Ammonia-oxidizing Communities Research Articles

Diverse Thaumarchaeota Dominate Subsurface Ammonia-oxidizing Communities in Semi-arid Floodplains in the Western United States.

Subsurface microbial communities mediate biogeochemical transformations that drive both local and ecosystem-level cycling of essential elements, including nitrogen. However, their study has been largely limited to the deep ocean, terrestrial mines, caves, and topsoils (<30cm). Here, we present regional insights into the microbial ecology of aerobic ammonia oxidation within the terrestrial subsurface of five semi-arid riparian sites spanning a 900-km N-S transect. We sampled sediments, profiled communities to depths of ≤ 10m, and compared them to reveal trends regionally within and surrounding the Upper Colorado River Basin (CRB). The diversity and abundance of ammonia-oxidizing microbial communities were evaluated in the context of subsurface geochemistry by applying a combination of amoA (encoding ammonia monooxygenase subunit A) gene sequencing, quantitative PCR, and geochemical techniques. Analysis of 898 amoA sequences from ammonia-oxidizing archaea (AOA) and bacteria (AOB) revealed extensive ecosystem-scale diversity, including archaeal amoA sequences from four of the five major AOA lineages currently found worldwide as well as distinct AOA ecotypes associated with naturally reduced zones (NRZs) and hydrogeochemical zones (unsaturated, capillary fringe, and saturated). Overall, AOA outnumber AOB by 2- to 5000-fold over this regional scale, suggesting that AOA may play a prominent biogeochemical role in nitrification within terrestrial subsurface sediments.

Understanding the Response of Nitrifying Communities to Disturbance in the McMurdo Dry Valleys, Antarctica.

Polar ecosystems are generally limited in nitrogen (N) nutrients, and the patchy availability of N is partly determined by biological pathways, such as nitrification, which are carried out by distinctive prokaryotic functional groups. The activity and diversity of microorganisms are generally strongly influenced by environmental conditions. However, we know little of the attributes that control the distribution and activity of specific microbial functional groups, such as nitrifiers, in extreme cold environments and how they may respond to change. To ascertain relationships between soil geochemistry and the ecology of nitrifying microbial communities, we carried out a laboratory-based manipulative experiment to test the selective effect of key geochemical variables on the activity and abundance of ammonia-oxidizing communities in soils from the McMurdo Dry Valleys of Antarctica. We hypothesized that nitrifying communities, adapted to different environmental conditions within the Dry Valleys, will have distinct responses when submitted to similar geochemical disturbances. In order to test this hypothesis, soils from two geographically distant and geochemically divergent locations, Miers and Beacon Valleys, were incubated over 2 months under increased conductivity, ammonia concentration, copper concentration, and organic matter content. Amplicon sequencing of the 16S rRNA gene and transcripts allowed comparison of the response of ammonia-oxidizing Archaea (AOA) and ammonia-oxidizing Bacteria (AOB) to each treatment over time. This approach was combined with measurements of 15NH4+ oxidation rates using 15N isotopic additions. Our results showed a higher potential for nitrification in Miers Valley, where environmental conditions are milder relative to Beacon Valley. AOA exhibited better adaptability to geochemical changes compared to AOB, particularly to the increase in copper and conductivity. AOA were also the only nitrifying group found in Beacon Valley soils. This laboratorial manipulative experiment provided new knowledge on how nitrifying groups respond to changes on key geochemical variables of Antarctic desert soils, and we believe these results offer new insights on the dynamics of N cycling in these ecosystems.

