Validation of numerical simulations and experiments on impulse characteristics induced by self-excited oscillation.

The high-frequency pulse flow, equivalent to the natural frequency of rocks, is generated by a self-excited oscillating cavity to achieve resonance rock-breaking. The flow field and oscillating mechanism of the self-excited oscillating cavity were simulated using the large eddy simulation method of Computational Fluid Dynamics (CFD). A field-scale testing apparatus was developed to investigate the impulse characteristics and verify the simulation results. The results show that the fluid at the outlet at the tool is deflected due to the pulse oscillation of the fluid. The size and shape of low-pressure vortices constantly change, leading to periodic changes in fluid impedance within the oscillating cavity. The impulse frequency reaches its highest point when the length-diameter ratio is 0.67. As the length-diameter ratio increases, the tool pressure loss also increases. Regarding the cavity thickness, the impulse frequency of the oscillating cavity initially decreases, then increases, and finally decreases again. Moreover, both the impulse frequency and pressure loss increase with an increase in displacement. The numerical simulation findings align with the experimental results, thus confirming the validity of the theoretical model. This research provides theoretical guidance for the practical application of resonance rock-breaking technology.

Graphene oxide stimulated low-temperature denitrification activity of microbial communities in lake sediments by enhancing anabolism and inhibiting cellular respiration.

Nitrate pollution in fresh water is becoming increasingly serious. In this study, the effects of temperature and graphene oxide materials on the potential functions of denitrification communities in lake sediments were investigated by metagenome. The addition of graphene oxide significantly affected the abundance of denitrification genes such as Nap, Nos, and enhanced the contribution of Pseudomonas, making low temperature and material addition conducive to the denitrification process. Module network implied that low temperature increased the centrality of denitrification in community functions. At low temperatures, graphene oxide enhanced community anabolism by stimulation organic carbon consumption and regulating the gene abundance in the citric acid cycle and the semi-phosphorylation Entner-Doudoroff, thus possibly stimulating extracellular polymeric substances (EPS) synthesis and secretion. In addition, graphene oxide may also regulate the transfer of reducing electrons from NADH to denitrifying enzymes by affecting the gene abundances of complex I and complex IV.

Frequency of change determines effectiveness of microbial response strategies.

Nature challenges microbes with change at different frequencies and demands an effective response for survival. Here, we used controlled laboratory experiments to investigate the effectiveness of different response strategies, such as post-translational modification, transcriptional regulation, and specialized versus adaptable metabolisms. For this, we inoculated replicated chemostats with an enrichment culture obtained from sulfidic stream microbiomes 16 weeks prior. The chemostats were submitted to alternatingly oxic and anoxic conditions at three frequencies, with periods of 1, 4 and 16 days. The microbial response was recorded with 16S rRNA gene amplicon sequencing, shotgun metagenomics, transcriptomics and proteomics. Metagenomics resolved provisional genomes of all abundant bacterial populations, mainly affiliated with Proteobacteria and Bacteroidetes. Almost all these populations maintained a steady growth rate under both redox conditions at all three frequencies of change. Our results supported three conclusions: (1) Oscillating oxic/anoxic conditions selected for generalistic species, rather than species specializing in only a single condition. (2) A high frequency of change selected for strong codon usage bias. (3) Alignment of transcriptomes and proteomes required multiple generations and was dependent on a low frequency of change.

Performance, microbial growth and community interactions of iron-dependent denitrification in freshwaters.

Iron-dependent denitrification is a safe and promising technology for nitrogen removal in freshwaters. However, the understanding of microbial physiology and interactions during the process was limited. Denitrifying systems inoculated with freshwater samples were operated with and without iron(II) at a low C/N ratio for 54 days. Iron addition improved nitrogen removal. Batch experiments confirmed that microbially mediated reaction rather than abiotic reaction dominated during the process. Metagenomics recovered genomes of the five most abundant microorganisms, which accounted for over 99% of the community in every triplicate of the iron-based system. Based on codon usage bias, all of them were fast-growing organisms. The total abundance of fast-growing organisms was 38% higher in the system with iron than in the system without iron. Notably, the most abundant organism Diaphorobacter did not have enzymes for asparagine and aspartate biosynthesis, whereas Rhodanobacter could not produce serine and cobalamin. Algoriphagus and Areminomonas lost synthesis enzymes for more amino acids and vitamins. However, they could always obtain these growth-required substances from another microorganism in the community. The two-partner relationship minimized the limitation on microbial reproduction and increased community stability. Our results indicated that iron addition improved nitrogen removal by supplying electron donors, promoting microbial growth, and building up syntrophic interactions among microorganisms with timely communications. The findings provided new insights into the process, with implications for freshwater remediation.

