nosZ Gene Research Articles

Evaluation of soil properties and bulk δ15N to assess decadal changes in floodplain denitrification following restoration

Stream and floodplain restoration is a popular billion‐dollar industry in the United States, with many restorations being conducted to satisfy water pollution regulations and nutrient reduction goals. The long‐term efficacy of these restorations is, however, not well studied, and key soil metrics that can be used for performance assessments have not been developed. We evaluated a chronosequence of 12 restoration sites spanning an age range of 0–22 years to assess changes in denitrification rates and associated soil parameters. Restored versus unrestored reaches were compared for denitrification rate and functional gene nosZ, bulk soil δ15N, soil organic carbon (SOC), soil organic matter (SOM), bulk density, and soil moisture. Denitrification, SOM, SOC, and soil moisture were all found to increase with site age at restored sites, with the largest increase for the 10–22 age category. Bulk density decreased with time, with a significant decrease in restored floodplain soils. Bulk soil δ15N was highest immediately after restoration, decreased with restoration age, and was not positively correlated with denitrification. This may reduce its potential as a proxy for denitrification. Overall, this study reveals that selected soil metrics (SOC, SOM, soil moisture, and bulk density) could serve as a valuable proxy for denitrification and could help assess the denitrification effectiveness of floodplain restorations at the decadal time scales. Ideally, the soil metrics should be combined with other short‐term assessment measures, such as those for stream and groundwaters, for a robust performance assessment of restored floodplains.

Biochar amendment affects the microbial genetic profile of the soil, its community structure and phospholipid fatty acid contents

Biochar (BC) amendment has been proposed as a promising strategy for mitigating greenhouse gas (GHG) emissions, specifically carbon dioxide (CO2), methane (CH4), and nitrous oxide (N2O). Conducting a meta-analysis to evaluate the impact of biochar on microbial genetic profile, community structure, and phospholipid fatty acid (PLFA) contents can aid in identifying key microbial groups involved in GHG production and consumption, and assessing the overall effectiveness of biochar in reducing GHG emissions. The present meta-analysis revealed that the addition of biochar resulted in a 22 % and 41 % reduction in pmoA and mcrA genes of methanogenic microorganisms, respectively. The mcrA/pmoA ratio significantly increased by 81 %. Gene abundances exhibited a positive response to biochar amendment, with increases observed in nifH, nirK, nirS, nosZ, and nosZ (nirS + nirK) genes by 13 %, 32 %, 37 %, 42 %, and 79 %, respectively. Moreover, biochar amendment influenced the microbial community structure accordingly. The concentration of PLFAs increased in response to BC treatment in the following order: A-bacteria (+49 %) < Fungi (+30 %) < Gram-pb (+21 %) < G-bacteria (+17 %) < Gram-nb (+11 %). These findings indicate that biochar amendment shapes the microbial community structure, further emphasizing its significance in enhancing soil fertility.

Dose Effect of Drinking Water Nitrate on Health, Feed Intake, Rumen Fermentation and Microbiota, and Nitrogen Excretion in Holstein Heifers for a Sustainable Water Use

The present study aimed to evaluate the potential hazardous effects of NO3− concentration in drinking water on health, feed intake, rumen fermentation and microbiota, and nitrogen excretion of Holstein heifers fed a high-concentrate diet for a sustainable water use. Twenty-four Holstein heifers were individually allocated and assigned to one of four treatments with increasing drinking water NO3− concentration: CTR, without NO3−; LOW, with 44 mg NO3−/L; MOD, with 110 mg NO3−/L; and HIGH, with 220 mg NO3−/L. The entire study lasted 168 days. Fortnightly water NO3− concentration and daily feed and water intake were recorded. Blood parameters, rumen pH, volatile fatty acids, NO3− and NO2− concentration, microbiota, and apparent total tract digestibility were determined at the beginning and at the end of the study. Most of the analyzed parameters were similar among treatments. Denitrifying bacteria population, estimated as nosZ gene copies, were greater in HIGH animals than in CTR animals at the end of the study. In conclusion, drinking water NO3− concentration up to 220 mg/L has no detrimental effect on health, feed intake, rumen fermentation, nor N excretion in dairy beef cattle for periods up to 168 days; moreover, denitrifying bacteria population increased, which are related with the neutralization of the greenhouse gas N2O.

