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Ammonia oxidizers are key players in the global nitrogen cycle and are responsible for the oxidation of ammonia to nitrite, which is further oxidized to nitrate by other microorganisms. Their activity can lead to adverse effects on some human-impacted environments, including water pollution through leaching of nitrate and emissions of the greenhouse gas nitrous oxide (N2O). Ammonia monooxygenase (AMO) is the key enzyme in microbial ammonia oxidation and shared by all groups of aerobic ammonia oxidizers. The AMO has not been purified in an active form, and much of what is known about its potential structure and function comes from studies on its interactions with inhibitors. The archaeal AMO is less well studied as ammonia oxidizing archaea were discovered much more recently than their bacterial counterparts. The inhibition of ammonia oxidation by aliphatic alcohols (C1-C8) using the model terrestrial ammonia oxidizing archaeon ' Candidatus Nitrosocosmicus franklandus' C13 and the ammonia oxidizing bacterium Nitrosomonas europaea was examined in order to expand knowledge about the range of inhibitors of ammonia oxidizers. Methanol was the most potent specific inhibitor of the AMO in both ammonia oxidizers, with half-maximal inhibitory concentrations (IC50) of 0.19 and 0.31 mM, respectively. The inhibition was AMO-specific in ' Ca. N. franklandus' C13 in the presence of C1-C2 alcohols, and in N. europaea in the presence of C1-C3 alcohols. Higher chain-length alcohols caused non-specific inhibition and also inhibited hydroxylamine oxidation. Ethanol was tolerated by ' Ca. N. franklandus' C13 at a higher threshold concentration than other chain-length alcohols, with 80 mM ethanol being required for complete inhibition of ammonia oxidation. [ABSTRACT FROM AUTHOR]

[Display omitted] • A microelectrolysis-enhanced membrane-aerated biofilm reactor was developed. • The removal performance of NH 4 +-N and sulfamethoxazole (SMX) was enhanced. • The expression of ammonia monooxygenase (AMO) was up-regulated (0.92 log 2). • AMO exposed additional binding sites for NH 4 +-N and SMX. • Up-regulated antibiotic resistance genes were eliminated by 90%. A self-corrosion microelectrolysis (SME)-enhanced membrane-aerated biofilm reactor (eMABR) was developed for the removal of pollutants and reduction of antibiotic resistance genes (ARGs). Fe2+ and Fe3+ formed iron oxides on the biofilm, which enhanced the adsorption and redox process. SME can induce microorganisms to secrete more extracellular proteins and up-regulate the expression of ammonia monooxygenase (AMO) (0.92 log 2). AMO exposed extra binding sites (ASP-69) for antibiotics, weakening the competition between NH 4 +-N and sulfamethoxazole (SMX). The NH 4 +-N removal efficiency in the S-eMABR (adding SMX and IC) increased by 44.87 % compared to the S-MABR (adding SMX). SME increased the removal performance of SMX by approximately 1.45 times, down-regulated the expressions of sul 1 (−1.69 log 2) and sul 2 (−1.30 log 2) genes, and controlled their transfer within the genus. This study provides a novel strategy for synergistic reduction of antibiotics and ARGs, and elucidates the corresponding mechanism based on metatranscriptomic and molecular docking analyses. [ABSTRACT FROM AUTHOR]

• 4-CP biodegradation was enhanced with reduced volatilization in nitrifying MABR. • Cometabolism pathway of 4-CP in nitrifying MABR was proposed. • Mechanism involved in AMO-mediated cometabolism of 4-CP was elucidated. • MD approaches helped to explain the reactive sites of 4-CP metabolism. • DFT calculation can well elucidate the biotransformation product of 4-CP. Ammonia monooxygenase (AMO)-mediated cometabolism of organic pollutants has been widely observed in biological nitrogen removal process. However, its molecular mechanism remains unclear, hindering its practical application. Furthermore, conventional nitrification systems encounter significant challenges such as air pollution and the loss of ammonia-oxidizing bacteria, when dealing with wastewater containing volatile organic pollutants. This study developed a nitrifying membrane-aerated biofilm reactor (MABR) to enhance the biodegradation of volatile 4-chlorophenol (4-CP). Results showed that 4-CP was primarily removed via Nitrosomonas nitrosa -mediated cometabolism in the presence of NH 4 +-N, supported by the increased nicotinamide adenine dinucleotide (NADH) and adenosine triphosphate (ATP) content, AMO activity and the related genes abundance. Hydroquinone, detected for the first time and produced via oxidative dechlorination, as well as 4-chlorocatechol was primary transformation products of 4-CP. Nitrosomonas nitrosa AMO structural model was constructed for the first time using homology modeling. Molecular dynamics simulation suggested that the ortho -carbon in the benzene ring of 4-CP was more prone to metabolismcompared to the ipso -carbon. Density functional theory calculation revealed that 4-CP was metabolized by AMO via H-abstraction-OH-rebound reaction, with a significantly higher rebound barrier at the ipso -carbon (16.37 kcal·mol-1) as compared to the ortho -carbon (6.7 kcal·mol-1). This study fills the knowledge gap on the molecular mechanism of AMO-mediated cometabolism of organic pollutants, providing practical and theoretical foundations for improving volatile organic pollutants removal through nitrifying MABR. [Display omitted] [ABSTRACT FROM AUTHOR]