Denitrification fractionates N and O isotopes of nitrate following a ratio independent of carbon sources in freshwaters.

The stable isotope technique has been used in tracking nitrogen cycling processes, but the isotopic characteristics are influenced by environmental conditions. To better understand the variability of nitrate isotopes in nature, we investigated the influence of organic carbon sources on isotope fractionation characteristics during microbial denitrification. Denitrifying cultures were inoculated with freshwater samples and enriched with five forms of organic compounds, that is, acetate, citrate, glucose, cellobiose, and leucine. Though the isotope enrichment factors of nitrogen and oxygen (15 ε and 18 ε) changed with carbon sources, 18 ε/15 ε always followed a proportionality near 1. Genome-centred metagenomics revealed the enrichment of a few populations, such as Pseudomonas, Enterobacter, and Atlantibacter, most of which contained both NapA- and NarG-type nitrate reductases. Metatranscriptome showed that both NapA and NarG were expressed but to different extents in the enrichments. Furthermore, isotopic data collected from a deep reservoir was analysed. The results showed δ18 O- and δ15 N-nitrate did not correlate in the surface water where nitrification was active, but 18 ε/15 ε followed a proportionality of 1.05 ± 011 in deeper waters (≥ 12 m) where denitrification controlled the nitrate isotope. The independence of 18 ε/15 ε from carbon sources provides an opportunity to determine heterotrophic denitrification and helps the interpretation of nitrate isotopes in freshwaters.

Life strategies and metabolic interactions of core microbes during thiosulphate-based denitrification.

Sulphur-driven denitrification is a low-cost process for the treatment of nitrate-contaminated water. However, a comprehensive understanding of core populations and microbial interactions of a sulphur-based denitrifying system is lacking. This study presents results from three replicated denitrifying systems amended with thiosulphate and operated under a low C/N ratio. Amplicon sequencing revealed gradual enrichments of a few abundant denitrifiers. Based on genome-centred metagenomics and metatranscriptomics, a core set of microbes was identified in the systems, with Pseudomonas 1 and Thauera 2 being the most abundant ones. Although the replicates showed different enrichments, generalized observations were summarized. Most core populations conserved energy from denitrification coupled with sulphur. Pseudomonas 1 and Thauera 2 were able to finish complete denitrification. Surprisingly, they were also able to synthesize almost all amino acids and vitamins. In contrast, less abundant members, including Pseudomonas 2, were relatively auxotrophic and required an exogenous supply of amino acids and vitamins. The high expression of enzymes involved in biosynthesis and transport systems indicated their syntrophic relationships. The genomic findings suggested life strategies and interactions of the core thiosulphate-based denitrifying microbiome, with implications for nitrate-polluted water remediation.

Sequentially modified carbon felt for enhanced p-nitrophenol biodegradation through direct interspecific electron transfer.

The widely applied aromatic nitration in modern industry leads to toxic p-nitrophenol (PNP) in environment. Exploring its efficient degradation routes is of great interests. In this study, a novel four-step sequential modification procedure was developed to increase the specific surface area, functional group, hydrophilicity, and conductivity of carbon felt (CF). The implementation of the modified CF promoted reductive PNP biodegradation, attaining 95.2 ± 0.8% of removal efficiency with less accumulation of highly toxic organic intermediates (e.g., p-aminophenol), compared to carrier-free and CF-packed biosystems. The constructed anaerobic-aerobic process with modified CF in 219-d continuous operation achieved further removal of carbon and nitrogen containing intermediates and partial mineralization of PNP. The modified CF promoted the secretion of extracellular polymeric substances (EPS) and cytochrome c (Cyt c), which were essential components to facilitate direct interspecies electron transfer (DIET). Synergistic relationship was deduced that glucose was converted into volatile fatty acids by fermenters (e.g., Longilinea and Syntrophobacter), which donated electrons to the PNP degraders (e.g., Bacteroidetes_vadinHA17) through DIET channels (CF, Cyt c, EPS) to complete PNP removal. This study proposes a novel strategy using engineered conductive material to enhance the DIET process for efficient and sustainable PNP bioremediation.

