Ammonia-oxidizing Communities Research Articles

Ammonia-oxidizing archaea and ammonium concentration as drivers of nitrification in a protected freshwater lake

Nitrification rates and ammonia-oxidizer functional gene abundance were measured in the water column of Lake Lacawac, Pennsylvania, USA, a freshwater lake on a nature sanctuary that allowed an investigation with minimal human impacts. Nitrification is a 2-step process consisting of ammonia oxidation followed by nitrite oxidation. Recent studies have shown that these 2 nitrification steps may be uncoupled and respond in different ways to environmental conditions. Additionally, the relative contribution of ammonia-oxidizing archea (AOA) vs ammonia-oxidizing bacteria (AOB) to nitrification rates varies widely across aquatic systems. To determine how nitrification rates are related to environmental parameters and the ammonia-oxidizing community in a nearly pristine environment, rates and gene abundance were measured over multiple seasons where in-situ environmental conditions varied. Rates of ammonia and nitrite oxidation were measured separately and summed to calculate total nitrification rates ranging from 1 to 568 nM/d. Ammonia oxidation rates generally outpaced nitrite oxidation rates, and rates of both ammonia and nitrite oxidation were higher at depth (10 m) compared with near-surface and mid water column. Ammonia oxidation, nitrite oxidation, and total nitrification rates were all strongly, positively correlated with in-situ [NH4+] (Kendall’s τ > 0.35, p < 0.02). AOB ammonia monooxygenase (amoA) gene copy numbers were generally greater than AOA amoA. However, AOB gene copy numbers were not correlated with any ammonia oxidation or total nitrification rates, whereas AOA abundance was positively correlated with both ammonia oxidation and total nitrification rates (Kendall’s τ > 0.41, p < 0.01). A Bayesian generalized additive model, which accounted for sampling month, indicated that total nitrification rates were best explained by AOA and [NH4+]. Thus, substrate concentration and AOA likely play key roles in regulating rates of nitrification in this small, nearly pristine freshwater lake. These reported relationships between rates of ammonia and nitrite oxidation (and, thus, total nitrification), in-situ environmental parameters, and the ammonia-oxidizer community in a protected environment establish a reference for evaluating the impact of a changing environment on mesotrophic lake water quality.

Biochar and lime amendments promote soil nitrification and nitrogen use efficiency by differentially mediating ammonia-oxidizer community in an acidic soil

Lime and biochar have been proposed as effective ameliorants for increasing soil fertility and nitrogen (N) cycling by regulating the ammonia-oxidizer community. However, the biological mechanisms of lime- and biochar-induced changes in the ammonia-oxidizer community, soil nitrification, crop productivity, and N use efficiency (NUE) remain largely unknown. Here, we established a 9-year field experiment to explore the ammonia-oxidizing archaeal (AOA) and bacterial (AOB) communities and measure potential nitrification activity (PNA), crop productivity, and NUE in response to biochar and lime amendments. We found that soil pH was significantly increased under lime amendment more than under biochar amendment, while soil organic carbon, total nitrogen, and ammonia nitrogen followed the opposite trend. Lime and biochar amendments significantly increased the abundance and diversity of AOA and AOB, and substantially shaped the AOA and AOB community compositions. The co-occurrence networks analysis indicated that potential keystone taxa had negative relationships with connected members in the network, which exhibited significant correlations with the abundance and diversity of AOA and AOB communities, PNA, crop productivity, and NUE. Structural equation modelling indicated that biochar amendment may mediate potential keystone taxa to increase the abundance and diversity of AOB community, and exhibit a stronger impact on PNA, crop productivity, and NUE than lime amendment. Collectively, our results highlight the ecological importance of lime- and biochar-induced effects on the ammonia-oxidizer community and soil N cycling dynamics.

Determining Factors for Nitrite Accumulation in an Acidic Nitrifying System: Influent Ammonium Concentration, Operational pH, and Ammonia-Oxidizing Community.

