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The effects of penicillin G on the number and activity of nitrite-oxidizing bacteria were investigated in laboratory-scale reactors and batch tests. At a concentration of 100 mg L(-1), addition of penicillin G for short periods did not significantly affect nitrite oxidation, while addition for more than 2 months suppressed nitrite oxidation. Fluorescence in situ hybridization with 16S ribosomal RNA-targeted probes revealed a slight decrease in the abundance of Nitrospira, while Nitrobacter was not affected by addition of penicillin G for more than 39 days. The resistance of nitrite-oxidizing bacteria to penicillin appeared to be positively affected by intermittent aeration only when accompanied by denitrification; otherwise, the aeration mode (continuous or intermittent aeration) did not significantly affect the abundance of Nitrobacter and Nitrospira.

Anammox (anaerobic ammonium oxidation) is an important process of nitrogen cycle, with great potential for the practical use in removing nitrogen from the wastewater containing high concentration ammonium. However, the presence of high concentration organic carbon source is considered unfavorable to anammox. Coupling anammox and denitrification under the presence of organic carbon source could be a useful technique for removing both nitrogen and carbon. This paper reviewed the mechanisms of anammox-denitrification coupling process, functional microbial groups, initiation of the coupling process and its control, and related environmental affecting factors. The future research prospects and potential applications of anammox-denitrification coupling process in wastewater treatment were discussed.

This study investigated the performance of one-step aerobic biological oxidation process and anoxic/aerobic two-step biological oxidation process treating modeled river water containing low carbon and rich ammonia. Biofilm microbial ecology was analyzed with multiple molecular technologies including PCR-DGGE, FISH/CLSM and FISH/FCM to investigate the succession of bacteria community and space distribution along with abundance of the main functional bacteria, and to research the micro-influential factors and the mechanism of different biological contact oxidation processes for their performance. Results showed that two-step contact oxidation process achieved higher removal percentage than that of the one-step process, with COD and NH4(+) -N removal enhanced about 10% and 32%-59%, respectively. A much thicker biofilm was obtained by the one-step process compared to the two-step process, and nitrobacterium was mainly distributed in the depth of 180-200 microm and 105-125 microm, respectively. PCR-DGGE results found that the two-step process demonstrated less microbial diversity than that of the one-step process, FISH/FCM results showed that ammonia oxidizing bacteria (AOB) and nitrite oxidizing bacteria (NOB) abundance increased in the two-step process with the increase of operation time, while that of the one-step process declined. Experiment results demonstrate that functional partitioning of the anoxic-aerobic two-step biological contact oxidation process could be in favor for harvesting nitrobacteria and other special bacteria in different reactor spaces, which can improve removal efficiency for organics and ammonia finally.

A biofilm with high nitrifying efficiency was converted into a nitritating and thereafter a nitritating-anammox biofilm in a moving-bed biofilm reactor at 26.5 (+/- 0.5) degrees C by means of a combination of intermittent aeration, low dissolved oxygen concentration, low hydraulic retention time, free ammonia and furthermore, also by elevated HCO3- concentration. Nitrite-oxidizing bacteria (NOB) were more effectively suppressed by an enhanced HCO3- concentration range of 1200-2350 mg/L as opposed to free-ammonia-based process control where NOBs recovered from inhibition; the respective total-nitrogen removal rates were 0.3 kg N/(m3 x d) and 0.2 kg N/(m3 x d). The biofilm modification strategies resulted in a shift in bacterial community as the NOB Nitrobacter spp. were replaced with NOB belonging to the genus Nitrospira spp. and were closely related to Candidatus Nitrospira defluvii. A community of anaerobic ammonium-oxidizing microorganisms -uncultured Planctomycetales bacterium clone P4 (closely related to Candidatus Brocadia fulgida)--was developed.

Previous studies showed that setting aerobic and anaerobic paragraph segments in the subsurface constructed wetlands (SFCWs) can improve the COD, NH4(+)-N, and TN removal rate, whereas the oxygen enrichment environment which produced by the artificial aeration could restrain the NO3(-)-N and NO2(-)-N removal process, and to a certain extent, inhibit the denitrification in SFCWs Therefore, in this research the structure and technology of SFCW with aerobic and anaerobic paragraph segments were optimized, by using the multi-point water inflow and setting the corresponding section for the extra pollutant removal. Results showed that with the hydraulic load of 0.06 m3 x (m2 x d)(-1), the COD, NH4(+)-N and TN removal efficiencies in the optimized SFCW achieved 91.6%, 100% and 87.7% respectively. COD/N increased to 10 speedily after the inflow supplement. The multi-point water inflow could add carbon sources, and simultaneously maximum utilization of wetland to remove pollutants. The optimized SFCW could achieve the purposes of purification process optimization, and provide theoretical basis and application foundation for improving the total nitrogen removal efficiency.

Large wastewater treatment plants have to deal not only with the influent wastewater, but also with the highly concentrated reject water coming from anaerobic digestion. The aim of this work was to verify the suitability of using encapsulated nitrifiers in polyvinyl alcohol carrier (so called Lentikats Biocatalyst) at temperatures between 5 and 30 °C. For laboratory nitrification batch tests synthetic wastewater with ammonia nitrogen (Namon) concentration 10-800 mg L(-1) was used. The system has been proved to operate at the temperature of 10 °C, but not at 5 °C. It was found that the highest specific nitrification rates were observed at 30 °C and with ammonia nitrogen concentrations above 250 mg L(-1), which means that separate treatment of reject water by using encapsulated biomass seems to be an effective tool for wastewater treatment plant optimization.

Non-optimal pH, dissolved oxygen concentration, the presence of toxic substances, or the influence of grazers are known to cause disturbances in nitrification. Because activated sludge is a mixture of different organisms, bacteria, and higher organisms, the stability of processes such as carbon removal, nitrification, denitrification, and dephosphatation depends on a range of interactions. These interactions occur both between and within trophic levels. Understanding of the ecology of microorganisms involved in bioprocesses is essential for effective control of startup and operation of a particular process. The aim of the study was to gain further insight into the dynamics of nitrifiers in activated sludge at various sludge ages while treating higher concentrations of ammonium. The results confirmed the importance of Nitrosococcus mobilis and Nitrobacter sp. as the dominant nitrifiers responsible for nitritation and nitratation, respectively, in the presence of unlimited ammonium. The size of the dominant bacteria colony was larger compared to the other species present and reached 25 microm. Problems with nitrification occurred in all high-ammonium loaded reactors. The dynamics of nitrifier population was monitored by oxygen uptake rate (OUR) using a test enabling the OUR measurement separately for ammonium-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB). The results reveal the hypersensitivity of nitrifiers to the substrate and products of incomplete nitrification.

