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A betaproteobacterial chemolithotrophic ammonia-oxidizing bacterium designated APG5T was isolated from supralittoral sand of the Edmonds City Beach, WA, USA. Growth was observed at 10–35 °C (optimum, 30 °C), pH 5–9 (optimum, pH 8) and ammonia concentrations as high as 100 mM (optimum, 1–30 mM NH4Cl). The strain grows optimally in a freshwater medium but tolerates up to 400 mM NaCl. It is most closely related to ‘Nitrosomonas ureae’ (96.7% 16S rRNA and 92.4% amoA sequence identity). The 3.75-Mbp of AGP5T draft genome contained a single rRNA operon and all necessary tRNA genes and has the lowest G+C content (43.5%) when compared to the previously reported genomes of reference strains in cluster 6 Nitrosomonas. Based on an average nucleotide identity of 82% with its closest relative (‘N. ureae’ Nm10T) and the suggested species boundary of 95–96%, a new species Nitrosomonas supralitoralis sp. nov. is proposed. The type strain of Nitrosomonas supralitoralis is APG5T (= NCIMB 14870T = ATCC TSD-116T). [ABSTRACT FROM AUTHOR]

Nitrogen species such as ammonia and nitrite are considered as major stressors in modern aquaculture practices. We developed enrichments of ammonia oxidising bacteria (AOB) and nitrite oxidising bacteria (NOB) for effective mitigation of nitrogenous wastes in the shrimp culture operations. The objective of this study was to understand the microbial community composition of AOB and NOB enrichments using the V3-V4 region of the 16S rDNA gene by Illumina MiSeq sequencing. The analysis revealed 2948 and 1069 OTUs at 97% similarity index and Shannon alpha diversity index of 7.64 and 4.85 for AOB and NOB enrichments, respectively. Comparative analysis showed that a total of 887 OTUs were common among AOB and NOB enrichments. The AOB and NOB enrichment were dominated by Eubacteria at 96% and 99.7% respectively. Proteobacterial phylum constituted 31.46% (AOB) and 39.75% (NOB) and dominated by α-Proteobacteria (20%) in AOB and γ-Proteobacteria (16%) in NOB. Among the species in AOB enrichment (2,948) two sequences were assigned to ammonia oxidising bacterial group belonging to Nitrosomonas, and Nitrosococcus genera and two belonged to archaeon group comprising Nitrosopumilus and Candidatus Nitrososphaeraea genera. The NOB enrichment was predominated by Nitrospiraceae and Thermodesulfovibrionaceae. Further, the data revealed the presence of heterotrophic bacteria contributing to the process of nitrification and form microcosm with the AOB and NOB. PICRUSt analysis predicted the presence of 24 different nitrogen cycling genes involved in nitrification, denitrification, ammonia and nitrogen transporter family, nitrate reduction and ammonia assimilation. The study confirms the presence of many lesser known nitrifying bacteria along with well characterised nitrifiers. [ABSTRACT FROM AUTHOR]

To understand the effects of biochar and urea on soil N availability and plant growth, we conducted a pot experiment growing barley (Hordeum vulgare L.) under six treatments: control (N0), soil with 30 g kg−1 biochar (N0B), soil with 0.23 g kg−1 urea (N1), soil with 0.23 g kg−1 urea and 30 g kg−1 biochar (N1B), soil with 0.46 g kg−1 urea (N2), and soil with 0.46 g kg−1 urea and 30 g kg−1 biochar (N2B). The nitrifying community abundance and compositions in rhizosphere and bulk soils were analyzed using quantitative polymerase chain reaction (qPCR) and amplicon-based Illumina Hiseq sequencing. Adding urea with biochar (N1B) produced the greatest increase in above- and belowground plant biomass, followed by doubling the amount of urea with biochar (N2B); both treatments raised pH (p < 0.001) and lowered extractable N in the rhizosphere (p < 0.05). N1B treatment produced the greatest increase in ammonia-oxidizing bacteria (AOB) amoA gene copies, presumably because the combined amendment raised soil pH, which favored AOB access to NH4+. Nitrifier sequences were selected after blasting with reported nitrifiers in NCBI (similarity ≥ 97%). Nitrosospira dominated AOB communities during the plant seedling stage; however, during the mature stage, Nitrosomonas dominated over Nitrosospira and the nitrite-oxidizing bacteria (NOB) community became diverse. Redundancy analysis indicated that nitrifying community composition was affected by multiple soil properties, including N availability (i.e., exchangeable NH4+ and NO3−) and soil chemistry (i.e., pH, dissolved organic C, and exchangeable base cations). Our research suggests a positive application of combining biochar with urea in improving N bioavailability and promoting plant growth in the acidic soil. [ABSTRACT FROM AUTHOR]

Ammonia-oxidizing bacteria (AOB) within the genus Nitrosomonas perform the first step in nitrification, ammonia oxidation, and are found in diverse aquatic and terrestrial environments. Nitrosomonas AOB were grouped into six defined clusters, which correlate with physiological characteristics that contribute to adaptations to a variety of abiotic environmental factors. A fundamental physiological trait differentiating Nitrosomonas AOB is the adaptation to either low (cluster 6a) or high (cluster 7) ammonium concentrations. Here, we present physiological growth studies and genome analysis of Nitrosomonas cluster 6a and 7 AOB. Cluster 6a AOB displayed maximum growth rates at ≤ 1 mM ammonium, while cluster 7 AOB had maximum growth rates at ≥ 5 mM ammonium. In addition, cluster 7 AOB were more tolerant of high initial ammonium and nitrite concentrations than cluster 6a AOB. Cluster 6a AOB were completely inhibited by an initial nitrite concentration of 5 mM. Genomic comparisons were used to link genomic traits to observed physiological adaptations. Cluster 7 AOB encode a suite of genes related to nitrogen oxide detoxification and multiple terminal oxidases, which are absent in cluster 6a AOB. Cluster 6a AOB possess two distinct forms of ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) and select species encode genes for hydrogen or urea utilization. Several, but not all, cluster 6a AOB can utilize urea as a source of ammonium. Hence, although Nitrosomonas cluster 6a and 7 AOB have the capacity to fulfill the same functional role in microbial communities, i.e., ammonia oxidation, differentiating species-specific and cluster-conserved adaptations is crucial in understanding how AOB community succession can affect overall ecosystem function. [ABSTRACT FROM AUTHOR]

Ammonia emission during composting results in anthropogenic odor nuisance and reduces the agronomic value of the compost due to the loss of nitrogen. Adjusting the operating parameters during composting is an emerging in situ odor control technique that is cheap and highly efficient. The effects of in situ NH3 emission control were investigated in this study by simultaneously adjusting key operating parameters (such as C/N ratio, aeration rate, and moisture content) during the composting processes (C1–C9). Results showed that the average NH3 emission concentrations for different treatments were in the order of C1 > C4 > C2 > C5 > C3 > C6 > C7 > C8 > C9. The total content of NH3 emission (21.02 g/kg) in C9 (C/N ratio = 35, aeration rate = 15 L/min, and moisture content = 60%) was much lower than that (65.95 g/kg) in C1 (C/N ratio = 15, aeration rate = 5 L/min, and moisture content = 60%). The nitrogen loss ratio was 27.36% for C1, while 16.15% for C9. The microbial diversity and abundance in C9 and C1 were compared using high-throughput sequencing. The relationship between NH3 emission, operating parameters, and the related functional microbial communities was also investigated. Results revealed that Nitrosospira, Nitrosomonas, Nitrobacter, Pseudomonas, Methanosaeta, Rhodobacter, Paracoccus, and Sphingobacterium were negatively related to NH3 emission. According to the above results, the optimal values for different operating parameters for the in situ NH3 control during kitchen waste composting were, respectively, moisture content of 70%, C/N ratio of 35, and aeration rate of 15 L/min, with the order of effectiveness from high to low being aeration rate > C/N > moisture. This information could be used as a valuable reference for the in situ NH3 emission control during kitchen waste composting. [ABSTRACT FROM AUTHOR]

