Complete ammonia oxidization in agricultural soils: High ammonia fertilizer loss but low N2 O production.

The contribution of agriculture to the sustainable development goals requires climate-smart and profitable farm innovations. Increasing the ammonia fertilizer applications to meet the global food demands results in high agricultural costs, environmental quality deterioration, and global warming, without a significant increase in crop yield. Here, we reported that a third microbial ammonia oxidation process, complete ammonia oxidation (comammox), is contributing to a significant ammonia fertilizer loss (41.9 ± 4.8%) at the rate of 3.53 ± 0.55 mg N kg-1 day-1 in agricultural soils around the world. The contribution of comammox to ammonia fertilizer loss, occurring mainly in surface agricultural soil profiles (0-0.2 m), was equivalent to that of bacterial ammonia oxidation (48.6 ± 4.5%); both processes were significantly more important than archaeal ammonia oxidation (9.5 ± 3.6%). In contrast, comammox produced less N2 O (0.98 ± 0.44 µg N kg-1 day-1 , 11.7 ± 3.1%), comparable to that produced by archaeal ammonia oxidation (16.4 ± 4.4%) but significantly lower than that of bacterial ammonia oxidation (72.0 ± 5.1%). The efficiency of ammonia conversion to N2 O by comammox (0.02 ± 0.01%) was evidently lower than that of bacterial (0.24 ± 0.06%) and archaeal (0.16 ± 0.04%) ammonia oxidation. The comammox rate increased with increasing soil pH values, which is the only physicochemical characteristic that significantly influenced both comammox bacterial abundance and rates. Ammonia fertilizer loss, dominated by comammox and bacterial ammonia oxidation, was more intense in soils with pH >6.5 than in soils with pH <6.5. Our results revealed that comammox plays a vital role in ammonia fertilizer loss and sustainable development in agroecosystems that have been previously overlooked for a long term.

Contribution of Ammonium-Induced Nitrifier Denitrification to N2O in Paddy Fields.

Paddy fields are one of the most important sources of nitrous oxide (N2O), but biogeochemical N2O production mechanisms in the soil profile remain unclear. Our study used incubation, dual-isotope (15N-18O) labeling methods, and molecular techniques to elucidate N2O production characteristics and mechanisms in the soil profile (0-60 cm) during summer fallow, rice cropping, and winter fallow periods. The results pointed out that biotic processes dominated N2O production (72.2-100%) and N2O from the tillage layer accounted for 91.0-98.5% of total N2O in the soil profile. Heterotrophic denitrification (HD) was the main process generating N2O, contributing between 53.4 and 96.6%, the remainder being due to ammonia oxidation pathways, which was further confirmed by metagenomics and quantitative polymerase chain reaction (qPCR) assays. Nitrifier denitrification (ND) was an important N2O production source, contributing 0-46.6% of total N2O production, which showed similar trends with N2O emissions. Among physicochemical and biological factors, ammonium content and the ratio of total organic matter to nitrate were the main driving factors affecting the contribution ratios of the ammonia oxidation pathways and HD pathway, respectively. Moisture content and pH affect norC-carrying Spirochetes and thus the N2O production rate. These findings confirm the importance of ND to N2O production and help to elucidate the impact of anthropogenic activities, including tillage, fertilization, and irrigation, on N2O production.

Nitrifiers Cooperate to Produce Nitrous Oxide in Plateau Wetland Sediments.

