Anammox Community Research Articles

Salinity-driven shifts in the activity, diversity, and abundance of anammox bacteria of estuarine and coastal wetlands

Anaerobic ammonium oxidation (anammox) plays a significant role in nitrogen removal in estuarine and coastal wetlands. However, the effects of changing salinity on anammox activity and anammox bacterial dynamics in these environments are not well understood. In this study, serial incubation experiments with a salinity gradient (0–40) were conducted to explore the responses of anammox bacterial activity, diversity and abundance to the changing salinity in the intertidal wetland of the Yangtze Estuary. Results show that activity and abundance of anammox bacteria firstly increased with the increase of salinity, but they were physiologically stressed by high-level salinity (>30) in a short-term incubation (<10 days). However, the treatment with salinity of 5 showed the maximal anammox activity and anammox bacterial abundance after a long-term incubation (60–120 days). In addition, Kuenenia (Kuenenia stuttgartiensis), Scalindua (Scalindua wagner, marina, and brodae), and three unknown anammox-like groups were observed, and anammox bacterial diversity increased along the salinity gradient. Anammox community structure varied slightly within the first 10-day incubation, but the dominant anammox bacterial group shifted from Kuenenia to Scalindua with increasing salinity after the long-term incubation. Overall, this study demonstrates the effects of salinity on anammox bacterial community and anammox activity, and suggests the importance of salinity in regulating the anammox process in estuarine and coastal wetlands with frequent salinity fluctuation.

Effects of HRT and Loading Rate on Performance of Carriers-Amended Anammox UASB Reactors.

Effects of hydraulic retention time (HRT) and nitrogen loading rate (NLR) on performance of anammox UASBs amended with spherical plastic and bamboo charcoal carriers were investigated. During the experimental period, the performance of reactors was continuously evaluated by monitoring ammonium-N, nitrite-N, and nitrate-N at HRT of 48, 36, 24, and 12 hours. With HRTs between 24 and 48 hours, the ammonium-N and nitrite-N removal efficiency was higher than 99%, while the total nitrogen (TN) removal efficiency was almost higher than 90%. When HRT decreased to 12 hours with NLR increasing to 0.16 kg N m-3 d-1, ammonium-N, and TN removal efficiency decreased to 87 and 80%, respectively. Moreover, phylogenetic analysis revealed anammox community still existed at the HRT of 12 hours and was most closely related to C. Brocadia sp.40. In addition, different types of carriers had no significant influence on the anammox community.

Faster through training: The anammox case

Anaerobic ammonium oxidizing (anammox) bacteria based technologies are widely applied for nitrogen removal from warm (25–40 °C) wastewater with high ammonium concentrations (∼1 gNH4–N L−1). Extension of the operational window of this energy and resource efficient process is restricted by the “supposed” low growth rate of the responsible microorganisms. Here we demonstrate that the maximum specific growth rate (μmax) of anammox bacteria can be increased to a μmax value of 0.33 d−1 by applying a novel selection strategy based on the maximization of the electron transfer capacity in a membrane bioreactor. This value is four times higher than the highest previously reported value. The microbial community was strongly dominated by anammox bacteria closely related (99%) to Candidatus Brocadia sp.40 throughout the experiment. The results described here demonstrate the remarkable capacity of a phylogenetically stable anammox community to adjust its growth rate in response to a change in the cultivation conditions imposed.

Study on anaerobic ammonium oxidation process coupled with denitrification microbial fuel cells (MFCs) and its microbial community analysis

Denitrifying MFC was successfully coupled with anaerobic ammonium oxidation process in this study. With the help of cathode electrons, the stoichiometry relationship Δ−NH3-N:Δ−NO2−-N:Δ+NO3−-N was approximate 1:1.37:0.03 during the stable operation, which demonstrated its further nitrite and nitrate reduction. According to microbial community analysis, Candidatus Brocadia sinica was the main anammox community, and Rhodopseudomonas palustris with electrochemical driven denitrifying ability, especially enriched in biofilm. Higher abundance of nirK genes in biofilm (compare to the control) and invariable amx genes in suspended sludge were responsible for its better nitrogen removal. The sludge and biofilm performed their own duties on anammox and denitrification, respectively, according to genes quantification. Under the totally autotrophic system, electron donors were “concentrated supplied” and easy to control, which avoided the suppression of anammox growth, making this autotrophic coupling process appears to be more promising.

