Improving dryland maize productivity and water efficiency with heterotrophic ammonia-oxidizing bacteria via nitrification and cytokinin activity

A two-year field experiment conducted under dryland conditions in semi-humid and drought-prone regions of China aimed to assess the effect of ammonia-oxidizing bacterial on maize water use efficiency and yield.A heterotrophic ammonia-oxidizing bacteria(HAOB)strain S2_8_1 was used.Six treatments were applied:(1)no irrigation+HAOB strain(DI),(2)no irrigation+blank culture medium(DM),(3)no irrigation control(DCK),(4)irrigation+HAOB(WI),(5)irrigation+blank culture medium(WM),and(6)irrigation control(WCK).Results revealed that HAOB treatment increased maize growth,yield,and water use efficiency over controls,regardless of whether the year was wet or dry.This improvement was attributed to the accelerated nitrification in the rhizosphere soil due to HAOB inoculation,which subsequently led to increased levels of leaf cytokinins.Overall,these findings suggest that HAOB inoculation holds promise as a strategy to boost water use efficiency and maize productivity in dryland agriculture.

Community analysis of ammonia-oxidizing bacteria in activated sludge of eight wastewater treatment systems

We investigated the communities of ammonia-oxidizing bacteria （AOB） in activated sludge collected from eight wastewater treatment systems using polymerase chain reaction （PCR） followed by terminal restriction fragment length polymorphism （T-RFLP）, cloning, and sequencing of the α-subunit of the ammonia monooxygenase gene （amoA）. The T-RFLP fingerprint analyses showed that different wastewater treatment systems harbored distinct AOB communities. However, there was no remarkable difference among the AOB T- RFLP profiles from different parts of the same system. The T-RFLP fingerprints showed that a full-scale wastewater treatment plant （WWTP） contained a larger number of dominant AOB species than a pilot-scale reactor. The source of influent affected the AOB community, and the WWTPs treating domestic wastewater contained a higher AOB diversity than those receiving mixed domestic and industrial wastewater. However, the AOB community structure was little affected by the treatment process in this study. Phylogenetic analysis of the cloned amoA genes clearly indicated that all the dominant AOB in the systems was closely related to Nitrosomonas spp. not to Nitrosospira spp. Members of the Nitrosomonas oligotropha and Nitrosomonas communis clusters were found in all samples, while members of Nitrosomonas europaea cluster occurred in some systems.

Impact of acetochlor on ammonia-oxidizing bacteria in microcosm soils

Acetochlor is an increasingly used herbicide on corn in North China. Currently, the effect of acetochlor on soil ammonia-oxidizing bacteria (AOB) communities is not well documented. Here, we studied the diversity and community composition of AOB in soil amended with three concentrations of acetochlor (50, 150, 250 mg/kg) and the control (0 mg acetochlor/kg soil) in a microcosm experiment by PCR-DGGE (polymerase chain reaction-denaturing gradient gel electrophoresis) and the phylogenetic analysis of excised ...

Distribution of ammonia-oxidizing Betaproteobacteria community in surface sediment off the Changjiang River Estuary in summer

The spatial distribution of ammonia-oxidizing Betaproteobacteria （βAOB） was investigated by FISH （fluorescence in situ hybridization） and DGGE （denaturing gradient get electrophoresis） techniques in the sediment off the Changjiang River Estuary. Sediment samples were collected from eight stations in June before the formation of hypoxia zone in 2006. The abundance of βAOB ranged from 1.87× 10^5 to 3.53×10^5 cells/g of sediment. βAOB abundance did not present a negative correlation with salinity, whereas salinity was implicated as the primary factor affecting nitrification rates. The DGGE profiles of PCR-amplified amoA gene fragments revealed that the βAOB community structure of sample S2 separated from other samples at the level of 40% similarity. The variations in composition ofβAOB were significantly correlated with the salinity, temperature, absorption ability of sediments and TOC. The statistical analysis indicates that theβAOB abundance was a main factor to influence nitrification rates with an influence ratio of 87.7% at the level of 40% biodiversity similarity. Considering the good correlation between βAOB abundance and nitrification estimates, the abundance and diversity of βAOB community could be expected as an indirect index of nitrification activity at the study sea area in summer.