The thawing of dormant plateau permafrost emits nitrous oxide (N2O) through wetlands; however, the N2O production mechanism in plateau wetlands is still unclear. Here, we used the 15N-18O double tracer technique and metagenomic sequencing to analyze the N2O production mechanism in the Yunnan-Kweichow and Qinghai-Tibet plateau wetlands during the summer of 2020. N2O production activity was detected in all 16 sediment samples (elevation 1020-4601 m: 2.55 ± 0.42-26.38 ± 3.25 ng N g-1 d-1) and was promoted by nitrifier denitrification (ND). The key functional genes of ND (amoA, hao, and nirK) belonged to complete ammonia oxidizing (comammox) bacteria, and the key ND species was the comammox bacterium Nitrospira nitrificans. We found that the comammox bacterial species N. nitrificans and the ammonia oxidizing bacterial (AOB) species Nitrosomonas europaea cooperate to produce N2O in the plateau wetland sediments. Furthermore, we inferred that environmental factors (elevation and total organic matter (TOM)) influence the cooperation pattern via N. nitrificans, thus affecting the N2O production activity in the plateau wetland sediments. Our findings advance the mechanistic understanding of nitrifiers in biogeochemical cycles and global climate change.

Abundance and Functional Importance of Complete Ammonia Oxidizers and Other Nitrifiers in a Riparian Ecosystem.

The discovery of complete ammonia oxidation (comammox) has altered our understanding of nitrification, which is the rate-limiting process in the global nitrogen cycle. However, understanding the ecological role of comammox or its contribution to nitrification in both natural and artificial ecosystems is still in its infancy. Here, we investigated the community distribution and function of comammox bacteria in riparian ecosystems and analyzed interactions between comammox and other nitrogen cycling microorganisms. The comammox bacterial abundance and rate were higher in summer than in winter and higher in nonrhizosphere soils than in the rhizosphere. Fringe soils in the riparian zone comprise a comammox hotspot, where the abundance (2.58 × 108 copies g-1) and rate (0.86 mg N kg-1 d-1) of comammox were not only higher than at other sampling sites but also higher than those of other ammonia oxidation processes. The comammox rate correlated significantly positively with relative abundance of the comammox species Candidatus Nitrospira nitrificans but not with that of the species Candidatus Nitrospira nitrosa. Analysis of comammox interaction with other ammonia-oxidizing processes revealed ammonia-oxidizing archaea to dominate interface soils, comammox to dominate in fringe soils, and anaerobic ammonium oxidation (anammox) to dominate in interface sediments of the riparian zone. These results indicate that comammox may constitute an important and currently underestimated process of microbial nitrification in riparian zone ecosystems.

Nitrate reduction in the reed rhizosphere of a riparian zone: From functional genes to activity and contribution.

The increased nitrogen (N) fertilizer usage caused substantial nitrate (NO3-) leaching into groundwater and eutrophication in downstream aquatic systems. Riparian zones positioned as the link interfaces of terrestrial and aquatic environments are effective in NO3- removal. However, the microbial mechanisms regulating NO3- reduction in riparian zones are still unclear. In this study, four microbial NO3- reduction processes were explored in fine-scale riparian soil horizons by isotopic tracing technique, qPCR of functional gene, high-throughput amplicon sequencing, and phylogenetic molecular ecological network analysis. Interestingly, anaerobic ammonium oxidation (anammox) contributed to NO3- removal of up to 48.2% only in waterward sediments but not in landward soil. Denitrification was still the most significant contributor to NO3- reduction (32.0-91.8%) and N-losses (51.7-100%). Additionally, dissimilatory nitrate reduction to ammonium (DNRA) played a key role in NO3- reduction (4.4-67.5%) and was even comparable to denitrification. Community structure analysis of denitrifying, anammox, and DNRA bacterial communities targeting the related functional gene showed that spatial heterogeneity played a greater role than both temporal and soil type (rhizosphere and non-rhizosphere soil) variability in microbial community structuring. Denitrification and DNRA communities were diverse, and their activities did not depend on gene abundance but were significantly related to organic matter, suggesting that gene abundance alone was insufficient in assessing their activity in riparian zones. Based on networks, DNRA plays a keystone role among the microbial NO3- reducers. As the last line of defense in the interception of terrestrial NO3-, these findings contribute to our understanding of NO3- removal mechanisms in riparian zones, and could potentially be exploited to reduce the diffusion of NO3- pollution.

Source, contribution and microbial N-cycle of N-compounds in China fresh snow.