Acidic nitrification is attracting wide attention because it can enable robust suppression of nitrite-oxidizing bacteria (NOB) in wastewater treatment. This study reports a comprehensive assessment of the novel acidic nitrification process to identify the key factors that govern stable nitrite accumulation. A laboratory-scale moving-bed biofilm reactor receiving low-alkalinity wastewater was continuously operated under acidic conditions (pH < 6) for around two years, including nine stages varying influent and operational conditions. The results revealed that nitrite accumulation was related to three factors, i.e., influent ammonium concentration, operating pH, and ammonia-oxidizing microbial community. These three factors impact nitrite accumulation by altering the in situ concentration of free nitrous acid (FNA), which is a potent inhibitor of NOB. The critical FNA concentration is approximately one part per million (ppm, ∼1 mg HNO2-N/L), above which nitrite accumulation is stably maintained in an acidic nitrifying system. The findings of this study suggest that stable nitrite accumulation via acidic ammonia oxidation can be maintained under a range of influent and operational conditions, as long as a ppm-level of FNA is established. Taking low-strength mainstream wastewater (40-50 mg NH4+-N/L) with limited alkalinity as an example, stable nitrite accumulation was experimentally demonstrated at a pH of 4.35, under which an in situ FNA of 2.3 ± 0.6 mg HNO2-N/L was attained. Under these conditions, Candidatus Nitrosoglobus became the only ammonia oxidizer detectable by 16S rRNA gene sequencing. The results of this study deepen our understanding of acidic nitrifying systems, informing further development of novel wastewater treatment technologies.

The Responses of Ammonia-Oxidizing Microorganisms to Different Environmental Factors Determine Their Elevational Distribution and Assembly Patterns.

The assembly mechanisms shaping the elevational patterns of diversity and community structure in ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) are not well understood. We investigated the diversities, co-occurrence network patterns, key drivers, and potential activities of AOA and AOB communities along a large altitudinal gradient. The α-diversity of the AOA communities exhibited a monotonically decreasing pattern with increasing elevation, whereas a sinusoidal pattern was observed for the AOB communities. The mean annual temperature was the single factor that most strongly influenced the α-diversity of the AOA communities; however, the interactions of plant richness, soil conductivity, and total nitrogen made comparable contributions to the α-diversity of the AOB communities. Moreover, the β-diversities of the AOA and AOB communities were divided into two distinct clusters by elevation, i.e., low- (1800-2600m) and high-altitude (2800-4100m) sections. These patterns were attributed mainly to the soil pH, followed by variations in plant richness along the altitudinal gradient. In addition, the AOB communities were more important to the soil nitrification potential in the low-altitude section, whereas the AOA communities contributed more to the soil nitrification potential in the high-altitude section. Overall, this study revealed the key factors shaping the elevational patterns of ammonia-oxidizing communities and might predict the consequences of changes in ammonia-oxidizing communities.

Improving Nitrogen Use Efficiency in Aerobic Rice Based on Insights Into the Ecophysiology of Archaeal and Bacterial Ammonia Oxidizers.