Effect of low temperature on contributions of ammonia oxidizing archaea and bacteria to nitrous oxide in constructed wetlands.

Constructed wetlands (CWs) have been widely used for ecological remediation of micro-polluted source water. Nitrous oxide (N2O) from CWs has caused great concern as a greenhouse gas. However, the contribution of ammonia oxidation driven by ammonia oxidizing archaea (AOA) and ammonia oxidizing bacteria (AOB) to N2O emission, especially at low temperature, was unknown. This study aimed to quantify the contributions of AOA and AOB to N2O through lab-scale subsurface CWs. The N2O emission flux of CW at 8 °C was 1.23 mg m-2·h-1, significantly lower than that at 25 °C (1.92 mg m-2·h-1). The contribution of ammonia oxidation to N2O at 8 °C (33.04%) was significantly higher than that at 25 °C (24.17%). The N2O production from AOA increased from 1.91 ng N·g-1 at 25 °C to 4.11 ng N·g-1 soil at 8 °C and its contribution increased from 23.38% to 30.18% (P < 0.05). Low temperature impaired functional gene groups and inhibited the activity of AOB, resulting in its declined contribution. Based on the transcriptional analysis, AOA was less affected by low temperature, thus stably contributing to N2O. Moreover, community diversity and relationships of AOA were enhanced at 8 °C, while AOB declined. The results confirmed the significant contribution of AOA and demonstrated molecular mechanisms (higher activity and community stability) of the increased contribution of AOA to N2O at low temperature.

Influences of carbon sources on N2O production during denitrification in freshwaters: Activity, isotopes and functional microbes.

Denitrification is one of the major sources of N2O in freshwaters. Diverse forms of organic compounds act as the electron donors for microbial denitrification. However, the influences of carbon sources on N2O production, N2O reduction, isotope fractionation and functional microbes during denitrification were largely unknown. In this study, five forms of carbon sources (i.e. acetate, citrate, glucose, cellobiose and leucine) were used to enrich denitrifiers in freshwater sediments. N2O conversion in the enrichments was investigated by a combination of inhibition technique, natural stable isotope method and metagenomics. Acetylene was effective in inhibiting N2O reduction without influencing the isotopic characteristics during N2O production. Glucose led to the least N2O production and reduction, in accordance with the lowest abundance of both NO and N2O reductases in this enrichment. δ18O and site preference value (SP, =δ15Nα-δ15Nß) of N2O were sensitive to discriminate the five carbon sources, except when comparing acetate and leucine. Isotopic values of N2O were not significantly different in these two enrichments due to the similarity of NO reductases - Pseudomonas-type cNorB. Specifically, the enrichment with cellobiose produced N2O with the lowest δ18O values (39.4‰±1.1‰), due to Alicycliphilus with both cNorB and qNorB. The enrichment with glucose led to the highest SP values (8.9‰±8.6‰), caused by Thiobacillus-type cNorB. Our results demonstrated the link between carbon sources, N2O production and reduction, isotopic signatures, microbial populations and enzymes during denitrification in freshwaters.

Distinct oxygen isotope fractionations driven by different electron donors during microbial nitrate reduction in lake sediments.

Microbial nitrate reduction can be driven by organic carbon oxidation, as well as by inorganic electron donors, such as reduced forms of sulfur and iron. An apparent inverse oxygen isotope fractionation effect was observed during nitrate reduction in sediment incubations from five sampling sites of a freshwater lake, Hongze Lake, China. Incubations with organic and inorganic electron donor additions were performed. Especially, the inverse oxygen isotope effect was intensified after glucose addition, whereas the incubations with sulfide and Fe2+ showed normal fractionation factors. Nitrate reductase encoding genes, napA and narG, were analysed with metagenomics. Higher napA/narG ratios were associated with higher oxygen fractionation factors. The most abundant clade (59%) of NapA in the incubation with glucose was affiliated with Rhodocyclales. In contrast, it only accounted for 8%-9% of NapA in the incubations with sulfide and Fe2+ . Differences in nitrate reductases might explain different oxygen isotope effects. Our findings also suggested that large variance of O-nitrate isotope fractionations might have to be considered in the interpretation of natural isotope records.

Using hierarchical stable isotope to reveal microbial food web structure and trophic transfer efficiency differences during lake melt season.

