An autotrophic nitrogen removal process:Short-cut nitrification combined with ANAMMOX for treating diluted effluent from an UASB reactor fed by landfill leachate

A combined process consisting of a short-cut nitrification （SN） reactor and an anaerobic ammonium oxidation upflow anaerobic sludge bed （ANAMMOX） reactor was developed to treat the diluted effluent from an upflow anaerobic sludge bed （UASB） reactor treating high ammonium municipal landfill leachate.The SN process was performed in an aerated upflow sludge bed （AUSB） reactor （working volume 3.05 L）,treating about 50% of the diluted raw wastewater.The ammonium removal efficiency and the ratio of NO 2 N to NOx-N in the effluent were both higher than 80%,at a maximum nitrogen loading rate of 1.47 kg/（m 3 ·day）.The ANAMMOX process was performed in an UASB reactor （working volume 8.5 L）,using the mix of SN reactor effluent and diluted raw wastewater at a ratio of 1：1.The ammonium and nitrite removal efficiency reached over 93% and 95%,respectively,after 70-day continuous operation,at a maximum total nitrogen loading rate of 0.91 kg/（m 3 ·day）,suggesting a successful operation of the combined process.The average nitrogen loading rate of the combined system was 0.56 kg/（m 3 ·day）,with an average total inorganic nitrogen removal efficiency 87%.The nitrogen in the effluent was mostly nitrate.The results provided important evidence for the possibility of applying SN-ANAMMOX after UASB reactor to treat municipal landfill leachate.

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

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

Potential Secretory Transporters and Biosynthetic Precursors of Biological Nitrification Inhibitor 1,9-Decanediol in Rice as Revealed by Transcriptome and Metabolome Analyses

Biological nitrification inhibitors(BNIs)are released from plant roots and inhibit the nitrification activity of microorganisms in soils,reducing NO_(3)^(‒)leaching and N2O emissions,and increasing nitrogenuse efficiency(NUE).Several recent studies have focused on the identification of new BNIs,yet little is known about the genetic loci that govern their biosynthesis and secretion.We applied a combined transcriptomic and metabolomic analysis to investigate possible biosynthetic pathways and transporters involved in the biosynthesis and release of BNI 1,9-decanediol(1,9-D),which was previously identified in rice root exudates.Our results linked four fatty acids,icosapentaenoic acid,linoleate,norlinolenic acid,and polyhydroxy-α,ω-divarboxylic acid,with 1,9-D biosynthesis and three transporter families,namely the ATP-binding cassette protein family,the multidrug and toxic compound extrusion family,and the major facilitator superfamily,with 1,9-D release from roots into the soil medium.Our finding provided candidates for further work on the genes implicated in the biosynthesis and secretion of 1,9-D and pinpoint genetic loci for crop breeding to improve NUE by enhancing 1,9-D secretion,with the potential to reduce NO_(3)^(‒)leaching and N2O emissions from agricultural soils.

Sludge fermentation liquid addition attained advanced nitrogen removal in low C/N ratio municipal wastewater through short-cut nitrification-denitrification and partial anammox

Biological nitrogen removal of wastewater with low COD/N ratio could be enhanced by the addition of wasted sludge fermentation liquid(SFL),but the performance is usually limited by the introducing ammonium.In this study,the process of using SFL was successfully improved by involving anammox process.Real municipal wastewater with a low C/N ratio of 2.8–3.4 was treated in a sequencing batch reactor(SBR).The SBR was operated under anaerobic-aerobic-anoxic(AOA)mode and excess SFL was added into the anoxic phase.Stable short-cut nitrification was achieved after 46d and then anammox sludge was inoculated.In the stable period,effluent total inorganic nitrogen(TIN)was less than 4.3 mg/L with removal efficiency of 92.3%.Further analysis suggests that anammox bacteria,mainly affiliated with Candidatus_Kuenenia,successfully reduced the external ammonia from the SFL and contributed approximately 28%–43%to TIN removal.Overall,this study suggests anammox could be combined with SFL addition,resulting in a stable enhanced nitrogen biological removal.