Antibiotics are mostly collected by sewage systems, but not completely removed within wastewater treatment plants. Their release to aquatic environment poses a great threat to public health. This study evaluated the removal of a widely used fluoroquinolone antibiotic, ciprofloxacin, in enriched nitrifying culture through a series of experiments by controlling ammonium concentrations and inhibiting functional microorganisms. The removal efficiency of ciprofloxacin at an initial concentration of 50 μg L−1 reached 81.86 ± 3.21% in the presence of ammonium, while only 22.83 ± 8.22% of ciprofloxacin was removed in its absence. A positive linear correlation was found between the ammonia oxidation rate (AOR) and ciprofloxacin biodegradation rate. These jointly confirmed the importance of the AOB-induced cometabolism in ciprofloxacin biodegradation, with adsorption and metabolic degradation pathways playing minor roles. The continuous exposure of AOB to ciprofloxacin led to decreases of ammonia monooxygenase (AMO) activities and AOR. The antibacterial effects of ciprofloxacin and its biodegradation products were further evaluated and the results revealed that biodegradation products of ciprofloxacin exhibited less toxicity compared to the parent compound, implying the potential application of cometabolism in alleviation of antimicrobial activity. The findings provided new insights into the AOB-induced cometabolic biodegradation of fluoroquinolone antibiotics. HIGHLIGHTS Cometabolic biodegradation was the major removal pathway of ciprofloxacin.; Contributions from metabolism by AOB and heterotrophs were insignificant.; AOR, AMO activities and amoA gene abundance decreased during exposure.; Cometabolic biodegradation products exhibited less toxicity than ciprofloxacin.;

• Bacterial ammonia monooxygenase genes and host were decreased after COVID-19 exposure. • The disturbance of ammonia oxidation recovered in the post-pandemic period. • Disinfectants destroy cell structure and cause oxidative stress of Nitrosomonas. • Sodium hypochlorite induce the downregulation of ammonia monooxygenase genes. • Sodium hypochlorite destroy the structure of ammonia monooxygenase protein. Anthropogenic activities significantly impact the elemental cycles in aquatic ecosystems, with the N-cycling playing a critical role in potential nutrient turnover and substance cycling. We hypothesized that measures to prevent COVID-19 transmission profoundly altered the nitrogen cycle in riverine ecosystems. To investigate this, we re-analyzed metagenomic data and identified 60 N-cycling genes and 21 host metagenomes from four urban reaches (one upstream city, Wuhan, and two downstream cities) along the Yangtze River. Our analyses revealed a marked decrease in the abundance of bacterial ammonia monooxygenase genes, as well as in the host, ammonia-oxidizing autotrophic Nitrosomonas , followed by a substantial recovery post-pandemic. We posited that discharge of sodium hypochlorite (NaOCl) disinfectant may be a primary factor in the reduction of N-cycling process. To test this hypothesis, we exposed pure cultures of Nitrosomonas europaea to NaOCl to explore the microbial stress response. Results indicated that NaOCl exposure rapidly compromised the cell structure and inhibited ammonia oxidation of N. europaea , likely due to oxidative stress damage and reduced expression of nitrogen metabolism-related ammonia monooxygenase. Using the functional tagging technique, we determined that NaOCl directly destroyed the ammonia monooxygenase protein and DNA structure. This study highlights the negative impacts of chlorine disinfectants on the function of aquatic ecosystems and elucidates potential mechanisms of action. [Display omitted] [ABSTRACT FROM AUTHOR]

Nitrosospira briensis C-128 is an ammonia-oxidizing bacterium isolated from an acid agricultural soil. N. briensis C-128 was sequenced with PacBio RS technologies at the DOE-Joint Genome Institute through their Community Science Program (2010). The high-quality finished genome contains one chromosome of 3.21 Mb and no plasmids. We identified 3073 gene models, 3018 of which are protein coding. The two-way average nucleotide identity between the chromosomes of Nitrosospira multiformis ATCC 25196 and Nitrosospira briensis C-128 was found to be 77.2 %. Multiple copies of modules encoding chemolithotrophic metabolism were identified in their genomic context. The gene inventory supports chemolithotrophic metabolism with implications for function in soil environments.

Recently, an activity-based labelling protocol for the in vivo detection of ammonia- and alkane-oxidizing bacteria became available. This functional tagging technique enabled targeted studies of these environmentally widespread functional groups, but it failed to capture ammonia-oxidizing archaea (AOA). Since their first discovery, AOA have emerged as key players within the biogeochemical nitrogen cycle, but our knowledge regarding their distribution and abundance in natural and engineered ecosystems is mainly derived from PCR-based and metagenomic studies. Furthermore, the archaeal ammonia monooxygenase is distinctly different from its bacterial counterparts and remains poorly understood. Here, we report on the development of an activity-based labelling protocol for the fluorescent detection of all ammonia- and alkane-oxidizing prokaryotes, including AOA. In this protocol, 1,5-hexadiyne is used as inhibitor of ammonia and alkane oxidation and as bifunctional enzyme probe for the fluorescent labelling of cells via the Cu(I)-catalyzed alkyne-azide cycloaddition reaction. Besides efficient activity-based labelling of ammonia- and alkane-oxidizing microorganisms, this method can also be employed in combination with deconvolution microscopy for determining the subcellular localization of their ammonia- and alkane-oxidizing enzyme systems. Labelling of these enzymes in diverse ammonia- and alkane-oxidizing microorganisms allowed their visualization on the cytoplasmic membranes, the intracytoplasmic membrane stacks of ammonia- and methane-oxidizing bacteria, and, fascinatingly, on vesicle-like structures in one AOA species. The development of this novel activity-based labelling method for ammonia- and alkane-oxidizers will be a valuable addition to the expanding molecular toolbox available for research of nitrifying and alkane-oxidizing microorganisms., Competing Interests: The authors declare no competing interests., (© The Author(s) 2024. Published by Oxford University Press on behalf of the International Society for Microbial Ecology.)