Abundance and Community Composition of Ammonia-Oxidizers in Paddy Soil at Different Nitrogen Fertilizer Rates

Ammonia oxidation, the first and rate-limiting step of nitrification, is carried out by both ammonia-oxidizing bacteria （AOB） and ammonia-oxidizing archaea （AOA）. However, the relative importance of AOB and AOA to nitrification in terrestrial ecosystems is not well understood. The aim of this study was to investigate the effect of the nitrogen input amount on abundance and community composition of AOB and AOA in red paddy soil. Soil samples of 10-20 cm （root layer soil） and 0-5 cm （surface soil） depths were taken from a red paddy. Rice in the paddy was fertilized with different rates of N as urea of N1 （75 kg N ha＂ yr-1）, N2 （150 kg N ha~ yrl）, N3 （225 kg N ha1 yrl） and CK （without fertilizers） in 2009, 2010 and 2011. Abundance and community composition of ammonia oxidizers was analyzed by real-time PCR and denaturing gradient gel electrophoresis （DGGE） based on amoA （the unit A of ammonia monooxygenase） gene. Archaeal amoA copies in N3 and N2 were significantly （P〈0.05） higher than those in CK and N1 in root layer soil or in surface soil under tillering and heading stages of rice, while the enhancement in bacterial amoA gene copies with increasing of N fertilizer rates only took on in root layer soil. N availability and soil NO3--N content increased but soil NH4＋-N content didn＇t change with increasing of N fertilizer rates. Otherwise, the copy numbers of archaeal amoA gene were higher （P〈0.05） than those of bacterial amoA gene in root lary soil or in surface soil. Redundancy discriminate analysis based on DGGE bands showed that there were no obvious differs in composition of AOA or AOB communities in the field among different N fertilizer rates. Results of this study suggested that the abundance of ammonia-oxidizers had active response to N fertilizer rates and the response of AOA was more obvious than that of AOB. Similarity in the community composition of AOA or AOB among different N fertilizer rates indicate that the community composition of ammonia-oxidizers was relatively stable in the paddy soil at least in short term for three years.

Seasonal and spatial distribution of ammonia-oxidizing microorganism communities in surface sediments from the East China Sea

Ammonia oxidation plays a significant role in the nitrogen cycle in marine sediments. Seasonal and spatial distribution of ammonia-oxidizing archaea （AOA） and betaproteobacteria （13-AOB） in surface sediments from the East China Sea （ECS） were investigated using ammonia monooxygenase ct subunit （amoA） gene. In order to characterize the community of AOA and 13-AOB, real-time quantitative polymerase chain reaction （qPCR） was carried out in this study, along with environmental parameters. The abundance of 13-AOB amoA gene （2.17x 10^6-4.54x10^7 copy numbers per gram wet weight sediment） was always greater than that ofAOA amoA gene （2.18x 105-9.89x 10^6 copy numbers per gram wet weight sediment） in all sampling stations. The qPCR results were correlated with environmental parameters. AOA amoA gene copy numbers in April were positively related to temperature and nitrite concentration （p〈0.05）. 13-AOB amoA gene copy numbers in August correlated negatively with salinity （p〈0.01）, and correlated positively with ammonium concentration （p〈0.05）. With the increase of salinity, the amoA gene copy ratio of AOB to AOA had a tendency to decrease, which suggested 13-AOB dominated in the area of high level ammonium and AOA preferred high salinity area.

Response of ammonia-oxidizing betaproteobacteria to short- term fertilization in a salt marsh in China

This study examines the impacts of short-term(6 months) fertilization on the community structure and abundance of ammonia-oxidizing betaproteobacteria(β-AOB) and the potential nitrification rate in sediment colonized by S uaeda heteroptera in a saltmarsh located in Shuangtai estuary, China. The sediment samples were collected from plots treated with different amounts of an N fertilizer(urea supplied at 0.1, 0.2, 0.4, and 0.8 g/kg(nitrogen content in dry sediment)), and with different forms of N fertilizers(urea,(NH4) 2 SO 4, and NH_4NO_3, each supplied at 0.2 g/kg). The fertilizers were applied 1–4 times during the plant-growing season in May, July, August and September of 2013. Untreated plots were included as a control. As revealed in denaturing gradient gel electrophoresis of the 16 S r RNA gene, the β-AOB community responded to both the amount and form of N. Real-time quantitative PCR indicated that both abundance and potential nitrification rate of β-AOB increased after N addition, regardless of concentration and form(except NH_4NO_3). These results provide evidence that short-term N application influences the sediment β-AOB community, β-AOB abundance and potential nitrification rate in a saltmarsh ecosystem.