Nitrogen removal and spatial distribution of denitrifier and anammox communities in a bioreactor for mine drainage treatment

Mine drainage water may contain high levels of nitrate (NO3−) due to undetonated nitrogen-based explosives. The removal of NO3− and nitrite (NO2−) in cold climates through the microbial process of denitrification was evaluated using a pilot-scale fixed-bed bioreactor (27 m3). Surface water was diverted into the above-ground bioreactor filled with sawdust, crushed rock, and sewage sludge. At hydraulic residence times of ca.15 h and with the addition of acetate, NO3− and NO2− were removed to below detection levels at a NO3− removal rate of 5–10 g N m−3 (bioreactor material) d−1. The functional groups contributing to nitrogen removal in the bioreactor were studied by quantifying nirS and nirK present in denitrifying bacteria, nosZI and nosZII genes from the nitrous oxide – reducing community, and a taxa-specific part of the16S rRNA gene for the anammox community. The abundances of nirS and nirK were almost 2 orders of magnitude greater than the anammox specific 16S rRNA gene, indicating that denitrification was the main process involved in nitrogen removal. The spatial distribution of the quantified genes was heterogeneous in the bioreactor, with trends observed in gene abundance as a function of depth, distance from the bioreactor inlet, and along specific flowpaths. There was a significant relationship between the abundance of nirS, nirK, and nosZI genes and depth in the bioreactor, such that the abundance of organisms containing these genes may be controlled by oxygen diffusion and substrate supply in the partially or completely water-saturated material. Among the investigated microbial functional groups, nirS and anammox bacterial 16S rRNA genes exhibited a systematic trend of decreasing and increasing abundance, respectively, with distance from the inlet, which suggested that the functional groups respond differently to changing environmental conditions. The greater abundance of nirK along central flowpaths may indicate that the bioreactor design favored preferential flow along these flowpaths, away from the sides of the bioreactor. An improved bioreactor design should consider the role of preferential flowpaths and the heterogeneous distribution of the genetic potential for denitrification, nitrous oxide reduction and anammox on bioreactor function.

Diversity and distribution of planktonic anaerobic ammonium-oxidizing bacteria in the Dongjiang River, China

Anaerobic ammonium-oxidizing (anammox) process has recently been recognized as an important pathway for removing fixed nitrogen (N) from aquatic ecosystems. Anammox organisms are widely distributed in freshwater environments. However, little is known about their presence in the water column of riverine ecosystems. Here, the existence of a diverse anammox community was revealed in the water column of the Dongjiang River by analyzing 16S rRNA and hydrazine oxidation (hzo) genes of anammox bacteria. Phylogenetic analyses of hzo genes showed that Candidatus Jettenia related clades of anammox bacteria were dominant in the river, suggesting the ecological microniche distinction from freshwater/estuary and marine anammox bacteria with Ca. Brocadia and Kuenenia genera mainly detected in freshwater/estuary ecosystems, and Ca. Scalindua genus mainly detected in marine ecosystems. The abundance and diversity of anammox bacteria along the river were both significantly correlated with concentrations of NH4+-N based on Pearson and partial correlation analyses. Redundancy analyses showed the contents of NH4+-N, NO3−-N and the ratio of NH4+-N to NO2−-N significantly influenced the spatial distributions of anammox bacteria in the water column of the Dongjiang River. These results expanded our understanding of the distribution and potential roles of anammox bacteria in the water column of the river ecosystem.

The first report of a microdiverse anammox bacteria community in waters of Colombian Pacific, a transition area between prominent oxygen minimum zones of the eastern tropical Pacific.