Heavy nitrogen application increases soil nitrification through ammonia-oxidizing bacteria rather than archaea in acidic tea (Camellia sinensis L.) plantation soil

Nitrogen (N) fertilizer is widely used in agricultural ecosystems and influences N transformation processes in the soil such as nitrification. However, whether nitrification is primarily dominated by ammonia-oxidizing bacteria (AOB) or archaea (AOA) under heavy N application is still under debate. In the present work, the effect of long-term (12 years) N fertilization on soil nitrification and the key influencing factors were investigated in acidic tea plantation soil that received four different rates of N application (0, 119, 285, and 569 kg N ha−1 yr−1). Nitrification potential was measured and partitioned using chemical inhibitors. The abundance of functional genes involved in ammonia oxidation was quantified using quantitative polymerase chain reaction (qPCR). Ammonia-oxidizing communities were identified by shotgun metagenome sequencing. Potential nitrification rate in tea plantation soil was mainly dominated by autotrophic nitrification (PNRA) (71–79%). PNRA and heterotrophic nitrification (PNRH) were both significantly increased by heavy N (569 kg ha−1) application. Moreover, PNRA was mainly due to the contribution of AOB (52–66%) in N-treated soils, and N569 significantly increased the AOB contribution without affecting the AOA contribution. N569 increased the functional gene abundance of AOB and TAO100 (a non-halophilic γ-AOB) but decreased that of AOA. The dominant AOB (Nitrosomonas, Nitrosospira, and Nitrosococcus), AOA (Nitrososphaera and Nitrosopumilus) and commamox (Nitrospira) groups were profoundly altered by long-term N application rates. Partial least squares regression showed that total nitrification (PNRT), PNRA, and PNRAOB were primarily explained by the functional gene abundance of nitrifiers whereas PNRH and PNRAOA were closely associated with soil and pruned litter properties. Moreover, structural equation modeling (SEM) revealed that long-term N application significantly and indirectly affected nitrification potential by directly influencing soil properties, pruned litter properties, and functional gene abundance. Understanding the relative contribution of AOA and AOB to nitrification may help to better regulate N fertilizer use in agricultural ecosystems.

Ammonia oxidizers in river sediments of the Qinghai-Tibet Plateau and their adaptations to high-elevation conditions

Ammonia-oxidizing bacteria (AOB) and archaea (AOA) as well as complete ammonia oxidizers (comammox) aerobically catalyze ammonia oxidation which plays essential roles in riverine nitrogen cycle. However, performances of these ammonia oxidizers in high-elevation river sediments have rarely been documented. This study investigated the abundance, community, and activity of ammonia oxidizers in five high-elevation rivers of the Qinghai-Tibet Plateau (QTP). Comammox were dominant ammonia oxidizers in 23% of studied samples and the clade B was principal comammox type. amoA gene abundances of AOA and AOB in these high-elevation rivers were comparable to those in low-elevation rivers. However, in contrast to most studied low-elevation rivers, AOB amoA gene abundance outnumbered AOA in 92% samples, which might be caused by the lower temperature and more intense solar radiation of the QTP. Potential nitrification rates (PNRs) ranged from 0.02 to 2.95 nmol-N h−1 g−1 dry sediment. Ammonia concentration was the limiting factor to PNRs at some sites, and when ammonia was not limiting, the PNR: ammonia ratio was greater at higher temperatures. There was no apparent variation in ammonia oxidizer community compositions along the elevation gradient due to the high elevation (2687 to 4223 m) of our entire study area. However, compared with low-elevation rivers, the lower temperature, huge diurnal temperature change, and lower nutrient conditions in the QTP rivers shaped distinctive communities for ammonia oxidizers; the unique community characteristics were significantly correlated to PNRs. These results suggest that ammonia oxidizers in the five high-elevation rivers have adapted to high-elevation conditions; more research should be conducted to study their adaptation mechanisms and their roles in riverine nitrogen cycle.