Simultaneous nitrification and autotrophic denitrification in fluidized bed reactors using pyrite and elemental sulfur as electron donors

In this study, simultaneous nitrification and autotrophic denitrification (SNAD) with either elemental sulfur or pyrite were investigated in fluidized bed reactors in mesophilic conditions. The reactor performance was evaluated at different ammonium (12-40 mg/L of NH4+-N), nitrate (35-45 mg/L of NO3--N), and dissolved oxygen (DO) (0.1-1.5 mg/L) concentrations, with a hydraulic retention time of 12 h. The pyrite reactor supported the SNAD process with a maximum nitrogen removal efficiency of 139.5 mg/(L·d) when the DO concentration was in the range of 0.8-1.5 mg/L. This range, however, limited the denitrification efficiency of the reactor, which decreased from 90.0% ± 5.3% in phases II-V to 67.9% ± 7.2% in phases VI and VII. Sulfate precipitated as iron sulfate (FeSO4/Fe2(SO4)3) and sodium sulfate (Na2SO4) minerals during the experiment. The sulfur reactor did not respond well to nitrification with a low and unstable ammonium removal efficiency, while denitrification occurred with a nitrate removal efficiency of 97.8%. In the pyrite system, the nitrifying bacterium Nitrosomonas sp. was present, and its relative abundance increased from 0.1% to 1.1%, while the autotrophic denitrifying genera Terrimonas, Ferruginibacter, and Denitratimonas dominated the community. Thiobacillus, Sulfurovum, and Trichlorobacter were the most abundant genera in the sulfur reactor during the entire experiment.

Effects of chlorothalonil and carbendazim on nitrification and denitrification in soils

The effects of chlorothalonil and carbendazim on nitrification and denitrification in six soils in upland and rice paddy environments were investigated. Laboratory aerobic （60% water holding capacity） and anaerobic （flooded） conditions were studied at 25℃ and fungicide addition rates of 5.5 mg/kg A. I. （field rate, FR）, 20 times （20FR） and 40 times （40FR） field rate, respectively. The results indicated that chlorothalonil at the field rate had a slight inhibitory effect on one soil only, and that soil did not nitrify much in the first place. But chlorothalonil at higher rates inhibited nitrification significantly in all soils. For soils JXP and JXU with a pH of less than 5.0, chlorothalonil almost completely stopped their nitrification at 20FR and 40FR during the whole 14 d incubation period. For soils HNP and HNU with a pH of greater than 8.0, chlorothalonil also significantly inhibit nitrification at 20FR and 40FR （p 〈 0.05）. However, NH4＋ that was added to the soil was also almost completely nitrified by the end of the incubation period in these two soils. The effects of chlorothalonil at 20FR and 40FR on the nitrification of JSP and JSU soils, with a pH of 5.4 and 7.2, respectively, were intermediate between the other soil types. Chlorothalonil had no effect on denitrification at the field rate and had little effect at the higher rates of application in some soils. Carbendazim had essentially no effect on nitrification and denitrification in soils assessed.

Temperature effect on aerobic denitrification and nitrification

Nitrogen loss without organic removal in biofilter was observed and its possible reason was explained. A lower hydraulic loading could improve aerobic denitrification rate. Aerobic denitrification was seriously affected by low temperature(below 10℃). However, nitrification rate remained high when the temperature dropped from 15℃ to 5℃. It seemed the autotrophic biofilm in BAF could alleviate the adverse effect of low temperature.