The abundance and taxonomic composition of ammonia‐oxidizing archaea (AOA) were assessed in paddy soils that had received more than 50 years of fertilization with and without inorganic N. The inorganic N fertilized treatments were: NPK and NPK + CO (nitrogen (N), phosphorus (P), potassium (K) and compost (CO)). The treatments without inorganic N were: CO, PK and control (unfertilized soils). Quantitative PCR (qPCR) analysis of the archaeal amoA gene showed no significant changes in AOA abundance following the long‐term application of inorganic N fertilizers. However, subsequent analysis of amoA gene sequencing data showed that inorganic N application significantly changed the AOA community composition and alpha diversity indices. Edge principal components analysis (PCA) showed varying contributions of distinct AOA lineages in separating samples according to the fertilization treatment. Addition of inorganic N favoured an increase in the abundance of AOA lineages belonging to Nitrososphaera, and the treatments without inorganic N additions formed a separate cluster dominated by Nitrosotalea lineage. The distinct response of the two AOA lineages points towards different community organization of soil AOA and strongly supports the concept of habitat partitioning in paddy soil ecosystems. Highlights: Lineages of AOA responded differently to different N management practices.Long‐term N fertilization selectively enriched AOA lineages, with C:N ratio a factor associated with such changesInorganic N favoured Nitrososphaera lineage abundanceRegardless of fertilizer status, Nitrosotalea lineage was dominant in the absence of inorganic N fertilization. [ABSTRACT FROM AUTHOR]

To date, little is known about the ecological significance of Comammox (COMplete AMMonia OXidizers) Nitrospira in the water column of freshwater lakes. Water samples collected along depth profiles were used to investigate the distribution of Comammox in 13 lakes characterized by a wide range of physicochemical properties. Several published primers, which target the α-subunit of the ammonia monooxygenase, generated non-specific PCR products or did not amplify target genes from lake water and other habitats. Therefore, a new primer set has been designed for specific detection of Comammox in lakes. The high specificity of the PCR assay was confirmed by sequencing analysis. Quantification of Comammox amoA genes in lake water samples based on droplet digital PCR (ddPCR) revealed very low abundances (not exceeding 85 amoA copies ml−1), which suggest that Comammox is of minor importance for the nitrification process in the water column of the study sites. Surprisingly, samples taken from the sediment/water-interface along an oxygen gradient in dimictic Piburger See showed Comammox abundances three to four magnitudes higher than in the pelagic realm of the lake, which indicates a preference of Comammox to a particle-attached lifestyle. [ABSTRACT FROM AUTHOR]

Ammonia monooxygenase is a copper-dependent membrane-bound enzyme that catalyzes the first step of nitrification in ammonia-oxidizing bacteria to convert ammonia to hydroxylamine, through the reductive insertion of a dioxygen-derived O atom in an N–H bond. This reaction is analogous to that carried out by particulate methane monooxygenase, which catalyzes the conversion of methane to methanol. The enzymatic activity of ammonia monooxygenase must be modulated to reduce the release of nitrogen-based soil nutrients for crop production into the atmosphere or underground waters, a phenomenon known to significantly decrease the efficiency of primary production as well as increase air and water pollution. The structure of ammonia monooxygenase is not available, rendering the rational design of enzyme inhibitors impossible. This study describes a successful attempt to build a structural model of ammonia monooxygenase, and its accessory proteins AmoD and AmoE, from Nitrosomonas europaea, taking advantage of the high sequence similarity with particulate methane monooxygenase and the homologous PmoD protein, for which crystal structures are instead available. The results obtained not only provide the structural details of the proteins ternary and quaternary structures, but also suggest a location for the copper-containing active site for both ammonia and methane monooxygenases, as well as support a proposed structure of a CuA-analogue dinuclear copper site in AmoD and PmoD. [ABSTRACT FROM AUTHOR]

Photoinhibition of ammonia oxidation occurs widely in aquatic environments and could suppress the nitrification rate, lead to the composition variation of inorganic nitrogen and influence the stability of aquatic ecosystems. Both ammonia-oxidizing bacteria (AOB) and archaea are sensitive to light. The extent of photoinhibition and the time required for recovery depend on light wavelength, intensity, photon quantity and strains. Strong evidence indicates that photoinhibition in AOB by visible light is mainly caused by irreversible damage to ammonia monooxygenase (AMO) and the degradation of AMO is beneficial to AOB recovery. This review discusses photoinhibition in metabolic pathways used by ammonia oxidizers. [ABSTRACT FROM AUTHOR]