Anaerobic ammonium oxidizers contribute to the removal of fixed nitrogen in oxygen-deficient marine ecosystems such as oxygen minimum zones (OMZ). Here we surveyed for the first time the occurrence and diversity of anammox bacteria in the Colombian Pacific, a transition area between the prominent South and North Pacific OMZs. Anammox bacteria were detected in the coastal and oceanic areas of the Colombian Pacific in low oxygen (< 22 μM), high nitrate (25–35 μM) and low nitrite (< 0.07 μM), and ammonium (< 1 μM) waters. In these waters, anammox bacteria were rich [∼ 7 operational taxonomic units (OTUs), 98% cut-off) and microdiverse (Shannon index H′ < 1.24), in comparison with the observed at the prominent OMZ of the Eastern Tropical South Pacific, Arabian Sea and Black Sea. Anammox bacteria-like sequences from the Colombian Pacific were grouped together with sequences retrieved from the distinct OMZ's marine subclusters (Peru, Northern Chile and Arabian Sea) within Candidatus ‘Scalindua spp’. Moreover, some anammox bacteria OTUs shared a low similarity with environmental phylotypes (86–94%). Our results indicated that a microdiverse anammox community inhabits the Colombian Pacific, generating new questions about the ecological and biogeochemical differences influencing its community structure.

Impacts of freshwater flushing on anammox community structure and activities in the New River Estuary, USA

Anaerobic ammonium oxidation (anammox) and denitrification are 2 microbial nitro- gen removal processes that may play an important role in controlling the intensity and duration of estuarine and coastal eutrophication. Sediment communities in the New River Estuary, North Car- olina were investigated to determine the dynamics of anammox activity and community structure in conjunction with environmental conditions. 15 N tracer incubation experiments with sediment slurries were used to measure anammox and denitrification rates and estimate anammox contri- bution to total N2 production. Molecular analyses targeting the hydrazine oxidoreductase (hzo) gene were conducted to examine the structure of anammox communities and quantify the abun- dance of anammox bacteria in sediments. Potential anammox rates ranged from 0.02 to 1.4 nmol N2 g �1 h �1 , with the highest potential activities observed during winter and spring when the estu- ary received large doses of nitrogen from the watershed. Anammox contributed up to 14.1% of total N2 production in upstream estuarine sediments and abundance of anammox communities ranged from 1.55 × 10 2 to 2.59 × 10 5 hzo gene copies g �1 sediment. Both activities and abundance of anammox communities were correlated with percent sediment organics (%organics) and the porewater concentrations of hydrogen sulfide. Based on hzo sequence analysis, anammox bacte- ria related to 'Candidatus Jettenia spp.' were widespread in estuarine sediments, which may be attributed to freshwater flushing and associated changes in environmental parameters as well as the geomorphology of the estuary. This is the first study to describe a dominance of 'Candidatus Jettenia spp.' in relation to %organics and hydrogen sulfide in an estuarine ecosystem driven by meteorological forcing.

Impact of volcanic ash on anammox communities in deep sea sediments

Subaerial explosive volcanism contributes substantial amounts of material to the oceans, but little is known about the impact of volcanic ash on sedimentary microbial activity. We have studied anammox communities in deep sea sediments near the volcanically active island of Montserrat, Lesser Antilles. The rates of anammox and denitrification in the sediments were measured using (15)N isotope pairing incubation experiments, while 16S rRNA genes were used to examine anammox community structures. The higher anammox rates were measured in sediment containing the lower accumulation of volcanic ash in the surface sediments, while the lowest activities were found in sediments with the highest ash deposit. 16S rRNA gene analysis revealed the presence of 'Candidatus Scalindua spp.' in the sediments. The lowest diversity of anammox bacteria was observed in the sediments with the highest ash deposit. Overall, this study demonstrates that the deposition of volcanic material in deep sea sediments has negative impacts on activity and diversity of the anammox community. Since anammox may account for up to 79% of N2 production in marine ecosystems, periods of extensive explosive volcanism in Earth history may have had a hitherto unrecognized negative impact on the sedimentary nitrogen removal processes.