Aims: Root exudates of rice (Oryza sativa L.) can inhibit nitrification in Nitrosomonas bioassays, and 1,9-decanediol was recently identified as an important new biological nitrification inhibitor (BNI) from rice. However, the release characteristics of 1,9-decanediol have not been studied. The present study was designed to identify the major factors influencing the release of 1,9-decanediol from rice roots.Methods: Rice plants were hydroponically grown in controlled environment chambers for 6 weeks, and root exudates were collected. Responses of exudate release to nitrogen form and concentration, pH, aeration, and bacterial inoculation were explored. The pH of root exudates, collected under different nitrogen-provision regimes, was determined, and 1,9-decanediol levels in exudates were monitored.Results: Ammonium (NH4+) and low pH in the root environment stimulated the release of 1,9-decanediol from rice roots. When only a part of the root system was exposed to NH4+, the secretion of 1,9-decanediol was triggered in the whole root system. Aeration of the root environment significantly enhanced 1,9-decanediol release. The presence of two major nitrifiers (Nitrosomonas europaea and Nitrosomonas stercoris) in the root medium stimulated release of 1,9-decanediol, whereas denitrifiers had no effect.Conclusions: Our results demonstrate that the release of 1,9-decanediol is enhanced by low to moderate concentrations of NH4+ (≤1.0 mM), low pH, and aeration of the rhizosphere. Our study provides the first evidence of significant 1,9-decanediol secretion induced by nitrifying bacteria. [ABSTRACT FROM AUTHOR]

Aquaculture ponds are simple and unique ecosystems, which are affected intensively by human activities. In this mini-review, we focus our attention on the distribution and community diversity of ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA) in pond water and sediments, as well as the possible ecological mechanisms involved. Moreover, we discuss the possibility of increasing the activity of ammonia-oxidizing organisms in order to improve the water quality in aquaculture ponds. Compared with eutrophic lakes, the significantly higher ammonia concentration in pond water does not lead to significantly higher AOB levels, and the abundance of AOA is too low to quantify accurately. Similar to eutrophic lakes, high abundances of AOA and AOB are present in the surface sediments at the same time, where the oxidation of ammonia is performed mainly by AOB. AOB and AOA exhibit significant seasonal variations in aquaculture ponds, which are affected by the temperature, pH, and dissolved oxygen. The dominant AOB species are Nitrosomonas and the Nitrosospira lineage in pond environments. Nitrososphaera or members of the Nitrososphaera-like cluster dominate the AOA species in surface sediments, whereas the Nitrosopumilus cluster dominates the deeper sediments. AOB and AOA can be enriched on artificial substrates suspended in the pond water, thereby potentially improving the water quality. [ABSTRACT FROM AUTHOR]

Nitrification at the site of a contaminant ammonium plume from a former coal carbonisation plant can be modelled with three competing bacterial populations of Nitrosomonas, Nitrobacter, and Brocadia anammoxidans. Oscillations of chemical species at the site can be explained by a reduced model of ammonium competition between Nitrosomonas and B. anammoxidans which effectively acts as an activator-inhibitor system. Stable oscillations occur in conditions of low nutrient (ammonium) supply and this causes a spatial travelling wave in a borehole profile when diffusion is introduced. [ABSTRACT FROM AUTHOR]

For the possible highest performance of single-stage combined partial nitritation/anammox (PNA) process, a continuous complete-mix granular reactor was operated at progressively higher nitrogen loading rate. The variations in bacterial community structure of granules were also characterized using high-throughput pyrosequencing, to give a detail insight to the relationship between reactor performance and functional organism abundance within completely autotrophic nitrogen removal system. In 172 days of operation, a superior total nitrogen (TN) removal rate over 3.9 kg N/(m3/day) was stable implemented at a fixed dissolved oxygen concentration of 1.9 mg/L, corresponding to the maximum specific substrate utilization rate of 0.36/day for TN based on the related kinetics modeling. Pyrosequencing results revealed that the genus Nitrosomonas responsible for aerobic ammonium oxidation was dominated on the granule surface, which was essential to offer the required niche for the selective enrichment of anammox bacteria (genus Candidatus Kuenenia) in the inner layer. And the present of various heterotrophic organisms with general functions, known as fermentation and denitrification, could not be overlooked. In addition, it was believed that an adequate excess of ammonium in the bulk liquid played a key role in maintaining process stability, by suppressing the growth of nitrite-oxidizing bacteria through dual-substrate competitions. [ABSTRACT FROM AUTHOR]

The proper use of selective ammonia-oxidizing archaea (AOA) and/or ammonia-oxidizing bacteria (AOB) inhibitors is critical to distinguish AOA and AOB contribution. In this research, three inhibitors including ampicillin, dicyandiamide (DCD), and allylthiourea (ATU) were examined mainly focusing on inhibiting dosage, adaptability, and effects. The results showed that the optimized inhibitory dosage of ampicillin, DCD, and ATU was separately 1.5 g L, 1 mM, and 25 μM. Among the three inhibitors, ATU exhibited the strongest and persistent inhibition effects and resulted in up to 90% inhibition in the AOB-enriched culture. The seemingly weakening inhibiting effects of ATU in the simulated river systems can be attributed to the involved role of AOA, the uneven spatial distribution of ATU, and protection by sediment structure in complex malodorous rivers. The high-throughput pyrosequencing analysis showed the AOB-related genus Nitrosomonas and Nitrosococcus were mostly affected by ATU in the enrichments and the river systems, respectively. The inhibition of ATU was realized mainly by reducing the abundance and activity of AOB. The decrease of the ratio of AOB/AOA amoA gene copy numbers after addition of ATU further confirmed the inhibiting effectiveness of ATU in complex microbial community of malodorous rivers. [ABSTRACT FROM AUTHOR]

Ammonia-oxidizing bacteria (AOB) play an important role in nitrification in estuaries. The aim of this study was to examine the spatial abundance, diversity, and activity of AOB in coastal sediments of the Liaohe Estuary using quantitative PCR, high-throughput sequencing of the amoA gene coding the ammonia monooxygenase enzyme active subunit, and sediment slurry incubation experiments. AOB abundance ranged from 8.54 × 10 to 5.85 × 10 copies g of wet sediment weight and exhibited an increasing trend from the Liaohe Estuary to the open coastal zone. Potential nitrification rates (PNRs) ranged from 0.1 to 336.8 nmol N g day along the estuary to the coastal zone. Log AOB abundance and PNRs were significantly positively correlated. AOB richness decreased from the estuary to the coastal zone. High-throughput sequencing analysis indicated that the majority of amoA gene sequences fell within the Nitrosomonas and Nitrosomonas-like clade, and only a few sequences were clustered within the Nitrosospira clade. This finding indicates that the Nitrosomonas-related lineage may be more adaptable to the specific conditions in this estuary than the Nitrosospira lineage. Sites with high nitrification rates were located in the southern open region and were dominated by the Nitrosomonas-like lineage, whereas the Nitrosospira lineage was found primarily in the northern estuary mouth sites with low nitrification rates. Thus, nitrification potentials in Liaohe estuarine sediments in the southern open region were greater than those in the northern estuary mouth, and the Nitrosomonas-related lineage might play a more important role than the Nitrosospira lineage in nitrification in this estuary. [ABSTRACT FROM AUTHOR]

