Estimates of Microbial Populations Involved in the N Cycle and their Activity in Water and Sediments of Fish Farming Ponds under Mono and Polyculture Systems in India

The population sizes of ammonifying, protein mineralizing, nitrogen fixing and nitrifying bacteria, and the rates of ammonification and nitrification (natural and potential) were measured in water and sediments of four fish ponds being used for traditional, mono- and polyculture systems of fish farming. Spatial differences in the microbial density in these ponds were related to the fish culturing practices adopted. The seasonal variation of ammonifying bacteria was found to be positively correlated with the NH₄-N level in the water. The natural and potential capacity to generate both nitrite and nitrate in these water bodies was strongly correlated with the concentrations of the different forms of inorganic nitrogen present. The rates of NO₂-N and NO₃-N formation occurring in these fish ponds were directly proportional to the amount of dissolved oxygen and pH of the environment, respectively.     

川西北高寒草地沙化过程中土壤氮素变化特征

草地沙化是我国最严重的环境问题之一,但关于草地沙化过程中氮素变化特征的研究报道多集中于干旱半干旱地区,而半湿润地区的相关报道还比较缺乏。通过野外调查,研究了川西北半湿润地区高寒沙质草地沙化过程中土壤氮素变化特征。结果表明,草地沙化对0—100cm土层土壤氮素具有显著影响,全氮、碱解氮、铵态氮(NH_4~+-N)、硝态氮(NO_3~--N)和微生物量氮(MBN)均呈现极显著下降的变化特征,极度沙化阶段较未沙化阶段分别减少了73.95%、77.72%、76.75%、79.77%和84.12%。其中,0—20cm土层变化最显著,全氮、碱解氮、NH_4~+-N、NO_3~--N和MBN含量分别减少了86.43%、83.52%、82.11%、88.82%和91.77%。随着土层深度增加,不同程度沙化草地土壤氮素含量及其变化量逐渐减少;草地沙化过程中,不同沙化阶段土壤氮素损失数量不尽相同,其中,以轻度沙化阶段氮素损失最严重,全氮、碱解氮、NH_4~+-N、NO-3-N和MBN含量分别降低了41.18%、35.17%、46.74%、43.46%和46.88%。草地沙化过程中,土壤全氮、碱解氮、NH_4~+-N、NO_3~--N和MBN含量与土壤粉粒、粘粒含量和植被群落盖度均呈极显著正相关特征,与土壤沙粒含量呈极显著负相关特征。研究区土壤氮素损失与风蚀选择性吹蚀土壤粉粒、粘粒及地表植物覆盖状况逐渐变差密切相关,因此该区域治沙的关键是采取措施降低风蚀对地表土壤吹蚀作用,提高沙化草地地表植被覆盖。同时,还应及时对沙化前期阶段及潜在沙化的草地进行生态治理,从而避免草地沙化继续恶化。     

Temperature sensitivity of soil carbon and nitrogen mineralization: impacts of nitrogen species and land use type

Background and aims

Climate warming, nitrogen (N) deposition and land use change are some of the drivers affecting ecosystem processes such as soil carbon (C) and N dynamics, yet the interactive effects of those drivers on ecosystem processes are poorly understood. This study aimed to understand mechanisms of interactive effects of temperature, form of N deposition and land use type on soil C and N mineralization.

Methods

We studied, in a laboratory incubation experiment, the effects of temperature (15 vs. 25 °C) and species of N deposition (NH₄ ⁺-N vs. NO₃ ^?-N) on soil CO₂ efflux, dissolved organic C (DOC) and N (DON), NH₄ ⁺-N, and NO₃ ^?-N concentrations using intact soil columns collected from adjacent forest and grassland ecosystems in north-central Alberta.

Results

Temperature and land use type interacted to affect soil CO₂ efflux, concentrations of DON, NH₄ ⁺-N and NO₃ ^?-N in most measurement times, with the higher incubation temperature resulted in the higher CO₂ efflux and NH₄ ⁺-N concentrations in forest soils and higher DON and NO₃ ^?-N concentrations in grassland soils. Temperature and land use type affected the cumulative soil CO₂ efflux, and DOC, DON, NH₄ ⁺-N and NO₃ ^?-N concentrations. The form of N added or its interaction with the other two factors did not affect any of the C and N cycling parameters.