Combined metabolomic and microbial community analyses reveal that biochar and organic manure alter soil C-N metabolism and greenhouse gas emissions

The use of biochar to reduce the gas emissions from paddy soils is a promising approach. However, the manner in which biochar and soil microbial communities interact to affect CO2, CH4, and N2O emissions is not clearly understood, particularly when compared with other amendments. In this study, high-throughput sequencing, soil metabolomics, and quantitative real-time PCR were utilized to compare the effects of biochar (BC) and organic manure (OM) on soil microbial community structure, metabolomic profiles and functional genes, and ultimately CO2, CH4, and N2O emissions. Results indicated that BC and OM had opposite effects on soil CO2 and N2O emissions, with BC resulting in lower emissions and OM resulting in higher emissions, whereas BC, OM, and their combined amendments increased cumulative CH4 emissions by 19.5 %, 31.6 %, and 49.1 %, respectively. BC amendment increased the abundance of methanogens (Methanobacterium and Methanocella) and denitrifying bacteria (Anaerolinea and Gemmatimonas), resulting in an increase in the abundance of mcrA, amoA, amoB, and nosZ genes and the secretion of a flavonoid (chrysosplenetin), which caused the generation of CH4 and the reduction of N2O to N2, thereby accelerating CH4 emissions while reducing N2O emissions. Simultaneously, OM amendment increased the abundance of the methanogen Caldicoprobacter and denitrifying Acinetobacter, resulting in increased abundance of mcrA, amoA, amoB, nirK, and nirS genes and the catabolism of carbohydrates [maltotriose, D-(+)-melezitose, D-(+)-cellobiose, and maltotetraose], thereby enhancing CH4 and N2O emissions. Moreover, puerarin produced by Bacillus metabolism may contribute to the reduction in CO2 emissions by BC amendment, but increase in CO2 emissions by OM amendment. These findings reveal how BC and OM affect greenhouse gas emissions by modulating soil microbial communities, functional genes, and metabolomic profiles.

Time course transcriptomic profiling suggests Crp/Fnr transcriptional regulation of nosZ gene in a N2O-reducing thermophile

Time course transcriptomic profiling suggests Crp/Fnr transcriptional regulation of nosZ gene in a N2O-reducing thermophile

Dual mechanism of membrane covering on GHG and NH3 mitigation during industrial-scale experiment on dairy manure composting: Inhibiting formation and blocking emissions

An industrial-scale experiment on dairy manure composting with the control group (Ctrl) and the membrane covering group (CM) was conducted to explore the effects of functional membrane covering on gas emissions, the conversion of carbon and nitrogen, and revealing the underlying mechanisms. Results indicated that CM achieved the synergistic effects on gas mitigation and improved compost product quality. CO2, CH4, N2O, and NH3 emissions were reduced by 81.8%, 87.0%, 82.6%, and 82.2%, respectively. The micro-aerobic condition formed in membrane covering compost pile together with the covering inhibiting effect dominated the mitigation effect. CM significantly downregulated the mcrA gene copies and the value of mcrA/pmoA (p < 0.01), which reduced CH4 emission. CM decreased the nirS and nirK gene copies and increased the nosZ gene copies to reduce N2O emission. Functional Annotation of Prokaryotic Taxa showed that membrane covering effectively amended part of carbon and nitrogen cycles, which stimulated the degradation of organic matter, accelerated compost maturity and reduced the gaseous emissions.

Unraveling the genetic potential of nitrous oxide reduction in wastewater treatment: insights from metagenome-assembled genomes.