Microbial Biofilms Formed in a Laboratory-Scale Anammox Bioreactor with Flexible Brush Carrier

This study is focused on investigation of biofilms formed in an anaerobic laboratory-scale bioreactor fed with medium for anammox bacteria oxidizing ammonia with nitrite. The mixed culture of anammox bacteria was enriched from the microbial community that sampled from the activated sludge of a denitrifying reactor at a wastewater treatment station located in the Sochi region, Russia. This community forms biofilms on the surface of the flexible polymer brush carriers, which are used for biomass immobilization in both laboratory and full-scale bioreactors. Anammox bacteria were discovered in the activated sludge community. The anammox community was enriched by incubation in an up-flow laboratory-scale anaerobic bioreactor with a flexible brush carrier. In the course of ~3 years, the loading rate of nitrogen substrates (ammonium and nitrite) increased from 100 to 5000 mg N L-1 day-1. The concentration of the substrates in the upper part of the reactor was 40 times less than in the influent. The pH values were 7.5 at the bottom and up to 9 in the upper part up of the reactor. Biofilms of two types developed in the reactor. Bunches of irregular spherical granules formed on the carrier filaments, while films of irregular thickness containing submerged spherical granules were formed on the walls of reactor. The anammox population was found to consist of at least three active species: a new strain of Candidatus “Jettenia asiatica” named “strain ecos” and two species of the genus Candidatus “Brocadia”. Other types of bacteria found in the community, including members of phylum Chloroflexi, were presumably involved in biofilm spatial organization.

Molecular diversity and distribution of anammox community in sediments of the Dongjiang River, a drinking water source of Hong Kong

The aim of this study was to characterize anaerobic ammonium oxidation (anammox) community in sediments of the Dongjiang River, a drinking water source of Hong Kong. The diversity and distribution of the anammox community were investigated based on a comparative analyses of 16S rRNA and hydrazine oxidation (hzo) genes of anammox bacteria. Candidatus Brocadia and two new anammox bacterial clusters were detected based on phylogenetic analysis of 16S rRNA genes. In contrast, the targeting of hzo genes indicated the presence of only Candidatus Jettenia with four different clusters. It was found that the sequence diversities of hzo genes were higher than those of the 16S rRNA genes. The abundance of anammox bacteria varied significantly among the sediment samples based on qPCR. Pearson correlation analysis indicated that nitrite concentration was the key factor influencing the abundance of anammox bacteria. The redundance analysis (RDA) confirmed that the combination of the contents of nitrite and nitrate, and the ratio of total nitrogen vs total carbon (TN/TC) had significant impact on the anammox bacterial community structure. The results revealed that the diverse anammox bacteria were present in sediments of the Dongjiang River, and the community structures were associated with varied environmental factors caused by urban pollutant invasion. This is the first report about the distribution of anammox bacterial community in sediments of the Dongjiang River, which provides helpful information of anammox niche specificity and influencing factors in the river ecosystem.

Biases in community structures of ammonia/ammonium-oxidizing microorganisms caused by insufficient DNA extractions from Baijiang soil revealed by comparative analysis of coastal wetland sediment and rice paddy soil

Repetitive extraction of DNAs from surface sediments of a coastal wetland in Mai Po Nature Reserve (MP) of Hong Kong and surface Baijiang soils from a rice paddy (RP) in Northeast China was conducted to compare the microbial diversity in this study. Community structures of ammonia/ammonium-oxidizing microorganisms in these samples were analyzed by PCR-DGGE technique. The diversity and abundance of ammonia-oxidizing archaea (AOA), ammonia-oxidizing bacteria (AOB), and anaerobic ammonium-oxidizing (anammox) bacteria were also analyzed based on archaeal and bacterial ammonia monooxygenase subunit A encoding (amoA) and anammox bacterial 16S rRNA genes, respectively. DGGE profiles of archaeal and bacterial amoA and anammox bacterial 16S rRNA genes showed a similar pattern among all five repetitively extracted DNA fractions from both MP and RP, except the anammox bacteria in RP, indicating a more diverse anammox community retrieved in the second to the fifth fractions than the first one. Both soil and marine group AOA were detected while soil and coastal group AOB and Scalindua-anammox bacteria were dominant in MP. Soil group AOA and marine group AOB were dominant in RP, while both Scalindua and Kuenenia species were detected in RP. Pearson correlation analysis showed that the abundance of archaeal and bacterial amoA and anammox bacterial 16S rRNA genes was significantly correlated with the DNA concentrations of the five DNA fractions from MP, but not from RP (except the archaeal amoA gene). Results suggest that anammox bacteria diversity may be biased by insufficient DNA extraction of rice paddy soil samples.