Nitrification potentials of Chinese tea orchard soils and their adjacent wasteland and forest soils

To investigate the nitrifying activities of different soil types, soil samples collected from 8-, 50- and 90-year old tea orchards, the adjacent wasteland, and 90-year old forest were measured for their nitrification potentials using the conventional soil incubation and the liquid incubation method. Among different soil types, the nitrification potential of soil in tea orchards was higher than that of wasteland and forest soils. The slurry shaken liquid incubation method was confirmed to be more accurate and have reliable results than the soil incubation. Interestingly, experimental result revealed that the generally applied pH value of 7.2 for the liquid media was not the optimal pH for these acid soils with a strong buffer capacity. This suggested that tea orchard soils may have nitrifiers requiring pHneutral condition for the best activity. Our data also showed that treatment with the commonly used nitrogen fertilizer urea significantly improved nitrification potential of the soils; such enhancement effect was stronger on all of three tea orchard soils than on wasteland and forest soils, and also stronger on the younger （8- and 50-year old） tea orchard soils than on the older one （90-year old）.

Effect of nitrification inhibitor DMPP on nitrogen leaching, nitrifying organisms, and enzyme activities in a rice-oilseed rape cropping system

DMPP （3,4-dimethylpyrazole phosphate） has been used to reduce nitrogen （N） loss from leaching or denitrification and to improve N supply in agricultural land. However, its impact on soil nitrifying organisms and enzyme activities involved in N cycling is largely unknown. Therefore, an on-farm experiment, for two years, has been conducted, to elucidate the effects of DMPP on mineral N （NH4^＋- N and NO3^--N） leaching, nitrifying organisms, and denitrifying enzymes in a rice-oilseed rape cropping system. Three treatments including urea alone （UA）, urea ＋ 1% DMPP （DP）, and no fertilizer （CK）, have been carded out. The results showed that DP enhanced the mean NH4^＋-N concentrations by 19.1%-24.3%, but reduced the mean NO3^--N concentrations by 44.9%-56.6% in the leachate, under a two-year rice-rape rotation, compared to the UA treatment. The population of ammonia oxidizing bacteria, the activity of nitrate reductase, and nitrite reductase in the DP treatment decreased about 24.5%-30.9%, 14.9%-43.5%, and 14.7%-31.6%, respectively, as compared to the UA treatment. However, nitrite oxidizing bacteria and hydroxylamine reductase remained almost unaffected by DMPP. It is proposed that DMPP has the potential to either reduce NO3^--N leaching by inhibiting ammonia oxidization or N losses from denitrification, which is in favor of the N conversations in the rice-oilseed rape cropping system.

Determination of respiration, gross nitrification and denitrification in soil profile using BaPS system

A facility of BaPS （Barometric Process Separation） was used to determine soil respiration, gross nitrification and denitrification in a winter wheat field with depths of 0-7, 7--14 and 14-21 cm. N2O production was determined by a gas chromatograph. Crop root mass and relevant soil parameters were measured. Results showed that soil respiration and gross nitrification decreased with the increase of soil depth, while denitrification did not change significantly. In comparison with no-plowing plot, soil respiration increased significantly in plowing plot, especially in the surface soil of 0-7 cm, while gross nitrification and denitrification rates were not affected by plowing. Cropping practice in previous season was found to affect soil gross nitrification in the following wheat-growing season. Higher gross nitrification rate occurred in the filed plot with preceding crop of rice compared with that of maize for all the three depths of 0-7, 7-14 and 14-21 cm. A further investigation indicated that the nitrification for all the cases accounted for about 76% of the total nitrogen transformation processes of nitrification and denitrification and the N2O production correlated with nitrification significantly, suggesting that nitrification is a key process of soil N2O production in the wheat field. In addition, the variations of soil respiration and gross nitrification were exponentially dependent on root mass （p〈0.00l）.