This study provides critical insights into the genetic diversity of nitrous oxide reductase (nosZ) genes and the microorganisms harboring them in wastewater treatment plants (WWTPs) by exploring 1,083 high-quality metagenome-assembled genomes (MAGs) from 23 Danish full-scale WWTPs. Despite the pivotal role of nosZ-containing organisms, their diversity remains largely unexplored in WWTPs. Our custom pipeline for detecting nosZ provides near-full-length genes with detailed information on secretory pathways and accessory nos genes. Using these genes as templates, we developed taxonomically diverse clade-specific primers that generate nosZ amplicons for phylogenetic annotation and gene-to-MAG linkage. This approach improves detection and expands the discovery of novel sequences, highlighting the prevalence of non-denitrifying N2O reducers and their potential as N2O sinks. These findings have the potential to optimize nitrogen removal processes and mitigate greenhouse gas emissions from WWTPs by fully harnessing the capabilities of the microbial communities.

Temperature differentially regulates estuarine microbial N2O production along a salinity gradient

Nitrous oxide (N2O) is atmospheric trace gas that contributes to climate change and affects stratospheric and ground-level ozone concentrations. Ammonia oxidizers and denitrifiers contribute to N2O emissions in estuarine waters. However, as an important climate factor, how temperature regulates microbial N2O production in estuarine water remains unclear. Here, we have employed stable isotope labeling techniques to demonstrate that the N2O production in estuarine waters exhibited differential thermal response patterns between nearshore and offshore regions. The optimal temperatures (Topt) for N2O production rates (N2OR) were higher at nearshore than offshore sites. 15N-labeled nitrite (15NO2-) experiments revealed that at the nearshore sites dominated by ammonia-oxidizing bacteria (AOB), the thermal tolerance of 15N-N2OR increases with increasing salinity, suggesting that N2O production by AOB-driven nitrifier denitrification may be co-regulated by temperature and salinity. Metatranscriptomic and metagenomic analyses of enriched water samples revealed that the denitrification pathway of AOB is the primary source of N2O, while clade II N2O-reducers dominated N2O consumption. Temperature regulated the expression patterns of nitrite reductase (nirK) and nitrous oxide reductase (nosZ) genes from different sources, thereby influencing N2O emissions in the system. Our findings contribute to understanding the sources of N2O in estuarine waters and their response to global warming.

Innovative approaches for enhanced nutrient (N and P) removal and biosecurity in marine recirculating aquaculture systems using biofloc and ultrafiltration combined systems

Marine recirculating aquaculture systems (marine RAS) are increasingly utilized for their high productivity and minimal environmental impact. However, effective nutrient management and biosecurity measures remain crucial for sustainable operation of marine RAS. This study successfully established a simultaneous nitrification and denitrification (SND) process using a biofloc technology (BFT)–ultrafiltration (UF) combined approach. A stable autotrophic marine nitrification system was rapidly initiated within 23 days using a ceramic ring (CR)–granular activated carbon (GAC)–UF process (CGUF), followed by the substitution of polyhydroxyalkanoates (PHA) for GAC filter to achieve rapid start-up of SND in 24 h (CG/PUF). Soluble reactive phosphate (SRP) was removed in-situ via biofloc adsorption. The co-existence of ammonia-oxidizing archaea (Candidatus_Nitrosopumilus) and denitrifying bacteria (Stenotrophomonas, Ruegeria and Ilumatobacter) synergistic promoted the SND start-up in CG/PUF process. Stenotrophomonas emerged as the keystone species which closely linked to amoA/B/C, hao and nosZ genes (p < 0.05). The CG/PUF process effectively enhanced the biosecurity of marine RAS, with 100 % elimination of potential pathogens (Vibrio and Tenacibaculum). UF membrane could maintain fluxes at 15 L∙ m−2∙ h−1 (LMH) for 180 days, with a 2-day intermittent hydraulic flushing keeping transmembrane pressure below 50 kPa. These findings provide insights for broader application of BFT–UF combined process in marine RAS.