Compositional Shifts in Ammonia-Oxidizing Microorganism Communities of Eight Geographically Different Paddy Soils —Biogeographical Distribution of Ammonia-Oxidizing Microorganisms

Soil nitrification is mediated by ammonia-oxidizing archaea (AOA) and bacteria (AOB), which occupy different specialized ecological niches. However, little is known about the diversification of AOA and AOB communities in a large geographical scale. Here, eight paddy soils collected from different geographic regions in China were selected to investigate the spatial distribution of AOA and AOB, and their potential nitrification activity (PNA). The result showed that the abundance of AOA was predominant over AOB, indicating that the rice fields favor the growth of AOA. PNA highly varied from 0.43 to 3.57 μg NOX-N·g·dry·soil·h-1, and was positively related with soil NH3 content, the abundance of AOA community, and negatively related with the diversity of AOB community (P amoA genes revealed remarkable differences in the compositions of AOA and AOB community. Phylogenetic analyses of amoA genes showed that Nitrosospiracluster-3-like and Nitrosomonas cluster 7-like AOB extensively dominated the AOB communities, and 54d9-like AOA within the soil group 1.1b predominated in AOA communities in paddy soils. Redundancy analysis suggested that the spatial variations of AOA community structure were influenced by soil TN content (P < 0.01), while no significant correlation between AOB community structure and soil properties was found. Findings highlight that ammonia oxidizers exhibit spatial variations in complex paddy fields due to the joint influence of soil variables associated with N availability.

Characterization of Bacterial Community,Ammonia-Oxidizing Bacteria,and Nitrospira During the Operation of a Commercial-Scale Recirculating Aquaculture System for Culturing Pufferfish Takifugu rubripes

We investigated the changes in communities of bacteria,ammonia-oxidizing bacteria,and Nitrospira during the operation of a pufferfish Takifugu rubripes recirculating aquaculture system by using high-throughput DNA sequencing.Differences in bacterial communities were observed at days 1-32,47-62 and 78-93 of biofilm development by using 16S rRNA gene pyrosequencing.The relative abundance of Proteobacteria(Gammaproteobacteria)increased,while that of Bacteroidetes(Flavobacteria)decreased.The proportions of Nitrosomonas and Nitrospina ranged from 0.02%to 0.30%and from 0.02%to 0.83%,respectively.Ammonia monooxygenase gene pyrosequencing revealed that the top three operational taxonomic units were related to Nitrosomonas aestuarii(17.5%-61.1%),uncultured beta proteobacterium clone B67S-54(1.9%-45.2%),and uncultured bacterium clone AZPa8(3.6%-24.7%).Nitrite oxidoreductase gene pyrosequencing revealed that the relative abundance of the dominant strain Nitrospira sp.Ecomares 2.1 increased,but that of the abundant species Nitrospira marina decreased.Our results demonstrated that the communities of bacteria,ammonia-oxidizing bacteria,and Nitrospira were changing during the operation of the pufferfish recirculating aquaculture system.

Distribution characteristics of ammonia-oxidizing bacteria in the Typha latifolia constructed wetlands using fluorescent in situ hybridization(FISH)

A molecular biology method, fluorescent in situ hybridization（FISH）, in which the pre-treatment was improved in allusion to the media of the constructed wetlands（CW）, e.g. the soil and the grit, was used to investigate the vertical distribution characteristics of ammonia-oxidizing bacteria（AOB） quantity and the relation with oxidation-reduction potential（ORP） in the Typha latifolia constructed wetlands under three different Ioadings in summer from May to September. Results showed that the quantity of the AOB decreased in the Typha latifolia CW with the increase of vertical depth. However, the AOB quantity was 2-4 times the quantity of the control in the root area. Additionally, ORP in the rhizosphere was found to be higher than other areas, which showed that Typha latifolia CW was in an aerobic state in summer when using simulated non-point sewage at the rural area of Taihu Lake in China and small town combined sewage.