The abundance and structural composition of nitrogen (N) transformation-related microbial communities under certain environmental conditions provide sufficient information about N cycle under different soil conditions. This study aims to explore the major challenge of low N use efficiency (NUE) and N dynamics in aerobic rice systems and reveal the agronomic-adjustive measures to increase NUE through insights into the ecophysiology of ammonia oxidizers. Water-saving practices, like alternate wetting and drying (AWD), dry direct seeded rice (DDSR), wet direct seeding, and saturated soil culture (SSC), have been evaluated in lowland rice; however, only few studies have been conducted on N dynamics in aerobic rice systems. Biological ammonia oxidation is majorly conducted by two types of microorganisms, ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB). This review focuses on how diversified are ammonia oxidizers (AOA and AOB), whose factors affect their activities and abundance under different soil conditions. It summarizes findings on pathways of N cycle, rationalize recent research on ammonia oxidizers in N-cycle, and thereby suggests adjustive agronomic measures to reduce N losses. This review also suggests that variations in soil properties significantly impact the structural composition and abundance of ammonia oxidizers. Nitrification inhibitors (NIs) especially nitrapyrin, reduce the nitrification rate and inhibit the abundance of bacterial amoA without impacting archaeal amoA. In contrast, some NIs confine the hydrolysis of synthetic N and, therefore, keep low NH4+-N concentrations that exhibit no or very slight impact on ammonia oxidizers. Variations in soil properties are more influential in the community structure and abundance of ammonia oxidizers than application of synthetic N fertilizers and NIs. Biological nitrification inhibitors (BNIs) are natural bioactive compounds released from roots of certain plant species, such as sorghum, and could be commercialized to suppress the capacity of nitrifying soil microbes. Mixed application of synthetic and organic N fertilizers enhances NUE and plant N-uptake by reducing ammonia N losses. High salt concentration promotes community abundance while limiting the diversity of AOB and vice versa for AOA, whereas AOA have lower rate for potential nitrification than AOB, and denitrification accounts for higher N2 production. Archaeal abundance, diversity, and structural composition change along an elevation gradient and mainly depend on various soil factors, such as soil saturation, availability of NH4+, and organic matter contents. Microbial abundance and structural analyses revealed that the structural composition of AOA was not highly responsive to changes in soil conditions or N amendment. Further studies are suggested to cultivate AOA and AOB in controlled-environment experiments to understand the mechanisms of AOA and AOB under different conditions. Together, this evaluation will better facilitate the projections and interpretations of ammonia oxidizer community structural composition with provision of a strong basis to establish robust testable hypotheses on the competitiveness between AOB and AOA. Moreover, after this evaluation, managing soils agronomically for potential utilization of metabolic functions of ammonia oxidizers would be easier.

Distribution and Activity of Ammonia-Oxidizers on the Size-Fractionated Particles in the Pearl River Estuary

To distinguish between the distribution and activity of ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA) in the Pearl River estuary (PRE), we investigated the DNA- and cDNA-based β-proteobacterial and archaeal amoA genes on three size-fractionated particles of &amp;gt;3.0 μm, 0.45–3.0 μm, and 0.22–0.45 μm. Results showed that AOB were more abundant in the freshwater with high concentrations of ammonium (NH4+) and low dissolved oxygen, whereas AOA were dominant in the NH4+-depleted seawater and sensitive to temperature. Obvious shifts in ammonia-oxidizing communities were found along the salinity gradient in the PRE. AOB clearly presented a particle-associated nature, as evidenced by higher relative abundance of amoA genes attached to the large particles (&amp;gt;3.0 μm) and their transcripts exclusively detected on this fraction. Moreover, higher transcriptional activity (indicated by the cDNA/DNA ratio) of AOB on the large particles, suggesting AOB were actively involved in ammonia oxidation despite their lower abundance in the mid- and lower estuarine regions. In contrast, AOA exhibited higher transcriptional activity on the 0.45–3.0 μm and 0.22–0.45 μm particles, implying the free-living strategy of these microbes. Together, these findings from field observations provide useful information on the ecological strategies of ammonia-oxidizing communities in response to different environmental conditions.

Ammonia-oxidizing archaea possess a wide range of cellular ammonia affinities

Nitrification, the oxidation of ammonia to nitrate, is an essential process in the biogeochemical nitrogen cycle. The first step of nitrification, ammonia oxidation, is performed by three, often co-occurring guilds of chemolithoautotrophs: ammonia-oxidizing bacteria (AOB), archaea (AOA), and complete ammonia oxidizers (comammox). Substrate kinetics are considered to be a major niche-differentiating factor between these guilds, but few AOA strains have been kinetically characterized. Here, the ammonia oxidation kinetic properties of 12 AOA representing all major cultivated phylogenetic lineages were determined using microrespirometry. Members of the genus Nitrosocosmicus have the lowest affinity for both ammonia and total ammonium of any characterized AOA, and these values are similar to previously determined ammonia and total ammonium affinities of AOB. This contrasts previous assumptions that all AOA possess much higher substrate affinities than their comammox or AOB counterparts. The substrate affinity of ammonia oxidizers correlated with their cell surface area to volume ratios. In addition, kinetic measurements across a range of pH values supports the hypothesis that—like for AOB—ammonia and not ammonium is the substrate for the ammonia monooxygenase enzyme of AOA and comammox. Together, these data will facilitate predictions and interpretation of ammonia oxidizer community structures and provide a robust basis for establishing testable hypotheses on competition between AOB, AOA, and comammox.