Microplastics and biochar interactively affect nitrous oxide emissions from tobacco planting soil

Biochar application to amend acidified tobacco-soils can enhance tobacco quality and reduce nitrous oxide (N2O) emissions. Microplastics from agricultural mulch are commonly found in cash-crop farmland soils and, together with biochar, affect soil N2O emissions. In this study, we applied three types of microplastics (polyethylene, PE; polylactic acid, PLA; polybutylene adipate terephthalate, PBAT) and rice biochar alone or in combination to acidified tobacco planting soil in central China to investigate their effects on soil N2O emissions, soil chemical properties, nitrogen-cycle-related functional genes, and microbial functional diversity during a 35-day laboratory incubation period. Significant increases in N₂O emissions were observed with PE and PLA, which raised emissions by 15.96 % and 21.52 %, respectively. Additionally, different microplastics affected soil N₂O emissions through distinct regulatory pathways. Co-application of microplastics and biochar suppressed N2O emissions compared to microplastics alone. Biochar mitigates N2O emissions mainly by increasing the abundance of the nosZ gene. It can remediate soil contaminated by microplastics and reduce their negative impacts on the soil environment. This study provides deeper insight into the effects of microplastics on soil nitrogen cycling and biochar-mitigated remediation of microplastic-contaminated soil.

NosZ II/nosZ I ratio regulates the N2O reduction rates in the eutrophic lake sediments

Nitrous oxide (N2O) is a more potent greenhouse gas with an atmospheric lifetime of 121 years, contributing significantly to climate change and stratospheric ozone depletion. Lakes are hotspots for N2O release due to the imbalance between N2O sources and sinks. N2O-reducing bacteria are the only biological means to mitigate N2O emission, yet their roles in lakes are not well studied. This study investigated the potential for N2O reduction, keystones of typical and atypical N2O-reducing bacterial communities, and their correlations with environmental factors in the sediments of Lake Taihu through microcosm experiments, high-throughput sequencing of the nosZ gene, and statistical modeling. The results showed that potential N2O reduction rates in sediments ranged from 13.71 to 76.95 μg N2O g−1 d−1, with lower rates in December compared to March and July. Correlation analysis indicated that the nosZ II/nosZ I ratio and the trophic lake index (TLI) were the primary factors influencing N2O reduction rates and N2O-reducing bacterial community structures. The genera Pseudogulbenkiania and Ardenticatena were identified as the most abundant typical and atypical N2O-reducing bacteria, respectively, and were also recognized as the keystone taxa. Quantitative real-time PCR (qPCR) results revealed that nosZ II was more abundant than nosZ I in the sediments. Partial least squares path modeling (PLS-PM) further demonstrated that atypical N2O-reducing bacteria had significant positive effects on N2O reduction process in the sediments (p < 0.05). Overall, this study highlights the crucial ecological roles of atypical N2O-reducing bacteria in the sediments of the eutrophic lake of Taihu, underscoring their potential in mitigating N2O emissions.

Evaluating the role of high N2O affinity complete denitrifiers and non-denitrifying N2O reducing bacteria in reducing N2O emissions in river