Variation of Potential Nitrification and Ammonia-Oxidizing Bacterial Community with Plant-Growing Period in Apple Orchard Soil

In this study, we investigated the potential nitrification and community structure of soil-based ammonia-oxidizing bacteria （AOB） in apple orchard soil during different growth periods and explored the effects of environmental factors on nitrification activity and AOB community composition in the soil of a Hanfu apple orchard, using a culture-dependent technique and denaturing gradient gel electrophoresis （DGGE）. We observed that nitrification activity and AOB abundance were the highest in November, lower in May, and the lowest in July. The results of statistical analysis indicated that total nitrogen （N） content, NH4＋-N content, NO3-N content, and pH showed significant correlations with AOB abundance and nitrification activity in soil. The Shannon-Winner diversity, as well as species richness and evenness indices （determined by PCR-DGGE banding patterns） in soil samples were the highest in September, but the lowest in July, when compared to additional sampled dates. The DGGE fingerprints of soil-based 16S rRNA genes in November were apparently distinct from those observed in May, July, and September, possessing the lowest species richness indices and the highest dominance indices among all four growth periods. Fourteen DGGE bands were excised for sequencing. The resulting analysis indicated that all AOB communities belonged to the 13-Proteobacteria phylum, with the dominant AOB showing high similarity to the Nitrosospira genus. Therefore, soil-based environmental factors, such as pH variation and content of NHa＋-N and NO3--N, can substantially influence the abundance of AOB communities in soil, and play a critical role in soil-based nitrification kinetics.

Linking bacterial and archaeal community dynamics to related hydrological,geochemical and environmental characteristics between surface water and groundwater in a karstic estuary

Subterranean estuaries(STEs)are characterized by the mixing of terrestrial fresh groundwater and seawater in coastal aquifers.Although microorganisms are important components of coastal groundwater ecosystems and play critical roles in biogeochemical transformations in STEs,limited information is available about how their community dynamics interact with hydrological,geochemical and environmental characteristics in STEs.Here,we studied bacterial and archaeal diversities and distributions with 16S rRNA-based Illumina MiSeq sequencing technology between surface water and groundwater in a karstic STE.Principal-coordinate analysis found that the bacterial and archaeal communities in the areas where algal blooms occurred were significantly separated from those in other stations without algal bloom occurrence.Canonical correspondence analysis showed that nutrients and salinity can explain the patterns of bacterial and archaeal community dynamics.The results suggest that hydrological,geochemical and environmental characteristics between surface water and groundwater likely control the bacterial and archaeal diversities and distributions in STEs.Furthermore,we found that some key species can utilize terrestrial pollutants such as nitrate and ammonia in STEs,indicating that these species(e.g.,Nitrosopumilus maritimus,Limnohabitans parvus and Simplicispira limi)may be excellent candidates for in situ degradation/remediation of coastal groundwater contaminations concerned with the nitrate and ammonia.Overall,this study reveals the coupling relationship between the microbial communities and hydrochemical environments in STEs,and provides a perspective of in situ degradation/remediation for coastal groundwater quality management.

Nitrogen transformation processes in soil along a High Arctic tundra transect

Soil nitrogen(N)transformation processes in the High Arctic tundra are poorly understood even though nitrogen is one of the main limiting nutrients.We analyzed soil samples collected along a High Arctic tundra transect to investigate spatial variability in key nitrogen transformation processes,functional gene abundances,ammonia-oxidizing archaea(AOA)community structures,and key nitrogen transformation regulators.The potential denitrification rates were higher than the nitrification rates in the soil samples,although nitrification may still regulate N2O emissions from tundra soil.The nutrient(total carbon,total organic carbon,total nitrogen,and NH_(4)^(+)-N)contents were important determinants of spatial variability in the potential denitrification rates of soil along the tundra transect.The total sulfurcontent was the main variable controlling potential nitrification processes,probably in association with sulfate-reducing bacteria.The nitrate content was the main variable affecting potential dissimilatory nitrate reduction to ammonium.AOA and ammonia-oxidizing bacteria amoA,nirS,and anammox 16S rRNA genes were found in all of the soil samples.AOA play more important roles than ammonia-oxidizing bacteria in soilnitrification.Anammox bacteria may utilize NO_(2)^(-)produced through nitrification.Phylogenetic analysis indicated that the AOA amoA sequences could be grouped into eight unique operational taxonomic units(OTUs)with a 97%sequence similarity and were affiliated with three group 1.1b Nitrososphaeraclusters.The results indicated that heterogeneous environmental factors(e.g.,the carbon and nitrogen contents of soil)along the High Arctic tundra transect strongly affected the nitrogen transformation rate and relevant functional gene abundances in soil.