Yak Excreta Application Alter Nitrification by Regulating the Ammonia-Oxidizing Bacterial Communities in Wetland Soils

Yak Excreta Application Alter Nitrification by Regulating the Ammonia-Oxidizing Bacterial Communities in Wetland Soils

Dominance of comammox Nitrospira in soil nitrification.

The first and limiting step of nitrification is catalyzed by ammonia-oxidizing archaea (AOA) and bacteria (AOB). Recently, complete ammonia oxidizers (comammox Nitrospira) have been discovered to perform complete nitrification in one cell, yet their role in soil nitrification is still unclear. This study investigated the abundance and contribution of aerobic ammonia oxidizers in typical soil habitats, and assessed the role of comammox Nitrospira in ammonia-oxidizing communities. The results showed that comammox Nitrospira were dominant in 70% of the samples and their abundance displayed a significant positive correlation with nitrification potential. The median amoA gene transcription level of comammox Nitrospira exceeded that of AOA and AOB in in-situ soils. The abundance of comammox Nitrospira was negatively correlated with soil pH, dominating in 84% of soil samples with pH<6.17. The results challenge the role of AOA and AOB in soils, highlighting the importance of comammox Nitrospira in soil nitrification, especially in acid soils. This work supports better understanding and regulation of the soil nitrogen cycle.

Robust Nitritation of Anaerobic Digester Centrate Using Dual Stressors and Timed Alkali Additions.

Nitrogen is commonly removed from wastewater by nitrification to nitrate followed by nitrate reduction to N2. Shortcut N removal saves energy by limiting ammonia oxidation to nitrite, but nitrite accumulation can be unstable. We hypothesized that repeated short-term exposures of ammonia-oxidizing communities to free ammonia (FA) and free nitrous acid (FNA) would stabilize nitritation by selecting against nitrite-oxidizing bacteria (NOB). Accordingly, we evaluated ammonium oxidation of anaerobic digester centrate in two bench-scale sequencing batch reactors (SBRs), seeded with the same inoculum and operated identically but with differing pH-control strategies. A single stressor SBR (SS/SBR) using pH set-point control produced HNO3, while a dual stressor SBR (DS/SBR) using timed alkalinity addition (TAA) produced HNO2 (ammonium removal efficiency of 97 ± 2%; nitrite accumulation ratio of 98 ± 1%). The TAA protocol was developed during an adaptation period with continuous pH monitoring. After adaptation, automated TAA enabled stable nitritation without set-point control. In the SS/SBR, repeatedly exposing the community to FA (8-10 h/exposure, one exposure/cycle) selected for FA-tolerant ammonia-oxidizing bacteria (Nitrosomonas sp. NM107) and NOB (Nitrobacter sp.). In the DS/SBR, repeatedly exposing the community to FA (2-4 h/exposure, three exposures/cycle) and FNA (4-6 h/exposure, two exposures/cycle) selected for FA- and FNA-resistant AOB (Nitrosomonas IWT514) and against NOB, stabilizing nitritation.

Comammox Nitrospira Species Dominate in an Efficient Partial Nitrification-Anammox Bioreactor for Treating Ammonium at Low Loadings.

Bacteria capable of complete ammonia oxidation (comammox) are widespread and contribute to nitrification in wastewater treatment facilities. However, their roles in partial nitrification-anaerobic ammonium oxidation (anammox) systems remain unclear. In this study, a bench-scale bioreactor with continuous stirring was operated for more than 1000 days with limited oxygen supply to achieve efficient nitrogen removal (70.1 ± 2.7%) at a low ammonium loading of 35.2 mg-N/L/day. High-throughput amplicon sequencing analysis of the comammox ammonia monooxygenase subunit A (amoA) gene revealed seven sequence types from two clusters in clade A of comammox Nitrospira. Quantitative polymerase chain reaction analyses suggested that the comammox species dominated the ammonia-oxidizing community, with an abundance as high as 89.2 ± 7.9% in total prokaryotic amoA copies. Multiple linear regression further revealed the substantial contribution of the comammox Nitrospira to ammonia oxidation in the bioreactor. The investigation with bioreactor and batch experiments consistently showed that activities of comammox Nitrospira were inhibited by free ammonia far more severely than other ammonia-oxidizing microbes. Overall, this study provided new insight into the ecology of comammox Nitrospira under hypoxic conditions and suggested comammox-associated partial nitrification-anammox as a potential method for treating low-strength ammonium-containing wastewater.