A comparative study was conducted to determine nitrification potentials and ammonia-oxidizing bacterial (AOB) community composition in 0-20 cm soil depth in adjacent native forest,natural grassland, and cropland soils on the Tibetan Plateau, by incubation experiment and by denaturing gradient gel electrophoresis (DGGE) of 16S rDNA, respectively. Cropland has the highest nitrification potential and nitrate concentration among the three land-use types (LUT), approximately 9 folds and more than 11 folds than that of the forests and grasslands, respectively. NO3(-) -N accounted for 70%-90% of inorganic N in cropland soil, while NH4(+) -N was the main form of inorganic N in forest and grassland soils. Nitrification potentials and nitrate concentrations showed no significant difference between native forest and grassland soils. The native forest showed the lowest nitrification potentials and the lowest AOB diversity and community composition among the three LUT. Conversions from natural grasslands to croplands remarkably decreased the AOB diversity and composition, but croplands remain high similarity in AOB community composition compared with grasslands. The minimal and the lowest diversity of AOB in native forests directly resulted to the lowest nitrification potentials compared to natural grasslands and croplands. From the fact of the highest nitrification potentials and nitrate concentrations in croplands indicated that there were the most substantial AOB with higher activity and priority. The results provide evidence that changes of land-use type can affect both soil nitrogen internal cycling process, the diversity, community and activity of AOB, which further affect soil environment quality and the long-term sustainability of ecosystems.

The dynamic succession of ammonia-oxidizing bacteria (AOB) community during the agricultural waste composting was investigated using the polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). The changes of pile temperature, pH, moisture content, soluble NH4(+) -N and NO3(-)-N were measured. DGGE results showed that the Shannon-Weaver index changed from 2.58 to 2.02 during the composting process, which reflected the diversity trends downward overall. Nitrosospira and Nitrosomonas were dominant species. Nitrosomonas eutropha,which was found throughout the whole composting process, was identified as a kind of well-tolerated species. Redundancy analysis (RDA) was applied to explore the correlation between the dynamic succession of AOB community and composting physic-chemical parameters. Compared with the slight change referring to its structure during the earlier phase of thermophilic period (4-9 d), the AOB community experienced a more significant change during the later phase of thermophilic period (9-12 d) as well as the following temperature falling phase (12-25 d). Significant relationships have been found between pile temperature, soluble NH4(+) -N (P < 0.01) and NO3(-)-N (P < 0.05) with the dynamic succession of AOB community, which suggested those three parameters were likely to influence, or be influenced by AOB during the agricultural waste composting.

The effect of pH and nitrite concentration on the activity of the nitrite oxidizing bacteria (NOB) in an activated sludge reactor has been determined by means of laboratory batch experiments based on respirometric techniques. The bacterial activity was measured at different pH and at different total nitrite concentrations (TNO₂). The experimental results showed that the nitrite oxidation rate (NOR) depends on the TNO₂ concentration independently of the free nitrous acid (FNA) concentration, so FNA cannot be considered as the real substrate for NOB. NOB were strongly affected by low pH values (no activity was detected at pH 6.5) but no inhibition was observed at high pH values (activity was nearly the same for the pH range 7.5-9.95). A kinetic expression for nitrite oxidation process including switch functions to model the effect of TNO₂ concentration and pH inhibition is proposed. Substrate half saturation constant and pH inhibition constants have been obtained., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

The approach and factors for realizing the rapid startup of nitrosation were researched at the low level of dissolved oxygen (DO) in sequencing batch reactor (SBR). The main parameters of the reactor were controlled as follows: DO were 0.15-0.40 mg/L, pH values kept from 7.52 to 8.30, temperature maintained at 22.3-27.1 degrees C, and time of aeration was 8 hours. The purpose of rapid startup for nitrosation was achieved after 57 cycles (36 d) with the alternative influent of high and low ammonium wastewater (the mean values were 245.28 mg/L and 58.08 mg/L respectively) in a SBR, and the nitrosation rate was even 100%. Factors of accumulation of nitrite were investigated and the effects of DO and pH were analyzed during the startup for nitrosation. The results showed that it could improve the efficiency of nitrosation when DO concentration was increased appropriately. The activity of nitrite oxidizing bacteria (NOB) was recovered gradually when DO was higher than 0.72 mg/L. The key factor of controlling nitrosation reaction was the concentration of free ammonia (FA), while the final factor was the concentration of DO. pH was a desired controlling parameter to show the end of nitrification in a SBR cycle, while DO concentration did not indicate the finishing of SBR nitrification accurately because it increased rapidly before ammonia nitrogen was oxidized absolutely.

Chlorite dismutase (Cld) is a unique heme enzyme catalyzing the conversion of ClO(2)(-) to Cl(-) and O(2). Cld is usually found in perchlorate- or chlorate-reducing bacteria but was also recently identified in a nitrite-oxidizing bacterium of the genus Nitrospira. Here we characterized a novel Cld-like protein from the chemolithoautotrophic nitrite oxidizer Nitrobacter winogradskyi which is significantly smaller than all previously known chlorite dismutases. Its three-dimensional (3D) crystal structure revealed a dimer of two identical subunits, which sharply contrasts with the penta- or hexameric structures of other chlorite dismutases. Despite a truncated N-terminal domain in each subunit, this novel enzyme turned out to be a highly efficient chlorite dismutase (K(m) = 90 μM; k(cat) = 190 s(-1); k(cat)/K(m) = 2.1 × 10(6) M(-1) s(-1)), demonstrating a greater structural and phylogenetic diversity of these enzymes than was previously known. Based on comparative analyses of Cld sequences and 3D structures, signature amino acid residues that can be employed to assess whether uncharacterized Cld-like proteins may have a high chlorite-dismutating activity were identified. Interestingly, proteins that contain all these signatures and are phylogenetically closely related to the novel-type Cld of N. winogradskyi exist in a large number of other microbes, including other nitrite oxidizers.

In August 15-28, 2009, a preliminary study was conducted on the spatial distribution characteristics of heterotrophic bacteria (HB), inorganic phosphate bacteria (IPB), organic phosphate bacteria (OPB), denitrifying bacteria (DB), and ammonifying bacteria (AB) in the hypoxic zone of Yangtze River Estuary. In the water surface, water bottom, and sediment surface of the zone, the average quantity of AB was the largest, being 307.52 x 10(4) cells x L(-1), 184.50 x 10(4) cells x L(-1), 199.97 x 10(2) cells x g(-1), followed by that of HB (87.35 x 10(4) cfu x L(-1), 86.85 x 10(4) cfu x L(-1), and 19.56 x 10(2) cfu x g(-1)), and of OPB (19.26 x 10(4) cfu x L(-1), 18.82 x 10(4) cfu x L(-1), and 19.56 x10(2) cfu x g(-1), respectively). IPB was only observed within the Yangtze Estuary, south passage of the Estuary, and Zhoushan inshore, and its average quantity in the water surface, water bottom, and sediment surface was 18.50 x 10(4) cfu x L(-1), 31.00 x 10(4) cfu x L(-1), and 7.17 x 10(2) cfu x g(-1) respectively. DB had a wide distribution, but its average quantity was low, being 3.94 x 10(4) cells x L(-1), 23.08 x 10(4) cells x L(-1), and 6.22 x 10(2) cells x g(-1) in the water surface, water bottom, and sediment surface, respectively. Salinity, NO3(-)-N, PO4(3-)-P, SiO3(2)-Si, and pH were the main factors affecting the distribution of HB, IPB, OPB, and DB in water body and sediment surface. The HB, IPB, and OPB in water bottom and sediment surface had significant positive correlation with water temperature; the HB and OPB in water bottom and the IPB in sediment surface were significantly positively correlated with dissolved oxygen (DO); while the AB in sediment surface was significantly negatively correlated with DO. Cluster analysis showed that hypoxia affected the bacterial community structure in sediment surface.