The importance and contribution of nitrogen compounds and the related microbial nitrogen cycling processes in fresh snow are not well understood under the current research background. We collected fresh snow samples from 21 cities that 80% are from China during 2016 and 2017. Principal component analysis showed that SO42- were in the first principal component, and N-compounds were the second. Furthermore, the main pollutant ions SO42- and NO3- were from anthropogenic sources, and SO42- contributed (61%) more to the pollution load than NO3- (29%), which were confirmed through a series of precipitation mechanism analysis. We selected five N-cycle processes (consist of oxidation and reduction processes) for molecular biology experiments, including Ammonia-oxidation process, Nitrite-oxidation process, Denitrification process, Anaerobic-ammoxidation process (Anammox) and Dissimilatory nitrate reduction to ammonium process (DNRA). Except ammonia-oxidizing archaeal (AOA) and bacterial (AOB) amoA genes (above 107 copies g-1), molecular assays of key functional genes in various nitrogen conversion processes showed a belowed detection limit number, and AOB abundance was always higher than AOA. The determination of the microbial transformation rate using the 15N-isotope tracer technique showed that the potential rate of five N-conversion processes was very low, which is basically consistent with the results from molecular biology studies. Taken together, our results illustrated that microbial nitrogen cycle processes are not the primary biological processes causing the pollution in China fresh snow.

Factors driving the distribution and role of AOA and AOB in Phragmites communis rhizosphere in riparian zone.

Ammonia oxidation, mainly driven by ammonia-oxidizing archaea (AOA) and bacteria (AOB), plays an important role in determining the rate of nitrification in riparian zones. However, the underlying factors driving the distribution and activity of AOA and AOB in riparian zones, especially in the rhizosphere of Phragmites communis remain unknown. This study revealed the dominance of AOA in ammonium oxidization with higher abundance and activity in both rhizosphere and bulk soil in summer and winter over AOB in riparian zones, based on molecular methods and double-inhibitors method. Phylogenetic analysis showed that 54d9 cluster and Nitrososphaera dominated the AOA community and Nitrosospira dominated the AOB, respectively. For the distribution of AOA and AOB, it was the spatial heterogeneity of physicochemical properties that had the most significant effect. Specifically, TOM & TC were the main physicochemical variables accounting for the difference in abundance and community composition of AOA, and TN had an important influence on AOB in the sediment/soil in riparian zones. For abundance and activity, seasonal heterogeneity and P. communis rhizosphere had a significant impact on the archaeal activity and abundance, respectively, but did not show significant influencing on AOB. These findings suggest that the small-scale environmental heterogeneities in riparian zones are important in shaping the community composition and abundance of AOA and AOB.

Robustness of anammox granular sludge treating low-strength sewage under various shock loadings: Microbial mechanism and little N2O emission.

With the increasing application of anammox for the treatment of high-strength industrial wastewater, application of anammox in municipal sewage has been gaining more attention. Sludge granulation in particular enhances the enrichment and retention of anammox bacteria in municipal sewage treatment systems. However, the performance of granular sludge under continuous and varying hydraulic loading shock remains little understood. In this study, the robustness of anammox granular sludge in treating low-strength municipal sewage under various shock loadings was investigated. Results showed that an upflow anaerobic sludge blanket (UASB) reactor with anammox granules performed well, with anammox specific activity up to 0.28 kg N/kg VSS/day and anti-loading shock capability up to 187.2 L/day during the 8-month testing period. The accumulation rate of N2O (<0.01 kg N/kg VSS/day) in the liquid phase was seven times higher than that of the gas phase, which could be mainly attributed to the incomplete denitrification and insufficient carbon source. However, only a small part of the produced N2O escaped into the atmosphere. High-throughput sequencing and molecular ecological network analyses also identified the bacterial diversity and community structure, indicating the potential resistance against loading shock. The composition and structural analyses showed that polysaccharides were an important functional component in the tightly bound extracellular polymeric substances (TB-EPS), which was the major EPS layer of anammox granules. Scanning electron microscopy (SEM) also showed that the gaps in between the anammox-clusters in the granules inhibit the flotation of the sludge and ensure efficient settling and retention of anammox granules.