Mitigating N2O emission by synthetic inhibitors mixed with urea and cattle manure application via inhibiting ammonia-oxidizing bacteria, but not archaea, in a calcareous soil.

Synthetic inhibitors and organic amendment have been proposed for mitigating greenhouse gas N2O emissions. However, their combined effect on the N2O emissions and ammonia-oxidizer (ammonia-oxidizing bacteria and archaea, AOB and AOA) communities remains unclear in calcareous soils under climate warming. We conducted two incubation experiments (25 and 35°C) to examine how N2O emissions and AOA and AOB communities responded to organic amendment (urea plus cattle manure, UCM), and in combination with urease (N-(n-butyl) thiophosphoric triamide, NBPT) and nitrification inhibitor (nitrapyrin). The treatments of UCM+nitrapyrin and UCM+nitrapyrin+NBPT significantly lowered total N2O emissions by average 64.5 and 71.05% at 25 and 35°C, respectively, compared with UCM treatment. AOB gene abundance and α-diversity (Chao1 and Shannon indices) were significantly increased by the application of urea and manure (P<0.05). However, relative to UCM treatment, nitrapyrin addition treatments decreased AOB gene abundance and Chao 1 index by average 115.4 and 30.4% at 25 and 35°C, respectively. PCA analysis showed that UCM or UCM plus nitrapyrin notably shifted AOB structure at both temperatures. However, fertilization had little effects on AOA community (P>0.05). Potential nitrification rate (PNR) was greatly decreased by nitrapyrin addition, and PNR significantly positively correlated with AOB gene abundance (P=0.0179at 25°C and P=0.0029at 35°C) rather than AOA (P>0.05). Structural equation model analysis showed that temperature directly increased AOA abundance but decrease AOB abundance, while fertilization indirectly influenced AOB community by altering soil NH4+, pH and SOC. In conclusion, the combined application of organic amendment, NBPT and nitrapyrin significantly lowered N2O emissions via reducing AOB community in calcareous soil even at high temperature. Our findings provide a solid theoretical basis in mitigating N2O emissions from calcareous soil under climate warming.

Exploring the communities of bacteria, fungi and ammonia oxidizers in rhizosphere of Fusarium-diseased greenhouse cucumber

Long-term greenhouse vegetable cultivation commonly incurs soil-borne diseases, which seriously threaten the sustainability of greenhouse production system. Rhizomicrobiome plays an important role in ensuring plant health and driving soil nutrient cycling. However, how rhizomicrobiome respond to the occurrence of soil-borne disease is still not completely clear. Increasing knowledge involved would benefit to the prevention of soil-borne disease in intensive greenhouse production by providing the insight into soil microbial ecology. Herein, with the help of a continuously cultivated greenhouse cucumber field, soil physicochemical and biochemical properties and the abundances (quantitative real-time PCR) and communities (high-throughput amplicon sequencing) of bacteria, fungi, ammonia-oxidizing archaea (AOA) and bacteria (AOB) in the rhizosphere of healthy and Fusarium-diseased cucumbers were investigated. Diseased cucumber rhizosphere had significantly higher available nutrient contents and acidity compared to healthy cucumber rhizosphere, but significantly lower microbial activity and nitrification capacity. Although there was no significant difference in the abundances of bacteria, fungi, AOA and AOB, diseased cucumber rhizosphere had more abundant Fusarium oxysporum. For bacteria, only community richness in diseased cucumber rhizosphere was significantly lower than that in healthy cucumber rhizosphere, whereas for fungi both community richness and Shannon diversity were significantly lower. Community structures of bacteria, fungi, AOA and AOB all significantly differed between healthy and diseased cucumber rhizosphere. Particularly, healthy cucumber rhizosphere had more abundant beneficial microbes (Bacillus and Penicillium) and putative biocontrol microbes (Mycothermus) as well as Nitrosospira briensis Clade-related AOB. These results revealed significantly different rhizomicrobiome between Fusarium wilt diseased and healthy cucumbers under long-term intensive greenhouse cultivation, and suggested that rhizomicrobiome may play an important role in the occurrence of Fusarium wilt disease of cucumber. The findings obtained here also provided potential directions towards the prevention of soil-borne diseases and the improvement of N turnover in diseased soil by manipulating rhizosphere microbiome.