The microbial food web (MFW) is a material and energy source in lake water ecosystems. Although it is crucial to determine its structure and function for water ecological health, MFW changes during lake melt period have not been well studied. In this study, the MFW was divided into three categories by analyzing its structure and trophic transfer efficiency using hierarchical C/N stable isotopes and eDNA sequencing techniques, including the detrital food web (DFC, 15 %), classical grazing food web (CFC, 60 %), and mixed trophic food web (MFC, 25 %). The trophic structure and type of MFW in ice-melting lakes are always in the process of succession and adaptation, which is in a relatively low trophic transfer efficiency stage under stable conditions (i.e. CFC), whereas the input of exogenous debris and organic pollutants may lead to an increase in MFW trophic transfer efficiency (i.e. MFC, DFC). The trophic transfer efficiency from the previous trophic level to protozoa and micrometazoa was 16.32 % and 20.77 % in DFC and 10.20 % and 29.43 % in MFC, respectively. Both are obviously higher than those of the CFC (11.69 % and 9.45 %, respectively). In terms of trophic structure, the community interaction and trophic cascade effect of DFC and MFC were enhanced but easily changed with environmental factors. In contrast, the core species and cascading effects of the CFC were clearer, and the MFW structure was relatively stable. Overall, this study reveals that the explosive increase in MFW trophic transfer efficiency induced by exogenous input during the lake melt period may subsequently lead to the destabilization of the microbial community structure and cause potential ecological risks. These are manifested in the absence of ecological trophic processes, the decrease in trophic structure complexity and stability, and the weakening of microecology self-adaptive regulation ability.

Archaeal and bacterial ecological strategies in sediment denitrification under the influence of graphene oxide and different temperatures.

As an emerging material, graphene oxide (GO) has been widely used in recent years and will inevitably enter into natural water bodies, and it may have an impact on lake microbial communities owing to its potential toxicity and denitrification-enhancing ability. This study simulated the effect of 0.1 g/L GO on denitrification in lake sediments under summer (28 °C) and winter temperatures (8 °C). GO promoted carbon source metabolism and denitrification. Phylogenetic bin-based null model analysis suggested that GO significantly altered the contribution of heterogeneous selection in bacterial and archaeal community assembly. The co-occurrence network indicated that bacterial communities responded to the enhancement of heterogeneous selection by strategies of enhancing positive correlation and shared niche, whereas archaeal communities adopted strategies of enhancing negative correlation and competition. Bacterial networks also emerged with more non-hub connector species that could drive changes in community structure. Our study contributed to the understanding of different ecological strategies adopted by bacterial and archaeal communities in response to changes in ecological selection driven by GO.

Effect of sulfur sources on the competition between denitrification and DNRA.

The fate of nitrogen is controlled by the competition between nitrate reduction pathways. Denitrification removes nitrogen in the system to the atmosphere, whereas dissimilatory nitrate reduction to ammonia (DNRA) retains nitrate in the form of ammonia. Different microbes specialize in the oxidation of different electron donors, thus electron donors might influence the outcomes of the competition. Here, we investigated the fate of nitrate with five forms of sulfur as electron donors. Chemoautotrophic nitrate reduction did not continue after the passages of the enrichments with sulfide, sulfite and pyrite. Nitrate reduction rate was the highest in the enrichment with thiosulfate. Denitrification was stimulated and no DNRA was observed with thiosulfate, while both denitrification and DNRA were stimulated with elemental sulfur. Metagenomes of the enrichments were assembled and binned into ten genomes. The enriched populations with thiosulfate included Thiobacillus, Lentimicrobium, Sulfurovum and Hydrogenophaga, all of which contained genes involved in sulfur oxidation. Elemental sulfur-based DNRA was performed by Thiobacillus (with NrfA and NirB) and Nocardioides (with only NirB). Our study established a link between sulfur sources, nitrate reduction pathways and microbial populations.

Denitrification and dissimilatory nitrate reduction to ammonia in long-term lake sediment microcosms with iron(II).