Anammox bacterial populations in deep marine hypersaline gradient systems

To extend the knowledge of anaerobic ammonium oxidation (anammox) habitats, bacterial communities were examined in two hypersaline sulphidic basins in Eastern Mediterranean Sea. The 2m thick seawater-brine haloclines of the deep anoxic hypersaline basins Bannock and L'Atalante were sampled in intervals of 10cm with increasing salinity. (15)N isotope pairing incubation experiments showed the production of (29)N2 and (30)N2 gases in the chemoclines, ranging from 6.0 to 9.2% salinity of the L'Atalante basin. Potential anammox rates ranged from 2.52 to 49.65nmolN2 L(-1) day(-1) while denitrification was a major N2 production pathway, accounting for more than 85.5% of total N2 production. Anammox-related 16S rRNA genes were detected along the L'Atalante and Bannock haloclines up to 24% salinity, and the amplification of the hydrazine synthase genes (hzsA) further confirmed the presence of anammox bacteria in Bannock. Fluorescence in situ hybridisation and sequence analysis of 16S rRNA genes identified representatives of the marine anammox genus 'Candidatus Scalindua' and putatively new operational taxonomic units closely affiliated to sequences retrieved in marine environments that have documented anammox activity. 'Scalindua brodae' like sequences constituted up to 84.4% of the sequences retrieved from Bannock. The anammox community in L'Atalante was different than in Bannock and was stratified according to salinity increase. This study putatively extends anammox bacterial habitats to extremely saline sulphidic ecosystems.

Influence of temperature and salinity on microbial structure of marine anammox bacteria

Anaerobic ammonium oxidation (anammox) is a type of biological oxidation mediated by a group of Planctomycete-like bacteria. Members of the genus Candidatus Scalindua are mainly found in marine environments, but not exclusively. This group is cultured using different inoculums and conditions; however, its optimal growth conditions are not clear. Additionally, little information is known about the factors that influence the activity and the selection of a population of marine anammox bacteria. This study was conducted to investigate the influence of temperature and salinity on the marine anammox community. To accomplish this, an up-flow fixed-bed column reactor was operated, and quantitative fluorescence in situ hybridization (FISH) with probes specific to dominant marine anammox bacteria was conducted. Anammox activity was observed at 20 and 30 °C, but not at 10 °C. A nitrogen removal rate of 0.32 kg TN m(-3) day(-1) was obtained at 20 °C. These results suggest that temperature affects the activity (nitrogen removal rate) of anammox bacteria, while salinity does not affect the activity in the marine anammox biofilm.

Dominance of Candidatus Scalindua species in anammox community revealed in soils with different duration of rice paddy cultivation in Northeast China