Nitrification is the process that converts ammonium to nitrate and thus links the regeneration of organic nitrogen to fixed nitrogen loss by denitrification. The first step, oxidation of ammonia to nitrite, is performed by a phylogenetically restricted group of proteobacteria (ammonia-oxidizing bacteria, AOB) and Crenarchaea (ammonia-oxidizing archaea, AOA). The second step is restricted to nitrite-oxidizing bacteria (NOB) as far as currently known. All three groups are assumed to be autotrophic and obligately aerobic, but the true extent of autotrophy and potential anaerobic pathways in these organisms is currently under investigation. Here, we describe methods for the measurement of nitrification rates in the marine environment, with a focus on seawater systems and stable isotopic tracer methods. The methods vary in analytical requirements but share the need for incubations, which must be optimized for different environments with different substrate concentrations. Recent advances in mass spectrometry now make it possible to minimize incubation artifacts and to achieve greatly improved sensitivity., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Understanding nitrification rates and their regulation continues as a key area of research for assessing human's increasing impact on the terrestrial N cycle. We review the organisms and processes responsible for nitrification in terrestrial systems. The control of nitrification by substrate availability is discussed with particular attention to the factors affecting ammonia/ammonium availability. The effects on nitrification rates of environmental controls including oxygen, water potential, temperature and pH are described. With this general understanding of the factors affecting nitrification rates as a basis, we present an in depth analysis of methods used to measure nitrification in terrestrial systems. Net, gross and potential nitrification rate measurements are explained including the use of isotopes and inhibitors to measure rates in soils. Methods for the estimation of nitrification kinetics and modeling are briefly described. Future challenges will require understanding the factors controlling nitrification across spatial scales from ecosystems to soil microsites if we are to sustainably manage reactive nitrogen in terrestrial environments., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Nitrobacteria, nitrification and its impact factors were discussed based on the method of on-site incubation and laboratory simulation in Liaohe estuary wetland sediments in June and August 2009, respectively. The results showed that the number of ammonia-oxidizing bacteria (AOB) ranged from 0.54 x 10(4) to 5.69 x 10(4) cells x g(-1) with an average of (2.21 +/- 2.32) x 10(4) cells x g(-1) in June and ranged from 1.90 x 10(4) to 7.90 x 10(4) cells x g(-1) with an average of (3.61 +/- 2.87) x 10(4) cells x g(-1) in August. The potential nitrification rates ranged from 9.72 to 16.45 mmol x (m2 x h)(-1) with an average of (12.54 +/- 3.14) mmol x (m2 x h)(-1) in June and ranged from 14.66 to 24.62 mmol x (m2 x h)(-1) with an average of (18.71 +/- 4.21) mmol x (m2 x h)(-1) in August. The net nitrification rates were 0.41 mmol x (m2 x h)(-1) in June and ranged from 0.20 to 0.53 mmol x (m2 x h)(-1) with an average of (0.35 +/- 0.16) mmol x (m2 x h)(-1) in August. The potential nitrification rates were apparently higher than the net nitrification rates. The AOB abundance and nitrification rates were all higher in August than in June. Ammonia concentration in the overlying water, pH, organic matter, total nitrogen content, total phosphorus content, ammonia nitrogen content and the number of AOB in sediments were the main environmental impact factors on nitrification by SPSS 13.0 statistical analysis (p < 0.05), and it also showed ammonia concentration in the overlying water, total phosphorus content and ammonia nitrogen content in sediments were key impact factors on nitrification by partial correlation analysis. The flux of transformed ammonium into nitrate by nitrification process was 1.14 x 10(5) kg per day, suggesting that nitrification was important on nitrogen cycle in Liaohe estuary wetland sediments.

With A/O phosphorus removal process of secondary effluent as raw water, the stable accumulation of nitrite was achieved under low DO. Total nitrogen loss, hydraulic retention time (HRT) and sludge recycle ratio(R) was separately studied on the effects of stable nitrosation. According the result, apparent nitrosation rate was affected by the concentration of COD influence, when COD < or = 50 mg/L, apparent nitrosation rate was reduced; when COD > 50 mg/L, it was increased. Extension and shortening of HRT has positive and negative effects on nitrosation efficiency, dynamic controlling was an effective method. The system stability would be undermined leading to the risk of nitrification by elevating sludge recycle ration. In addition, low DO concentration did not reduce nitrosation rate. Under the condition of HRT = 6 h, R = 0.5, T = 22-24 degrees C, the average ammonia nitrogen removal rate reached 83%, the ammonia nitrogen removal load reached 0.28 kg/(kg x d), the nitrite accumulation ratio was close to 100%.

Air-drying is always accompanied by soil moisture loss. The different influences of air-drying on soil nitrification might due to the different nitrification responses to moisture changing of acid subtropical soils. So, after applying 0 and 150 mg x kg(-1) of ammonium bicarbonate, a 35-day incubation study was conducted to determine the nitrification potential of four acid subtropical soils under 5 soil moisture levels, namely 30% water-holding capacity (WHC), 45% WHC, 60% WHC, 75% WHC and 90% WHC. Four soils, designated QR, QU, SR and SU, derived from Quaternary red earth and Tertiary red sandstone, were collected from rice and upland field. The results indicated that the soil nitrification was significantly influenced by the moisture content (p < 0.01), and the nitrification sensitivities to soil moisture content varied with land-use types. For the treatments without ammonium input, the ranges of nitrification ratio were 11% and 8% in upland soils, QU and SU respectively, which were obviously lower than those in paddy soils (35% for QR, and 20% for SR). The ammonium input would increase the ranges of nitrification ratio which were 56%, 26%, 31%, and 26% for soil QR, QU, SR, and SU, respectively. And the ammonium input would accelerate soil acidification under high moisture levels. In a word, the land-use type presents a significant influence on the nitrification response to moisture content, which might lead to the difference of air-drying effect.

The feasibility of heterotrophic nitrification-aerobic denitrification bacteria applied in shortcut nitrification system was studied. Four heterotrophic nitrification-aerobic denitrification strains mixed with halotolerant activated sludge was added into SBR in order to test their bioaugmentation ability for shortcut nitrification system, which was treating for sewage containing sea water, and the difference between bioaugmentation system and original system was compared. The results showed that the maximum accumulation of NO2(-) -N in bioaugmentation system was 34.92% lower than that in original system, and the time of maximum accumulation of NO2(-) -N was 2 hours earlier than that in original system. The TN and COD was continuously decreasing in the later phase of nitrification in bioaugmentation system, and finally the removal rate of TN and COD were 15.24% and 5.39% higher than that in original system respectively, as well as the removal rate of NH4(+) -N and the nitrosation rate were 6.85% and 14.47% higher than that in original system. And the pH was 0.46 higher than that in original system, whereas the ORP was 25.84 mV lower. It was considered that the function of heterotrophic nitrification-aerobic denitrification bacteria should strengthen the performance of bioaugmentation system. When the seawater content raised to 70%, the stability of bioaugmentation system was better than that in original system, and the current that transforming shortcut nitrification to complete nitrification was restrained by heterotrophic nitrification-aerobic denitrification bacteria effectively. The number of heterotrophic nitrification-aerobic denitrification bacteria was changed when bioaugmentation system and original system ran in different phase and the bacteria had a great loss with the discharge of activated sludge. These results may provide a theoretical reference about the feasibility that the heterotrophic nitrification-aerobic denitrification bacteria applied in shortcut nitrification system.