Freshwater rivers are hotspots of N2O greenhouse gas emissions. Dissolved organic carbon (DOC) is the dominant electron donor for microbial N2O reduction, which can reduce N2O emission through enriching high N2O affinity denitrifiers or enriching non-denitrifying N2O-reducing bacteria (N2ORB), but the primary regulatory pathway remains unclear. Here, field study indicated that high DOC concentration in rivers enhanced denitrification rate but reduced N2O flux by improving nosZ gene abundance. Then, four N2O-fed membrane aeration biofilm reactors inoculated with river sediments from river channel, estuary, adjacent lake, and a mixture were continuously performed for 360 days, including low, high, and mixed DOC stages. During enrichment stages, the (nirS+nirK)/nosZ ratio showed no significant difference, but the community structure of denitrifiers and N2ORB changed significantly (p < 0.05). In addition, N2ORB strains isolated from different enrichment stages positioned in different branches of the phylogenetic tree. N2ORB strains isolated during high DOC stage showed significant higher maximum N2O-reducing capability (Vmax: 0.6 ± 0.4 ×10−4 pmol h−1 cell−1) and N2O affinity (a0: 7.8 ± 7.7 ×10−12 L cell−1 h−1) than strains isolated during low (Vmax: 0.1 ± 0.1 ×10−4 pmol h−1 cell−1, a0: 0.7 ± 0.4 ×10−12 L cell−1 h−1) and mixed DOC stages (Vmax: 0.1 ± 0.1 ×10−4 pmol h−1 cell−1, a0: 0.9 ± 0.9 ×10−12 L cell−1 h−1) (p < 0.05). Hence, under high DOC concentration conditions, the primary factor in reducing N2O emissions in rivers is the enrichment of complete denitrifiers with high N2O affinity, rather than non-denitrifying N2ORB.

Converse Responses of Biochar Application on N2O Emissions in Soils at Different pH Values in a Subtropical Citrus Orchard

The aim of this study was to explore the effect of biochar on N2O emissions in soils with different pH levels. Soils with five pH levels (4.0, 5.1, 5.8, 6.6, and 7.2) were incubated in two conditions, with 0% biochar (CK) and 1% biochar (BC), for 23 days. N2O emissions were measured at nine time points, and soil chemical properties, AOA-amoA, AOB-amoA, nirK, nirS, and nosZ, were analyzed. Partial least squares path modelling (PLS-PM) was used to assess the effect of nitrification and denitrification pathways on potential N2O emissions. The results showed that biochar reduced N2O emissions in highly acidic soil (pH 4.0) but increased emissions in soils with pH values ranging from 5.1 to 7.2. In highly acidic soils, decreased N2O emission was associated with increased soil pH (p &lt; 0.05) and decreased dissolved organic carbon content (p &lt; 0.05), leading to higher nosZ gene abundance (p &lt; 0.05). Meanwhile, in acidic to neutral soils, biochar application increased soil pH (6.6–11.7%), dissolved organic nitrogen (5.9–29.5%), dissolved organic carbon (8.6–41.0%), stimulated AOB-amoA, nirK, nirS gene abundance (p &lt; 0.05), and thus increased N2O emissions. The results verified the influence of nitrification and denitrification genes on N2O production in soils with different pH values. In conclusion, biochar had different effects on N2O emissions based on soil pH, highlighting the need to consider pH when using biochar to mitigate N2O emissions in subtropical citrus orchards.

The electrochemical mechanism of biochar for mediating the product ratio of N2O/(N2O + N2) in the denitrification process

The biochar electrochemical properties and surface functional groups significantly impact N2O production and reduction during denitrification process. However, its effects on N2O emissions during the denitrification process and its electrochemical mechanisms remain unclear. The study examined the impact of pristine and oxidized biochar combined with two types of nitrogen fertilizers on the N2O/(N2O + N2) ratio and N2O emissions in an incubation experiment with seven treatments: (1) CK (no application of chemical fertilizer); (2) N1 (applying (NH4)2SO4); (3) N1B ((NH4)2SO4 + pristine biochar); (4) N1BO ((NH4)2SO4 + oxidized biochar); (5) N2 (applying KNO3); (6) N2B (KNO3 + pristine biochar); (7) N2BO (KNO3 + oxidized biochar). The study found that in comparison with applying nitrogen fertilizer alone, combining pristine biochar decreased soil N2O concentration by 7.1 %–85.8 %, while combining oxidized biochar increased it by 15.7 %–125.6 %. Applying pristine biochar reduced N2O/(N2O + N2) ratio by 10.4 %–86.2 %, whereas applying oxidized biochar increased it by 12.9 %–121.6 %. The application of pristine biochar increased the nosZ gene abundance and decreased the (nirS + nirK)/nosZ ratio, which contributed to reducing N2O to N2. Compared with oxidized biochar, the oxygen-containing functional groups of pristine biochar decreased by 46.6 %, and it possessed a higher specific surface area (23.01 m2 g−1) and electrical conductivity (0.003 mS cm−1). The correlation analysis showed that DOC and inorganic nitrogen were the key environmental factors affecting N2O emissions. Additionally, the electrical conductivity, specific capacitance, and oxygen-containing functional groups of the biochar were identified as the main factors driving N2O emissions. The SEM analysis suggested that the indirect influence of biochar electrochemical properties on N2O emissions was greater than its direct influence. Our work provides fresh perspectives on reducing soil N2O emissions and establishes a theoretical foundation for the subsequent preparation of biochar materials with enhanced N2O reduction capabilities.