Mineralization and nitrification: Archaea dominate ammonia-oxidising communities in grassland soils

In grasslands, N mineralization and nitrification are important processes and are controlled by several factors, including the in situ microbial community composition. Nitrification involves ammonia oxidising archaea (AOA) and bacteria (AOB) and although AOA and AOB co-exist in soils, they respond differently to environmental characteristics and there is evidence of AOA/AOB niche differentiation. Here, we investigated temporal variation in N mineralization and nitrification rates, together with bacterial, archaeal and ammonia-oxidiser communities in grassland soils, on different geologies: clay, Greensand and Chalk. Across geologies, N mineralization and nitrification rates were slower in the autumn than the rest of the year. Turnover times for soil ammonium pools were <24 h, whilst several days for nitrate. In clay soils, bacterial, archaeal, AOA, and AOB communities were clearly distinct from those in Chalk and Greensand soils. Spatially and temporally, AOA were more abundant than AOB. Notably, Nitrososphaera were predominant, comprising 37.4% of archaeal communities, with the vast majority of AOA found in Chalk and Greensand soils. AOA abundance positively correlated with nitrate concentration, whereas AOB abundance correlated with ammonium and nitrite concentrations, suggesting that these N compounds may be potential drivers for AOA/AOB niche differentiation in these grassland soils.

Effects of environmental variables on abundance of ammonia-oxidizing communities in sediments of Luotian River, China.

Ammonia-oxidizing communities play important functional roles in the nitrification. However, environmental stresses can significantly affect this process by controlling the abundant communities of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) communities. In this study, we examined the abundance variations of ammonia-oxidizing communities using quantitative polymerase chain reaction (qPCR) and terminal-restriction fragment length polymorphism (T-RFLP) in a typical subtropical river, Luotian County, South Dabie Mountains, China. Clone libraries were conducted to evaluate the community structure and abundance of AOA and AOB in sediments. Results showed that Nitrososphaera sp and Nitrosopumilus sp were the most dominant AOA. The abundance of the AOA and AOB amoA gene ranged from 5.28 × 108 gene copies (g-soil−1) to 2.23 × 108 gene copies (g-soil−1) and 5.45 × 108 gene copies (g-soil−1) to 3.30 × 107 gene copies (g-soil−1), respectively. Five environmental variables, namely, ORP, DO, NOn}{}{}_{3}^{-}, Temp, and NHn}{}{}_{4}^{+} were played a major function in microbial communities of AOA and AOB in sediments. The T-RFLP profiles of AOA showed that 488 and 116 bp T-RFs were dominated. Overall, the results of this study showed that anthropogenic activities andenvironmental stress in rivers can alter the structure and function of microbes in their variable environment.

Vegetation-Dependent Response to Drought in Salt Marsh Ammonia-Oxidizer Communities.

We investigated the impacts of drought on ammonia-oxidizing archaea (AOA) and bacteria (AOB) in a salt marsh and compared the response to the total bacterial community. We analyzed abundance and community composition of amoA genes by QPCR and TRFLP, respectively, in three vegetation zones in 2014 (pre-drought), 2016 (drought), and 2017 (post-drought), and analyzed bacterial 16S rRNA genes by QPCR, TRFLP, and MiSeq analyses. AOA and AOB abundance in the Spartina patens zone increased significantly in 2016, while abundance decreased in the tall S. alterniflora zone, and showed little change in the short S. alterniflora zone. Total bacterial abundance declined annually in all vegetation zones. Significant shifts in community composition were detected in 2016 in two of the three vegetation zones for AOA and AOB, and in all three vegetation zones for total bacteria. Abundance and community composition of AOA and AOB returned to pre-drought conditions by 2017, while bacterial abundance continued to decline, suggesting that nitrifiers may be more resilient to drought than other bacterial communities. Finding vegetation-specific drought responses among N-cycling microbes may have broad implications for changes in N availability and marsh productivity, particularly if vegetation patterns continue to shift as predicted due to sea level rise.