To understand how to optimize performance of a partially nitrifying plant, the dynamics of Nitrospira and Nitrobacter abundance were studied over a 1 year period using quantitative polymerase chain reaction (qPCR) and their relative contributions to nitrite oxidation assessed including the affects of temperature and dissolved oxygen (DO). Correlation coefficients linking shifts in the community composition of nitrite-oxidizing bacteria (NOB) to operational or environmental variables indicated Nitrospira was significantly and negatively correlated to nitrite concentrations (r = -0.45, P < 0.01) and DO (r = -0.46, P < 0.01), while temperature showed a strong positive correlation (r = 0.59, P < 0.0001). However, the Nitrobacter portion of the total NOB populations showed a positive correlations with DO (r = 0.38, P < 0.01) and hydraulic retention time (HRT) (r = 0.33, P < 0.05), as well as being negatively correlated with temperature (r = -0.49, P < 0.001) suggesting specific niche adaptations within the NOB community. Nitrospira was dominant being better adapted to the low DO and shorter sludge retention times (SRT) of this plant, while Nitrobacter increased in abundance during the winter months, when temperatures were lower and DO concentrations higher. Principal component analysis (PCA) results supported these findings by the close proximity of Nitrospira and temperature biplots of PC1 and PC2 as well as grouping Nitrobacter, NO(2)(-)-N, HRT, and DO in the loadings together. The clustering of samples from specific dates also exhibited a strong seasonality., ((c) 2010 Elsevier Ltd. All rights reserved.)

A set of automatically titrimetric system to monitor the hydrogen ion variation during biological wastewater treatment process in a batch bioreactor was developed, which consists of a batch bioreactor, data auto-acquisition and preservation unit, and titrant autodosing unit. The accuracy of measurement for the system was evaluated by measuring the stoichiometric ratio of hydrogen ion production amount to ammonium consumption amount of ammonium oxidation in an activated sludge system. The ratios measured in a 1L bioreactor with NH4+ -N concentrations of 1.67, 3.33, 8.33, 16. 66 and 30.00 mg/L as N respectively were very close to the theoretical value, and the relative errors were among 2.09%-6.34%. However, the relative errors in bioreactors of 1, 2, 3 and 4 L with NH4+ -N concentration of 16.66 mg/L as N were among 2.09%- -18.57%, and increased significantly with accretion of the volume of bioreactor. The buffers of bicarbonate and the ammonium, especially the titrimetric dynamic effects in a larger bioreactor are the primarily factors resulting in errors. This study provides an important approach for monitoring hydrogen ion variation in the process of biological wastewater treatment by titrimetry.

Three submerged membrane bioreactors (MBRs) were operated continuously for 230 days by feeding with synthetic inorganic wastewater (NH(4)(+)-N, 100 mg L(-1)) under different solids retention times (SRTs. M(30d), 30 days; M(90d), 90 days; M(infinite), no sludge purge) to examine the influence of SRT on nitrification performance and microbial characteristics. All the reactors could oxidize NH(4)(+)-N to NO(3)(-)-N effectively without accumulation of NO(2)(-)-N. M(30d) with the shortest SRT showed significantly higher specific ammonium oxidizing rate (SAOR, 0.22 kg NH(4)(+)-N kg(-1) MLSS day(-1)) and specific nitrate forming rate (SNFR, 0.13 kg NO(3)(-)-N kg(-1) MLSS day(-1)) than the other two MBRs (0.12-0.14 kg NO(3)(-)-N kg(-1) MLSS day(-1) and 0.042-0.068kg NO(3)(-)-N kg(-1) MLSS day(-1), respectively). Short SRT led to low extracellular polymeric substances (EPS) concentration and long operating cycle. The nitrite oxidizing bacteria (NOB) ratios by both the fluorescence in situ hybridization (FISH) (3.6% for M(30d) and 2.1-2.2% for M(90d) and M(infinite)) and MPN (1.4 x 10(7)cells g(-1) MLSS for M(30d) and 6.2 x 10(5) and 2.7 x 10(4)cells g(-1) MLSS for M(90d) and M(infinite)) analyses showed that M(30d) favored the accumulation of NOB, which was in accordance with the SNFR result. However, the ammonia oxidizing bacteria (AOB) ratios (3.5%, 3.2% and 4.9% for M(30d), M(90d) and M(infinite)) were not in accordance with the SAOR result. PCR-DGGE, clone library and FISH results showed that the fast-growing Nitrosomonas and Nitrobacter sp. were the dominant AOB and NOB, respectively for M(30d), while considerable slow-growing Nitrosospira and Nitrospira sp. existed in M(infinite), which might be an important reason why M(infinite) had a low SAOR and SNFR., (Copyright (c) 2010 Elsevier Ltd. All rights reserved.)

Populations and activities of nitrifier can be significantly influenced by air-drying. A 35-day incubation study was conducted to determine the effects of air-drying on nitrification potential of four acid subtropical soils after applying 0 and 150 mg x kg(-1) of ammonium bicarbonate. Four soils, designated QR, QU, SR and SU, derived from Quaternary red earth and Tertiary red sandstone, were collected from rice and upland field. The results indicated that, without ammonium input, the responses of nitrification in upland soils to air-drying were insignificant (p > 0.05). The nitrification ratio in fresh and air-dried samples of soil QU were 48% and 54% respectively, and 76% and 78% for SU; however, air-drying did have a significant effect on the nitrification in paddy soils (p < 0.01). Nitrification ratios were 40% and 89% for fresh and air-dried samples of QR, 76% and 94% for SR, and there was a lag-phase of the nitrification in air-dried paddy soils. Ammonium input would accelerate nitrification and make the nitrification ratios of fresh samples significantly higher than those of air-dried ones. In generally, land-use type presents a significant impact on the response of nitrification to air-drying effect, and there was a significant interaction of land-use type and fertilization.

Autotrophic granular sludge was developed in an SBR reactor using inorganic carbonal substrate. The variation of ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) during the densification process and their effects on granulation have been evaluated. It was found that the autotrophic bacteria granular sludge was compact with the density reached up to 1.06 g/mL, and the rod-like bacteria predominated in granules on the evidence of scanning electron microscopic (SEM) results. Ammonia, nitrite and nitrate in the effluent were 4.5-15.2 mg/L, 10.2-20.3 mg/L and 17.9-30.1 mg/L, respectively, and the ammonia removal efficiency was 78% -92%. By evaluating the profile of various types of nitrogen and their conversion rates, it was found that short settling time was the main factor that enriched the AOB at the beginning of this experiment, and the granulation did not correlate with AOB. On the contrary, nitrification rate well correlated with granulation, and evidence demonstrated that the formation of granulation was in favorate of immoblization of NOB and the metabolite of NOB stabilized granules, therefore granules and NOB mutually enhanced. Additionally, it was found that the autotrophic denitrification was gradually increased with the process of granulation.