Ammonia monooxygenase (AMO) is a key nitrogen transforming enzyme belonging to the same copper-dependent membrane monooxygenase family (CuMMO) as the particulate methane monooxygenase (pMMO). The AMO from ammonia oxidising archaea (AOA) is very divergent from both the AMO of ammonia oxidising bacteria (AOB) and the pMMO from methanotrophs and little is known about the structure or substrate range of the archaeal AMO. This study compares inhibition by C2-C8 linear 1-alkynes of AMO from two phylogenetically distinct strains of AOA, "Candidatus Nitrosocosmicus franklandus" C13 and "Candidatus Nitrosotalea sinensis" Nd2, with AMO from Nitrosomonas europaea and pMMO from Methylococcus capsulatus (Bath). An increased sensitivity of the archaeal AMO to short-chain32 length alkynes (=C5) appeared to be conserved across AOA lineages. Similarities in C2-C8 alkyne inhibition profiles between AMO from AOA and pMMO from M. capsulatus suggested that the archaeal AMO has a narrower substrate range compared to that of N. europaea AMO. Inhibition of AMO from "Ca. Nitrosocosmicus franklandus" and N. europaea by the aromatic alkyne phenylacetylene was also investigated. Kinetic data revealed that the mechanism by which phenylacetylene inhibits "Ca. Nitrosocosmicus franklandus" and N. europaea is different, indicating differences in the AMO active site between AOA and AOB. Phenylacetylene was found to be a specific and irreversible inhibitor of AMO from "Ca. Nitrosocosmicus franklandus" which does not compete with NH3 for binding at the active site. [ABSTRACT FROM AUTHOR]

Approximately two percent of the world's energy is consumed in the production of ammonia from hydrogen and nitrogen gas. Ammonia is used as a fertilizer ingredient for agriculture and distributed in the environment on an enormous scale to promote crop growth in intensive farming. Only 30–50 % of the nitrogen applied is assimilated by crop plants; the remaining 50–70 % goes into biological processes such as nitrification by microbial metabolism in the soil. This leads to an imbalance in the global nitrogen cycle and higher nitrous oxide emissions (a potent and significant greenhouse gas) as well as contamination of ground and surface waters by nitrate from the nitrogen‐fertilized farmland. This Review gives a critical overview of the current knowledge of soil microbes involved in the chemistry of ammonia nitrification, the structures and mechanisms of the enzymes involved, and phytochemicals capable of inhibiting ammonia nitrification. [ABSTRACT FROM AUTHOR]

To mitigate the toxicity of ammonia in aquaculture systems, marine and brackish water ammoniaoxidizing bacterial consortia have been developed and are used for activation of nitrifying bioreactors integrated to recirculating aquaculture systems. To shed more light on to these biological entities, diversity of both the consortia were analyzed based on random cloning of 16S rRNA gene and ammonia-oxidizing bacterial specific amoA gene sequences. The dendrograms of representative clones on the basis of amplified ribosomal DNA restriction analysis generated 22 and 19 clusters for marine and brackish water nitrifying consortia, respectively. Phylogenetic analysis demonstrated the presence of various autotrophic nitrifiers belonging to α-, β- and γ-Proteobacteria, anaerobic ammonia oxidizers, heterotrophic denitrifiers, Bacteroidetes, and Actinobacteria. Distribution patterns of the organisms within the two consortia were determined using the software Geneious and diversity indices were investigated using Mega 5.0, VITCOMIC and Primer 7. The abundance of ammonia oxidizers was found in the order of 2.21±0.25 x 109 copies/g wet weight of marine consortium and 6.20±0.23 x 107 copies/g of brackish water consortium. Besides, marine ammoniaoxidizing consortium exhibited higher mean population diversity and Shannon Wiener diversity than the brackish water counterparts. [ABSTRACT FROM AUTHOR]

Published PCR primers targeting the ammonia monooxygenase gene (amoA) were applied to samples from activated sludge systems operated with low dissolved oxygen (DO) to quantify total and clade-level Nitrospira that perform complete ammonium oxidation (comammox); however, we found these existing primers resulted in significant artifact-associated non-target amplification. This not only overestimated comammox amoA copies but also resulted in numerous false positive detections in the environmental samples tested, as confirmed by gel electrophoresis. Therefore, instead of attempting to quantify comammox diversity, we focused on accurately quantifying the candidate comammox species. We designed specific and sensitive primers targeting 3 candidate species: Candidatus (Ca.) Nitrospira nitrosa, Ca. N. inopinata, and Ca. N. nitrificans. The primers were tested with amoA templates of these candidate species and used to quantify comammox at the species level in low DO activated sludge systems. We found that comammox related to Ca. N. nitrosa were present and abundant in the majority of samples from low DO bioreactors and were not detected in samples from a high DO system. In addition, the greatest abundance of Ca. N. nitrosa was found in bioreactors operated with a long solids retention time. Ca. N. inopinata and Ca. N. nitrificans were only detected sporadically in these samples, indicating a minor role of these comammox in nitrification under low DO conditions. [ABSTRACT FROM AUTHOR]

Abstract Although the adverse impacts of emerging nanoparticles (NPs) on the biological nitrogen removal (BNR) process have been broadly reported, the adaptive responses of NP-impaired nitrifiers and the related mechanisms have seldom been addressed to date. Here, we systematically explored the adaption and recovery capacities of the ammonia oxidizer Nitrosomonas europae a under chronic TiO 2 NP exposure and different dissolved oxygen (DO) conditions at the physiological and transcriptional levels in a chemostat reactor. N. europae a cells adapted to 50 mg/L TiO 2 NP exposure after 40-d incubation and the inhibited cell growth, membrane integrity, nitritation rate, and ammonia monooxygenase activity all recovered regardless of the DO concentrations. Transmission electron microscope imaging indicated the remission of the membrane distortion after the cells' 40-d adaption to the NP exposure. The microarray results further suggested that the metabolic processes associated with the membrane repair were pivotal for cellular adaption/recovery, such as the membrane efflux for toxicant exclusion, the structural preservation or stabilization, and the osmotic equilibrium adjustment. In addition, diverse metabolic and stress-defense pathways, including aminoacyl-tRNA biosynthesis, respiratory chain, ATP production, toxin-antitoxin 'stress-fighting', and DNA repair were activated for the cellular adaption coupled with the metabolic activity recovery, probably via recovering the energy production/conversion efficiency and mediating the non-photooxidative stress. Finally, low DO (0.5 mg/L) incubated cells were more susceptible to TiO 2 NP exposure and required more time to adapt to and recover from the stress, which was probably due to the stimulation limitation of the oxygen-dependent energy metabolism with a lower oxygen supply. The findings of this study provide new insights into NP contamination control and management adjustments during the BNR process. Graphical abstract Image 1 Highlights • Cells adapted to chronic NP stress and all the metabolic activities recovered. • Membrane repair associated regulations were pivotal for cellular adaption. • Diverse metabolic and stress-defense pathways were activated for cell recovery. • Low DO condition compromised NP impaired cells' resistance and adaption capacities. • Stimulation limitation of respiratory chain accounted for the compromised capacity. [ABSTRACT FROM AUTHOR]