Co-incorporation of wheat straw and hairy vetch reduced soil N2O emission via regulating nitrifier and denitrifier structure on the Qinghai plateau

Introducing green manure is considered a sustainable practice in wheat production on the Qinghai plateau, an ecologically fragile area. However, the regulatory mechanisms governing N2O emission through green manure and wheat straw co-incorporation in nitrifiers and denitrifiers communities remain unclear. Herein, a microcosm experiment was conducted, including no-addition (CK), hairy vetch (VS), wheat straw (WS), and the mixture of hairy vetch and wheat straw (VWS), to investigate soil N2O emission and functional microbes communities. Compared with CK, cumulative N2O emission decreased by 19.1 % and 74.8 % in WS and VWS, whereas it increased by 60.4 % in VS. Residues incorporation increased absolute abundances of denitrifier (narG, nirK, and nosZ genes) by 39 %–139 % than CK. However, the contribution of structures of functional microbes on N2O emission overweighed their absolute abundances. The reduced N2O emission was mainly associated with the decreased relative abundance of M5 and M7 (key ecological cluster, gained from co-occurrence network of narG- and nirK-types denitrifier, dominant genera were Pseudomonas and Bradyrhizobium), and with the increased M13 (from nosZ-type denitrifier, dominant genus was Tardiphaga). In N2O emission peak (the 6th day of incubation), VWS reduced the relative abundance of Pseudomonas by 9.9 % and 11.2 % and Bradyrhizobium by 39.7 % and 49.2 % compared with CK and VS, respectively. Additionally, VWS increased the relative abundance of Tardiphaga by 134.1 % and 172.1 % compared with CK and WS. Our results suggest that wheat straw and hairy vetch co-incorporation is an effective practice to mitigate N2O emission via regulating key denitrifier taxa on the Qinghai Plateau.

Discovery of Candidatus Nitrosomaritimum as a New Genus of Ammonia-Oxidizing Archaea Widespread in Anoxic Saltmarsh Intertidal Aquifers.

Ammonia-oxidizing archaea (AOA) are widely distributed in marine and terrestrial habitats, contributing significantly to global nitrogen and carbon cycles. However, their genomic diversity, ecological niches, and metabolic potentials in the anoxic intertidal aquifers remain poorly understood. Here, we discovered and named a novel AOA genus, Candidatus Nitrosomaritimum, from the intertidal aquifers of Yancheng Wetland, showing close metagenomic abundance to the previously acknowledged dominant Nitrosopumilus AOA. Further construction of ammonia monooxygenase-based phylogeny demonstrated the widespread distribution of Nitrosomaritimum AOA in global estuarine-coastal niches and marine sediment. Niche differentiation among sublineages of this new genus in anoxic intertidal aquifers is driven by salinity and dissolved oxygen gradients. Comparative genomics revealed that Candidatus Nitrosomaritimum has the genetic capacity to utilize urea and possesses high-affinity phosphate transporter systems (phnCDE) for surviving phosphorus-limited conditions. Additionally, it contains putative nosZ genes encoding nitrous-oxide (N2O) reductase for reducing N2O to nitrogen gas. Furthermore, we gained first genomic insights into the archaeal phylum Hydrothermarchaeota populations residing in intertidal aquifers and revealed their potential hydroxylamine-detoxification mutualism with AOA through utilizing the AOA-released extracellular hydroxylamine using hydroxylamine oxidoreductase. Together, this study unravels the overlooked role of priorly unknown but abundant AOA lineages of the newly discovered genus Candidatus Nitrosomaritimum in biological nitrogen transformation and their potential for nitrogen pollution mitigation in coastal environments.