Influences of nitrification inhibitor 3,4-dimethyl pyrazole phosphate on nitrogen and soil salt-ion leaching

An undisturbed heavy clay soil column experiment was conducted to examine the influence of the new nitrification inhibitor, 3,4- dimethylpyrazole phosphate （DMPP）, on nitrogen and soil salt-ion leaching. Regular urea was selected as the nitrogen source in the soil. The results showed that the cumulative leaching losses of soil nitrate-N under the treatment of urea with DMPP were from 57.5% to 63.3% lower than those of the treatment of urea without DMPP. The use of nitrification inhibitors as nitrate leaching retardants may be a proposal in regulations to prevent groundwater contaminant. However, there were no great difference between urea and urea with DMPP treatments on ammonium-N leaching. Moreover, the soil salt-ion leaching losses of Ca^2＋, Mg^2＋, K^＋, and Na^＋ were reduced from 26.6% to 28.8%, 21.3% to 27.8%, 33.3% to 35.5%, and 21.7% to 32.1%, respectively. So, the leaching losses of soil salt-ion were declined for nitrification inhibitor DMPP addition, being beneficial to shallow groundwater protection and growth of crop. These results indicated the possibility of ammonium or ammonium producing compounds using nitrification inhibitor DMPP to control the nitrate and nutrient cation leaching losses, minimizing the risk of nitrate pollution in shallow groundwater.

Simultaneous nitrification and denitrification in step feeding biological nitrogen removal process

The simultaneous nitrification and denitrification in step-feeding biological nitrogen removal process were investigated under different influent substrate concentrations and aeration flow rates. Biological occurrence of simultaneous nitrification and denitrification was verified in the aspect of nitrogen mass balance and alkalinity. The experimental results also showed that there was a distinct linear relationship between simultaneous nitrification and denitrification and DO concentration under the conditions of low and high aeration flow rate. In each experimental run the floc sizes of activated sludge were also measured and the results showed that simultaneous nitrification and denitrification could occur with very small size of floc.

Organic matter and concentrated nitrogen removal by shortcut nitrification and denitrification from mature municipal landfill leachate

An UASB＋Anoxic/Oxic （A/O） system was introduced to treat a mature landfill leachate with low carbon-to-nitrogen ratio and high ammonia concentration. To make the best use of the biodegradable COD in the leaehate, the denitrifieation of NOx^--N in the reeireulation effluent from the elarifier was carried out in the UASB. The results showed that most biodegradable organic matters were removed by the denitrifieation in the UASB. The NH4^＋-N loading rate （ALR） of A/O reactor and operational temperature was 0.28- 0.60 kg NH4^＋-N/（m^3-d） and 17-29℃ during experimental period, respectively. The short-cut nitrification with nitrite accumulation efficiency of 90%-99% was stabilized during the whole experiment. The NH4^＋-N removal efficiency varied between 90% and 100%. When ALR was less than 0.45 kg NH4^＋-N/（m^3.d）, the NH4^＋-N removal efficiency was more than 98%. With the influent NH4^＋-N of 1200-1800 mg/L, the effluent NH4^＋-N was less than 15 mg/L. The shortcut nitrification and denitrifieation can save 40% carbon source, with a highly efficient denitrifieation taking place in the UASB. When the ratio of the feed COD to feed NH4^＋-N was only 2-3, the total inorganic nitrogen （TIN） removal efficiency attained 67%-80%. Besides, the sludge samples from A/O reactor were analyzed using FISH. The FISH analysis revealed that ammonia oxidation bacteria （AOB） accounted for 4% of the total eubaeterial population, whereas nitrite oxidation bacteria （NOB） accounted only for 0.2% of the total eubaeterial population.