Different ammonia oxidizers are responsible for nitrification in two neutral paddy soils

Human intervention in agriculture, such as farming and fertilization, could result in major changes of ammonia availability that shapes niche-specific occupied communities of soil ammonia oxidizers. The aim of this study was to investigate nitrification and active ammonia oxidizers in N-rich and N-limited soils, which were induced by different management practices. Here, we used DNA-based stable isotope probing (DNA-SIP) microcosm to decipher active ammonia-oxidizing archaea (AOA) and bacteria (AOB) phylotypes involved in ammonia oxidation in the cultivated and uncultivated soils. The results showed that net nitrification rate in the cultivated soil was significantly higher than in the uncultivated soil (6.19 and 1.40 mg N kg−1dry soil d-1, respectively). Growth of soil AOB and AOA during incubation occurred in the cultivated soil, while in the uncultivated soil, only AOA significantly increased after 56 days. Combining DNA-SIP and sequencing results, we gathered evidence of ammonia oxidation by Nitrososphaera-like AOA, in addition to active AOB, within the classic Nitrosospira group in the cultivated soil. For the uncultivated soil, ammonia oxidation was driven by Nitrososphaera-like and Nitrosopumilus-like AOA, rather than AOB. Our results demonstrate the potential of ammonium concentration for shaping the communities of active AOA and AOB, which imply that ammonia-dependent “diversification” may be important strategies for niche differentiation of sympatric AOA/AOB under different field management conditions.

Leaf leachates have the potential to influence soil nitrification via changes in ammonia‐oxidizing archaea and bacteria populations

AbstractSoil nitrogen (N) transformation is mainly controlled by microorganisms. Different plant species have specific effects on soil ammonia‐oxidizing archaea (AOA) and bacteria (AOB). Furthermore, plant secondary metabolites have dramatic influences on soil N transformation and soil ammonia‐oxidizers. However, no study has directly linked the changes in soil AOA and AOB communities to N transformation due to different plant species' leaf leachates, including comparing invasive versus native plants. We selected three invasive species, Wedelia trilobata, Ipomoea cairica and Mikania micrantha, and two native species, Wedelia chinensis and Merremia hederacea, and incubated soil with high and low concentrations of leaf leachates from the five species. Soil N transformation and ammonia oxidizer communities (based on quantitative PCR and 16S rRNA high‐throughput sequencing) were determined. Leaf leachates significantly affected soil N transformation and soil AOA and AOB abundance, and the effects were dependent on the plant species and leachate concentration. The leachate of W. chinensis increased soil net nitrification rates (NNR), whereas that of M. micrantha increased soil nitrate (NO3−) and NNR. The leachate of M. micrantha increased AOB amoA genes, whereas that of M. hederacea decreased AOA amoA genes. At the higher concentration, all leachates showed inhibitory effects on the relative abundance of all AOA taxonomic groups. Phenolics, flavonoids and organic carbon in leaf leachates had a negative correlation with soil NO3−, NNR and the amoA genes of AOA and AOB, whereas NH4+ and NO3− in leachates showed a positive correlation. Among the five species, the invasive species M. micrantha had the strongest positive effect on soil N transformation and AOB abundance. The altered soil ammonia oxidizer communities and modified N process rates induced by leaf leachates provide an explanatory mechanism for the differential effects of plant species on N cycling.Highlights Link changes in soil AOA and AOB to N cycling due to leaf leachates. Specific plants can dramatically alter soil ammonia‐oxidizer presence and abundance. Effects of invasive plants on soil N and AOB are mainly driven by M. micrantha. High concentrations of plant leachates reduced the relative abundance of AOA groups.

Effects of elevated temperature and elevated CO2 on soil nitrification and ammonia-oxidizing microbial communities in field-grown crop