Partial nitrification (PN) has been considered as one of the promising processes for pretreatment of ammonium-rich wastewater. In this study, a kind of novel carriers with enhanced hydrophilicity and electrophilicity was implemented in a moving bed biofilm reactor (MBBR) to start up PN process. Results indicated that biofilm formation rate was higher on modified carriers. In comparison with the reactor filled with traditional carriers (start-up period of 21 days), it took only 14 days to start up PN successfully with ammonia removal efficiency and nitrite accumulation rate of 90 and 91%, respectively, in the reactor filled with modified carriers. Evident changes of spatial distributions and community structures had been detected during the start-up. Free-floating cells existed in planktonic sludge, while these microorganisms trended to form flocs in the biofilm. High-throughput pyrosequencing results indicated that Nitrosomonas was the predominant ammonia-oxidizing bacterium (AOB) in the PN system, while Comamonas might also play a vital role for nitrogen oxidation. Additionally, some other bacteria such as Ferruginibacter, Ottowia, Saprospiraceae, and Rhizobacter were selected to establish stable footholds. This study would be potentially significant for better understanding the microbial features and developing efficient strategies accordingly for MBBR-based PN operation. [ABSTRACT FROM AUTHOR]

Nitrification activities and microbial populations of ammonium oxidizing bacteria (AOB) and nitrite oxidizing bacteria (NOB) were investigated in 10 full-scale biological nutrient removal wastewater treatment plants in Xi'an, China. Aerobic batch tests were used to determine the nitrifying activities while fluorescence in situ hybridization was used to quantify the fractions of AOB and NOB in the activated sludge. The results showed that nitrifying bacteria accounted for 1-10% of the total population. Nitrosomonas and Nitrospira were the dominant bacteria for AOB and NOB respectively. Moreover, the average percentage of AOB was 1.27% and that of NOB was 4.02%. The numerical ratios of NOB/AOB varied between 1.72 and 5.87. The average ammonium uptake rate and nitrite uptake rate were 3.25 ± 0.52 mg (NH -N)/g(VSS) h and 4.49 ± 0.49 mg (NO -N)/g(VSS) h, respectively. Correspondingly, the activity of NOB was 1.08-2.00 times higher than that of AOB. Thus, NOB was the dominating bacteria in nitrifying communities. The year-round data of Dianzicun (W6) also expressed a similar trend. Since NOB had higher activities than that of AOB, a large nitrite oxidation pool could be formed, which guaranteed that no nitrite would be accumulated. Therefore, stable nitrification could be achieved. A conceptual model was proposed to describe the population variation of AOB and NOB in a nitrifying community. [ABSTRACT FROM AUTHOR]

The nitrogen cycle has been expanded with the recent discovery of Nitrospira strains that can conduct complete ammonium oxidation (commamox). Their importance in the nitrogen cycle within engineered ecosystems has not yet been analyzed. In this research, the community structure of the Bacteria domain of six full-scale activated sludge systems and three autotrophic nitrogen removal systems in the Netherlands and China has been investigated through tag-454-pyrosequencing. The phylogenetic analyses conducted in the present study showed that just a few of the Nitrospira sequences found in the bioreactors were comammox. Multivariate redundancy analysis of nitrifying genera showed an outcompetition of Nitrosomonas and non-comammox Nitrospira. Operational data from the bioreactors suggested that comammox could be favored at low temperature, low nitrogen substrate, and high dissolved oxygen. The non-ubiquity and low relative abundance of comammox in full-scale bioreactors suggested that this phylotype is not very relevant in the nitrogen cycle in wastewater treatment plants. [ABSTRACT FROM AUTHOR]

NO emission in the anaerobic ammonium oxidation (anammox) process is of growing concern. In this study, effects of substrate concentrations on NO emissions were investigated in an anammox reactor. Extremely high NO emissions of 1.67 % were led by high NH-N concentrations. Results showed that NO emissions have a positive correlation with NH-N concentrations in the anammox reactor. Reducing NH-N concentrations by recycling pump resulted in decreasing NO emissions. In addition, further studies were performed to identify a key biological process that is contributed to NO emissions from the anammox reactor. Based on the results obtained, Nitrosomonas, which can oxidize ammonia to nitrite, was deemed as the main sources of NO emissions. [ABSTRACT FROM AUTHOR]

The effects of free ammonia (FA) and dissolved oxygen (DO) on nitrite accumulation in the treatment of high ammonium wastewater and on the evolution of the microbial community were investigated. Under high DO conditions (3.75 ± 0.49 mg/L), FA as high as 10.61 ± 2.89 mg NH/L maintained stable nitrite accumulation rate (NAR) of 84 % with NH-N load of 2.05 kg N/(m day) at sludge retention time (SRT) of 15-18 days. After 56 days of operation, Proteobacteria and Nitrosomonas were the dominant phylum and genus, respectively; Nitrosomonas increased from 21.14 to 54.57 %. By contrast, under relative low DO and low FA, nitrite-oxidizing bacteria (NOB) were nearly eliminated (NOB/AOB of 0; ammonium-oxidizing bacteria (AOB)), and NAR of 94 % was achieved with lower NH-N load of 0.48 kg N/(m day). DO correlated with AOB and NOB abundance, and FA decreased NOB activity and the NOB/AOB ratio. In conclusion, high FA and high DO conditions are optimal for efficient nitrite accumulation. [ABSTRACT FROM AUTHOR]

Nitrogen pollutants in low-organic carbon wastewater are difficult to biodegrade. Therefore, the Fe(0)-carbon-based bio-carrier (FCBC) was firstly used as hydrogen producer in a biological-aerated filter (BAF) to make up for the lack of organic carbon in biological nitrogen removal. Physical and chemical properties of FCBC were detected and compared in this study. The nitrogen removal rate for low COD/TN ratio wastewater, nitrogen transformation process, and microbial communities in the FCBC filled in BAF were investigated. Results showed that the nitrogen removal rates was 0.38-0.41 kg N m day in the FCBC filled BAF and reached 0.62 kg N m day within the filter depth of 60-80 cm, under the conditions of the dissolved oxygen 3.5 ± 0.2 mg L and the inlet pH 7.2 ± 0.1. Hydrogenophaga (using hydrogen as electron donor), Sphaerotilus (absorbing [Fe]), Nitrospira (nitrificaion), and Nitrosomonas (ammonia oxidation) were found to be the predominant genera in the reactor. The reaction schemes in the FCBC filled in BAF was calculated: hydrogen and [Fe] were produced by Fe(0)-C galvanic cells in the FCBC, ammonia was oxidized into nitrate by Nitrosomonas and Nitrospira genera, hydrogen was used as electron donors by Hydrogenophaga genus to reduce nitrate into N, and [Fe] was partly absorbed by Sphaerotilus and diverted via sludge discharging. [ABSTRACT FROM AUTHOR]

It has been demonstrated that an anaerobic digestion process cannot attain an efficient removal of several amino acids, with methionine being one of the most persistent of these. Thus, the effect that methionine amino acid has over the partial-nitritation process with fixed-biofilm configuration in terms of performance and bacterial community dynamics has been investigated. With respect to the performance with no addition, 100 mg/L methionine loading decreased ammonium oxidation efficiency in 60 % and 100 % at concentrations of 300 and 500 mg/L methionine, respectively. Bacterial biomass sharply increased by 30, 65, and 230 % with the addition of 100, 300, and 500 mg/L methionine, respectively. Bacterial community analysis showed that methionine addition supported the proliferation of a diversity of heterotrophic genera, such as Lysobacter and Micavibrio, and reduced the relative abundance of ammonium oxidizing genus Nitrosomonas. This research shows that the addition of methionine affects the performance of the partial-nitritation process. In this sense, amino acids can pose a threat for the of partial-nitritation process treating anaerobic digester supernatant at full-scale implementation. [ABSTRACT FROM AUTHOR]