Removal of organic matter and nitrogen from distillery wastewater by a combination of methane fermentation and denitrification/nitrification processes

The distillery wastewater of Guangdong Jiujiang Distillery, which is characteristic of containing high organic matters and rich total nitrogen, was treated by a combination of methane fermentation and denitrification/nitrification processes. 80% of COD in the raw wastewater was removed by methane fermentation at the COD volumetric loading rate of 20 kg COD/（m^3·d） using the expanded granule sludge bed （EGSB） process. However, almost all the organic nitrogen in the raw wastewater was converted into ammonia by ammonification there. Ammonia and volatile fatty acids （VFA） remaining in the anaerobically treated wastewater were simultaneously removed utilizing VFA as an electron donor by denitrification occurring in the other EGSB reactor and nitrification using PEG-immobilized nitrifying bacteria with recirculation process. An aerobic biological contact oxidization reactor was designed between denitrification/nitrification reactor for further COD removal. With the above treatment system, 18000-28000 mg/L of COD in raw wastewater was reduced to less than 100 mg/L. Also, ammonia in the effluent of the system was not detected and the system had a high removal rate for 900-1200 mg/L of TN in the raw wastewater, only leaving 400 mg/L of nitrate nitrogen.

Nitrogen removal influence factors in A/O process and decision trees for nitrification/denitrification system

In order to improve nitrogen removal in anoxic/oxic(A/O) process effectively for treating domestic wastewaters, the influence factors, DO(dissolved oxygen), nitrate recirculation, sludge recycle, SRT(solids residence time), influent COD/TN and HRT(hydraulic retention time) were studied. Results indicated that it was possible to increase nitrogen removal by using corresponding control strategies, such as, adjusting the DO set point according to effluent ammonia concentration; manipulating nitrate recirculation flow according to nitrate concentration at the end of anoxic zone. Based on the experiments results, a knowledge-based approach for supervision of the nitrogen removal problems was considered, and decision trees for diagnosing nitrification and denitrification problems were built and successfully applied to A/O process.

Nitrogen additions inhibit nitrification in acidic soils in a subtropical pine plantation: effects of soil pH and compositional shifts in microbial groups

Plantation forests play a pivotal role in carbon sequestration in terrestrial ecosystems, but enhanced nitrogen(N) deposition in these forests may affect plantation productivity by altering soil N cycling. Hence,understanding how simulated N deposition affects the rate and direction of soil N transformation is critically important in predicting responses of plantation productivity in the context of N loading. This study reports the effects of N addition rate(0, 40, and 120 kg N ha^(-1) a^(-1)) and form(NH_4Cl vs. NaNO_3) on net N mineralization and nitrification estimated by in situ soil core incubation and on-soil microbial biomass determined by the phospholipid fatty acid(PLFA) method in a subtropical pine plantation. N additions had no influences on net N mineralization throughout the year. Net nitrification rate was significantly reduced by additions of both NH_4Cl(71.5) and NaNO_3(47.1%) during the active growing season, with the stronger inhibitory effect at high N rates. Soil pH was markedly decreased by 0.16 units by NH_4Cl additions. N inputs significantly decreased the ratio of fungal-to-bacterial PLFAs on average by 0.28(49.1%) in November. Under NH_4Cl additions, nitrification was positively related with fungal biomass and soil pH. Under NaNO_3 additions,nitrification was positively related with all microbial groups except for bacterial biomass. We conclude that simulated N deposition inhibited net nitrification in the acidic soils of a subtropical plantation forest in China,primarily due to accelerated soil acidification and compositional shifts in microbial functional groups. These findings may facilitate a better mechanistic understanding of soil N cycling in the context of N loading.

Cultivation of aerobic granules for simultaneous nitrification and denitrification by seeding different inoculated sludge

Cultivation of aerobic granules for simultaneous nitrification and denitrification in two sequencing batch airlift bioreactors was studied. Conventional activated floc and anaerobic granules served as main two inoculated sludge in the systems. Morphological variations of sludge in the reactors were observed. It was found that the cultivation of aerobic granules was closely associated with the kind of inoculated sludge. Round and regular aerobic granules were prevailed in both reactors, and the physical characteristics of the aerobic granules in terms of settling ability, specific gravity, and ratio of water containing were distinct when the inoculate sludge was different. Aerobic granules formed by seeding activated floc are more excellent in simultaneous nitrification and denitrification than that by aerobic granules formed from anaerobic granules. It was concluded that inoculated sludge plays a crucial role in the cultivation of aerobic granules for simultaneous nitrification and denitrification.