Rising global air temperature and atmospheric CO2 are expected to have considerable effects on soil nutrient cycling and plant productivity. Soil nitrification controlled by ammonia-oxidizing bacteria and archaea (AOB and AOA) communities plays a key role in contributing to plant nitrogen (N) availability; however, response of soil nitrification and functional microbial communities to climate change and subsequent consequences for crop yields remain largely unknown. Cotton productivity is a function of temperature and N availability under well-watered conditions. In general, cotton growth responds positively to elevated CO2, but simultaneous warming may offset benefits of rising CO2. In this study, cotton was used as a model system to elucidate the short-term response of soil nitrification and ammonia-oxidizing communities to elevated temperature and elevated CO2 using field-based environmentally-controlled chambers. Elevated temperature (ambient + 1.1 °C) altered the AOA community, while elevated temperature and elevated CO2 (ambient + 132 ppm) significantly increased soil nitrification rate and shifted AOB and AOA communities, but these effects depended on cotton developmental stages. Ammonia-oxidizing community abundance and structure were statistically correlated with nitrifying activity. Our findings suggest that climate change will positively affect soil nitrifying communities, leading to an increase in process rates and subsequent N availability, which is directly linked to crop productivity.

Community shift of microbial ammonia oxidizers in air-dried rice soils after 22 years of nitrogen fertilization

In this study, we show that composition shifts of ammonia oxidizer communities imposed by a 22-year field fertilization regime could be well retained in both fresh and air-dried soils. The abundance and composition of ammonia-oxidizing bacteria (AOB) and archaea (AOA) were measured in fresh soils, which received no fertilization (CK), chemical fertilization (NPK), and chemical plus organic matter fertilization (NPK/OM) for 22 years. The air-drying treatment of fresh soil was also conducted for pairwise analysis. We found that in fresh soils DGGE fingerprints of AOB showed significant changes under both NPK and NPK/OM treatments when compared with control (CK) and that microbial shift was almost identical in air-dried soils. Long-term nutrient fertilization did not affect AOA communities in either air-dried or fresh soils. Compared to CK treatment, real-time PCR indicated that AOB amoA genes increased significantly in fresh soils of NPK (59-fold) and NPK/OM (48-fold) plots and in air-dried NPK and NPK/OM soils by 22-fold and 19-fold respectively. Our results demonstrate that community shifts of AOB in fresh soils under chronic N fertilization could be well preserved in air-dried soils, despite the apparent decline in absolute abundance of ammonia oxidizers. These results suggest that air-dried soil could be a useful resource for deciphering the adaptive strategy of ammonia oxidizers under N enrichment when the significant changes of community composition occurred in fresh soils.

Purple Sulfur Bacteria Dominate Microbial Community in Brazilian Limestone Cave.

The mineralogical composition of caves makes the environment ideal for inhabitation by microbes. However, the bacterial diversity in the cave ecosystem remains largely unexplored. In this paper, we described the bacterial community in an oxic chamber of the Sopradeira cave, an iron-rich limestone cave, in the semiarid region of Northeast Brazil. The microbial population in the cave samples was studied by 16S rDNA next-generation sequencing. A type of purple sulfur bacteria (PSB), Chromatiales, was found to be the most abundant in the sediment (57%), gravel-like (73%), and rock samples (96%). The predominant PSB detected were Ectothiorhodospiraceae, Chromatiaceae, and Woeseiaceae. We identified the PSB in a permanently aphotic zone, with no sulfur detected by energy-dispersive X-ray (EDX) spectroscopy. The absence of light prompted us to investigate for possible nitrogen fixing (nifH) and ammonia oxidizing (amoA) genes in the microbial samples. The nifH gene was found to be present in higher copy numbers than the bacterial-amoA and archaeal-amoA genes, and archaeal-amoA dominated the ammonia-oxidizing community. Although PSB dominated the bacterial community in the samples and may be related to both nitrogen-fixing and ammonia oxidizing bacteria, nitrogen-fixing associated gene was the most detected in those samples, especially in the rock. The present work demonstrates that this cave is an interesting hotspot for the study of ammonia-oxidizing archaea and aphotic PSB.