Both ammonia-oxidizing bacteria (AOB) and archaea (AOA) can play important roles in ammonia biotransformation in ecosystems. However, the factors regulating the distribution of these microorganisms in lacustrine ecosystems remain essentially unclear. The present study investigated the effects of geographic location on the distribution of sediment AOA and AOB in 13 freshwater lakes on the Yunnan Plateau (China). The spatial dissimilarity in the abundance and structure of sediment AOA and AOB communities was observed in these plateau lakes. AOA abundance was usually less than AOB abundance, and the AOA/AOB ratio was positively correlated with water depth. Nitrososphaera-like AOA occurred in most of the studied lakes and were dominant in two lakes. Nitrosospira was the dominant AOB species in most of the lakes, while Nitrosomonas showed high abundance only in three lakes. In addition, geographic location was found to affect lake sediment AOB community structure. [ABSTRACT FROM AUTHOR]

Under the increasing pressure of human activities, Hangzhou Bay has become one of the most seriously polluted waters along China's coast. Considering the excessive inorganic nitrogen detected in the bay, in this study, the impact of an effluent from a coastal industrial park on ammonia-oxidizing microorganisms (AOMs) of the receiving area was interpreted for the first time by molecular technologies. Revealed by real-time PCR, the ratio of archaeal amoA/bacterial amoA ranged from 5.68 × 10 to 4.79 × 10 in the activated sludge from two wastewater treatment plants (WWTPs) and 0.54-3.44 in the sediments from the effluent receiving coastal area. Analyzed by clone and pyrosequencing libraries, genus Nitrosomonas was the predominant ammonia-oxidizing bacteria (AOB), but no ammonia-oxidizing archaea (AOA) was abundant enough for sequencing in the activated sludge from the WWTPs; genus Nitrosomonas and Nitrosopumilus were the dominant AOB and AOA, respectively, in the coastal sediments. The different abundance of AOA but similar structure of AOB between the WWTPs and nearby coastal area probably indicated an anthropogenic impact on the microbial ecology in Hangzhou Bay. [ABSTRACT FROM AUTHOR]

Purpose: The aim of this study was to elucidate the impact of plant on the activity, abundance, and the community composition of the ammonia oxidizers, including both ammonia-oxidizing archaea (AOA) and bacteria (AOB). Moreover, the relationship between AOA and AOB in mangrove sediment was also analyzed. Materials and methods: Sediment used for microcosm experiments was collected in the mangrove wetland. The native plant species Kandelia obovata and invasive species Spartina alterniflora were selected. Nitrifying activity was determined by assaying the potential nitrification rate (PNR). Abundances of gene and transcript were measured via real-time quantitative PCR (qPCR). Terminal restriction fragment length polymorphism (T-RFLP) was used to analyze the nitrifier community structures. Clone libraries were constructed for further phylogenetic analysis. Results and discussion: PNR and abundances of both AOA and AOB were greatly enhanced in the vegetated sediments. S. alterniflora showed a greater promoting effect on nitrification activity, indicating the potential of exotic invasion on perturbing the nitrogen balance. Abundance of AOB transcript was hundreds of times higher than that of AOA. Moreover, ammonia-oxidizing communities were distinctly grouped responding to vegetation with two plant species. Along with direct impact of plants, these variations are as well related to the different sediment properties. Phylogenetic analysis revealed that both AOA and AOB communities formed apparent clusters. The latter contained two Nitrosomonas and Nitrosospira clusters. The Nitrosomonas genus is predominant over Nitrosospira genus, which might be due to the better adaption of Nitrosomonas to the flooded habitats in sediments. Conclusions: Wetland plants were found to influence the activity, abundance, and community structure of ammonia oxidizers. Moreover, AOB is suggested to be more significant than AOA in nitrogen cycling in mangrove wetlands. [ABSTRACT FROM AUTHOR]

Community structures of ammonia-oxidizing microorganisms were investigated using PCR primers designed to specifically target the ammonia monooxygenase α-subunit ( amoA) gene in the sediment of Jinshan Lake. Relationships between the abundance and diversity of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB), and physicochemical parameters were also explored. The AOA abundance decreased sharply from west to east; however, the AOB abundance changed slightly with AOB outnumbering AOA in two of the four sediment samples (JS), JS3 and JS4. The AOA abundance was significantly correlated with the NH-N, NO-N, and TP. No significant correlations were observed between the AOB abundance and environmental variables. AOB had a higher diversity and richness of amoA genes than AOA. Among the 76 archaeal amoA sequences retrieved, 57.89, 38.16, and 3.95 % fell within the Nitrosopumilus, Nitrososphaera, and Nitrososphaera sister clusters, respectively. The 130 bacterial amoA gene sequences obtained in this study were grouped with known AOB sequences in the Nitrosomonas and Nitrosospira genera, which occupied 72.31 % and 27.69 % of the AOB group, respectively. Compared to the other three sample sites, the AOA and AOB community compositions at JS4 showed a large difference. This work could enhance our understanding of the roles of ammonia-oxidizing microorganisms in freshwater lake environment. [ABSTRACT FROM AUTHOR]

In this study, the effect of the organic micropollutant and known inhibitor of nitrification, p-cresol, was investigated on the metabolism of the ammonia oxidizing bacteria (AOB) Nitrosomonas eutropha C91 using MS-based quantitative proteomics. Several studies have demonstrated that AOB are capable of biotransforming a wide variety of aromatic compounds making them suitable candidates for bioremediation, yet the underlying molecular mechanisms are poorly described. The effect of two different concentrations of the aromatic micropollutant p-cresol (1 and 10 mg L) on the metabolism of N. eutropha C91, relative to a p-cresol absent control, was investigated. Though the rate of nitrification in N. eutropha C91 appeared essentially unaffected at both concentrations of p-cresol relative to the control, the expressional pattern of the proteins of N. eutropha C91 changed significantly. The presence of p-cresol resulted in the repressed expression of several key proteins related to N-metabolism, seemingly impairing energy production in N. eutropha C91, contradicting the observed unaltered rates of nitrification. However, the expression of proteins of the TCA cycle and proteins related to xenobiotic degradation, including a p-cresol dehydrogenase, was found to be stimulated by the presence of p-cresol. This indicates that N. eutropha C91 is capable of degrading p-cresol and that it assimilates degradation intermediates into the TCA cycle. The results reveal a pathway for p-cresol degradation and subsequent entry point in the TCA cycle in N. eutropha C91. The obtained data indicate that mixotrophy, rather than cometabolism, is the major mechanism behind p-cresol degradation in N. eutropha C91. [ABSTRACT FROM AUTHOR]

Nitrification is an integral part of biological nitrogen removal processes and usually the limiting step in wastewater treatment systems. Since nitrification is often considered not feasible at temperatures higher than 40 °C, warm industrial effluents (with operating temperatures higher than 40 °C) need to be cooled down prior to biological treatment, which increases the energy and operating costs of the plants for cooling purposes. This study describes the occurrence of thermophilic biological nitrogen removal activity (nitritation, nitratation, and denitrification) at a temperature as high as 50 °C in an activated sludge wastewater treatment plant treating wastewater from an oil refinery. Using a modified two-step nitrification-two-step denitrification mathematical model extended with the incorporation of double Arrhenius equations, the nitrification (nitrititation and nitratation) and denitrification activities were described including the cease in biomass activity at 55 °C. Fluorescence in situ hybridization (FISH) analyses revealed that Nitrosomonas halotolerant and obligatehalophilic and Nitrosomonas oligotropha (known ammonia-oxidizing organisms) and Nitrospira sublineage II (nitrite-oxidizing organism (NOB)) were observed using the FISH probes applied in this study. In particular, this is the first time that Nitrospira sublineage II, a moderatedly thermophilic NOB, is observed in an engineered full-scale (industrial) wastewater treatment system at temperatures as high as 50 °C. These observations suggest that thermophilic biological nitrogen removal can be attained in wastewater treatment systems, which may further contribute to the optimization of the biological nitrogen removal processes in wastewater treatment systems that treat warm wastewater streams. [ABSTRACT FROM AUTHOR]