Despite their role in soil functioning, the ecology of nitrite-oxidizing bacteria, NOB, and their response to disturbances such as those generated by agricultural practices are scarcely known. Over the course of 17 months, we surveyed the potential nitrite oxidation, PNO, the abundance of the Nitrobacter- and Nitrospira-like NOB (by quantitative PCR) and the community structure of the Nitrobacter-like NOB (by PCR-DGGE and cloning-sequencing targeting the nxrA gene) in soils for four treatments: after establishment of tillage on a previously no-tillage system, after cessation of tillage on a previously tillage system, and on control tillage and no-tillage systems. Key soil variables (moisture, organic carbon content and gross mineralization--i.e. ammonification--measured by the 15N dilution technique) were also surveyed. PNO was always higher for the no-tillage than tillage treatments. Establishment of tillage led to a strong and rapid decrease in PNO whereas cessation of tillage did not change PNO even after 17 months. PNO was strongly and positively correlated to the abundance of Nitrobacter-like NOB and was also strongly related to gross mineralization, a proxy of N-availability; in contrast, PNO was weakly and negatively correlated to the abundance of Nitrospira-like NOB. Selection of a dominant population was observed under no-tillage, and PNO was loosely correlated to the community structure of Nitrobacter-like NOB. Our results demonstrate that Nitrobacter-like NOB are the key functional players within the NOB community in soils with high N availability and high activity level, and that changes in PNO are due to shifts between Nitrospira-like and Nitrobacter-like NOB and to a weaker extent by shifts of populations within Nitrobacter-like NOB.

The cell numbers of ammonia-oxidising bacteria (AOBs), Nitrospira and Nitrobacter in activated sludge used to treat wastewater from a thermal power plant in Japan were examined for nine months using a real-time PCR quantification technique. AOB cell numbers ranged 2.8 x 10(10)-2.3 x 10(11) cell/L. The amoA clone analysis showed that the only Nitrosomonas halophila was responsible for ammonia oxidation over the period. Nitrospira were in the range of 2.6 x 10(9)-2.4 x 10(10) cell/L and Nitrobacter were less than 1% as common as Nitrospira. Meanwhile, maximum nitrification rates, maximum ammonia- and nitrite-oxidation rates obtained from aerobic batch tests, ranged 0.5-1.3 mmol-N/L h and 1.0-2.5 mmol-N/L h, respectively. No clear correlations were observed between the cell numbers of AOBs or Nitrospira and their maximum rates, because the maximum cell-specific ammonia- and nitrite-oxidation rates varied remarkably over the ranges of 1.1-11.9 and 2.4-21.6 fmol-N/cell h, respectively. To explore the factors controlling maximum cell-specific nitrification rates, the relationship to influent nitrogen loads per AOB or Nitrospira cell numbers was investigated. Fairly good correlations were obtained. Considering the effluent ammonia and nitrite concentrations were zero and only Nitrosomonas halophila had a role in ammonia oxidation over the period, we conclude that the amount of nitrogen oxidised per AOB or Nitrospira cell numbers likely controls maximum cell-specific ammonia- or nitrite-oxidation rates, respectively.

The influence factors and environmental effects of nitrification process were discussed based on the measurements of nitrifier quantities and nitrification rates at eight sites off Yangtze River Estuary in June 2006. Using FISH (Fluorescence in situ hybridization) method, the nitrifying bacteria were counted as 1.87 x 10(5) -3.53 x 10(5) cells/g wet weight and exhibited salt tolerance to some extent. The nitrification rates, measured by acetylene inhibition technique, ranged from 101.3 to 514.3 micromol/(m2 x h) and had a decreased gradient from nearshore to offshore sediment with two high value regions located at the mouth of Yangtze River and Hangzhou Bay respectively. The nitrifier quantity was a main factor to effect nitrification rates at high-salinity sea area, with an influence ratio of 87.7%. Fluxes of transformed nitrogen and consumed oxygen by nitrification process were 4.68 x 10(5) kg and 6.07 x 10(4) mol per day respectively, suggesting that nitrification would be an important factor for the distribution of DIN species and DO at bottom water in early summer.

Taking simulated domestic sewage as treatment object, the characteristics of separate-stage phosphorus, organic substrate and nitrogen removal were studied in two-stage SBR by contrast experiment and mechanism analysis under the normal temperature, its efficiency-predominance was analyzed. The results indicate that the heterotrophic PAOs and nitrobacteria can be dominant growth in individual reactor respectively, under the effluent quality is more superior, the treatment efficiency is one time higher than that in the single SBR by controlling the sludge age (the phosphorus removal stage 5-7 d, the nitrogen removal stage about 50 d). The two-stage SBR can relieve the attack of organic load to nitrification process effectively, its nitrogen removal stage (SBR2) can still maintain stable nitrification rate under the higher COD concentration and the final effluent of the system is easy and stable to reach national standards (TP < or = 0.5 mg x L(-1)). In addition, the nitrogen removal stage (SBR2) of two-stage SBR not only can maintain nitrobacteria predominant, but also can cultivate heterotrophic bacteria which can remove difficult degraded organic substrate, that results in lower COD concentration than in the single's at the end of nitrification.

According to nitrogen-containing wastewater, effects of salinity variation on nitrification and denitrification in internal circulation sequencing batch biofilm reactor (SBBR) and sequencing batch reactor (SBR), and the process of recovery as decreased salinity were investigated. The results show that nitrosation process is effected in internal circulation SBBR and SBR as the increase of salinity, and as the salinity is less than 1.0 x 10(4) mg x L(-1), the speed of ammonia-oxidize is quicker in the SBR than in the SBBR, but the state is opposite as the salinity beyond 1.5 x 10(4) mg x L(-1). As the salinity increased to 4.0 x 10(4) mg x L(-1), the process of ammonia-oxidize was restrained strongly both in SBBR and SBR. The nitrite accumulation is attained continually as the salinity at 1.5 x 10(4) mg x L(-10 and 2.5 x 10(4) mg x L(-1) in the SBBR and SBR respectively. In the SBBR, the removal rate of TN was about 60% as the salinity was less than 1.5 x 10(4) mg x L(-1), and the SND was restrained strongly as the salinity beyond 3.0 x 10(4) mg x L(-1).

In a bid to identify suitable microbial indicators of compost stability, the process evolution during windrow composting of poultry manure (PM), green waste (GW) and biowaste was studied. Treatments were monitored with regard to abiotic factors, respiration activity (determined using the SOUR test) and functional microflora. The composting process went through typical changes in temperature, moisture content and microbial properties, despite the inherent feedstock differences. Nitrobacter and pathogen indicators varied as a monotonous function of processing time. Some microbial groups have shown a potential to serve as fingerprints of the different process stages, but still they should be examined in context with respirometric tests and abiotic parameters. Respiration activity reflected well the process stage, verifying the value of respirometric tests to access compost stability. SOUR values below 1 mg O(2)/g VS/h were achieved for the PM and the GW compost.

The community of nitrifiers cultivated with reject water were investigated by fluorescent in situ hybridization (FISH) performed with 16S rRNA-targeted oligonucleotide probes, and the kinetic properties such as temperature correction factor tauN and the half saturation coefficient K(N) were also investigated. The results showed that the percentage of ammonia oxidizing bacteria (AOB) and nitrite oxidizing bacteria (NOB) in the activated sludge was 15.7% +/- 3.7% and 12.9% +/- 3.2%, Nitrosomonas europaea-Nitrosococcus mobilis lineage were the dominant AOB, Nitrobacter spp. were the dominant NOB. The temperature correction factors of ammonium utilized rate (AUR) and nitrite utilized rate (NUR) were 1.092 and 1.061 respectively; the K(N) of AOB and NOB were (1.60 +/- 0.29) mg/L (NH4(+) -N), (2.78 +/- 0.30) mg/L (NO2(-) -N) respectively.