Conclusions

Temperature and land use type were dominant factors affecting soil C loss, with the soil C in grassland soils more stable and resistant to temperature changes. The lack of short-term effects of the deposition of different N species on soil C and N mineralization suggest that maybe there was a threshold for the N effect to kick in and long-term experiments should be conducted to further elucidate the species of N deposition effects on soil C and N cycling in the studied systems.     

Foliar N application reduces soil NO<Stack><Subscript>3</Subscript><Superscript>−</Superscript></Stack>-N leaching loss in apple orchards

A comparison of the effects of foliar and soil N application was made in field-grown mature fruiting Gala/M9 apple trees (Malus domestica Borkh) in 2001 and 2002 growing seasons under Pacific Northwest growing conditions in southern British Columbia, Canada. The trees, six years old at the start of the experiment, were treated: (1) with 5 g/l urea sprays supplied every two weeks (7 times) from mid May to mid August (total about 50 g N/tree/year), (2) with the same amount of N applied to the soil with the same timing and quantity as for the foliar treatment, and (3) with no N (control). Leaf color (as SPAD readings) and N concentrations (mg/g), and soil NH₄⁺-N and NO₃^–-N were measured periodically throughout the two seasons. Leached NO₃^–-N was monitored monthly via an anion exchange probe from June to October in 2001 and from May to November in 2002. Shoot length was measured in October and N concentration of one-year-old wood and roots was determined in December of each growing s eason. Soil N application significantly increased shoot length relative to control or foliar N application. Leaf color, leaf N, and N concentration of one-year-old wood and roots were similarly increased relative to control by both soil and foliar N application. These treatments also increased fruit yield relative to control. There was no significant difference in yield and fruit quality between soil and foliar N applications. Soil N application increased soil NH₄⁺-N and NO₃^–-N content in the root zone, and also increased the NO₃^– leaching loss below the root zone especially late in the growing season. Our results suggested that tree N status and yield and fruit quality could be maintained by multiple urea sprays during the growing season in apple orchards, and foliar N application will reduce the risk of soil NO₃^–-N leaching.     

氮肥配施生化抑制剂组合对黄泥田土壤氮素淋溶特征的影响

揭示尿素类肥料添加生化抑制剂组合后,在黄泥田土壤中硝态氮(NO~-_3-N)和铵态氮(NH~+_4-N)的淋溶损失规律。采用室内土柱淋溶培养试验,研究脲酶抑制剂N-丁基硫代磷酰三胺(NBPT)和硝化抑制剂2-氯-6-(三氯甲基)吡啶(CP)单独添加及配合施用对尿素和尿素硝铵(300 kg N/hm~2)中氮(N)素在土体中淋溶损失的影响。结果表明:尿素和尿素硝铵处理淋溶液中NH~+_4-N和NO~-_3-N浓度均呈先升后降的变化趋势,而出峰时间不一。NH~+_4-N和NO~-_3-N淋失量随着时间的延长,处理间差异逐渐变大。NBPT处理可以减缓尿素水解,有效抑制NH~+_4-N生成,延缓其出峰时间,减少NH~+_4-N流失;CP处理可以有效抑制NH~+_4-N向NO~-_3-N转化,减少NO~-_3-N流失。与单独添加NBPT和CP处理相比,两者配施对N素淋溶损失有明显的协同抑制效果在黄泥田土壤中,既能减缓尿素水解,保持土壤中较高NH~+_4-N含量,又能降低淋溶液中NO~-_3-N浓度。培养结束时(第72天),UAN处理中NO~-_3-N、NH~+_4-N、矿质态N淋失总量及硝化率较U处理高34.39%、5.32%、31.72%和15.71%。U+NBPT、U+CP和U+NBPT+CP处理较U处理分别显著降低NO~-_3-N淋失总量达15.58%、114.77%和73.45%;UAN+NBPT、UAN+CP和UAN+NBPT+CP处理较UAN处理分别显著降低达15.88%、54.87%和37.46%。不同处理NO~-_3-N淋失总量大小表现为:UAN UAN+NBPT U UAN+NBPT+CP U+NBPT UAN+CP U+NBPT+CP U+CP CK。在一定施肥量条件下,NBPT和CP单独施用或配施均可降低黄泥田土壤中NO~-_3-N累积淋失量。对各处理淋溶液中NO~-_3-N淋失量(y)随时间(x)的变化进行拟合,其中以线性方程(y=ax+b)的拟合度较高,且各抑制剂处理a、b值均存在明显差异。总体认为,在黄泥田土壤中施用CP及其与NBPT配施可以显著降低土壤NO~-_3-N淋溶损失,减少N素淋失风险,提高肥料利用率。     