Nitrate is an abundant pollutant in aquatic environments. Competition between the nitrate reduction processes, denitrification, which converts nitrate into nitrogen gas, and dissimilatory nitrate reduction to ammonia (DNRA), which converts nitrate into ammonia, decides whether an ecosystem removes or retains nitrogen. The presence of iron was previously reported to stimulate DNRA while sometimes inhibiting denitrification in in-situ studies, but long-term effect of iron(II) inputs on the competition is unknown. Here we inoculated long-term microcosms with sediments from two freshwater lakes. During 540 days of incubations, the microcosms with nitrate and Fe(II) additions of both lakes were able to sustain high nitrate reduction rates. Lepidocrocite was produced as a product of iron oxidation. We found both denitrification and DNRA were stimulated by nitrate and iron in the absence of external organic carbon addition. Phylogenetic analysis of denitrification genes, nirK and nirS, and DNRA genes, nirB and nrfA, was performed with metagenomic sequencing results. Enrichment was shown for reported Fe(II)-dependent nitrate reducers associated with nirS and nirB. Most of these bacteria are affiliated with Betaproteobacteria. From 16S rRNA gene analysis, Betaproteobacteria was enriched as well. In parallel, heterotrophic denitrifiers and methanotrophic DNRA archaea increased in abundance. Our results suggested heterotrophic and Fe(II)-dependent nitrate reducers both contributed to denitrification and DNRA in long-term microcosm incubations provided with iron.

Increase of N2O production during nitrate reduction after long-term sulfide addition in lake sediment microcosms.

Microbial denitrification is a main source of nitrous oxide (N2O) emissions which have strong greenhouse effect and destroy stratospheric ozone. Though the importance of sulfide driven chemoautotrophic denitrification has been recognized, its contribution to N2O emissions in nature remains elusive. We built up long-term sulfide-added microcosms with sediments from two freshwater lakes. Chemistry analysis confirmed sulfide could drive nitrate respiration in long term. N2O accumulated to over 1.5% of nitrate load in both microcosms after long-term sulfide addition, which was up to 12.9 times higher than N2O accumulation without sulfide addition. Metagenomes were extracted and sequenced during microcosm incubations. 16 S rRNA genes of Thiobacillus and Defluviimonas were gradually enriched. The nitric oxide reductase with c-type cytochromes as electron donors (cNorB) increased in abundance, while the nitric oxide reductase receiving electrons from quinols (qNorB) decreased in abundance. cnorB genes similar to Thiobacillus were enriched in both microcosms. In parallel, enrichment was observed for enzymes involved in sulfur oxidation, which supplied electrons to nitrate respiration, and enzymes involved in Calvin Cycle, which sustained autotrophic cell growth, implying the coupling relationship between carbon, nitrogen and sulfur cycling processes. Our results suggested sulfur pollution considerably increased N2O emissions in natural environments.

The promotion and inhibition effect of graphene oxide on the process of microbial denitrification at low temperature.

This study found that graphene oxide (GO) improved microbial denitrification at low temperatures (~12 °C), and the optimal concentration was 10 mg/L as the removal rate of NO3-N increased by 17%. At the optimal concentration, GO improved the electron transport system activity of the microbes and enhanced the activity of nitrate reductase and nitrite reductase while exhibited low microbial toxicity. The addition of GO increased the content of tightly bound extracellular polymeric substances (EPS). The results of fluorescence spectrometer indicated that GO accelerated the renewal of bound EPS (B-EPS). Fourier Transform infrared spectroscopy (FTIR) results showed that GO affected the secondary structure of the protein in B-EPS, making B-EPS more hydrophobic and promoting microbial aggregation. B-EPS affected by GO can promote the electron transfer process of microorganisms. However, high concentration (>25 mg/L) of GO may inhibit denitrification by competing for electrons, which was not conducive to denitrification thermodynamically.

Long-term sulfide input enhances chemoautotrophic denitrification rather than DNRA in freshwater lake sediments.

Partitioning between nitrate reduction pathways, denitrification and dissimilatory nitrate reduction to ammonium (DNRA) determines the fate of nitrate removal and thus it is of great ecological importance. Sulfide (S2-) is a potentially important factor that influences the role of denitrification and DNRA. However, information on the impact of microbial mechanisms for S2- on the partitioning of nitrate reduction pathways in freshwater environments is still lacking. This study investigated the effects of long-term (108 d) S2- addition on nitrate reduction pathways and microbial communities in the sediments of two different freshwater lakes. The results show that the increasing S2- addition enhanced the coupling of S2- oxidation with denitrification instead of DNRA. The sulfide-oxidizing denitrifier, Thiobacillus, was significantly enriched in the incubations of both lake samples with S2- addition, which indicates that it may be the key genus driving sulfide-oxidizing denitrification in the lake sediments. During S2- incubation of the Hongze Lake sample, which had lower inherent organic carbon (C) and sulfate (SO42-), Thiobacillus was more enriched and played a dominant role in the microbial community; while during that of the Nansi Lake sample, which had higher inherent organic C and SO42-, Thiobacillus was less enriched, but increasing abundances of sulfate reducing bacteria (Desulfomicrobium, Desulfatitalea and Geothermobacter) were observed. Moreover, sulfide-oxidizing denitrifiers and sulfate reducers were enriched in the Nansi Lake control treatment without external S2- input, which suggests that internal sulfate release may promote the cooperation between sulfide-oxidizing denitrifiers and sulfate reducers. This study highlights the importance of sulfide-driven denitrification and the close coupling between the N and S cycles in freshwater environments, which are factors that have often been overlooked.