Impacts of Cu and sulfadiazine on soil potential nitrification and diversity of ammonia-oxidizing archaea and bacteria

ABSTRACTSoil potential nitrification rate (PNR) and diversity of ammonia-oxidizing microbes were investigated in spiked soils with Cu and Sulfadiazine (SDZ). An obvious decrease of PNR was observed with the increase of Cu and SDZ concentrations in the soil. Real-time fluorescence quota PCR result showed that AOA and AOB were slightly stimulated at the gene level in both contaminated soils. Sequential analysis indicated that 200 mg kg−1 Cu could improve AOA diversity but reduce AOB diversity, but 5 mg kg−1 SDZ caused a decrease of both AOA and AOB diversity. Microbial community’s analysis also found that 200 mg kg−1 Cu and 5 mg kg−1 SDZ had different influence on the populations of AOA and AOB. It could be concluded that Cu and SDZ might have a different ecological effect mechanism on soil potential nitrification and ammonia-oxidizing microbial communities.

Variations of Abundance and Community Structure of Ammonia Oxidizers and Nitrification Activity in Two Paddy Soils Polluted by Heavy Metals

While microbial nitrogen transformations are sensitive indicators of trace metal toxicity in soils, studies that quantify the impacts of heavy metal pollution in polluted rice soils on microbial communities and their activities remain limited. We examined changes in the abundance, composition and activity of ammonia oxidizing communities in two paddy fields that have been polluted by metal mining and smelting activities for more than three decades. The results showed a shift in the community structure of ammonia oxidizing archaea (AOA) and, to a lesser extent, of ammonia oxidizing bacteria (AOB) under metal pollution in the soils. All the retrieved AOB sequences in this study belonged to the genus Nitrosospira. Among them, the species in Cluster 3 a.1 seemed to be more sensitive to heavy metal pollution. Both AOB abundance and nitrification activity were not affected by heavy metal pollution in the two sites; whereas, AOA abundance increased. Our results suggested an effect of metal pollutants on communities of ammonia oxidizers, the degree of which varied in accordance with the amount of metal pollution. Therefore, it is difficult to quantify the relationship between the AOB/AOA communities and nitrification activity in the polluted soil.

The efficacy of 3,4-dimethylpyrazole phosphate on N2O emissions is linked to niche differentiation of ammonia oxidizing archaea and bacteria across four arable soils

The nitrification inhibitor, 3,4-dimethylpyrazole phosphate (DMPP), can be used to reduce N2O emissions from agricultural ecosystems. However, the effectiveness of DMPP varies among soils, and this is due to both abiotic (e.g. soil properties) and biotic factors (e.g. ammonia oxidizers and denitrifier communities). Understanding the nature of these effects is necessary to improve the efficacy of DMPP, therefore encouraging wider adoption and environmental benefits. In particular, soil microbial properties associated with variation in efficacy remain largely unknown. In this study four contrasting arable soils (a grey desert soil, an alluvial paddy soil, a loess-formed paddy soil, and a red soil), were characterized based on DMPP inhibition of N2O emissions and associated microbial functional guilds. DMPP significantly inhibited nitrification and N2O emissions, with an average inhibitory rate ranging from 41.7% in a red soil to 90.0% in a grey desert soil. Ammonia oxidizing bacteria (AOB) and archaea (AOA) exhibited contrasting response patterns to DMPP addition. Notably, suppression of N2O emissions by DMPP only occurred alongside fluctuations in AOB abundance. However, when AOB were inhibited, AOA abundance increased. Soil-dependent response patterns to DMPP were observed for ammonia oxidizers and denitrifiers in terms of community structure. Partial least squares path modeling (PLS-PM) found that abiotic factors, particularly pH, and biological factors such as ammonia oxidizer communities, were closely linked to N2O emissions. Our findings provide evidence that: (i) DMPP effectively inhibits nitrification through inhibiting the abundance of AOB across soil types; (ii) releasing AOA from the competition with AOB allows AOA to efficiently grow and multiply, even under high ammonium conditions; and (iii) abiotic factors play a more important role than biotic factors in soil N2O emissions.