Nitrogen cycling bacteria, including ammonifying, nitrobacteria, nitrosobacteria and denitrifying bacteria were screened, carrier was made and immobilized nitrogen cycling bacteria (INCB) was prepared. The results demonstrated that ammonifying, nitrobacteria, nitrosobacteria and denitrifying bacteria were increased markedly in the experimental areas and root zone of aquatic plants by releasing of INCB. The results also showed that the average removal efficiencies for total N (TN), NH4(+) -N, NO3(-) -N and NO2(-) -N were 44.70%, 67.17%, 31.79% and 74.21%, respectively. Furthermore, NH4(+) -N, total N (TN) reached the National Standard II and IV for surface water, respectively. With INCB, local lake water quality could improve. Therefore, the technique of INCB could play an important role for remedying and rehabilitating in desertification water.

Paddy soil samples were collected in layers (0-5, 5-12, and 12-20 cm) during rice growth period to investigate the characteristics of the N forms and N-transforming bacteria in the soil profile under different tillage patterns (no-tillage with straw returning, NTS; conventional tillage with straw returning, CTS; no-tillage, NT; and conventional tillage, CT). In the whole rice growth period, ammonifying bacteria in 0-5 cm soil layer had the highest number under NTS, and nitrosobacteria in 0-5 cm and 5-12 cm soil layers were more abundant but in 12-20 cm soil layer were lesser under CT than under NT. Nitrosobacteria and denitrobacteria in 0-20 cm soil layer were lesser under NTS than under CTS. At elongating and ripening stages, anaerobic N-fixing bacteria in 0-5 cm soil layer were more abundant under NT than under CT. In the whole rice growth period, the alkali-hydrolyzable N and total N contents in 0-5 cm soil layer were significantly higher but in 5-12 cm and 12-20 cm soil layers were lower under NT than under CT, and the NH4(+)-N and NO3(-)-N contents in 0-20 cm soil layer were higher under NTS but in 12-20 cm soil layer had no significant differences between NT and CT. Correlation analysis and multiple polynomial regression analysis further revealed that there were significant relationships between soil NH4(+)-N and soil ammonifying bacteria, nitrosobacteria and denitrobacteria, and between soil alkali-hydrolyzable N and soil anaerobic N-fixing bacteria. Among the test tillage patterns, NTS could be the more desirable one for the N supply and fertility maintenance of paddy soil.

An expanded granular sludge bed (EGSB) reactor inoculated simultaneously with aerobic ammonium oxidation sludge and anaerobic ammonium oxidation sludge were start-up to enrich completely autotrophic nitrogen removal granular sludge. Total nitrogen (TN) removal rate reached 0.101 kg x (m3 x d)(-1). Based on hypothesis of boundary layer, the transfer process between granular sludge and bulk liquid was modified, which was coupled with substance transfer process in granular sludge and aerobic ammonium oxidation, anaerobic ammonium oxidation, nitrite oxidation process, and completely autotrophic nitrogen removal model was found. The model was validated with the experimental results. According to simulation results, the operation of the reactor was optimized, TN removal efficiency and TN removal rate were increased from 52% to 61% and 0.103 kg x (m3 x d)(-1) to 0.114 kg x (m3 x d)(-1) respectively.

The 16S rDNA-based molecular technique was applied to investigate the functional microbial community of a membrane-aerated biofilm bioreactor (MABR) that was used for completely autotrophic nitrogen removal over nitrite (CANON). The relationships among two kinds of key bacteria responsible for CANON: aerobic ammonia-oxidizing bacteria (AOB) and ANAMMOX bacteria, and their possible distributions in the MAB were discussed based on the microbial community analysis. FISH analysis showed the existence of two visible active layers in experimental MAB. One was the partial nitrifying layer located in the region of oxygen-rich membrane-biofilm interface, dominated by NSO190-positive AOB. The other was the ANAMMOX active layer located in the region of anoxic liquid-biofilm interface, dominated by PLA46 and AMX820-positive ANAMMOX microorganisms. As a result, the AOB as well as ANAMMOX bacteria were present and active in experimental MABR, and the cooperation between AOB and ANAMMOX bacteria was considered to be responsible for CANON.

At normal temperature of 19 degrees C +/- 1 degree C and seed sludge from the system of conventional nitrogen removal via nitrate composed of both ammonia oxidizing bacteria (AOB) and nitrite oxidizing bacteria (NOB), the competition and optimization of AOB and NOB was investigated in a lab-scale SBR treating domestic wastewater. The reactor was operated under the aeration rate of 40 L/h with the average DO concentration below 1.0 mg/L, and combined with the control of aeration time, i.e., aeration was stopped before the pH "ammonia valley" was detected. After AOB became pre-dominant nitrifying bacteria (nitrite accumulation rate NO2(-)-N/NOx(-)-N reached 50%), the aerobic durations were gradually prolonged from 3 h to 4 h and 5 h to improve the ammonia removal efficiencies and enhance AOB dominant. By the above operational pattern, nitrogen removal via nitrite was successfully achieved with NO2(-)-N/NOx(-)-N over 95%. Fluorescence in-situ hybridization (FISH) analysis showed that AOB was accounted for 9.97% of total biomass. The on-line control of aerobic duration was an effective method to maintain the partial nitrification and the NH4(+)-N removal efficiency was above 97%. The research results also proved that at normal temperatures, for the seed sludge from the conventional nitrogen removal via nitrate, AOB would not become dominant nitrifying bacteria only by the aeration time control without the DO restriction.

This study evaluated nitrification performance and microbial ecology of nitrifying sludge in two full-scale wastewater treatment plants (WWTPs) including a municipal WWTP treating 20mgNL(-1) of ammonium and a swine WWTP treating 220mgNL(-1) of ammonium. These two plants differed in both wastewater characteristics and operating parameters, such as influent COD, TKN, ammonium, hydraulic retention time, and solids retention time, even though both plants achieve >85% nitrification efficiency. By employing molecular techniques, including terminal restriction fragment length polymorphism, cloning-sequencing and phylogenetic analyses targeting the 16S ribosomal RNA and group specific ammonia-monooxygenase functional gene (amoA), microbial community structures of nitrifying sludge and their significance to nitrification performance were evaluated. The results reveal that for the municipal WWTP Nitrosomonas marina-like AOB (ammonia-oxidizing bacteria) and Nitrospira-like NOB (nitrite-oxidizing bacteria) were the ubiquitously dominant nitrifiers, while Nitrosomonas europaea-, Nitrosomonas oligotropha-, and Nitrosospira-like AOB and Nitrobacter- and Nitrospira-like NOB were the major nitrifying populations found in the swine WWTP. The observed dissimilar nitrifying populations prevailing in these two plants may be related to niche differentiation concerning ammonium concentrations, system operation, and salinity. Moreover, our results suggest that the swine nitrifying sludge, involving relatively diverse AOB and NOB populations that perform the same task but with distinct growth and survival characters, may allow communities to maintain nitrifying capabilities when conditions change such as sudden increases in ammonium concentrations as examined with nitrification kinetic batch tests.