The Response of Estuarine Ammonia-Oxidizing Communities to Constant and Fluctuating Salinity Regimes.

Aerobic nitrification is a fundamental nitrogen biogeochemical process that links the oxidation of ammonia to the removal of fixed nitrogen in eutrophicated water bodies. However, in estuarine environments there is an enormous variability of water physicochemical parameters that can affect the ammonia oxidation biological process. For instance, it is known that salinity can affect nitrification performance, yet there is still a lack of information on the ammonia-oxidizing communities behavior facing daily salinity fluctuations. In this work, laboratory experiments using upstream and downstream estuarine sediments were performed to address this missing gap by comparing the effect of daily salinity fluctuations with constant salinity on the activity and diversity of ammonia-oxidizing microorganisms (AOM). Activity and composition of AOM were assessed, respectively by using nitrogen stable isotope technique and 16S rRNA gene metabarcoding analysis. Nitrification activity was negatively affected by daily salinity fluctuations in upstream sediments while no effect was observed in downstream sediments. Constant salinity regime showed clearly higher rates of nitrification in upstream sediments while a similar nitrification performance between the two salinity regimes was registered in the downstream sediments. Results also indicated that daily salinity fluctuation regime had a negative effect on both ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA) community’s diversity. Phylogenetically, the estuarine downstream AOM were dominated by AOA (0.92–2.09%) followed by NOB (0.99–2%), and then AOB (0.2–0.32%); whereas NOB dominated estuarine upstream sediment samples (1.4–9.5%), followed by AOA (0.27–0.51%) and AOB (0.01–0.23%). Analysis of variance identified the spatial difference between samples (downstream and upstream) as the main drivers of AOA and AOB diversity. Our study indicates that benthic AOM inhabiting different estuarine sites presented distinct plasticity toward the salinity regimes tested. These findings help to improve our understanding in the dynamics of the nitrogen cycle of estuarine systems by showing the resilience and consequently the impact of different salinity regimes on the diversity and activity of ammonia oxidizer communities.

Surface ammonium loading rate shifts ammonia-oxidizing communities in surface water-fed rapid sand filters.

Nitrification is important in drinking water treatment plants (DWTPs) for ammonia removal and is widely considered as a stepwise process mediated by ammonia- and nitrite-oxidizing microorganisms. The recent discovery of complete ammonia oxidizers (comammox) has challenged the long-held assumption that the division of metabolic labor in nitrification is obligate. However, little is known about the role of comammox Nitrospira in DWTPs. Here, we explored the relative importance of comammox Nitrospira, canonical ammonia-oxidizing archaea (AOA) and bacteria (AOB) in 12 surface water-fed rapid sand filters (RSFs). Quantitative PCR results showed that all the three ammonia-oxidizing guilds had the potential to dominate nitrification in DWTPs. Spearman's correlation and redundancy analysis revealed that the surface ammonium loading rate (SLR) was the key environmental factor influencing ammonia-oxidizing communities. Comammox Nitrospira were likely to dominate the nitrification under a higher SLR. PCR and phylogenetic analysis indicated that most comammox Nitrospira belonged to clade A, with clade B comammox Nitrospira almost absent. This work reveals obvious differences in ammonia-oxidizing communities between surface water-fed and groundwater-fed RSFs. The presence of comammox Nitrospira can support the stability of drinking water production systems under high SLR and warrants further investigation of their impact on drinking water quality.