Ammonia-oxidizing bacterial and archaeal communities in tropical bioaugmented zero water exchange shrimp production systems

Ammonia oxidation is an important process in the removal of ammonia generated from feed and metabolic wastes in aquaculture systems. Considering the biogeochemical importance of ammonia oxidation in bioaugmented zero water exchange aquaculture systems, the diversity and abundance of bacterial and archaeal ammonia-oxidizing communities were analyzed in three selected ponds at different time intervals during the culture period, to unravel the key environmental factors influencing their distribution in the system. The diversity and abundance of ammonia-oxidizing bacteria (AOB) and archaea (AOA) in three tropical bioaugmented zero water exchange (ZWE) shrimp culture systems were analyzed using ammonia monooxygenase A (amoA) gene from the sediment metagenome during different phases of culture. The environmental factors associated with the variability in bacterial and archaeal amoA gene abundance and diversity were elucidated using RDA and Pearson correlation analysis. Ammonia-oxidizing archaea (AOA), Nitrosopumilus sp., Nitrosospharea sp., and ammonia-oxidizing bacteria (AOB), Nitrosomonas sp., were the dominant ammonia-oxidizing communities in the ZWE ponds studied. AOA shared 41 OTUs, and the maximum distribution was influenced by dissolved oxygen in the system, whereas AOB shared 4 OTUs. The copy numbers amoA gene determined using qPCR showed that the AOA amoA gene was 10- to 100-fold abundant than AOB amoA gene. Gene abundance of AOA was positively related to total organic carbon (TOC) and salinity of sediments, and the temperature had a negative impact on bacterial amoA gene abundance. The dissolved oxygen and TOC had a negative and redox potential a positive impact on the diversity of AOA, whereas pH had a negative impact on the diversity of AOB. The ammonia-oxidizing archaeal communities dominated the bioaugmented zero water exchange aquaculture systems compared to bacteria based on the abundance and diversity analysis using amoA gene sequence-based OTU analysis and gene copy numbers. Dissolved oxygen, total organic carbon, and Eh of the sediments contributed to the distribution and abundance of AOA group in the ZWE ponds. This study points to the importance of environmental management in these culture systems for maintaining ammonia-oxidizing populations for optimal ammonia removal. The relative contribution of the archaea and bacteria to ammonia oxidation in these systems is to be further resolved along with that of anammox and comammox bacteria, which would help to develop appropriate biostimulation or bioaugmentation strategies for the management of these sustainable aquaculture production systems.

Soil aggregate size and long-term fertilization effects on the function and community of ammonia oxidizers

Long-term field fertilization trials have suggested that reduced chemical nitrogen (N) plus organic fertilization can effectively reduce N loss without sacrificing crop yield, while the knowledge of how organic fertilizers regulate soil microorganisms and their function in N transformation are limited. In this study, the response of net nitrification rate and the ammonia-oxidizer community within soil aggregates to long-term combined organic N and reduced chemical N fertilization was evaluated to understand the underlying mechanism of the practice in mitigating soil N loss. The fertilization experiment included an unfertilized control; chemical N fertilizer (N), superphosphate (P) and potassium sulfate (K) fertilizer (NPK); NPK plus straw (NPKS); and NPK plus manure (NPKM). The results showed that the large macro-aggregates mass (>2 mm) in soil significantly increased from 34.1% in NPK to 47.2% in NPKS (P < 0.05). NPKS and NPKM both had positive effects on soil moisture retention, total N (TN), soil organic carbon (SOC), ammonium (NH4+-N) and nitrate (NO3−-N) accumulation, particularly within micro- (<0.25 mm) and small macro-aggregates. Compared with the NPK treatment, soil net nitrification rate (NNR) and ammonia-oxidizing bacterial (AOB) abundance decreased by 67.1% and 40.7% respectively under NPKS (P < 0.05), and the decrease mainly appeared in large macro-aggregate and micro-aggregates. The net nitrification rate was significantly correlated with ammonia-oxidizing archaea (AOA) abundance only in small macro-aggregates (r = 0.642, n = 12, P < 0.05). In contrast, NRR was positively correlated with AOB abundance within small macro- and micro-aggregate size classes (r values ranged from 0.654 to 0.813, P < 0.05). The community structure of AOB varied among different fertilization treatments, while AOA community differentiation was mainly dependent on aggregate size. The shift of the AOA and AOB communities corresponded with high moisture and lower ammonia content in the large macro-aggregates and were potentially responsible for the suppressed nitrification activity in the NPKS treatment. These results indicated that reduced chemical N plus organic fertilization is beneficial for increasing N concentration in large macro-aggregates.