Enhancement of nitrogen removal in constructed wetlands through boron mud and elemental sulfur addition: Regulation of sulfur and oxygen cycling

Constructed wetlands (CWs) utilizing elemental sulfur (S0) as an electron donor are effective for nitrate (NO3–-N) removal from low C/N wastewater. However, the limited bioavailability of chemical S0 (chem-S0) due to low water solubility and inadequate mass transfer impedes complete denitrification. In this study, boron mud waste and S0 (S0&BM) were processed and combined to convert chem-S0 to biological S0 (bio-S0) in CWs. The employment of S0&BM improved NO3–-N and total nitrogen (TN) removal rates by 15.36 % and 10.55 % compared to S0 alone. The 15N isotope tracing experiment unveiled the pathways of N2O production and demonstrated that S0&BM facilitated full denitrification, resulting in a higher percentage of nitrogen being degraded in a non-polluting form (54.70 %). Releasing trace amounts of boron and magnesium in S0&BM mitigated the excessive acidification caused by the sulfur autotrophic process and promoted biofilm formation, thereby pronouncing vertical oxygen fluctuations. Meanwhile, oxygen vacancies generated in the S0&BM promoted the closed-loop sulfur cycle, starting from the oxidation of chem-S0 to sulfate (SO42-), the important process of SO42--reducing bacteria using partial organic compounds as electron donors to reduce SO42- to sulfide (S2-) was realized, and finally the S2- was oxidized to bio-S0 with better properties. S0&BM enriched functional microbial communities in the biofilm related to nitrogen and sulfur transformation. Functional gene analysis showed an increase in the narG, nirS, norB, and nosZ genes supporting denitrification. The dsrA and sqr genes responsible for the sulfur cycle exhibited the highest abundance. This realization allowed for the utilization of waste materials for treatment, enhancing the energy efficiency of carbon reduction and pollution reduction in CWs.

Insights into nitrogen biogeochemical cycling in mangrove wetland from Genome-Resolved metagenomic sequencing

The critical role of nitrogen (N) in sustaining the vitality of mangrove wetlands is widely recognized, yet research elucidating complex microbiome structures and functional genes driving N biogeochemical cycles are markedly limited. This study leverages high-throughput and metagenomic sequencing to investigate N biogeochemical cycling from the Dongzhai Harbor mangrove wetland. Results reveal that Proteobacteria (29.95–49.26%), Chloroflexi (15.68–35.33%), and Acidobacteria (5.18–12.46%) are the predominant phyla, with Deltaproteobacteria (3.71–7.95%), Gammaproteobacteria (0.83–8.73%), and Alphaproteobacteria (1.73–14.13%) leading at the order level. Microbial diversity and the prevalence of functional genes diminish with increasing sediment depth, indicating that N transformation predominantly occurs in surface sediments. The identified N metabolic pathways in mangrove sediments encompass N fixation (nifD, nifK, nifH genes), nitrification (amoA, amoB, amoC, hao, nxrA and nxrB genes), denitrification (narG, nirK, nirS, norB and nosZ genes), dissimilatory nitrate reduction to ammonium (hzo、hzsA and hzsB genes), and organic N mineralization (gdhA and ureC genes). Organic N mineralization and N fixation are most pronounced in mangrove sediments, with N fixation exhibiting greater intensity in the mangrove than mudflat zone. Salinity, TOC, TN, and SO42- predominantly foster N cycling facilitated by microbial activities, while Fe appears to impede it. These insights offer a scientific foundation for the ecological restoration and conservation of mangrove wetlands.