Effect of organic carbon on nitrification effciency and community composition of nitrifying biofilms

The effects of organic carbon/inorganic nitrogen （C/N） ratio on the nitrification processes and the community shifts of nitrifying biofilms were investigated by kinetic comparison and denaturing gradient gel electrophoresis （DGGE） analysis. The results showed that the nitrification rate decreased with an increasing organic concentration. However, the effect became weak when the carbon concentration reached a sufficiently high level. Denitrification was detected after organic carbon was added. The 12 h ammonium removal rate ranged from 85% to 30% at C/N = 0.5, 1, 2, 4, 8, and 16, as compared to the control （C/N = 0）. The loss of nitrogen after 24 h at C/N = 0.5, 1, 2, 4, 8, and 16 was 31%, 18%, 24%, 65%, 59%, and 62%, respectively. The sequence analysis of 16S rRNA gene fragments revealed that the dominant populations changed from nitrifying bacteria （Nitrosomonas europaea and Nitrobacter sp.） to denitrifying bacteria （Pseudomonas sp., Acidovorax sp. and Comamonas sp.） with an increasing C/N ratio. Although at high C/N ratio the denitrifying bacteria were the dominant populations, nitrifying bacteria grew simultaneously. Consequently, nitrification process coexisted with denitrification.

Effect of forest thinning on soil net nitrogen mineralization and nitrification in a Cryptomeria japonica plantation in Taiwan

We investigated the effect of forest thinning on soil nitrogen mineralization, nitrification and transformation in a Cryptomeria japoni-ca plantation at high elevation to provide basic data for forest manage-ment. We chose four study plots for control, light, medium and heavy thinning treatment, and three sub-plots for buried bag studies at similar elevations in each treatment plot to measure the net N mineralization and nitrification rates in situ. The contents of soil inorganic N （ammonium and nitrate） were similar between treatments, but all varied with season, reaching maxima in September 2003 and 2004. The seasonal maximum net Nmin rates after four treatments were 0.182, 0.246, 0.303 and 0.560 mg？kg-1？d-1 in 2003, and 0.242,0.258,0.411 and 0.671 mg？kg-1？d-1in 2004, respectively. These estimates are approximate with the lower annual rates of N mineralization for this region. Forest thinning can enhance net N mineralization and microbial biomass carbon. The percentage of annual rates of Nmin for different levels of forest thinning compared with the control plot were 13.4%, 59.8%and 154.2%in 2003, and 0.1%, 58.8%and 157.7%in 2004 for light, medium, and heavy thinning, respectively. These differences were related to soil moisture, temperature, precipita-tion, and soil and vegetation types. Well-planned multi-site comparisons, both located within Taiwan and the East-Asia region, could greatly im-prove our knowledge of regional patterns in nitrogen cycling.

Wastewater Treatment in a Hybrid Biological Reactor (HBR): Nitrification Characteristics

Objective To investigate the nitrifying characteristics of both suspended- and attached- biomass in a hybrid bioreactor. Methods The hybrid biological reactor was developed by introducing porous ceramic particles into the reactor to provide the surface for biomass attachment. Microorganisms immobilized on the ceramics were observed using scanning electron microscopy (SEM). All chemical analyses were performed in accordance with standard methods. Results The suspended- and attached-biomass had approximately the same nitrification activity. The nitrifying kinetic was independent of the initial biomass concentration, and the attached-biomass had a stronger ability to resist the nitrification inhibitor. Conclusion The attached biomass is superior to suspended-biomass for nitrifying wastewater, especially that containing toxic organic compounds. The hybrid biological reactor consisting of suspended- and attached-biomass is advantageous in such cases.