Methods for characterizing nitrifying bacteria within biofilms are of key importance to understand and optimize the nitrification kinetics of attached growth treatment facilities. In this work, we propose an analytical protocol based upon environmental scanning electron microscopy (ESEM) and confocal laser scanning microscopy (CSLM) in combination with fluorescent in situ hybridization (FISH) to characterize the structure of nitrifying biofilm as it remains attached to the original reactor substratum. This protocol minimizes the loss of mass and distortion of in situ perspective commonly associated with traditionally applied microscopic techniques and thereby enables a more accurate estimation of the nitrifying biomass within biofilm attached to the substratum. The use of ESEM eliminates the destructive preparatory procedures associated with traditional scanning electron microscopy and thus the loss of mass and shrinking of the samples. ESEM is used in this study to evaluate the percent coverage of the substratum with biofilm and the biofilm thickness. CLSM-FISH is used to determine cell counts in the biofilm and to characterize the undisturbed substratum/biofilm interface. By hybridizing and analyzing the nitrifying biofilm using CLSM as it remains attached to the substratum, the loss of material and distortion of in situ perspective associated with the biofilm detachment process is minimized. Moreover, by conducting the CLSM analysis directly on the nitrifying biofilm as it remains attached to the substratum it is shown that cell counts at the substratum/biofilm interface differ significantly from that located above the interface.

A packed bed bioreactor (PBBR) was developed for rapid establishment of nitrification in brackish water hatchery systems in the tropics. The reactors were activated by immobilizing ammonia-oxidizing (AMONPCU-1) and nitrite-oxidizing (NIONPCU-1) bacterial consortia on polystyrene and low-density polyethylene beads, respectively. Fluorescence in situ hybridization demonstrated the presence of autotrophic nitrifiers belong to Nitrosococcus mobilis, lineage of beta ammonia oxidizers and nitrite oxidizer Nitrobacter sp. in the consortia. The activated reactors upon integration to the hatchery system resulted in significant ammonia removal (P < 0.01) culminating to its undetectable levels. Consequently, a significantly higher percent survival of larvae was observed in the larval production systems. With spent water the reactors could establish nitrification with high percentage removal of ammonia (78%), nitrite (79%) and BOD (56%) within 7 days of initiation of the process. PBBR is configured in such a way to minimize the energy requirements for continuous operation by limiting the energy inputs to a single stage pumping of water and aeration to the aeration cells. The PBBR shall enable hatchery systems to operate under closed recirculating mode and pave the way for better water management in the aquaculture industry.

In order to investigate shortcut nitrification, an intermittent-aeration SBR was employed to treat the domestic wastewater containing low concentration of NH4+ . The results shows that under the SRTs of 10, 5, 2.5 and 1.25 d, NO2- produced in effluent were built up to 18, 19, 14 and 5 mg/L, respectively, with accumulation ratio of 73%, 85%, 91% and 78% respectively. As to the continuous-aeration SBR, however, the NO2- accumulation ratios were only 14%, 21%, 31% and 34%. In addition, the corresponding removals of NH4- in the intermittent-aeration SBR were maintained at a comparable level in contrast to those for the continuous-aeration SBR, i.e., 97%, 95%, 76% and 39% vs. 92%, 97%, 71% and 47%. By exploring the growth kinetics of nitrifiers, ammonia-oxidizing bacteria (AOB) in the intermittent-aeration system was capable of increasing its biomass in the reactor by an elevation in growth yield (Y(AOB)), which as a result compensated the decreased specific substrate utilization rate at low DO level and further led to the unchanged microm value and NH4+ oxidation rate. On the contrary, nitrite-oxidizin bacteria (NOB) was lack of the function of compensability, and it hence lowered the microm value and NO2-oxidation rate, and therefore resulted in an accumulation of NO2- in the effluent.

Manmade ecosystems differ from their prototype biosphere by the principle of control. The Earth Biosphere is sustainable by stochastic control and very large time constants. By contrast, in a closed ecosystem such as the micro-ecological life support system alternative (MELiSSA system) developed by the European Space Agency for space exploration, a deterministic control is a prerequisite of sustainable existence. MELiSSA is an integrated sum of interconnected biological subsystems. On one hand, all unit operations in charge of the elementary functions constitutive of the entire life support system are studied until a thorough understanding and mathematical modelling. On the other hand, the systemic approach of complex, highly branched systems with feedback loops is performed. This leads to study in the same perspective, with the same degree of accuracy and with the same language, waste degradation, water recycling, atmosphere revitalisation and food production systems prior to the integration of knowledge-based control models. This paper presents the mathematical modelling of the MELiSSA system and the interface between the control strategy of the entire system and the control of the bioreactors.

Nitrification is a key step in the global nitrogen cycle. Classically, nitrifying bacteria are chemoautotroph. Recently, bacteria that have the ability of heterotrophic nitrification have been intensively studied as potential microorganisms that may be used to overcomeproblems inherent in the conventional method. This review gives a broad overview of the current status of heterotrophic nitrification including the heterotrophic species which nitrify actively, optimal condition for nitrification, heterotrophic nitrification pathway, enzymatic system and isolation methods. Also, the promising prospects of heterotrophic nitrification are especially introduced.

A sequencing batch airlift reactor (SBAR) was conducted to study the performance of physical characteristics and respirometric activities during reactivation of aerobic granules after a 2-month storage time. Results showed that the color of the aerobic granules had completely become dark, the size and settleability did hardly change during the storage period. The color turned to be brownish-yellowish when the reactor was restarted for 2 weeks. The MLSS, granule size, and settleability increased rapidly along with reactivation. The heterotrophic activity resumed 86% within 1 day and was fully reactivated after 5 days. Then the COD removal rate was stabilized above 80%. Higher aeration rate and longer cycle time facilitated the reactivation of nitrifiers. When aeration rate was increased from 0.05 m3 x h(-1) to 0.10 m3 x h(-1) at 41st day, activities of ammonia-oxidizing bacteria and nitrite-oxidizing bacteria were improved from 88% and 82% to 122% and 92%, respectively. The removal rate of ammonium-nitrogen promptly reached above 96%. Nitrite-oxidizing bacteria were fully reactivated after the cycle time was prolonged from 4 h to 6 h.

The nitrite oxidizing Alphaproteobacterium, Nitrobacter winogradskyi, primarily conserves energy from the oxidation of nitrite (NO(2)(-))to nitrate (NO(3)(-)) through aerobic respiration. Almost 20 years ago, NO-dependent NADH formation was reported to occur in both aerobic and anaerobic cell suspensions of N. winogradskyi strain 'agilis', suggesting that NO oxidation might contribute to energy conservation by Nitrobacter. Recently, the N. winogradskyi Nb-255 genome was found to contain a gene (Nwin&#95;2648) that encodes a putative copper-containing nitrite reductase (NirK), which may reduce NO(2)(-) to NO. In this study, the putative nirK was found to be maximally transcribed under low O(2) (between zero and 4% O(2)) in the presence of NO(2)(-). Transcription of nirK was not detected under anaerobic conditions in the absence of NO(2)(-) or in the presence of NO(3)(-) and pyruvate. Although net production of NO could not be detected from either aerobically grown or anaerobically incubated cells, exogenous NO was consumed by viable cells and concomitantly inhibited NO(2)(-)-dependent O(2) uptake in a reversible, concentration dependent manner. Both NO(2(-)-dependent O(2) uptake and NO consumption were inhibited by 1 mM cyanide suggesting involvement of cytochrome oxidase with NO consumption. Abiotic consumption of NO was measured, yet, both the rates and kinetics of NO transformation in buffer alone, or by heat killed, or cyanide-treated cells differed from those of viable cells. In light of this new information, a modified model is proposed to explain how NirK and NO manage electron flux in Nitrobacter.