Microbial pathways for nitrogen loss in an upland soil.

The distribution and importance of anaerobic ammonium oxidation (anammox) and nitrite-dependent anaerobic methane oxidation (n-damo) have been identified in aquatic ecosystems; their role in agricultural upland soils however has not yet been well investigated. In this study, we examined spatio-temporal distributions of anammox and n-damo bacteria in soil profiles (300 cm depth) from an agricultural upland. Monitoring nitrogen (N) conversion activity using isotope-tracing techniques over the course of one year showed denitrification (99.0% N-loss in the winter and 85.0% N-loss in the summer) predominated over anammox (1.0% N-loss in the winter and 14.4% N-loss in the summer) and n-damo (0.6% N-loss in the winter) in surface soils (0-20 cm). While below 20 cm depth, N-loss was dominated by anammox (79.4 ± 14.3% in the winter and 65.4 ± 12.5% in the summer) and n-damo was not detected. Phylogenetic analysis showed that Candidatus Brocadia anammoxidans dominated the anammox community in the surface soil and Candidatus Brocadia fulgida dominated below 20 cm depth. Dissimilatory nitrate reduction to ammonium (DNRA), another nitrite reduction process, was found to play a limited role (4.9 ± 3.5%) in the surface soil compared with denitrification; below 80 cm DNRA rates were much higher than rates of anammox and denitrification. Ammonium oxidation was the main source of NO2- above 80 cm (70.9 ± 23.3%), the key influencing factor on anammox rates, and nitrate reduction (100%) was the main NO2- source below 80 cm. Considering the anammox, n-damo and denitrification rates as a whole in the sampled soil profile, denitrification is still the main N-loss process in upland soils.

Microbial Nitrogen Cycle Hotspots in the Plant-Bed/Ditch System of a Constructed Wetland with N2O Mitigation.

Artificial microbial nitrogen (N) cycle hotspots in the plant-bed/ditch system were developed and investigated based on intact core and slurry assays measurement using isotopic tracing technology, quantitative PCR and high-throughput sequencing. By increasing hydraulic retention time and periodically fluctuating water level in heterogeneous riparian zones, hotspots of anammox, nitrification, denitrification, ammonium (NH4+) oxidation, nitrite (NO2-) oxidation, nitrate (NO3-) reduction and DNRA were all stimulated at the interface sediments, with the abundance and activity being about 1-3 orders of magnitude higher than those in nonhotspots. Isotopic pairing experiments revealed that in microbial hotspots, nitrite sources were higher than the sinks, and both NH4+ oxidation (55.8%) and NO3- reduction (44.2%) provided nitrite for anammox, which accounted for 43.0% of N-loss and 44.4% of NH4+ removal in riparian zones but did not involve nitrous oxide (N2O) emission risks. High-throughput analysis identified that bacterial quorum sensing mediated this anammox hotspot with B.fulgida dominating the anammox community, but it was B. anammoxidans and Jettenia sp. that contributed more to anammox activity. In the nonhotspot zones, the NO2- source (NO3- reduction dominated) was lower than the sink, limiting the effects on anammox. The in situ N2O flux measurement showed that the microbial hotspot had a 27.1% reduced N2O emission flux compared with the nonhotspot zones.

Manure fertilization alters the population of ammonia-oxidizing bacteria rather than ammonia-oxidizing archaea in a paddy soil.