Coexistence of an autotrophic ammonia-oxidizing bacterium ( Nitrosomonas sp. RA) and heterotrophic bacteria was consistently observed when cultured in an inorganic medium without any external supply of organic carbon. The present study was undertaken to understand the association between autotrophs and the associated heterotrophs for which a system containing active autotrophs and heterotrophs controlled by Hg addition was developed. The study revealed interdependence of heterotrophs and Nitrosomonas sp. RA for growth under iron-limited condition. Growth of Nitrosomonas sp. RA was supported by siderophores produced by the associated heterotroph, Pusillimonas sp., thereby complementing its high iron requirement while the organics (such as pyruvate) excreted by Nitrosomonas sp. RA during its autotrophic growth supported the survival of heterotrophs in the inorganic medium. The study thus sheds light on the nature of the mutual interactions between heterotrophs and autotrophs that play a role in the ammonia-oxidizing system involved in wastewater treatment. [ABSTRACT FROM AUTHOR]

The autotrophic nitrifying bacterium Nitrosomonas europaea does not synthesize 2-oxoglutarate (α-ketoglutarate) dehydrogenase under aerobic conditions and so has an incomplete citric acid cycle. l-malate ( S-malate) dehydrogenase (MDH) from N. europaea was predicted to show similarity to the NADP-dependent enzymes from chloroplasts and was separated from the NAD-dependent proteins from most other bacteria or mitochondria. MDH activity in a soluble fraction from N. europaea ATCC 19718 was measured spectrophotometrically and exhibited simple Michaelis-Menten kinetics. In the reductive direction, activity with NADH increased from pH 6.0 to 8.5 but activity with NADPH was consistently lower and decreased with pH. At pH 7.0, the K for oxaloacetate was 20 μM; the K for NADH was 22 μM but that for NADPH was at least 10 times higher. In the oxidative direction, activity with NAD increased with pH but there was very little activity with NADP. At pH 7.0, the K for l-malate was 5 mM and the K for NAD was 24 μM. The reductive activity was quite insensitive to inhibition by l-malate but the oxidative activity was very sensitive to oxaloacetate. MDH activity was not strongly activated or inhibited by glycolytic or citric acid cycle metabolites, adenine nucleotides, NaCl concentrations, or most metal ions, but increased with temperature up to about 55 °C. The reductive activity was consistently 10-20 times higher than the oxidative activity. These results indicate that the l-malate dehydrogenase in N. europaea is similar to other NAD-dependent MDHs (EC 1.1.1.37) but physiologically adapted for its role in a reductive biosynthetic sequence. [ABSTRACT FROM AUTHOR]

A bacterial culture of Nitrosomonas sp. was isolated from a nitrifying biofilm to construct a biosensor for ammonium nitrogen (NH−N) measurements in high ammonia wastewaters. The pure culture of microorganisms was immobilized into agarose gel matrix to attain a stable biosensor with a long service life. Biosensors were calibrated using (NH)SO solution and a steady-state method. Subsequently, several experiments with synthetic and industrial wastewaters were conducted. A linear range up to 20 mg/L of NH−N, and sensitivities between 0.030 and 0.036 were gained with biosensors. During 14 days of stable service life of the Nitrosomonas sp. biosensors, variation of the signal was less than 7%. Response times of biosensors were 15 ∼ 25 min, while recovery times were up to 25 min. Measurements with high ammonia content synthetic and industrial wastewaters were conducted, and 8.3 and 5.6% over estimation of NH−N was gained, respectively, compared with results of Nessler method. In spite of the small overestimation, the biosensor based on a pure culture of Nitrosomonas sp. and calibrated with (NH)SO is suitable for the analysis of NH−N in high ammonia content wastewaters. [ABSTRACT FROM AUTHOR]

In this study, we investigated during 400 days the microbial community variations as observed from 16S DNA gene DGGE banding patterns from an aerobic granular sludge pilot plant as well as the from a full-scale activated sludge treatment plant in Epe, the Netherlands. Both plants obtained the same wastewater and had the same relative hydraulic variations and run stable over time. For the total bacterial population, a similarity analysis was conducted showing that the community composition of both sludge types was very dissimilar. Despite this difference, general bacterial population of both systems had on average comparable species richness, entropy, and evenness, suggesting that different bacteria were sharing the same functionality. Moreover, multi-dimensional scaling analysis revealed that the microbial populations of the flocculent sludge system moved closely around the initial population, whereas the bacterial population in the aerobic granular sludge moved away from its initial population representing a permanent change. In addition, the ammonium-oxidizing community of both sludge systems was studied in detail showing more unevenness than the general bacterial community. Nitrosomonas was the dominant AOB in flocculent sludge, whereas in granular sludge, Nitrosomonas and Nitrosospira were present in equal amounts. A correlation analysis of process data and microbial data from DGGE gels showed that the microbial diversity shift in ammonium-oxidizing bacteria clearly correlated with fluctuations in temperature. [ABSTRACT FROM AUTHOR]

A pilot-scale ultrafiltration membrane bioreactor (MBR) was used for the aerobic treatment of urban wastewater in four experimental stages influenced by seasonal temperature and different sets of operation conditions. The structure of the ammonia-oxidizing bacteria (AOB) community was profiled by temperature gradient gel electrophoresis (TGGE), based on the amplification and separation of partial ammonia-monoxygenase subunit A ( amoA) genes. Canonical correspondence analysis revealed that temperature, hydraulic retention time and percentage of ammonia removal had a significant effect on the fingerprints of AOB communities. Phylogenetic analysis conducted on amoA/AmoA sequences of reamplified TGGE bands showed, however, that closely related ammonia-oxidizing populations inhabited the sludge of the MBR in all experimental stages. Nitrosomonas cluster 7 populations ( N. europaea- N. eutropha cluster) prevailed under all conditions tested, even when the MBR was operated under complete biomass retention or at low temperatures, suggesting that the high ammonia concentrations in the system were determinant to select r-strategist AOB. [ABSTRACT FROM AUTHOR]

The crude oil degradability and plasmid profile of autotrophic nitrifying bacteria, Nitrosomonas and Nitrobacter species, isolated from mangrove sediment in the Niger Delta of Nigeria were studied. The effects of temperature, pH and optical density on the utilization of different carbon sources by the bacteria were also investigated. Results showed that nitrifying bacteria could utilize kerosene, diesel oil, jet fuel and engine oil as carbon sources. None utilized hexane and xylene but moderate growth was observed in benzene, phenol and toluene. However, their ability to utilized crude oil varied both in rates of utilization and in growth profiles. Mixed culture of the isolates degrades 52 % of crude oil introduced into the medium followed by Nitrosomonas sp. with 40 % degradation. The least was Nitrobacter sp. with 20 % degradation. The ability of the autotrophs to degrade crude oil was found to be plasmid-mediated through curing experiment and electrophoresis. The size of the plasmid involved was estimated to be 23 kb. The high crude oil utilization of the mixed culture implies that nitrifying bacteria isolated from contaminated ecosystem are excellent crude oil degraders and can be harnessed for bioremediation purposes. [ABSTRACT FROM AUTHOR]