Microbial coupling mechanisms of nitrogen removal in constructed wetlands: A review.

Nitrogen removal through microorganisms is the most important pathway in constructed wetlands (CWs). In this review, we summarize the microbial coupling mechanisms of nitrogen removal, which are the common methods of nitrogen transformation. The electron pathways are shortened and consumption of oxygen and energy is reduced during the coupling of nitrogen transformation functional microorganisms. The highly efficient nitrogen removal mechanisms are cultivated from the design conditions in CWs, such as intermittent aeration and tidal flow. The coupling of microorganisms and substrates enhances nitrogen removal mainly by supplying electrons, and plants affect nitrogen transformation functional microorganisms by the release of oxygen and exudates from root systems as well as providing carriers for microbial attachment. In addition, inorganic elements such as Fe, S and H act as electron donors to drive the autotrophic denitrification process in CWs.

Responses of AOA and AOB activity and DNA/cDNA community structure to allylthiourea exposure in the water level fluctuation zone soil.

Ammonia oxidation is mainly performed by ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB). Allylthiourea (ATU) has been found to specifically inhibit ammonia oxidation. However, the effect of ATU on AOA and AOB transcription has been infrequently studied. In the present study, we examined the responses of AOA and AOB activity and DNA/cDNA community structure to ATU exposure. The ammonia oxidation activity in the 100-mg/L ATU group was 4.3% of that in the control group after 7 days. When exposed to ATU, the gene abundance of AOA was favored compared with that of AOB, and there were no statistically significant differences in the abundance of AOB amoA in DNA and cDNA between the two groups. Compared with the control group, the gene abundance of AOA significantly increased by 5.23 times, while the transcription of AOA significantly decreased by 0.70 times. Moreover, the transcriptional ratio of AOA in the ATU group was only 0.05 times as high as that in the control group. ATU selectively affected AOB and completely inhibited Nitrosomonas europaea and Bacterium amoA.22.HaldeII.kultur at the genetic level. Under ATU exposure, all AOA clusters were transcribed, but three AOB clusters were not transcribed. Our results indicated that the ammonia oxidation potential of the soil of water level fluctuation areas, based on ATU inhibition, was associated mainly with AOA amoA gene abundance and AOB community shifts in DNA and cDNA.

The abundance and community structure of active ammonia-oxidizing archaea and ammonia-oxidizing bacteria shape their activities and contributions in coastal wetlands.

Aerobic ammonia oxidation, an important part of the global nitrogen cycle, is thought to be jointly driven by ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA) in coastal wetlands. However, the activities and contributions of AOA and AOB in coastal wetlands have remained largely unknown. Here, we investigated the oxidation capability of AOA and AOB in four types of typical coastal wetlands (paddy, estuary, shallow and reed wetland) in the Bohai region in China using DNA-based stable-isotope probing (DNA-SIP), quantitative PCR and high-throughput sequencing techniques. We found that the community structure of AOB varied substantially, and the AOA structure was more stable across different coastal wetlands. The rate of AOA was 0.12, 0.84, 0.45 and 0.93 µg N g-1 soil d-1 in paddy, estuary, shallow and reed wetlands, and the rate of AOB was 5.61, 10.72, 0.74 and 1.16 µg N g-1 soil d-1, respectively. We found that the contribution of AOA gradually increased from paddy to estuary to shallow wetland and finally to reed wetland, with values of 2.03%, 7.25%, 37.53% and 44.51%, respectively. Our results provide new insight into the mechanisms of the differences in activities and the contributions of AOA and AOB in different coastal wetlands, and our findings may contribute to further understanding of the global nitrogen cycle.