Manure fertilizers are widely used in agriculture and highly impacted the soil microbial communities such as ammonia oxidizers. However, the knowledge on the communities of archaeal versus bacterial ammonia oxidizers in paddy soil affected by manure fertilization remains largely unknown, especially for a long-term influence. In present work, the impact of manure fertilization on the population of ammonia oxidizers, related potential nitrification rates (PNRs) and the key factors manipulating the impact were investigated through studying two composite soil cores (long-term fed with manure fertilization versus undisturbed). Moreover, soil incubated with NH(4)(+) for 5 weeks was designed to verify the field research. The results showed that the copy numbers of bacterial amoA gene in the manure fed soil were significant higher than those in the unfed soil (p < 0.05), suggesting a clear stimulating effect of long-term manure fertilization on the population of ammonia-oxidizing bacteria (AOB). The detected PNRs in the manure fed soil core (14-218 nmol L(-1) N g(-1) h(-1)) were significant higher than those in the unfed soil core (5-72 nmol L(-1) N g(-1) h(-1) ; p < 0.05). Highly correlations between the PNRs and the bacterial amoA gene copies rather than archaeal amoA gene were observed, indicating strong nitrification capacity related to bacterial ammonia oxidizers. The NH(4)(+) -N significantly correlated to the abundance of AOB (p < 0.01) and explained 96.1% of the environmental variation, showing the NH(4)(+) -N was the main factor impacting the population of AOB. The incubation experiment demonstrated a clear increase of the bacterial amoA gene abundance (2.0 × 10(6) to 8.4 × 10(6) g(-1) d.w.s. and 1.6 × 10(4) to 4.8 × 10(5) g(-1) d.w.s.) in both soil but not for the archaeal amoA gene, in agreement with the field observation. Overall, our results suggested that manure fertilization promoted the population size of bacterial ammonia oxidizers rather than their archaeal counterparts whether in long-term or short-term usage and the NH(4)(+) -N was the key impact factor.

Exploring the mechanisms of Qingdu Fang for the treatment of cervical HR-HPV using UPLC-QTOF-MS, network pharmacology, and cell experimentation.

Background: Qingdu Fang (QDF) is a traditional Chinese herbal formula with remarkable clinical effect in the treatment of HR-HPV, but its mechanism remains unclear. In this study, UPLC-QTOF-MS was used to detect its components, network pharmacology was used to explore the traditional Chinese medicine monomers and their related targets for the treatment of HR-HPV in QDF. Molecular docking and in vitro experiments were performed to verify the results. Methods: QDF constituents and active compounds were identified using UPLC-QTOF-MS analysis. TCMSP and GeneCard databases were used to identify active components, targets, and potential therapeutic targets in HR-HPV. PPI network was constructed using the String database to analyze protein-protein interactions. Cytoscape3.7.2 was used to construct PPI networks, while GO enrichment and KEGG pathway analyses with R. The effect of QDF on H8 cell proliferation was measured using the CCK-8 method, and apoptosis and cell cycle was assessed with flow cytometry. The effects of QDF on PI3K/AKT pathway were detected by Western blotting. Results: A total of 27 compounds were identified on QDF by UPLC-QTOF-MS. Base on Network pharmacology,a total of 254 target genes are involved in the action of QDF on cervical HR-HPV. PPI analysis suggested that TP53, JUN, AKT1, STAT3, TNF and IL6 were potential targets for QDF treatment of HR-HPV. Molecular docking shows that two compounds have strong binding activity with AKT1. CCK-8 and morphological observation have shown that QDF inhibits H8 cell proliferation in a dose-dependent manner. Flow cytometry experiments suggest that QDF induces apoptosis and cell cycle arrest in H8 cells. Western blotting experiments reveal that QDF inhibits the PI3K/AKT signaling pathway. Conclusion: QDF has a multi-faceted therapeutic approach for HR-HPV, targeting inflammation, oxidation, and apoptosis. It induces apoptosis in H8 cells by inhibiting the PI3K/AKT pathway.

Diverse metabolism drives comammox in continental-scale agricultural streams: Important ammonia oxidation but low N2O production.