The anaerobic ammonium-oxidizing (anammox) bacteria play an important role in the oxygen-limited zone for nitrogen cycling, but their roles in agricultural ecosystems are still poorly understood. In this study, soil samples were taken from the rhizosphere and non-rhizosphere and from surface (0–5 cm) and subsurface (20–25 cm) layers with 1, 4, and 9 years of rice cultivation history on the typical albic soil of Northeast China to examine the diversity and distribution of anammox bacteria based on 16S rRNA gene and hydrazine oxidoreductase encoding gene (hzo). By comparing these soil samples, no obvious difference was observed in community composition between the rhizosphere and non-rhizosphere or the surface and subsurface layers. Surprisingly, anammox bacterial communities of these rice paddy soils were consisted of mainly Candidatus Scalindua species, which are best known to be dominant in marine and pristine environments. The highest diversity was revealed in the 4-year paddy soil based on clone library analysis. Phylogenetic analysis of 16S rRNA gene and deduced HZO from the corresponding encoding gene showed that most of the obtained clones are grouped together with Candidatus Scalindua sorokinii, Candidatus Scalindua brodae, and Candidatus Scalindua spp. of seawater. The obtained clone sequences from all samples are distributed in two subclusters that contain sequences from environmental samples only. Tentative new species were also discovered in this paddy soil. This study provides the first evidence on the existence of anammox bacteria with limited diversity in agricultural ecosystems in Northern China.Electronic supplementary materialThe online version of this article (doi:10.1007/s00253-012-4036-x) contains supplementary material, which is available to authorized users.

Induced cooperation between marine nitrifiers and anaerobic ammonium-oxidizing bacteria by incremental exposure to oxygen

In oxygen-limited marine ecosystems cooperation between marine nitrifiers and anaerobic ammonium-oxidizing (anammox) bacteria is of importance to nitrogen cycling. Strong evidence for cooperation between anammox bacteria and nitrifiers has been provided by environmental studies but little is known about the development of such communities, the effects of environmental parameters and the physiological traits of their constituents. In this study, a marine laboratory model system was developed. Cooperation between marine nitrifiers and anammox bacteria was induced by incremental exposure of a marine anammox community dominated by Scalindua species to oxygen in a bioreactor set-up under high ammonium (40 mM influent) conditions. Changes in the activities of the relevant functional groups (anammox bacteria, aerobic ammonia oxidizers and nitrite oxidizers) were monitored by batch tests. Changes in community composition were followed by Fluorescence in situ Hybridization (FISH) and by amplification and sequencing of 16S rRNA and amoA genes. A co-culture of Scalindua sp., an aerobic ammonia-oxidizing Nitrosomonas-like species, and an aerobic (most likely Nitrospira sp.) nitrite oxidizer was obtained. Aerobic ammonia oxidizers became active immediately upon exposure to oxygen and their numbers increased 60-fold. Crenarchaea closely related to the ammonia-oxidizer Candidatus ‘Nitrosopumilus maritimus’ were detected in very low numbers and their contribution to nitrification was assumed negligible. Activity of anammox bacteria was not inhibited by the increased oxygen availability. The developed marine model system proved an effective tool to study the interactions between marine anammox bacteria and nitrifiers and their responses to changes in environmentally relevant conditions.

Diversity and abundance of anammox bacterial community in the deep-ocean surface sediment from equatorial Pacific

The community structure and diversity of anaerobic ammonium oxidation (anammox) bacteria in the surface sediments of equatorial Pacific were investigated by phylogenic analysis of 16S rRNA and hydrazine oxidoreductase (hzo) genes and PCoA (principal coordinates analysis) statistical analysis. Results indicated that 16S rRNA and hzo sequences in the P2 (off the center of western Pacific warm pool) and P3 (in the eastern equatorial Pacific) sites all belong to the Candidatus "Scalindua", the dominate anammox bacteria in the low-temperature marine environment proved by previous studies. However, in the P1 site (in center of warm pool of western Pacific), large part of 16S rRNA gene sequences formed a separated cluster. Meanwhile, hzo gene sequences from P1 sediment also grouped into a single cluster. PCoA analysis demonstrated that the anammox community structure in the P1 has significant geographical distributional difference from that of P2, P3, and other marine environments based on 16S rRNA and hzo genes. The abundances of anammox bacteria in surface sediments of equatorial Pacific were quantified by q-PCR analysis of hzo genes, which ranged from 3.98 × 10(3) to 1.17 × 10(4) copies g(-1) dry sediments. These results suggested that a special anammox bacteria phylotypes exist in the surface sediment of the western Pacific warm pool, which adapted to the specific habitat and maybe involved in the nitrogen loss process from the fixed inventory in the habitat.