The discovery of complete ammonia oxidizers (comammox) refutes the century-old paradigm that nitrification requires the activity of two types of microbes. Determining the distribution and abundance of comammox in various environments is important for revealing the ecology of microbial nitrification within the global nitrogen cycle. In this study, the ubiquity and diversity of comammox were analyzed for samples from different types of environments, including soil, sediment, sludge, and water. The results of a two-step PCR using highly degenerate primers (THDPPCR) and quantitative real-time PCR (qPCR) supported the relatively high abundance of comammox in nearly half of all samples tested, sometimes even outnumbering canonical ammonia-oxidizing bacteria (AOB). In addition, a relatively high proportion of comammox in tap and coastal water samples was confirmed via analysis of metagenomic data sets in public databases. The diversity of comammox was estimated by comammox-specific partial nested PCR amplification of the ammonia monooxygenase subunit A (amoA) gene, and phylogenetic analysis of comammox AmoA clearly showed a split of clade A into clades A.1 and A.2, with the proportions of clades A.1, A.2, and B differing among the various environmental samples. Moreover, compared to the amoA genes of AOB and ammonia-oxidizing archaea (AOA), the comammox amoA gene exhibited higher diversity indices. The ubiquitous distribution and high diversity of comammox indicate that they are likely overlooked contributors to nitrification in various ecosystems. IMPORTANCE The discovery of complete ammonia oxidizers (comammox), which oxidize ammonia to nitrate via nitrite, refutes the century-old paradigm that nitrification requires the activity of two types of microbes and redefines a key process in the biogeochemical nitrogen cycle. Understanding the functional relationships between comammox and other nitrifiers is important for ecological studies on the nitrogen cycle. Therefore, the diversity and contribution of comammox should be considered during ecological analyses of nitrifying microorganisms. In this study, a ubiquitous and highly diverse distribution of comammox was observed in various environmental samples, similar to the distribution of canonical ammonia-oxidizing bacteria. The proportion of comammox was relatively high in coastal water and sediment samples, whereas it was nearly undetectable in open-ocean samples. The ubiquitous distribution and high diversity of comammox indicate that these microorganisms might be important contributors to nitrification. [ABSTRACT FROM AUTHOR]

17α-ethinylestradiol (EE2) has received increasing attention as an emerging and difficult-to-remove emerging contaminant in recent years. Ammonia-oxidizing bacteria (AOB) have been reported to be effective in EE2 removal, and ammonia monooxygenase (AMO) is considered as the primary enzyme for EE2 removal. However, the molecular mechanism underlying the transformation of EE2 by AOB and AMO is still unclear. This study investigated the molecular mechanism of EE2 degradation using a combination of experimental and computational simulation methods. The results revealed that ammonia nitrogen was essential for the co-metabolism of EE2 by AOB, and that NH 3 bound with Cu C (one active site of AMO) to induce a conformational change in AMO, allowing EE2 to bind with the other active site (Cu B), and then EE2 underwent biological transformation. These results provide a theoretical basis and a novel research perspective on the removal of ammonia nitrogen and emerging contaminants (e.g., EE2) in wastewater treatment. [Display omitted] • The molecular mechanism underlying the transformation of EE2 by AMO was revealed. • Ammonia nitrogen was essential for the co-metabolism of EE2 by AMO. • NH 3 bound with Cu C in AMO and induced a conformational change of AMO. • EE2 bound with Cu B in AMO and underwent biotransformation after NH 3 binding. [ABSTRACT FROM AUTHOR]

In their natural habitats, microorganisms are often exposed to periods of starvation if their substrates for energy generation or other nutrients are limiting. Many microorganisms have developed strategies to adapt to fluctuating nutrients and long-term starvation. In the environment, ammonia oxidizers have to compete with many different organisms for ammonium and are often exposed to long periods of ammonium starvation. We investigated the effect of ammonium starvation on ammonia-oxidizing archaea (AOA) and bacteria (AOB) enriched from freshwater lake sediments. Both AOA and AOB were able to recover even after almost two months of starvation; however, the recovery time differed. AOA and AOB retained their 16S rRNA (ribosomes) throughout the complete starvation period. The AOA retained also a small portion of the mRNA of the ammonia monooxygenase subunit A (amoA) for the complete starvation period. However, after 10 days, no amoA mRNA was detected anymore in the AOB. These results indicate that AOA and AOB are able to survive longer periods of starvation, but might utilize different strategies.