Agriculture receives approximately 25 % of the annual global nitrogen input, 37 % of which subsequently runs off into adjacent low-order streams and surface water, where it may contribute to high nitrification and nitrous oxide (N2O). However, the mechanisms of nitrification and the pathways controlling N2O production in agricultural streams remain unknown. Here, we report that the third microbial ammonia oxidation process, complete ammonia oxidation (comammox), is widespread and contributes to important ammonia oxidation with low ammonia-N2O conversion in both basin- and continental-scale agricultural streams. The contribution of comammox to ammonia oxidation (21.5 ± 2.3 %) was between that of bacterial (68.6 ± 2.7 %) and archaeal (9.9 ± 1.8 %) ammonia oxidation. Interestingly, N2O production by comammox (18.5 ± 2.1 %) was higher than archaeal (10.5 ± 1.9 %) but significantly lower than bacterial (70.2 ± 2.6 %) ammonia oxidation. The first metagenome-assembled genome (MAG) of comammox bacteria from agricultural streams further revealed their potential extensive diverse and specific metabolism. Their wide habitats might be attributed to the diverse metabolism, i.e. harboring the functional gene of nitrate reduction to ammonia, while the lower N2O would be attributed to their lacking biological function to produce N2O. Our results highlight the importance of widespread comammox in agricultural streams, both for the fate of ammonia fertilizer and for climate change. However, it has not yet been routinely included in Earth system models and IPCC global assessments. Synopsis Widespread but overlooked comammox contributes to important ammonia oxidation but low N2O production, which were proved by the first comammox MAG found in agricultural streams.

Ammonium-derived nitrous oxide is a global source in streams.

Global riverine nitrous oxide (N2O) emissions have increased more than 4-fold in the last century. It has been estimated that the hyporheic zones in small streams alone may contribute approximately 85% of these N2O emissions. However, the mechanisms and pathways controlling hyporheic N2O production in stream ecosystems remain unknown. Here, we report that ammonia-derived pathways, rather than the nitrate-derived pathways, are the dominant hyporheic N2O sources (69.6 ± 2.1%) in agricultural streams around the world. The N2O fluxes are mainly in positive correlation with ammonia. The potential N2O metabolic pathways of metagenome-assembled genomes (MAGs) provides evidence that nitrifying bacteria contain greater abundances of N2O production-related genes than denitrifying bacteria. Taken together, this study highlights the importance of mitigating agriculturally derived ammonium in low-order agricultural streams in controlling N2O emissions. Global models of riverine ecosystems need to better represent ammonia-derived pathways for accurately estimating and predicting riverine N2O emissions.

Nitrous oxide reductase gene (nosZ) and N2O reduction along the littoral gradient of a eutrophic freshwater lake.

Lake littoral zones are characterized by heterogeneity in the biogeochemistry of nutrient elements. This study aimed to explore the relationship between the nitrous oxide reductase gene (nosZ)-encoding denitrifier community composition/abundance and N2O reduction. Five samples (deep sediment, near-transition sediment, transition site, near-transition land and land soil) were collected along a littoral gradient of eutrophic Baiyangdian Lake, North China. To investigate the relationship between the nosZ-encoding denitrifier community structure and N2O reduction, the nosZ-encoding denitrifier community composition/abundance, potential denitrification rate (DNR) and potential N2O production rate (pN2O) were investigated using molecular biological technologies and laboratory incubation experiments. The results showed that the average DNR of sediments was about 25 times higher than that of land soils, reaching 282.5 nmol N/(g dry weight (dw) x hr) and that the average pN2O of sediments was about 3.5 times higher than that of land soils, reaching 15.7 nmol N/(g dw x hr). In the land area, the nosZ gene abundance showed a negative correlation with the N2O/(N2O + N2) ratio, indicating that nosZ gene abundance dominated N2O reduction both in the surface soils of the land area and in the soil core of the transition site. Phylogenetic analysis showed that all the nosZ sequences recovered from sediment clustered closely with the isolates Azospirillum largimobile and Azospirillum irakense affiliated to Rhodospirillaceae in alpha-Proteobacteria, while about 92.3% (12/13) of the nosZ sequences recovered from land soil affiliated to Rhizobiaceae and Bradyrhizobiaceae in alpha-Proteobacteria. The community composition of nosZ gene-encoding denitrifiers appeared to be coupled with N2O reduction along the littoral gradient.