Aeration phase length control and step-feed of wastewater are used to achieve nitrogen removal from wastewater via nitrite in sequencing batch reactors (SBR). Aeration is switched off as soon as ammonia oxidation is completed, which is followed by the addition of a fraction of the wastewater that the SBR receives over a cycle to facilitate denitrification. The end-point of ammonia oxidation is detected from the on-line measured pH and oxygen uptake rate (OUR). The method was implemented in an SBR achieving biological nitrogen and phosphorus removal from anaerobically pre-treated abattoir wastewater. The degree of nitrite accumulation during the aeration period was monitored along with the variation in the nitrite oxidizing bacteria (NOB) population using fluorescence in situ hybridization (FISH) techniques. It is demonstrated that the nitrite pathway could be repeatedly and reliably achieved, which significantly reduced the carbon requirement for nutrient removal. Model-based studies show that the establishment of the nitrite pathway was primarily the result of a gradual reduction of the amount of nitrite that is available to provide energy for the growth of NOB, eventually leading to the elimination of NOB from the system., (2008 Wiley Periodicals, Inc.)

Nitrobacter hamburgensis X14 is a facultative lithoautotroph that conserves energy from the oxidation of nitrite (NO(-)2) and fixes carbon dioxide (CO(2)) as its sole source of carbon. The availability of the N. hamburgensis X14 genome sequence initiated a re-examination of its mixotrophic and organotrophic potential, as genes encoding three flavin-dependent oxidases were identified that may function to oxidize lactate, providing energy and carbon for growth. The response of N. hamburgensis to D- and L-lactate in the presence (mixotrophy) and absence (organotrophy) of NO(-)2 was examined. L-lactate did not support organotrophic growth or stimulate mixotrophic growth. In contrast, D-lactate enhanced the growth rate and yield of N. hamburgensis in the presence of NO(-)2, and served as the sole carbon and energy source for growth in the absence of NO(-)2 with ammonium as the sole nitrogen source. Lithoautotrophically grown cells immediately consumed D-lactate, suggesting that a lactate metabolic pathway is constitutively expressed. Nevertheless, a physiological adaptation to lactate occurred, as D-lactate-grown cells consumed and assimilated lactate at a faster rate than NO(-)2-grown cells, and the D-lactate-dependent O(2) uptake rate was significantly greater in cells grown either organotrophically or mixotrophically compared with cells grown lithoautotrophically. Although D-lactate was assimilated and metabolized to CO(2) in the presence or absence of NO(-)2, exposure to atmospheric CO(2) or the addition of 0.75 mM sodium carbonate was required for mixotrophic growth and for optimum organotrophic growth on D-lactate.

At the high level of dissolved oxygen (DO) in sequencing batch biofilm reactor (SBBR), the approach and mechanism for realizing shortcut nitrification were researched. Landfill leachate was used as handling of object, the mainly environment parameters of the reactor were controlled as follow: DO 5 mg/L, pH 7.0, temperature 25 degrees C, adopted all drainage mode and 12-hour cycle influent. Through mathematical derivation and modeling analysis, determined free ammonia (FA), CO2 and HNO2 as the direct control factors, whereas the influent cycle time was the indirect one, shortcut nitrification was achieved effectively in SBBR. When the volume load of ammonia (NH4(+) -N) was 0.52 kg/(m3 x d) and NaHCO3 was 1.5 mg/L in the reactor, the shortcut nitrification effect was apparent as NH4(+) -N conversion rate was 89% and NO2(-) -N accumulation rate achieved 83% at the same time. With adequate oxygen supply, the key factors of achieving NO2(-) -N accumulation is FA concentration, and as the carbon source of ammonia-oxidizing bacteria, CO2 can upgrade the reactor performance further.

A nitrifying sequencing batch reactor, operated under alternating anoxic/aerobic conditions achieved twice the nitrification rates of its strictly aerobic counterpart. Microbial populations in both reactors were examined with fluorescent in situ hybridization (FISH) and kinetic batch studies to determine the effects of ammonium, nitrite, and oxygen. FISH revealed a dominance of rapid nitrifiers like Nitrosomonas and Nitrobacter (79.5% of the nitrifying population) in the alternating reactor, compared with the dominance of slower nitrifiers like Nitrosospira and Nitrospira (78.2%) in the strictly aerobic reactor. Nitrifiers in the aerobic reactor operated at maximum rates and were negatively affected by ammonium or nitrite, whereas nitrifying rates in the alternating reactor were proportional to ammonium or nitrite concentrations. The alternating conditions were more favorable because they selected for faster nitrifiers due to their oxidation, growth, and decay rates. The findings are of importance to the design engineers, as the reactors are typically designed based on nitrifiers' growth rate determined in strictly aerobic conditions.

Sequencing batch reactor, treating simulated municipal wastewater for N removal and operated at near the minimum SRT (solids retention time) for nitrification (in reactor 2), was seeded daily with nitrifying biomass accumulated with reject water (in reactor 1) came from Xi' an Dengjiacun wastewater treatment plant. Comparison of the nitrification ability, the characteristics of the activated sludge before seeding and after seeding was made. The experiment result shows that, the maximum nitrification rate of the activated sludge in the reactor 1 reached 81.4 mg/(L x h). The effluent concentration of NH4+ -N of the reactor 2 decreased at a rate of 0.992 mg/(L x d) (R2 = 0.903), and the maximum nitrification rate increased 1.36 times. Once seeding was terminated, the effluent concentration of NH4+ -N increased at a rate of 1.956 mg/(L x d) (R2 = 0.999), and nitrification caused by seeding was not completely disappeared after seeding was stopped for 34 d. The amount and varieties of protozoa and the value of SVI in the reactor seeded with nitrifiers increased during the initial time of seeding, but were recovered when the reactor 2 reached stable state.

The hybrid membrane biofilm process (HMBP) is a novel approach to achieving total nitrogen removal from wastewater. It incorporates air-supplying, hollow-fiber membranes into a conventional activated sludge tank, where bulk aeration is suppressed. A nitrifying biofilm grows on the membranes and exports nitrite and nitrate to the anoxic bulk liquid, where suspended heterotrophic bacteria denitrify using influent BOD as an electron donor. Bench scale tests were used to assess the performance of the HMBP for a variety of nitrogen and BOD loading rates. For a nitrogen loading of 1.6 gN m(-2), the nitrification flux remained at approximately 1.0 gN m(-2) d(-1) for BOD loadings ranging from 4 to 17 gBOD m(-2) d(-1) day(-1). Full denitrification was achieved when sufficient BOD was available in the influent. Microsensor measurements indicated nitrite was the dominant form of oxidized nitrogen produced by the biofilm, showing that shortcut nitrogen removal was taking place. Fluorescence in-situ hybridization (FISH) tests on the biofilm revealed a unique stratification, with three distinct regions: AOB and NOB near the membrane, strictly AOB at intermediate depths, and AOB and heterotrophs at the outer edge of the biofilm., (IWA Publishing 2008.)