Rice-crayfish integrated system enhances global warming potential via increasing methane emission mainly driven by continuous deep flooding

The conversion from rice monoculture (RM) to rice-crayfish integrated system (RCIS) could change soil physicochemical properties and microorganisms related to greenhouse gas (GHG) emissions. Nevertheless, it is still unclear the responses of GHG emissions and global warming potential (GWP) from paddy fields to RCIS. Here, we conducted a field experiment to investigate the changes in the emissions of methane (CH4) and nitrous oxide (N2O), GWP, soil physicochemical properties and the associated microbial abundances (methanogen, methanotrophs, denitrifier and nitrifier) under RCIS in Gaoyou City, Jiangsu Province, China from 2022 to 2023. There were three treatments, including RM, rice monoculture with continuous deep flooding (RMF) and RCIS. Compared with RM, RCIS and RMF all significantly increased CH4 emission and decreased N2O emission. But the effectiveness of increasing CH4 emission and decreasing N2O emission was larger under RCIS than RMF. The increased CH4 emission was due to the significantly higher mcrA gene abundance and ratio of mcrA/pmoA under RCIS. While the significantly higher nosZ gene abundance and ratio of nosZ/(amoA+nirK+nirS) under RCIS were responsible for its reduced N2O emission. Furthermore, the increases in gene abundances of mcrA and nosZ under RCIS were closely correlated with the significantly lower redox potential and pH as well as the significantly higher contents of dissolved organic carbon, ammonium and nitrate in soil. Averaged across two years, GWP under RCIS and RMF were 4.6-fold and 3.4-fold that under RM, respectively. This revealed that continuous deep flooding had a greater effectiveness in increasing GWP than crayfish culture under RCIS. Notably, the contribution rates of CH4 emission to the total GWP decreased in the order of RCIS (98.6%) > RMF (97.5%) > RM (86.2%). Besides, greenhouse gas intensity under RCIS was 5.1-fold that of RM due to its enhanced GWP and reduced rice grain yield. In summary, RCIS could enhance GWP from paddy fields through increasing CH4 emission mainly caused by the continuous deep flooding.

New insights for simultaneous nitrogen removal and greenhouse gas reduction by biological-electrolysis constructed wetland in the treatment of aquaculture wastewater under varied C/N

Aiming to address the challenges of poor nitrogen (N) removal and greenhouse gas (GHG) emissions in aquaculture wastewater treatment, biological-electrolysis constructed wetland (BECW) and constructed wetland (CW) were established to comparatively evaluate the N removal efficiency, the greenhouse gas (GHG) emissions, and specific microbial gene abundances under various influent C/N ratios. Increasing influent C/N ratio markedly enhanced the removal rates of COD, NH4+-N, NO3−-N, and TN, while markedly reduced N2O fluxes in CW and BECW. The integration of an electric field with CW (BECW) significantly compensated for the shortcomings of the poor NO3−-N removal efficiency and high N2O emissions when C/N < 6. However, this compensating effect gradually weakened with the continue increase of C/N. BECW simultaneously achieved the high COD (89.26 ± 6.08 %), NO3−N (97.03 ± 1.22 %), and TN (83.17 ± 3.19 %) removal rates, and low GHG emissions (486.00 ± 36.61 mg/m2/h) at C/N = 6. The BECW significantly decreased the abundance of mcrA and pmoA genes while increased the abundance of nirK, nirS, and nosZ genes, and the ratios of pmoA/mcrA and nosZ/(nirS + nirK) across all C/N condition. The emission flux of CH4 in BECW was positively correlated with C/N and the mcrA abundance but significantly negatively correlated with pmoA abundance and pmoA/mcrA. The emission flux of N2O was positively correlated with nirS and nirK abundance but significantly negatively correlated with C/N, NO3−-N removal rates, nosZ abundance, and the index of nosZ/(nirS + nirK). This research offers an innovative approach to get both low GHG emissions and high NO3−-N removal rate in aquaculture wastewater treatment.