High-rate nitrogen removal by the Anammox process with a sufficient inorganic carbon source

This study focused on high-rate nitrogen removal by the anaerobic ammonium oxidation (Anammox) process with a sufficient inorganic carbon (IC) source. Experiments were carried out in an up-flow column Anammox reactor fed with synthetic inorganic wastewater for 110 days. The IC source was added into the influent tank in the form of bicarbonate. The results confirmed the positive impact of inorganic matter on stimulating Anammox activity. After the addition of sufficient IC, the nitrogen removal rate sharply increased from 5.2 to 11.8 kg-Nm(-3)day(-1) within only 32 days. NO(2)-N inhibition was not observed even at NO(2)-N concentrations greater than 460 mgN/L, indicating the enriched Anammox consortium adapted to high NO(2)-N concentrations. The ratio of NO(2)-N removal, NO(3)-N production and NH(4)-N removal for the reactor was correspondingly changed from 1.21:0.21:1 to 1.24:0.18:1. Simultaneously, the sludge volume index of the Anammox granules decreased markedly from 36.8 to 21.5 mL/g, which was attributed to the implementation of proper operational strategy. In addition, DNA analysis revealed that a shift from the KSU-1 strain to the KU2 strain occurred in the Anammox community.

Phylogenetic analysis of a microbial community involved in anaerobic oxidation of ammonium nitrogen

The phylogenetic diversity of a microbial community involved in anaerobic oxidation of ammonium nitrogen in the DEAMOX process was studied. Analysis of clone libraries containing 16S rRNA gene inserts of Bacteria, (including Planctomycetes) and Archaea revealed the presence of nucleotide sequences of the microorganisms involved in the main reactions of the carbon, nitrogen, and sulfur cycles, including nitrifying, denitrifying, and ANAMMOX bacteria. In the bacterial clone library, 16S rRNA gene sequences of representatives of the phyla Proteobacteria, Bacteroidetes, Chloroflexi, Firmicutes, Verrucomicrobia, Lentisphaerae, Spirochaetales, and Planctomycetes, as well as of some new groups, were detected. In the archaeal clone library, nucleotide sequences of methanogens belonging to the orders Methanomicrobiales, Methanobacteriales, and Methanosarcinales were found. It is possible that both ANAMMOX bacteria and bacteria of the genus Nitrosomonas are involved in anaerobic ammonium oxidation in the DEAMOX reactor. Many sequences were similar to those from the clone libraries obtained previously from the ANAMMOX community of marine sediments. It is also probable that the DEAMOX reactions occur in natural ecosystems (in marine and freshwater sediments and the oceanic water column), thereby providing for the coupling of the nitrogen and sulfur cycles.

Biogeographical distribution of diverse anaerobic ammonium oxidizing (anammox) bacteria in Cape Fear River Estuary

Anaerobic ammonium oxidation (anammox) specific PCR method was developed to examine diversity and distribution of anammox bacteria in sediments collected from three different sites at Cape Fear River Estuary, North Carolina, where environmental parameters vary greatly over the year. Abundance and activities of anammox bacteria in these sediments were measured using the quantitative PCR (Q-PCR) method and (15)N isotope tracer incubations. Different anammox bacterial communities composed with Brocadia, Kuenenia, Jettenia or Scalindua were found among sites along the estuarine gradient. Seasonal variations of anammox community structures were observed along the estuary based on terminal restriction fragment length polymorphism (T-RFLP) analysis of 16S rRNA genes. Correlation analysis suggested that salinity variation influenced the diversity and distribution of different anammox bacteria in the estuary. Q-PCR assays of anammox bacteria showed temporal and spatial variations of their abundances, which were highly correlated to salinity variation. (15)N isotope tracer incubations measured different anammox rates and its per cent contribution to total N(2) production among sites. The highest anammox rate was found at the site where Scalindua organisms dominated with the highest anammox bacterial abundance. Thus, we demonstrated a biogeographical distribution of diverse anammox bacteria influenced by salinity, and provide evidence to link anammox abundance and activities in estuarine sediments.