Abstract Ionic liquids (ILs) were widely applied because of their excellent properties. The present investigation studied the toxicity of the IL 1-octyl-3-methylimidazolium hexafluorophosphate ([Omim]PF 6) to the soil microbial population and community diversity with dose (1.0, 2.0, 4.0, 6.0, and 8.0 mg kg−1) and exposure time (7, 10, and 13 d). The results show the IL was stable during the entire experimental period. The Biolog-ECO plate results indicated that the average well color development (AWCD) in the 6.0 and 8.0 mg kg−1 treatments was lower than these in the other treatments. The diversity indices of the Biolog analysis were significantly reduced. The abundance of the ammonia-oxidizing archaea (AOA-) and the ammonia-oxidizing bacteria (AOB-) ammonia monooxygenase (amoA) genes was measured by the real-time polymerase chain reaction (RT-PCR). In the treatments of 4.0, 6.0 and 8.0 mg kg−1, the abundance of amoA genes of the AOA- and AOB- were inhibited by IL [Omim]PF 6. Graphical abstract Image Highlights • The levels of amoA genes was used to study the toxicity of [Omim]PF 6 in brown soil. • Functional diversity of soil microbes was used to study the toxicity of [Omim]PF 6. • The concentrations of investigated IL in soil relatively maintained stable. • The investigated IL has potential risks if they were released to soil by accident. [ABSTRACT FROM AUTHOR]

The extensive release of cerium dioxide nanoparticles (CeO 2 NPs) causes increasing potential health and environmental risks. In this study, acute and chronic responses of macrophyte and microorganisms were evaluated following exposure to CeO 2 NPs in constructed wetlands (CWs). It was found that most of the CeO 2 NPs were retained in the CW substrate, with minor amounts accumulated in plant tissues. Photosynthetic activity experiment indicated that environmentally relevant concentration (1 mg/L) of CeO 2 NPs enhanced the net photosynthesis rate of plants, while chronic exposure to a high concentration (50 mg/L) of CeO 2 NPs inhibited photosynthesis. The variations of the malondialdehyde (MDA) content and antioxidant enzyme activities in plant tissues revealed that acute exposure to CeO 2 NPs induced oxidative stress to plants, while there was no obvious chronic effect. Moreover, the biomass increased marginally under exposure to 1 mg/L CeO 2 NPs, whereas exposure to 50 mg/L CeO 2 NPs reduced the total biomass of plants by 29%. The release of lactate dehydrogenase (LDH) suggested that chronic exposure to CeO 2 NPs inhibited microbial viability. Metagenome and enzymatic activities analyses further revealed that CeO 2 NPs significantly changed the microbial community structure and inhibited the activities of dehydrogenase, urease, ammonia monooxygenase, and phosphatase. This resulted in the decline of contaminant removal efficiency. Overall, acute and chronic exposure to CeO 2 NPs had multiple effects on macrophyte and microorganisms in CWs, which significantly impacted the treatment performance of the ecological system. [ABSTRACT FROM AUTHOR]

Planktonic archaea are thought to play an important role in ammonia oxidation in marine environments. Data on the distribution, abundance, and diversity of ammonia oxidizers in the coastal sea-surface microlayer (SML) are lacking, despite previous reports of high abundance of Thaumarchaeota in the SML of estuaries and freshwater lakes. Here, we failed to detect the presence of ammonia-oxidizing bacteria in any of our samples taken from a semi-enclosed marine inlet in Japan. Therefore, we shifted our focus to examine the archaeal community composition as well as the Thaumarchaeota marine group I (MG-I) and ammonia monooxygenase subunit A (amoA) gene copy numbers and composition in the SML and corresponding underlying water (UW, 20 cm). amoA gene copy numbers obtained by quantitative PCR were consistent with the typical values observed in the surface waters of oceanic and coastal environments where nitrification activity has been detected, but the copy numbers were two- to three-fold less than those reported from the surface layers and UW of high mountain lakes. Both amoA and MG-I 16S rRNA gene copy numbers were significantly negatively correlated with chlorophyll-a and transparent exopolymer particle concentrations in the SML. Communities of archaea and ammonia-oxidizing archaea in SML samples collected during low wind conditions (≤5 m s–1) differed the most from those in UW samples, whereas the communities in SML samples collected during high wind conditions were similar to the UW communities. In the SML, low ratios of amoA to MG-I 16S rRNA genes were observed, implying that most of the SML Thaumarchaeota lacked amoA. To our knowledge, our results provide the first comparison of ammonia-oxidizing communities in the coastal SML with those in the UW. [ABSTRACT FROM AUTHOR]

Forest ecosystems have great ecological values in mitigation of climate change and protection of biodiversity of flora and fauna; re-forestry is commonly used to enhance the sequestration of atmospheric CO2 into forest storage biomass. Therefore, seasonal and spatial dynamics of the major microbial players in nitrification, ammonia-oxidizing archaea (AOA) and bacteria (AOB), in acidic soils of young and matured revegetated forests were investigated to elucidate the changes of microbial communities during forest restoration, and compared to delineate the patterns of community shifts under the influences of environmental factors. AOA were more abundant than AOB in both young and matured revegetated forest soils in both summer and winter seasons. In summer, however, the abundance of amoA-AOA decreased remarkably (p < 0.01), ranging from 1.90 (± 0.07) × 108 copies per gram dry soil in matured forest to 5.04 (± 0.43) × 108 copies per gram dry soil in young forest, and amoA-AOB was below detection limits to obtain any meaningful values. Moreover, exchangeable Al3+ and organic matter were found to regulate the physiologically functional nitrifiers, especially AOA abundance in acidic forest soils. AOB community in winter showed stronger correlation with the restoration status of revegetated forests and AOA community dominated by Nitrosotalea devanaterra, in contrast, was more sensitive to the seasonal and spatial variations of environmental factors. These results enrich the current knowledge of nitrification during re-forestry and provide valuable information to developmental status of revegetated forests for management through microbial analysis. [ABSTRACT FROM AUTHOR]