Methane-generating ammonia oxidizing nitrifiers within bio-filters in aquaculture tanks

The discovery of aerobic and anammox bacteria capable of generating methane in bio-filters in freshwater aquaculture systems is generating interest in studies to understand the activity, diversity, distribution and roles of these environmental bacteria. In this study, we used microbial enrichment of bio-filters to assess their effect on water quality. Profiles of ammonia-oxidizing bacterial communities generated using nested PCR methods and DGGE were used to assess the expression of 16S rRNA genes using DNA sequencing. Five dominant ammonia-oxidizing bacterial strains–clones; KB.13, KB.15, KB.16, KB.17 and KB.18—were isolated and identified by phylogenetic analysis as environmental samples closely related to genera Methylobacillus, Stanieria, Nitrosomonas, and Heliorestis. The methyl ammonia-oxidizing microbes thereby found suggest a biochemical pathway involving electron donors and carbon sources, and all strains were functional in freshwater aquaculture systems. Environmental parameters including TN (2.69–20.43); COD (9.34–31.47); NH4+-N (0.44–11.78); NO2−N (0.00–3.67); NO3−N (0.05–1.82), mg/L and DO (1.47–10.31 µg/L) assessed varied in the ranges in the different tanks. Principal component analysis revealed that these water quality parameters significantly influenced the ammonia oxidizing microbial community composition. Temperature rises to about 40 °C significantly affected environmental characteristics—especially DO, TN and NH4+-N—and directly or indirectly affected the microbial communities. Although the nested PCR design was preferred due to its high sensitivity for amplifying specific DNA regions, a more concise method is recommended, as an equimolar mixture of degenerate PCR primer pairs, CTO189f-GC and CTO654r, never amplified only 16S rRNA of ammonia-oxidizing bacteria.

Ammonia oxidizers in the sea-surface microlayer of a coastal marine inlet.

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.

Impacts of long-term nitrogen addition, watering and mowing on ammonia oxidizers, denitrifiers and plant communities in a temperate steppe

Global changes in nitrogen (N) deposition, precipitation patterns and land use could have an impact on the biogeochemical N cycle mediated by microorganisms. Microbial responses to global changes have been generally evaluated in short-term studies, while the long-term effects are not well understood. In this study, the interactive effects of multiple global change factors on N-cycling microorganisms and their interactions with plants were investigated in a 9-year field experiment with N addition, watering and mowing in a temperate steppe in northern China. Quantitative PCR, terminal restriction fragment length polymorphism (T-RFLP) and clone library were used to analyze the abundance and community structure of ammonia oxidizers and denitrifiers, respectively. Bacterial 16S rRNA gene abundance significantly increased under mowing, but the functional gene abundances including amoA, nirS, nirK and nosZ genes were not affected. Watering significantly affected the ammonia-oxidizing archaea (AOA) community and N addition showed a strong effect on soil ammonia-oxidizing bacteria (AOB) and nosZ-containing denitrifier community structure. No significant interactive effects of mowing, N addition and watering on communities of soil ammonia oxidizers and denitrifiers were observed. Structure equation models suggested that plant biomass and community were significantly correlated with microbial communities and activities. These findings suggest that there are complex interactions between plants and soil microorganisms in grassland ecosystems.