Hot moment of N2O emissions in seasonally frozen peatlands.

Since the start of the Anthropocene, northern seasonally frozen peatlands have been warming at a rate of 0.6 °C per decade, twice that of the Earth's average rate, thereby triggering increased nitrogen mineralization with subsequent potentially large losses of nitrous oxide (N2O) to the atmosphere. Here we provide evidence that seasonally frozen peatlands are important N2O emission sources in the Northern Hemisphere and the thawing periods are the hot moment of annual N2O emissions. The flux during the hot moment of thawing in spring was 1.20 ± 0.82 mg N2O m-2 d-1, significantly higher than that during the other periods (freezing, -0.12 ± 0.02 mg N2O m-2 d-1; frozen, 0.04 ± 0.04 mg N2O m-2 d-1; thawed, 0.09 ± 0.01 mg N2O m-2 d-1) or observed for other ecosystems at the same latitude in previous studies. The observed emission flux is even higher than those of tropical forests, the World's largest natural terrestrial N2O source. Furthermore, based on soil incubation with 15N and 18O isotope tracing and differential inhibitors, heterotrophic bacterial and fungal denitrification was revealed as the main source of N2O in peatland profiles (0-200 cm). Metagenomic, metatranscriptomic, and qPCR assays further revealed that seasonally frozen peatlands have high N2O emission potential, but thawing significantly stimulates expression of genes encoding N2O-producing protein complexes (hydroxylamine dehydrogenase (hao) and nitric oxide reductase (nor)), resulting in high N2O emissions during spring. This hot moment converts seasonally frozen peatlands into an important N2O emission source when it is otherwise a sink. Extrapolation of our data to all northern peatland areas reveals that the hot moment emissions could amount to approximately 0.17 Tg of N2O yr-1. However, these N2O emissions are still not routinely included in Earth system models and global IPCC assessments.

Anammox bacterial abundance, activity, and contribution in riparian sediments of the Pearl River estuary.

The hypothesis of an anammox hotspot in river riparian zones was put forward based on our investigation on freshwater ecotones for over 25 years and previous anammox research. Here we used a complementary array of methods including isotope-pairing technique, quantitative PCR assays, and 16S rRNA and hydrazine synthase gene (hzsB) clone libraries to document the spatiotemporal evidence for a high abundance zone of anammox bacteria in river riparian sediment with observed abundance of 1.3-12 × 10(6) (summer) and 1.4-20 × 10(8) (winter) hydrazine synthase gene copies g(-1), which is the highest abundance in natural environments recorded so far. Meanwhile high anammox bacterial biodiversity were detected with 'Brocadia' and 'Kuenenia' dominating. However, the high anammox bacterial abundances were not related with high activities and contributions for nitrogen gas generation. The anammox activities ranged from 0.07 to 0.15 nmol N cm(-3) h(-1) (summer) to 1.0-2.6 nmol N cm(-3) h(-1) (winter) with high temporal heterogeneity. The retrieval of archaeal and bacterial amoA sequences indicated that nitrifying microbes might be the major source of nitrite for anammox bacteria in winter, while in summer the anaerobic nitrate reduction is more likely the main source. On the basis of (15)N tracing technology, it was estimated that a total loss of 0.67-9.62 g N m(-2) yr(-1) is linked to anammox in the riparian zone while denitrification contributed 96.2-170.3 g N m(-2) yr(-1) in Pearl River riparian sediments.

Towards a more labor-saving way in microbial ammonium oxidation: A review on complete ammonia oxidization (comammox).