Summary: Thaumarchaea are ubiquitous in marine habitats where they participate in carbon and nitrogen cycling. Although metatranscriptomes suggest thaumarchaea are active microbes in marine waters, we understand little about how thaumarchaeal gene expression patterns relate to substrate utilization and activity. Here, we report the global transcriptional response of the marine ammonia‐oxidizing thaumarchaeon ‘Candidatus Nitrosopelagicus brevis’ str. CN25 to ammonia limitation using RNA‐Seq. We further describe the genome and transcriptome of Ca. N. brevis str. U25, a new strain capable of urea utilization. Ammonia limitation in CN25 resulted in reduced expression of transcripts coding for ammonia oxidation proteins, and increased expression of a gene coding an Hsp20‐like chaperone. Despite significantly different transcript abundances across treatments, two ammonia monooxygenase subunits (amoAB), a nitrite reductase (nirK) and both ammonium transporter genes were always among the most abundant transcripts, regardless of growth state. Ca. N. brevis str. U25 cells expressed a urea transporter 139‐fold more than the urease catalytic subunit ureC. Gene coexpression networks derived from culture transcriptomes and 10 thaumarchaea‐enriched metatranscriptomes revealed a high degree of correlated gene expression across disparate environmental conditions and identified a module of coexpressed genes, including amoABC and nirK, that we hypothesize to represent the core ammonia oxidation machinery. [ABSTRACT FROM AUTHOR]

The toxicity of ionic liquids (ILs) on soil organisms has aroused wide attention due to their high-solubility. The present investigation focused on the toxicity of 1-octyl-3-methylimidazolium nitrate ([C 8 mim]NO 3 ) on the microbial populations (bacteria, fungi, and actinomycetes), soil enzyme (urease, dehydrogenase, acid phosphatase, and β-glucosidase) activities, microbial community diversity using terminal restriction fragment length polymorphism (T-RFLP), and abundance of the ammonia monooxygenase (amoA) genes of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) using quantitative real-time polymerase chain reaction (q-PCR) in brown soil at each trial with doses of 0, 1.0, 5.0, and 10.0 mg/kg on days 10, 20, 30, and 40. The contents of [C 8 mim]NO 3 in soil were measured using high performance liquid chromatography with recoveries of 84.3% to 85.2%, and changed less than 10% during the experimental period. A significant decrease was observed from the bacteria, fungi and actinomycetes populations at 10.0 mg/kg, at which the urease activity was inhibited and the β-glucosidase activity was stimulated on days 20, 30, and 40. In addition, [C 8 mim]NO 3 inhibited the dehydrogenase activity at 10 mg/kg on days 30 and 40 and the acid phosphatase activity on day 20. The diversity of the soil microbial community and the gene abundance of AOA- and AOB- amoA were also inhibited. Furthermore, the present investigation provided more scientific information for the toxicity evaluation of ILs in soil. [ABSTRACT FROM AUTHOR]

With the phase-out of persistent, bioaccumalative, and toxic perfluoroalkyl carboxylic acids (PFCAs), it is needed to explore the potential release of PFCAs from precursors being emitted into the environment. Biotransformation of fluorotelomer alcohols (FTOHs) via biological processes in wastewater treatment plants (WWTPs) leads to discharge of PFCAs into receiving waters. However, the commonly existed microbial activity that can impact on FTOHs biodegradation in WWTPs remains unclear. The objective of present research was to explore the relationship between ammonia-oxidation process and the enhanced PFCAs generation from FTOHs biodegradation under aerobic activated sludge. The obtained results indicate that the cometabolism process performed by nitrifying microorganisms (NMs) was responsible for enhanced PFCAs generation. Among NMs, the ammonia-oxidation bacteria that can express non-specific enzyme of ammonia monooxygenases resulted in the enhanced PFCAs generation from FTOHs. Meanwhile, the different addition amount of ammonia contributed to different defluorination efficiency of FTOHs. The present study further correlated the enhanced PFCAs generation from FTOHs biodegradation with ammonia-oxidation process, which can provide practical information on effective management of PFCAs generation in WWTPs. [ABSTRACT FROM AUTHOR]

Ammonia-oxidising archaea (AOA) are ubiquitous and abundant in nature and play a major role in nitrogen cycling. AOA have been studied intensively based on the amoA gene (encoding ammonia monooxygenase subunit A), making it the most sequenced functional marker gene. Here, based on extensive phylogenetic and meta-data analyses of 33,378 curated archaeal amoA sequences, we define a highly resolved taxonomy and uncover global environmental patterns that challenge many earlier generalisations. Particularly, we show: (i) the global frequency of AOA is extremely uneven, with few clades dominating AOA diversity in most ecosystems; (ii) characterised AOA do not represent most predominant clades in nature, including soils and oceans; (iii) the functional role of the most prevalent environmental AOA clade remains unclear; and (iv) AOA harbour molecular signatures that possibly reflect phenotypic traits. Our work synthesises information from a decade of research and provides the first integrative framework to study AOA in a global context. [ABSTRACT FROM AUTHOR]