A recent study showed that ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA) coexist in the process of cattle manure composting. To investigate their physiological characteristics, liquid cultures seeded with fermenting cattle manure compost were incubated at various temperatures (37°C, 46°C, or 60°C) and ammonium concentrations (0.5, 1, 4, or 10 mM NH-N). The growth rates of the AOB and AOA were monitored using real-time polymerase chain reaction analysis targeting the bacterial and archaeal ammonia monooxygenase subunit A genes. AOB grew at 37°C and 4 or 10 mM NH-N, whereas AOA grew at 46°C and 10 mM NH-N. Incubation with allylthiourea indicated that the AOB and AOA grew by oxidizing ammonia. Denaturing gradient gel electrophoresis and subsequent sequencing analyses revealed that a bacterium related to Nitrosomonas halophila and an archaeon related to Candidatus Nitrososphaera gargensis were the predominant AOB and AOA, respectively, in the seed compost and in cultures after incubation. This is the first report to demonstrate that the predominant AOA in cattle manure compost can grow and can probably oxidize ammonia under moderately thermophilic conditions. [ABSTRACT FROM AUTHOR]

The importance of iron to the metabolism of the ammonia-oxidizing bacterium Nitrosomonas europaea is well known. However, the mechanisms by which N. europaea acquires iron under iron limitation are less well known. To obtain insight into these mechanisms, transcriptional profiling of N. europaea was performed during growth under different iron availabilities. Of 2,355 N. europaea genes on DNA microarrays, transcripts for 247 genes were identified as differentially expressed when cells were grown under iron limitation compared to cells grown under iron-replete conditions. Genes with higher transcript levels in response to iron limitation included those with confirmed or assigned roles in iron acquisition. Genes with lower transcript levels included those encoding iron-containing proteins. Our analysis identified several potentially novel iron acquisition systems in N. europaea and provided support for the primary involvement of a TonB-dependent heme receptor gene in N. europaea iron homeostasis. We demonstrated that hemoglobin can act as an iron source under iron-depleted conditions for N. europaea. In addition, we identified a hypothetical protein carrying a lipocalin-like domain that may have the ability to chelate iron for growth in iron-limited media. [ABSTRACT FROM AUTHOR]

The phylogenetic diversity and abundance of ammonia-oxidizing beta-proteobacteria (beta-AOB) was analyzed along an anthropogenic pollution gradient from the coastal Pearl River Delta to the South China Sea using the ammonia monooxygenase subunit A ( amoA) gene. Along the gradient from coastal to the open ocean, the phylogenetic diversity of the dominant genus changed from Nitrosomonas to Nitrosospira, indicating the niche specificity by these two genera as both salinity and anthropogenic influence were major factors involved. The diversity of bacterial amoA gene was also variable along the gradient, with the highest in the deep-sea sediments, followed by the marshes sediments and the lowest in the coastal areas. Within the Nitrosomonas-related clade, four distinct lineages were identified including a putative new one (A5-16) from the different sites over the large geographical area. In the Nitrosospira-related clade, the habitat-specific lineages to the deep-sea and coastal sediments were identified. This study also provides strong support that Nitrosomonas genus, especially Nitrosomonas oligotropha lineage (6a) could be a potential bio-indicator species for pollution or freshwater/wastewater input into coastal environments. A suite of statistical analyses used showed that water depth and temperature were major factors shaping the community structure of beta-AOB in this study area. [ABSTRACT FROM AUTHOR]

The ammonia-oxidizing bacterial community (AOB) was investigated in two types of laboratory-scale bioreactors performing partial oxidation of ammonia to nitrite or nitrate at high (80 mM) to extremely high (428 mM) concentrations of ammonium bicarbonate. At all conditions, the dominant AOB was affiliated to the Nitrosomonas europaea lineage as was determined by fluorescence in situ hybridization and polymerase chain reaction in combination with denaturing gradient gel electrophoresis. Molecular analysis of the mixed populations, based on the 16S rRNA and cbbL genes, demonstrated the presence of two different phylotypes of Nitrosomonas, while microbiological analysis produced a single phylotype, represented by three different morphotypes. One of the most striking features of the AOB populations encountered in the bioreactors was the domination of highly aggregated obligate microaerophilic Nitrosomonas, with unusual cellular and colony morphology, commonly observed in nitrifying bioreactors but rarely investigated by cultural methods. The latter is probably not an adaptation to stressful conditions created by high ammonia or nitrite concentrations, but oxygen seems to be a stressful factor in these bioreactors. [ABSTRACT FROM AUTHOR]

A lab-scale partial nitrifying sequencing batch biofilm reactor was a successful start-up. Denaturing gradient gel electrophoresis (DGGE) was used to investigate the bacterial community dynamics in three periods together with inocula sludge at ambient temperature. The DGGE profiles of bacteria and Shannon-Wiener index ( H′) results showed that high free ammonia (FA) concentration referred to lower diversity in the bioreactor system. Cluster analysis indicated that microorganism in period III was similar with inocula sludge and was different from that in periods I and II. Similar results also appeared in ammonia-oxidizing bacteria (AOB) community structure and nitrite-oxidizing bacteria (NOB) community structure, and at least four AOB species and two NOB species were present in period III, respectively. Phylogenetic analysis of amoA gene sequences showed that Nitrosomonas eutropha cluster was predominant in all the three periods. With lower ammonium loads, three new operational taxonomic units formed and consisted Nitrosomonas sp . Cluster. This article demonstrated that microbial community, AOB, and NOB diversity were related with FA concentration closely at ambient temperature. [ABSTRACT FROM AUTHOR]

In this study, dideoxy sequencing and 454 high-throughput sequencing were used to analyze diversities of the ammonia monooxygenas e ( amoA) genes and the 16S rRNA genes of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) in six municipal wastewater treatment plants. The results showed that AOB amoA genes were quite diverse in different wastewater treatment plants while the 16S rRNA genes were relatively conserved. Based on the observed complexity of amoA and 16S rRNA genes, most of the AOB can be assigned to the Nitrosomonas genus, with Nitrosomonas ureae, Nitrosomonas oligotropha, Nitrosomonas marina, and Nitrosomonas aestuarii being the four most dominant species. From the sequences of the AOA amoA genes, most AOA observed in this study belong to the CGI.1b group, i.e., the soil lineage. The AOB amoA and 16S rRNA genes were quantified by quantitative PCR and 454 high-throughput pyrosequencing, respectively. Although the results from the two approaches show some disconcordance, they both indicated that the abundance of AOB in activated sludge was very low. [ABSTRACT FROM AUTHOR]

In recent years, natural and synthetic estrogens have been recognized as endocrine disruptors in aquatic organisms. Although natural and synthetic estrogens are known to be degraded by microbes, only limited information about their degradation pathways is available. Here, we studied the degradation pathways of a natural estrogen, 17β-estradiol, by the nitrifying microorganism Nitrosomonas europaea, and we determined whether the degradation products of 17β-estradiol had estrogenic activity. To identify the degradation products, we subjected the culture solution to solid-phase extraction, and the extract was analyzed by gas chromatography-mass spectrometry. The potential estrogenic activity of the degradation products was investigated by means of a yeast two-hybrid assay. 1,3,5(10),16-Estratetraen-3-ol (estratetraenol) was newly identified as a degradation intermediate produced by dehydration of 17β-estradiol. Estratetraenol was also degraded by N. europaea, and its degradation rate was faster than that of 17β-estradiol. The two-hybrid assay confirmed that estratetraenol acted as a ligand for the estrogen receptor; estratetraenol thus has potential estrogenic activity. N. europaea eliminated the estrogenic activity derived from 17β-estradiol. This paper is the first to report dehydration as a mechanism of microbial estrogen degradation. [ABSTRACT FROM AUTHOR]