Substrate regulation leads to differential responses of microbial ammonia-oxidizing communities to ocean warming

In the context of continuously increasing anthropogenic nitrogen inputs, knowledge of how ammonia oxidation (AO) in the ocean responds to warming is crucial to predicting future changes in marine nitrogen biogeochemistry. Here, we show divergent thermal response patterns for marine AO across a wide onshore/offshore trophic gradient. We find ammonia oxidizer community and ambient substrate co-regulate optimum temperatures (Topt), generating distinct thermal response patterns with Topt varying from ≤14 °C to ≥34 °C. Substrate addition elevates Topt when ambient substrate is unsaturated. The thermal sensitivity of kinetic parameters allows us to predict responses of both AO rate and Topt at varying substrate and temperature below the critical temperature. A warming ocean promotes nearshore AO, while suppressing offshore AO. Our findings reconcile field inconsistencies of temperature effects on AO, suggesting that predictive biogeochemical models need to include such differential warming mechanisms on this key nitrogen cycle process.

Hurricane Disturbance Stimulated Nitrification and Altered Ammonia Oxidizer Community Structure in Lake Okeechobee and St. Lucie Estuary (Florida).

Nitrification is an important biological link between oxidized and reduced forms of nitrogen (N). The efficiency of nitrification plays a key role in mitigating excess N in eutrophic systems, including those with cyanobacterial harmful algal blooms (cyanoHABs), since it can be closely coupled with denitrification and removal of excess N. Recent work suggests that competition for ammonium (NH4+) between ammonia oxidizers and cyanoHABs can help determine microbial community structure. Nitrification rates and ammonia-oxidizing archaeal (AOA) and bacterial (AOB) community composition and gene abundances were quantified in Lake Okeechobee and St. Lucie Estuary in southern Florida (United States). We sampled during cyanobacterial (Microcystis) blooms in July 2016 and August 2017 (2 weeks before Hurricane Irma) and 10 days after Hurricane Irma made landfall. Nitrification rates were low during cyanobacterial blooms in Lake Okeechobee and St. Lucie Estuary, while low bloom conditions in St. Lucie Estuary coincided with greater nitrification rates. Nitrification rates in the lake were correlated (R2 = 0.94; p = 0.006) with AOA amoA abundance. Following the hurricane, nitrification rates increased by an order of magnitude, suggesting that nitrifiers outcompeted cyanobacteria for NH4+ under turbid, poor light conditions. After Irma, AOA and AOB abundances increased in St. Lucie Estuary, while only AOB increased in Lake Okeechobee. AOA sequences clustered into three major lineages: Nitrosopumilales (NP), Nitrososphaerales (NS), and Nitrosotaleales (NT). Many of the lake OTUs placed within the uncultured and uncharacterized NS δ and NT β clades, suggesting that these taxa are ecologically important along this eutrophic, lacustrine to estuarine continuum. After the hurricane, the AOA community shifted toward dominance by freshwater clades in St. Lucie Estuary and terrestrial genera in Lake Okeechobee, likely due to high rainfall and subsequent increased turbidity and freshwater loading from the lake into the estuary. AOB community structure was not affected by the disturbance. AOA communities were consistently more diverse than AOB, despite fewer sequences recovered, including new, unclassified, eutrophic ecotypes, suggesting a wider ecological biogeography than the oligotrophic niche originally posited. These results and other recent reports contradict the early hypothesis that AOB dominate ammonia oxidation in high-nutrient or terrestrial-influenced systems.

Land use pattern effects after 30 years of shifting cropland to fallow land on soil ammonia-oxidizer community

Sympatric differentiation of ammonia-oxidizing archaea and bacteria may occur due to differences in ammonia availability during agricultural activities such as fertilization, which shift key biological drivers of soil nitrification processes. Restoring native habitat from intensive agricultural activities (e.g. converting cropland into fallow land) reduces fertilizer inputs, which has the potential to change active ammonia-oxidizers and thus nitrification rates. We incubated soils from fields under 30 years fallow and under continuous cropping to understand shifts in ammonia-oxidizers and nitrification processes. Thirty years of land fallow practices sharply decreased the abundance and activity of soil ammonia-oxidizing bacteria compared to continuous cropping. Nitrification in cropland was driven mainly by ammonia-oxidizing bacteria while ammonia-oxidizing archaea were the main drivers of nitrification in fallow soil. Conversion of cropland into fallow land shifted the dominant active ammonia-oxidizers from ammonia-oxidizing bacteria to archaea, and the low nitrification rates in fallow soil were caused by the lack of active ammonia-oxidizing bacteria rather than N substrate availability.