In the Anthropocene, nitrogen pollution is becoming an increasing challenge for both mankind and the Earth system. Microbial nitrogen cycling begins with aerobic nitrification, which is also the key rate-limiting step. For over a century, it has been accepted that nitrification occurs sequentially involving ammonia oxidation, which produces nitrite followed by nitrite oxidation, generating nitrate. This perception was changed by the discovery of comammox Nitrospira bacteria and their metabolic pathway. In addition, this also provided us with new knowledge concerning the complex nitrogen cycle network. In the comammox process, ammonia can be completely oxidized to nitrate in one cell via the subsequent activity of the enzyme complexes, ammonia monooxygenase, hydroxylamine dehydrogenase, and nitrite oxidodreductase. Over the past five years, research on comammox made great progress. However, there still exist a lot of questions, including how much does comammox contribute to nitrification? How large is the diversity and are there new strains to be discovered? Do comammox bacteria produce the greenhouse gas N2O, and how or to which extent may they contribute to global climate change? The above four aspects are of great significance on the farmland nitrogen management, aquatic environment restoration, and mitigation of global climate change. As large number of comammox bacteria and pathways have been detected in various terrestrial and aquatic ecosystems, indicating that the comammox process may exert an important role in the global nitrogen cycle.

Anammox bacterial abundance, biodiversity and activity in a constructed wetland.

An integrated approach to document high anammox activity and biodiversity in a constructed wetland (CW) was performed and showed that substantial anammox activity could mitigate undesirable N(2)O emission. The enhanced anammox bacterial abundance, biodiversity and activity were achieved by supplementing activated sludge to the CW. Up to 3.38 × 10(7) gene copies g(-1) dry soil of anammox bacteria were enriched in the CW. The activity measured by isotope pairing technique increased from 1.6 nmol N g(-1) sludge h(-1) in the original activated sludge to 18 nmol N g(-1) soil h(-1) in the CW, with the specific cellular activity increased from 5.1 to 12.8 fmol cell(-1) d(-1). Up to 33% of produced N(2) could be attributed to anammox process in the CW, with the remainder being due to denitrification. Phylogenetic analysis of anammox bacterial 16S rRNA genes indicated a shift of community from single Candidatus "Brocadia fulgida" in sludge to multiple "Jettenia", "Brocadia", and "Anammoxoglobus" species in the CW. With static chambers and control experiments, the CW with supplemented sludge had a 30% reduced N(2)O emission flux compared with the tests without adding biomass during an 8 month testing period.

Quantitative analyses of ammonia-oxidizing Archaea and bacteria in the sediments of four nitrogen-rich wetlands in China.

With the rapid development of ammonia-synthesizing industry, the ammonia-nitrogen pollution in wetlands acting as the sink of point and diffuse pollution has been increased dramatically. Most of ammonia-nitrogen is oxidized at least once by ammonia-oxidizing prokaryotes to complete the nitrogen cycle. Current research findings have expanded the known ammonia-oxidizing prokaryotes from the domain Bacteria to Archaea. However, in the complex wetlands environment, it remains unclear whether ammonia oxidation is exclusively or predominantly linked to Archaea or Bacteria as implied by specific high abundance. In this research, the abundance and composition of Archaea and Bacteria in sediments of four kinds of wetlands with different nitrogen concentration were investigated by using quantitative real-time polymerase chain reaction, cloning, and sequencing approaches based on amoA genes. The results indicated that AOA distributed widely in wetland sediments, and the phylogenetic tree revealed that archaeal amoA functional gene sequences from wetlands sediments cluster as two major evolutionary branches: soil/sediment and sediment/water. The bacteria functionally dominated microbial ammonia oxidation in different wetlands sediments on the basis of molecule analysis, potential nitrification rate, and soil chemistry. Moreover, the factors influencing AOA and AOB abundances with environmental indicator were also analyzed, and the results addressed the copy numbers of archaeal and bacterial amoA functional gene having the higher correlation with pH and ammonia concentration. The pH had relatively great negative impact on the abundance of AOA and AOB, while ammonia concentration showed positive impact on AOB abundance only. These findings could be fundamental to improve understanding of the importance of AOB and AOA in nitrogen and other nutrients cycle in wetland ecosystems.