Nitrosomonas europaea has a single three-gene operon ( nitABC) encoding an iron ABC transporter system (NitABC). Phylogenetic analysis clustered the subunit NitB with Fe-ABC transporter permease components from other organisms. The N. europaea strain deficient in nitB ( nitB::kan) grew well in either Fe-replete or Fe-limited media and in Fe-limited medium containing the catecholate-type siderophore, enterobactin or the citrate-based dihydroxamate-type siderophore, aerobactin. However, the nitB::kan mutant strain was unable to grow in Fe-limited media containing either the hydroxamate-type siderophores, ferrioxamine and ferrichrome or the mixed-chelating type siderophore, pyoverdine. Exposure of N. europaea cells to a ferrichrome analog coupled to the fluorescent moiety naphthalic diimide (Fhu-NI) led to increase in fluorescence in the wild type but not in nitB::kan mutant cells. Spheroplasts prepared from N. europaea wild type exposed to Fhu-NI analog retained the fluorescence, while spheroplasts of the nitB::kan mutant were not fluorescent. NitABC transports intact Fe-ferrichrome complex into the cytoplasm and is an atypical ABC type iron transporter for Fe bound to ferrioxamine, ferrichrome or pyoverdine siderophores into the cytoplasm. The mechanisms to transport iron in either the Fe or Fe forms or Fe associated with enterobactin or aerobactin siderophores into the cell across the cytoplasmic membrane are as yet undetermined. [ABSTRACT FROM AUTHOR]

The influences of suspended particles (SPs) on NH4 + was significantly affected by the SPs concentration under the conditions typically present in the TGR. The amount of ammonia adsorbed per unit weight of suspended particles was inverse proportional to the concentration of suspended particles. However, the influences of the particle size and the organic matter concentration existing in SPs were insignificant under the experimental conditions. The effects of suspended particles on nitritation were determined by the use of ammonia-oxidizing bacteria (AOB) strain SW16, identified as Nitrosomonas nitrosa, which was isolated from sediment samples of the TGR. Suspended particle concentration in water–sediment solution played an important role in the nitritation process. The rate of nitritation enhanced with the increase of the suspended particle concentration. It was found that the critical factor controlling ammonia oxidizing rate was the AOB biomass resulting from the AOB growth rate. Moreover, results demonstrated that both particle size and organic matter content showed little effect on the nitritation process under the experimental conditions. [ABSTRACT FROM AUTHOR]4 + was significantly affected by the SPs concentration under the conditions typically present in the TGR. The amount of ammonia adsorbed per unit weight of suspended particles was inverse proportional to the concentration of suspended particles. However, the influences of the particle size and the organic matter concentration existing in SPs were insignificant under the experimental conditions. The effects of suspended particles on nitritation were determined by the use of ammonia-oxidizing bacteria (AOB) strain SW16, identified as Nitrosomonas nitrosa, which was isolated from sediment samples of the TGR. Suspended particle concentration in water–sediment solution played an important role in the nitritation process. The rate of nitritation enhanced with the increase of the suspended particle concentration. It was found that the critical factor controlling ammonia oxidizing rate was the AOB biomass resulting from the AOB growth rate. Moreover, results demonstrated that both particle size and organic matter content showed little effect on the nitritation process under the experimental conditions. [ABSTRACT FROM AUTHOR]

Nonylphenol (NP) is an estrogenic pollutant which is widely present in the aquatic environment. Biodegradation of NP can reduce the toxicological risk. In this study, aerobic biodegradation of NP in river sediment was investigated. The sediment used for the microcosm experiments was aged polluted with NP. The biodegradation of NP in the sediment occurred within 8 days with a lag phase of 2 days at 30°C. During the biodegradation, nitro-nonylphenol metabolites were formed, which were further degraded to unknown compounds. The attached nitro-group originated from the ammonium in the medium. Five subsequent transfers were performed from original sediment and yielded a final stable population. In this NP-degrading culture, the microorganisms possibly involved in the biotransformation of NP to nitro-nonylphenol were related to ammonium-oxidizing bacteria. Besides the degradation of NP via nitro-nonylphenol, bacteria related to phenol-degrading species, which degrade phenol via ring cleavage, are abundantly present. [ABSTRACT FROM AUTHOR]

Halophilic nitrifiers belonging to the genus Nitrosomonas and Nitrospira were enriched from seawater and marine sediment samples of the North Sea. The maximal ammonia oxidation rate (AOR) in batch enrichments with seawater was 15.1 mg N L−1 day−1. An intermediate nitrite accumulation was observed. Two fixed-bed reactors for continuous nitrification with either polyethylene/clay sinter lamellas (FBR A) or porous ceramic rings (FBR B) were run at two different ammonia concentrations, three different ammonia loading rates (ALRs), ± pH adjustment, and at an increased upflow velocity. A better overall nitrification without nitrite accumulation was observed in FBR B. However, FBR A revealed a higher AOR and nitrite oxidation rate of 6 and 7 mg N L−1 h−1, compared to FBR B with 5 and 5.9 mg N L−1 h−1, respectively. AORs in the FBRs were at least ten times higher than in suspended enrichment cultures. Whereas a shift within the ammonia-oxidizing population in the genus Nitrosomonas at the subspecies level occurred in FBR B with synthetic seawater at an increasing ALR and a decreasing pH, the nitrite oxidizing Nitrospira population apparently did not change. [ABSTRACT FROM AUTHOR]

Due to variations in the production levels, a full-scale sequencing batch reactor (SBR) for post-treatment of tannery wastewater was exposed to low and high ammonia load periods. In order to study how these changes affected the N-removal capacity, the microbiology of the reactor was studied by a diverse set of techniques including molecular tools, activity tests, and microbial counts in samples taken along 3 years. The recover capacity of the biomass was also studied in a lab-scale reactor operated with intermittent aeration without feeding for 36 days. The results showed that changes in the feeding negatively affected the nitrifying community, but the nitrogen removal efficiencies could be restored after the concentration stress. Species substitution was observed within the nitrifying bacteria, Nitrosomonas europaea and Nitrobacter predominated initially, and after an ammonia overload period, Nitrosomonas nitrosa and Nitrospira became dominant. Some denitrifiers, with nirS related to Alicycliphilus, Azospirillum, and Marinobacter nirS, persisted during long-term reactor operation, but the community fluctuated both in composition and in abundance. This fluctuating community may better resist the continuous changes in the feeding regime. Our results showed that a nitrifying–denitrifying SBR could be operated with low loads or even without feeding during production shut down periods. [ABSTRACT FROM AUTHOR]

The ammonia oxidizers Nitrosomonas europaea and Nitrosomonas eutropha are able to grow chemoorganotrophically under anoxic conditions with pyruvate, lactate, acetate, serine, succinate, α-ketoglutarate, or fructose as substrate and nitrite as terminal electron acceptor. The growth yield of both bacteria is about 3.5 mg protein (mmol pyruvate)−1 and the maximum growth rates of N. europaea and N. eutropha are 0.094 d−1 and 0.175 d−1, respectively. In the presence of pyruvate and CO2 about 80% of the incorporated carbon derives from pyruvate and about 20% from CO2. Pyruvate is used as energy and only carbon source in the absence of CO2 (chemoorganoheterotrophic growth). CO2 stimulates the chemoorganotrophic growth of both ammonia oxidizers and the expression of ribulose bisphosphate carboxylase/oxygenase is down-regulated at increasing CO2 concentration. Ammonium, although required as nitrogen source, is inhibitory for the chemoorganotrophic metabolism of N. europaea and N. eutropha. In the presence of ammonium pyruvate consumption and the expression of the genes aceE, ppc, gltA, odhA, and ppsA (energy conservation) as well as nirK, norB, and nsc (denitrification) are reduced. [ABSTRACT FROM AUTHOR]