铁矿区内重金属对土壤氨氧化微生物群落组成的影响

以密云水库上游某铁矿区为研究对象，采用荧光定量PCR和变性梯度凝胶电泳（DGGE）分析了矿区内不同采样点的土壤中氨氧化微生物的数量和群落结构的变化，结果表明，土样中氨氧化细菌（AOB）和氨氧化古菌（AOA）的数量变化范围分别为3.01×107～1.08×109copies/g干土和8.65×107～2.69×109copies/g 干土.重金属含量与氨氧化微生物数量的相关性分析以及氨氧化微生物群落结构的冗余分析结果表明，该矿区内重金属污染改变了土壤中的氨氧化微生物的数量和结构.Cu污染对AOA的数量起到了显著抑制作用（r=-0.653＊， P〈0.05），但是对AOB则没有明显作用；Zn污染对尾矿库区域土壤的AOA/AOB比值影响显著（r=-0.606＊， P〈0.05）；Cd污染改变了AOB的种群分布，降低了AOB 的多样性水平.土壤中 Cr 长期干扰并没有改变氨氧化微生物的数量和结构，但是明显得抑制了氨氧化速率，表明重金属污染在一定程度上也影响了土壤生态系统的氮循环.

Characteristics of the microbial communities in the integrated vertical-flow constructed wetlands

Microorganisms play an important role in removing pollutants from constructed wetlands. We investigated the microbial characteristics in a novel integrated vertical-flow constructed wetland （IVCW）, which has been in operation in Wuhan, China since 1998. We used phospholipid fatty acid （PLFA） and amoA gene to analyze the structure and diversity of the microbial community within the IVCW. PLFA results suggested that the amount of bacterial PLFA was significantly higher than that of fungal PLFA, but the total microbial biomass represented by PLFA index was low in the system. Microbial spatial distribution showed significantly higher bacterial （both G^＋ and G^-） and fungal biomass in the surface than in the subsurface layers. The ratios of monounsaturated to branched PLFA demonstrated that an anaerobic layer sandwiched by two aerobic layers existed in the IVCW, consistent with the redox potential results. Analysis of the amoA revealed the presence of Nitrosomonas-like sequences in the surface substrate of the downflow chamber and apparent diversities of ammonia-oxidizing bacteria in the system. These results suggest that microorganisms, despite their relatively low biomass, have inhabited the IVCW, and the results will offer some valuable information on microbe to system designers and managers.

An innovative membrane bioreactor and packed-bed biofilm reactor combined system for shortcut nitrification-denitrification

An innovative shortcut biological nitrogen removal system, consisting of an aerobic submerged membrane bioreactor （MBR） and an anaerobic packed-bed biofilm reactor （PBBR）, was evaluated for treating high strength ammonium-bearing wastewater. The system was seeded with enriched ammonia-oxidizing bacteria （AOB） and operated without sludge purge with a decreased hydraulic retention time （HRT） through three phases. MBR was successful in both maintaining nitrite ratio over 0.95 and nitrification efficiency higher than 98% at HRT of 24 h, and PBBR showed satisfactory denitrification efficiency with very low effluent nitrite and nitrate concentration （both below 3 mg/L）. By examining the nitrification activity of microorganism, it was found that the specifc ammonium oxidization rate （SAOR） increased from 0.17 to 0.51 g N/（g VSS.d） and then decreased to 0.22 g N/（g VSS.d） at the last phase, which resulted from the accumulation of extracellular polymers substances （EPS） and inert matters enwrapping around the zoogloea. In contrast, the average specific nitrite oxidization rate （SNOR） is 0.002 g N/（g VSS.d）, only 1% of SAOR. Because very little Nitrobactor has been detected by fluorescence in situ hybridization （FISH）, it is confirmed that the stability of high nitrite accumulation in MBR is caused by a large amount of AOB.

Irrigation water salinity and N fertilization:Effects on ammonia oxidizer abundance, enzyme activity and cotton growth in a drip irrigated cotton field

Use of saline water in irrigated agriculture has become an important means for alleviating water scarcity in arid and semi-arid regions. The objective of this field experiment was to evaluate the effects of irrigation water salinity and N fertilization on soil physicochemical and biological properties related to nitrification and denitrification. A 3×2 factorial design was used with three levels of irrigation water salinity（0.35, 4.61 and 8.04 d S m-1） and two N rates（0 and 360 kg N ha^（-1））. The results indicated that irrigation water salinity and N fertilization had significant effects on many soil physicochemical properties including water content, salinity, p H, NH_4-N concentration, and NO_3-N concentration. The abundance（i.e., gene copy number） of ammonia-oxidizing archaea（AOA） was greater than that of ammonia-oxidizing bacteria（AOB） in all treatments. Irrigation water salinity had no significant effect on the abundance of AOA or AOB in unfertilized plots. However, saline irrigation water（i.e., the 4.61 and 8.04 d S m-1 treatments） reduced AOA abundance, AOB abundance and potential nitrification rate in N fertilized plots. Regardless of N application rate, saline irrigation water increased urease activity but reduced the activities of both nitrate reductase and nitrite reductase. Irrigation with saline irrigation water significantly reduced cotton biomass, N uptake and yield. Nitrogen application exacerbated the negative effect of saline water. These results suggest that brackish water and saline water irrigation could significantly reduce both the abundance of ammonia oxidizers and potential nitrification rates. The AOA may play a more important role than AOB in nitrification in desert soil.

