastewater treatment.

Iron-scrap and pyrite mediated autotrophic denitrification in constructed wetland for enhancing nitrate-polluted groundwater remediation: Performance and mechanism

Nitrate (NO3−-N) is ubiquitously present in groundwater, constituting a growing threat to human health. In this study, iron-scrap and pyrite-based constructed wetlands (CWs) were constructed to investigate the treatment performance and removal mechanisms in purifying nitrate-contaminated groundwater. Results showed that the NO3−-N removal efficiency of iron-scrap-based CWs (87.1%) was higher than that of pyrite-based CWs (59.5%), and the higher N2O emission (28.46 μg/(m2•h)) was also obtained in CW with iron-scrap. Nevertheless, total nitrogen removal did not exhibit a significant difference between two CWs due to the accumulation of ammonium nitrogen (NH4+-N) in iron-scrap-based CWs. Additionally, sulfate was produced in CW with pyrite, as a by-product, and caused oxidative stress damage to plants. Metagenomic analysis showed that the addition of iron-scrap could increase microbial diversity and enrich autotrophs denitrifies (e.g., Ferritrophicum), which explained the higher NO3−-N removal reasonably. Additionally, the abundance of dissimilatory nitrate reduction bacterium was enhanced in iron-scrap-based CW, explaining the NH₄⁺-N accumulation. Further metabolic analysis confirmed the occurrence of Fe cycle in CW with iron-scrap, and functional genes related to NO3−-N removal and TCA cycle were up-regulated, offering a rationale for the improved NO3−-N removal in CW with iron-scrap. This study offers comprehensive insights into the NO3−-N removal and its mechanism through iron mediated CWs in controlling groundwater pollution.

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.

Deciphering linkages between DON and the microbial community for nitrogen removal using two green sorption media in a surface water filtration system

The presence of dissolved organic nitrogen (DON) in stormwater treatment processes is a continuous challenge because of the intertwined nature of its decomposition, bioavailability, and biodegradability and its unclear molecular characteristics. In this paper, 21 T Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) in combination with quantitative polymerase chain reaction was applied to elucidate the molecular change of DON and microbial population dynamics in a field-scale water filtration system filled with two specialty adsorbents for comparison in South Florida where the dry and wet seasons are distinctive annually. The adsorbents included CPS (clay-perlite and sand sorption media) and ZIPGEM (zero-valent iron and perlite-based green environmental media). Our study revealed that seasonal effects can significantly influence the dynamic characteristics and biodegradability of DON. The microbial population density in the filter beds indicated that three microbial species in the nitrogen cycle were particularly thrived for denitrification, dissimilatory nitrate reduction to ammonium, and anaerobic ammonium oxidation via competition and commensalism relationships during the wet season. Also, there was a decrease in the compositional complexity and molecular weight of the DON groups (CnHmOpN1, CnHmOpN2, CnHmOpN3, and CnHmOpN4), revealed by the 21 T FT-ICR MS bioassay, driven by a microbial population quantified by polymerase chain reaction from the dry to the wet season. These findings indirectly corroborate the assumption that the metabolism of microorganisms is much more vigorous in the wet season. The results affirm that the sustainable materials (CPS and ZIPGEM) can sustain nitrogen removal intermittently by providing a suitable living environment in which the metabolism of microbial species can be cultivated and enhanced to facilitate physico-chemical nitrogen removal across the two types of green sorption media.

Enhancing nitrogen removal performance using immobilized aerobic denitrifying bacteria by modified polyvinyl alcohol/sodium alginate (PVA/SA)

Aerobic denitrification has emerged as a promising and efficient method for nitrogen removal from wastewater. However, the direct application of aerobic denitrifying bacteria has faced challenges such as low nitrogen removal efficiency, bacterial loss, and poor stability. To address these issues, this study developed a novel microbial particle carrier using NaHCO3-modified polyvinyl alcohol (PVA)/sodium alginate (SA) gel (NaHCO3-PVA/SA). This carrier exhibits several advantageous properties, including excellent mass transfer efficiency, favorable biocompatibility, convenient film formation, abundant biomass, and exceptional pollutant treatment capacity. The carrier was modified with 0.3% NaHCO3, 8.0% PVA, and 1.0% SA, resulting in a remarkable 3.4-fold increase in the average pore diameter and a 12.8% improvement in mass transfer efficiency. This carrier was utilized to immobilize the aerobic denitrifying bacterium Stutzerimonas stutzeri W-2 to enhance nitrogen removal (NaHCO3-PVA/SA@W-2), resulting in a NO3−-N removal efficiency of 99.06%, which was 21.39% higher than that without modification. Compared with the non-immobilized W-2, the degradation efficiency was improved by 43.70%. After five reuses, the NO3−-N and TN removal rates remained at 99% and 93.01%, respectively. These results provide a solid foundation for the industrial application of the modified carrier as an effective tool for nitrogen removal in large-scale wastewater treatment processes.

Effects of soil water content at freezing, thaw temperature, and snowmelt infiltration on N2O emissions and denitrifier gene and transcript abundance during a single freeze-thaw event

Climate change-related warming and increased precipitation may alter winter snow cover and thawing events, and therefore, may carry significant consequences for nitrous oxide (N2O) production pathways such as denitrification, and the abundance and expression of denitrifying microorganisms. We used a soil microcosm study to investigate the combined effect of soil thaw temperature, initial water filled pore space (WFPS) prior to soil freezing, and snowmelt infiltration simulated by the addition of water on N2O emission and denitrification rates, soil respiration rate, and the abundance and transcription of denitrifying (nirK, nirS, and nosZ) bacteria during a single freeze-thaw event. Soil respiration rate was primarily controlled by an increase in soil thaw temperature, whereas soil N2O emission and denitrification rates were generally greater in soils with a higher initial WFPS and soil thaw temperature. In contrast, snowmelt infiltration generally had a negligible effect on these rates, which may be related to pre-existing soil conditions that were already conducive to denitrification. Unexpectedly, the nosZ transcript/nosZ gene abundance ratio was lower in soils thawed at 8.0 °C compared to 1.5 °C; however, this may have resulted in a lower N2O reduction, thus explaining the greater levels of N2O emitted from soils thawed at 8.0 °C. Overall, this study demonstrated that increased N2O production during a single freeze-thaw event was primarily linked to antecedent conditions of high initial WFPS, soil thaw temperature, and a synergistic interplay between these two environmental parameters, and provides evidence that an increase in annual temperature and precipitation, along with the timing of precipitation, may further stimulate N2O production pathways.

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+.