Transcriptional fusions with gfp driven by the promoter region of mbla (NE2571) in pPRO/mbla4 and clpB (NE2402) in pPRO/clpb7 were used to transform the ammonia-oxidizing bacterium Nitrosomonas europaea (ATCC 19718). The two genes were chosen because their transcript levels were found at much higher levels in N. europaea in response to oxidation of chloroform and chloromethane. In N. europaea transformed with pPRO/mbla4, green fluorescent protein (GFP)-dependent fluorescence increased from 3- to 18-fold above control levels in response to increasing chloroform concentrations (7 to 28 μM), and from 8- to 10-fold in response to increasing hydrogen peroxide concentrations (2.5–7.5 mM). The GFP-dependent fluorescence of N. europaea transformed with pPRO/clpb7 also showed an increase of 6- to 10-fold in response to chloroform (28–100 μM) but did not respond to H2O2. Our data provide proof of concept that biosensors can be fabricated in ammonia-oxidizing bacteria using “sentinel” genes that up-regulate in response to stress caused either by co-oxidation of chlorinated solvents or by the presence of H2O2. The fabricated biosensors had a consistent concentration-dependent response to chloroform; however, these did not respond to other chlorinated compounds that cause similar cellular stress. [ABSTRACT FROM AUTHOR]

The inhibitory effect of Cd on nitrification was investigated in a continuous-flow system with enriched nitrifying bacteria. The maximum specific ammonium utilization rate and the half-saturation constant were found as 671 mg NH4-N/g VSS day and 0.48 mg/l, respectively. In the case of continuous Cd input at 1 and 2.5 mg/l, nitrification was inhibited by 30% and 47%, respectively. Inhibition ranged from 20% to 40% and no further increase in inhibition was exhibited in new runs except at 10 mg/l influent Cd. At 10 mg/l influent Cd, specific ammonium utilization and nitrate production rates were inhibited by 90%. On the contrary, a serious nitrite accumulation was not observed during this period. When Cd feeding was stopped, recovery from inhibition was observed after 37 day which was seen by the improvement in ammonium utilization and nitrate production rates. A shift in microbial population from the initial Nitrosomonas sp. to the Cd-tolerant Nitrosospira sp. was observed in the recovery period from severe Cd inhibition. After the domination of Nitrosospira species, redosing at 10 mg/l and then at 15 mg/l did not affect the performance as before. [ABSTRACT FROM AUTHOR]

Abstract  Two distinct nitrifying bacterial consortia, namely an ammonia oxidizing non-penaeid culture (AMONPCU-1) and an ammonia oxidizing penaeid culture (AMOPCU-1), have been mass produced in a nitrifying bacterial consortia production unit (NBCPU). The consortia, maintained at 4°C were activated and cultured in a 2 l fermentor initially. At this stage the net biomass (0.105 and 0.112 g/l), maximum specific growth rate (0.112 and 0.105/h) and yield coefficients (1.315 and 2.08) were calculated respectively, for AMONPCU-1 and AMOPCU-1 on attaining stationary growth phase. Subsequently on mass production in a 200 l NBCPU under optimized culture conditions, the total amounts of NH4 +–N removed by AMONPCU-1 and AMOPCU-1 were 1.948 and 1.242 g/l within 160 and 270 days, respectively. Total alkalinity reduction of 11.7–14.4 and 7.5–9.1 g/l were observed which led to the consumption of 78 and 62 g Na2CO3. The yield coefficient and biomass of AMONPCU-1 were 0.67 and 125.3 g/l and those of AMOPCU-1 were 1.23 and 165 g/l. The higher yield coefficient and growth rate of AMOPCU-1 suggest better energy conversion efficiency and higher CO2 fixation potential. Both of the consortia were dominated by Nitrosomonas-like organisms. The consortia may find application in the establishment of nitrification within marine and brackish water culture systems. [ABSTRACT FROM AUTHOR]

The intracellular location of the membrane-bound ammonia monooxygenase (AMO) in all genera of ammonia oxidizing bacteria ( Nitrosomonas, Nitrosococcus and Nitrosospira) was determined by electron microscopic immunocytochemistry. Polyclonal antibodies recognizing the two subunits, AmoA- and AmoB-proteins, were used for post-embedding labeling. Ultrathin sections revealed that the AmoB-protein was located in all genera on the cytoplasmic membrane. In cells of Nitrosomonas and Nitrosococus additional but less AmoB-labeling was found on the intracytoplasmic membrane (ICM). In contrast to the detection of AmoB-protein, the AmoA-antibodies failed to detect the AmoA-protein. Based on quantitative immunoblots the extent of ICM in Nitrosomonas eutropha was correlated with the amount of AmoA in the cells. The highest extent of ICM and amount of AmoA was found at low ammonium substrate concentrations. [ABSTRACT FROM AUTHOR]

The soil nitrifying bacterium Nitrosomonas europaea has shown the ability to transform cometabolically naphthalene as well as other 2- and 3-ringed polycyclic aromatic hydrocarbons (PAHs) to more oxidized products. All of the observed enzymatic reactions were inhibited by acetylene, a selective inhibitor of ammonia monooxygenase (AMO). A strong inhibitory effect of naphthalene on ammonia oxidation by N. europaea was observed. Naphthalene was readily oxidized by N. europaea and 2-naphthol was detected as a major product (85%) of naphthalene oxidation. The maximum naphthol production rate was 1.65 nmole/mg protein-min in the presence of 240 μM naphthalene and 10 mM NH4 +. Our results demonstrate that the oxidation between ammonia and naphthalene showed a partial competitive inhibition. The relative ratio of naphthalene and ammonia oxidation, depending on naphthalene concentrations, demonstrated that the naphthalene was oxidized 2200-fold slower than ammonia at lower concentration of naphthalene (15 μM) whereas naphthalene was oxidized only 100-fold slower than ammonia oxidation. NH4 +- and N2H4-dependent O2 uptake measurement demonstrated irreversible inhibitory effects of the naphthalene and subsequent oxidation products on AMO and HAO activity. [ABSTRACT FROM AUTHOR]

Autotrophic ammonia-oxidizing bacteria use the essential enzyme ammonia monooxygenase (AMO) to transform ammonia to hydroxylamine. The amo operon consists of at least three genes, amoC, amoA, and amoB; amoA encodes the subunit containing the putative enzyme active site. The use of the amo genes as functional markers for ammonia-oxidizing bacteria in environmental applications requires knowledge of the diversity of the amo operon on several levels: (1) the copy number of the operon in the genome, (2) the arrangement of the three genes in an individual operon, and (3) the primary sequence of the individual genes. We present a database of amo gene sequences for pure cultures of ammonia-oxidizing bacteria representing both the β- and the γ-subdivision of Proteobacteria in the following genera: Nitrosospira (6 strains), Nitrosomonas (5 strains) and Nitrosococcus (2 strains). The amo operon was found in multiple (2–3) nearly identical copies in the β-subdivision representatives but in single copies in the γ-subdivision ammonia oxidizers. The analysis of the deduced amino acid sequence revealed strong conservation for all three Amo peptides in both primary and secondary structures. For the amoA gene within the β-subdivision, nucleotide identity values are approximately 85% within the Nitrosomonas or the Nitrosospira groups, but approximately 75% when comparing between these groups. Conserved regions in amoA and amoC were identified and used as primer sites for PCR amplification of amo genes from pure cultures, enrichments and the soil environment. The intergenic region between amoC and amoA is variable in length and may be used to profile the community of ammonia-oxidizing bacteria in environmental samples. Electronic supplementary material to this paper can be obtained by using the Springer LINK server located at http://dx.doi.org/10.1007/s00203-001-0369-z. [ABSTRACT FROM AUTHOR]