Microbial nitrification is a key process in the nitrogen cycle in the continental shelf ecosystems. The genotype compositions and abundance of the ammonia monooxygenase gene, amoA, derived from ammonia-oxidizing archaea (AOA) and bacteria (AOB) in two size fractions (2–10 and 0.2–2 µm), were investigated in the East China Sea (ECS) in May 2008 using PCR-denaturing gradient gel electrophoresis (DGGE) and quantitative PCR (qPCR). Four sites were selected across the continental shelf edge: continental shelf water (CSW), Kuroshio branch water (KBW), transition between CSW and KBW (TCSKB) and coastal KBW (CKBW). The gene copy numbers of AOA-amoA were higher than those of AOB-amoA in ECS. The relative abundance of amoA to the total 16S rRNA gene level reached approximately 15% in KBW and CKBW for the free-living fraction of AOA, whereas the level was less than 0.01% throughout ECS for the AOB. A cluster analysis of the AOA-amoA-DGGE band pattern showed distinct genotype compositions in CSW in both the size fractions and in the surface of the TCSKB and KBW. Sequences of the DGGE bands were assigned to two clades. One of the clades exclusively consisted of sequences derived from the 2–10-µm fraction. This study revealed that AOA-amoA abundance dominated over AOB-amoA throughout the ECS, whereas the genotype composition of AOA-amoA were distributed heterogeneously across the water masses. Additionally, this is the first report showing the distribution of AOA-amoA genotypes characteristic to particle-associated AOA in the offshore of the East China Sea. [ABSTRACT FROM AUTHOR]

Simultaneous nitrification and denitrification (SND) is a promising single-reactor biological nitrogen-removal method. Activated sludge with and without iron scrap supplementation (Sludge-Fe and Sludge-C, respectively) was acclimated under aerobic condition. The total nitrogen (TN) content of Sludge-Fe substantially decreased from 25.0 ± 1.0 to 11.2 ± 0.4 mg/L, but Sludge-C did not show the TN-removal capacity. Further investigations excluded a chemical reduction of NO3–-N by iron and a decrease of NH4+-N by microbial assimilation, and the contribution of SND was verified. Moreover, the amount of aerobic denitrifiers, such as bacteria belonging to the genera Thauera, Thermomonas, Rhodobacter, and Hyphomicrobium, was considerably enhanced, as observed through Miseq Illumina sequencing method. The activities of the key enzymes ammonia monooxygenase (AMO) and nitrite oxidoreductase (NXR), which are associated with nitrification, and periplasmic nitrate reductase (NAP) and nitrite reductase (NIR), which are related to denitrification, in Sludge-Fe were 1.23-, 1.53-, 3.60-, and 1.55-fold higher than those in Sludge-C, respectively. In Sludge-Fe, the quantity of the functional gene NapA encoding enzyme NAP, which is essential for aerobic denitrification, was significantly promoted. The findings indicate that SND is the primary mechanism underlying the removal of TN and that iron scrap can robustly stimulate SND under aerobic environment. [ABSTRACT FROM AUTHOR]

Ammonia-oxidizing bacteria (AOB) and archaea (AOA) are the main nitrifiers which are well studied in natural environments, and AOA frequently outnumber AOB by orders especially in acidic conditions, making AOA the most promising ammonia oxidizers. The phylogeny of AOA revealed in related studies, however, often varied and hardly reach a consensus on functional phylotypes. The objective of this study was to compare ammonia-oxidizing communities by amoA gene and transcript based on both genomic DNA and RNA in extremely acidic forest soils (pH <4.5). Our results support the numerical and functional dominance of AOA over AOB in acidic soils as bacterial amoA gene and transcript were both under detection limits and archaeal amoA, in contrast, were abundant and responded to the fluctuations of environmental factors. Organic matter from tree residues was proposed as the main source of microbial available nitrogen, and the potential co-precipitation of dissolved organic matter (DOM) with soluble Al species in acidic soil matrix may further restrict the amount of nitrogen sources required by AOB besides NH/NH equilibrium. Although AOA were better adapted to oligotrophic environments, they were susceptible to the toxicity of exchangeable Al. Phylotypes affiliated to Nitrososphaera, Nitrososphaera sister group, and Nitrosotalea were detected by amoA gene and transcript. Nitrosotalea devantaerra and Nitrososphaera sister group were the major AOA. Compared to the genomic DNA data, higher relative abundances of Nitrososphaera and Nitrososphaera sister group were recognized in amoA transcript inferred AOA communities, where Nitrosotalea relative abundance was found lower, implying the functional activities of Nitrososphaera sister group and Nitrososphaera were easily underestimated and Nitrosotalea did not attribute proportionally to nitrification in extremely acidic soils. Further comparison of the different AOA community compositions and relative abundance of each phylotypes revealed by amoA genes and transcripts make it possible to identify the functional AOA species and assess their ecological role in extremely acidic soils. [ABSTRACT FROM AUTHOR]

Knowledge of the molecular ecology and environmental determinants of ammonia-oxidizing organisms is critical to understanding and predicting the global nitrogen (N) and carbon cycles, but an incomplete biochemical picture hinders in vitro studies of N-cycling enzymes. Although an integrative structural and dynamic characterization at the atomic scale would advance our understanding of function tremendously, structural knowledge of key N-cycling enzymes from ecologically relevant ammonia oxidizers is unfortunately extremely limited. Here, we discuss the challenges and opportunities for examining the ecology of ammonia-oxidizing organisms, particularly uncultivated Thaumarchaeota, through (meta)genome-driven structural biology of the enzymes ammonia monooxygenase (AMO) and nitrite reductase (NirK). [ABSTRACT FROM AUTHOR]