Environmental Factors, More Than Spatial Distance, Explain Community Structure of Soil Ammonia-Oxidizers in Wetlands on the Qinghai-Tibetan Plateau.

In wetland ecosystems, ammonia oxidation highly depends on the activity of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB), which are, therefore, important for studying nitrogen cycling. However, the ammonia-oxidizer communities in the typical high-elevation wetlands are poorly understood. Here, we examined ammonia-oxidizer communities in soils from three wetland types and 31 wetland sites across the Qinghai–Tibetan Plateau. The amoA gene of AOA and AOB was widespread across all wetland types. Nitrososphaera clade (Group I.1b) overwhelmingly dominated in AOA community (90.36%), while Nitrosospira was the principal AOB type (64.96%). The average abundances of AOA and AOB were 2.63 × 104 copies g−1 and 9.73 × 103 copies g−1. The abundance of AOA amoA gene was higher in riverine and lacustrine wetlands, while AOB amoA gene dominated in palustrine wetlands. The environmental conditions, but not spatial distance, have a dominant role in shaping the pattern of ammonia-oxidizer communities. The AOA community composition was influenced by mean annual temperature (MAT) and mean annual precipitation (MAP), while MAT, conductivity and plant richness, pH, and TN influenced the AOB community composition. The net nitrification rate had a significant correlation to AOB, but not AOA abundance. Our results suggest a dominant role for climate factors (MAT and MAP) in shaping community composition across a wide variety of wetland sites and conditions.

Differential Ecosystem Function Stability of Ammonia-Oxidizing Archaea and Bacteria following Short-Term Environmental Perturbation.

Rapidly expanding conversion of tropical forests to oil palm plantations in Southeast Asia leads to soil acidification following intensive nitrogen fertilization. Changes in soil pH are predicted to have an impact on archaeal ammonia-oxidizing archaea (AOA), ammonia-oxidizing bacteria (AOB), and complete (comammox) ammonia oxidizers and, consequently, on nitrification. It is therefore critical to determine whether the predicted effects of pH on ammonia oxidizers and nitrification activity apply in tropical soils subjected to various degrees of anthropogenic activity. This was investigated by experimental manipulation of pH in soil microcosms from a land-use gradient (forest, riparian, and oil palm soils). The nitrification rate was greater in forest soils with native neutral pH than in converted acidic oil palm soils. Ammonia oxidizer activity decreased following acidification of the forest soils but increased after liming of the oil palm soils, leading to a trend of a reversed net nitrification rate after pH modification. AOA and AOB nitrification activity was dependent on pH, but AOB were more sensitive to pH modification than AOA, which demonstrates a greater stability of AOA than AOB under conditions of short-term perturbation. In addition, these results predict AOB to be a good bioindicator of nitrification response following pH perturbation during land-use conversion. AOB and/or comammox species were active in all soils along the land-use gradient, even, unexpectedly, under acidic conditions, suggesting their adaptation to native acidic or acidified soils. The present study therefore provided evidence for limited stability of soil ammonia oxidizer activity following intensive anthropogenic activities, which likely aggravates the vulnerability of nitrogen cycle processes to environmental disturbance.IMPORTANCE Physiological and ecological studies have provided evidence for pH-driven niche specialization of ammonia oxidizers in terrestrial ecosystems. However, the functional stability of ammonia oxidizers following pH change has not been investigated, despite its importance in understanding the maintenance of ecosystem processes following environmental perturbation. This is particularly true after anthropogenic perturbation, such as the conversion of tropical forest to oil palm plantations. This study demonstrated a great impact of land-use conversion on nitrification, which is linked to changes in soil pH due to common agricultural practices (intensive fertilization). In addition, the different communities of ammonia oxidizers were differently affected by short-term pH perturbations, with implications for future land-use conversions but also for increased knowledge of associated global nitrous oxide emissions and current climate change concerns.