Shifts in community structure and function of ammoniaoxidizing archaea in biological soil crusts along a revegetation chronosequence in the Tengger Desert

Metagenomic studies have demonstrated the existence of ammonia-oxidizing archaea(AOA) and revealed they are responsible for ammoxidation in some extreme environments. However, the changes in compositional structure and ammonia-oxidation capacity of AOA communities in biological soil crusts(BSCs) of desert ecosystems remain poorly understood.Here, we utilized Illumina MiSeq sequencing and microbial functional gene array(GeoChip 5.0) to assess the above changes along a 51-year revegetation chronosequence in the Tengger Desert, China. The results showed a significant difference in AOA-community richness between 5-year-old BSCs and older ones. The most dominant phylum during BSC development was Crenarchaeota, and the corresponding species were ammonia-oxidizing_Crenarchaeote and environmental_samples_Crenarchaeota. Network analysis revealed that the positive correlations among dominant taxa increased, and their cooperation was reinforced in AOA communities during BSC succession. Redundancy analysis showed that the dominant factor influencing the change in AOA-community structure was soil texture. GeoChip 5.0 indicated that the amoA gene abundances of AOA and ammonia-oxidizing bacteria(AOB) were basically the same, demonstrating that AOA and AOB played an equally important role during BSCs development. Our study of the long-term succession of BSC demonstrated a persistent response of AOA communities to revegetation development in desert ecosystems.

Environmental similarity is more important than distance in the community structuring processes of ammonia-oxidizing archaea in agricultural soils

Ammonia-oxidizing archaea(AOA) are important in converting ammonia into nitrate in soils. While many aspects of their community structure have been studied, the relative importance of stochastic versus deterministic processes has poorly been understood. We compared AOA communities across the North China Plain, targeting the amoA gene. A phylogenetic null modelling approach was used to calculate the beta nearest taxon index to quantify the influence of stochastic and deterministic processes. We found that spatial distance between samples predicted the perceived processes involved in community structuring, with stochastic processes dominating at local scales. At greater distances, stochasticity became weaker. However, soil pH, which was also the strongest determinant of AOA community, was a much stronger predictor of community structuring, leaving the distance effect redundant as an explanation of community structuring processes. The communities of AOA differing by less than 1 pH unit differed mainly stochastically in terms of operational taxonomic unit composition. At larger pH differences, deterministic processes based on heterogeneous selection between clades became increasingly dominant. It appears that AOA community composition is largely determined by the environment. However, very similar pH environments are the exception. In environments with very close pH values, stochastic effects dominantly cause differences in community composition, whether spatially near or far.

Functional relationship between ammonia-oxidizing bacteria and ammonia-oxidizing archaea populations in the secondary treatment system of a full-scale municipal wastewater treatment plant

The abundance of ammonia-oxidizing bacteria and archaea and their amo A genes from the aerobic activated sludge tanks,recycled sludge and anaerobic digesters of a full-scale wastewater treatment plant(WWTP)was determined.Polymerase chain reaction and denaturing gradient gel electrophoresis were used to generate diversity profiles,which showed that each population had a consistent profile although the abundance of individual members varied.In the aerobic tanks,the ammonia-oxidizing bacterial(AOB)population was more than 350 times more abundant than the ammonia-oxidizing archaeal(AOA)population,however in the digesters,the AOA population was more than 10 times more abundant.Measuring the activity of the amo A gene expression of the two populations using RT-PCR also showed that the AOA amo A gene was more active in the digesters than in the activated sludge tanks.Using batch reactors and dd PCR,amo A activity could be measured and it was found that when the AOB amo A activity was inhibited in the anoxic reactors,the expression of the AOA amo A gene increased fourfold.This suggests that these two populations may have a cooperative relationship for the oxidation of ammonia.