Microbial populations, ammonification and nitrification in soil treated with urea and inorganic salts

Soil fertilization with urea at rates of 0.2 and 0.5 mg N/g soil was toxic for total counts of bacteria and fungi except with cellulolytic fungi where growth was promoted by addition of urea after 90-d incubation. Also, the population numbers of both bacteria and fungi were significantly decreased when soil was amended with CaCl₂ and K₂SO₄ applied at two levels (50 and 100 μmol/g soil). Some alleviation of the toxic effect of either urea or the inorganic salts was observed when they were applied in combination. Fungal species composition was found to be affected in the treated soil. The most tolerant fungi wereAlternaria alternata, Aspergillus flavus, A. fumigatus, A. niger, A. terreus, Eurotium amstelodami, E. chevalieri, Nectria hœmatococca, Pœcilomyces variotii andStachybotrys chartarum. Other species of fungi were either sensitive to all treatments or sensitive to some and tolerant to others. Ammonium nitrogen was found to be more accumulated in soil treated with urea mixed with inorganic salts compared with soil treated only with urea whereas NO ₃ ⁻ -N (and NO ₂ ⁻ -N) was decreased. The overall effect of the addition of inorganic salts on total mineralized nitrogen, however was promotive. Soil pH was increased in ureatreated soil but was depressed by application of CaCl₂ or K₂SO₄ to values lower than that of untreated soil. The results may be of agronomical interest in overcoming problems encountered with urea application as N fertilizer.     

Suppression of nitrate formation within an exotic conifer plantation

Summary Nitrate-N losses to stream waters and soil inorganic N pools, nitrifying potentials and NO₃-N production rates were measured in 2 adjacent watersheds, one used as pasture and the other planted in exotic conifer forest (Pinus radiata D. Don). Estimated NO₃-N loss to stream waters draining the pine and pasture watersheds were 0.6kg ha⁻¹ y⁻¹ and 7.6 kg ha⁻¹ y⁻¹ respectively. Ammonium-N pool sizes were not significantly different between soils in the two watersheds but NO₃−N pools and nitrifying potentials were always lower in the pine watershed soil samples. Laboratory incubation experiments indicated that suppression of NO₃−N formation in pine watershed soils required the presence of live tree roots and was not due to the direct action of allelopathic chemicals on nitrifiers.     

Transformation of urea and ammonium sulphate in different soils

Summary The transformation of urea and ammonium sulphate in Ladwa sandy loam and Balsamand sand was studied in laboratory. Urea took at least one week in sandy loam and 2 weeks in sandy soils to hydrolyse completely. The process of hydrolysis was faster in finer soil with high organic matter than in coarse soil having low organic matter. There was no nitrification upto 3 days in sandy loam and upto 7 days in sandy soils, respectively, but there was immobilization of NO₃-N during these initial periods. The NO₃-N content at the end of incubation period (35 days) was more in case of urea than in case of ammonium sulphate treated samples in sandy loam soil and reverse was true in sandy soil. The hydrolysis of urea did not follow zero or first order kinetics as proposed in previous studies.     

天山林区不同类型群落土壤氮素对冻融过程的动态响应

季节性冻融过程对北方温带森林土壤氮素的转化与流失具有重要影响,但不同类型群落对冻融过程响应的差异尚不明确。通过在林地、草地、灌丛上设置系列监测样地,采用原位培养的方法,利用林冠遮挡形成的自然雪被厚度差异,监测分析了冻融期天山林区不同群落表层土壤(0—15 cm)的氮素动态及净氮矿化速率间的差异。结果表明:(1)不同类型群落土壤的铵态氮(NH+4-N)含量、微生物量氮(MBN)含量基本与土壤(5 cm)温度呈正相关,深冻期林地土壤铵态氮含量低于其他群落类型而硝态氮含量高于其他群落类型;(2)硝态氮(NO-3-N)为天山林区季节性冻融期间土壤矿质氮的主体,占比达78.4%。灌丛土壤硝态氮流失风险较大,融化末期较融化初期灌丛土壤硝态氮含量下降了64.6%;(3)冻融时期对整体氮素矿化速率影响显著,群落类型对氨化速率影响显著;(4)天山林区土壤氮素在冻结期主要以氮固持为主。通过揭示不同类型群落土壤氮素对冻融格局的响应,能够助益于对北方林区冬季土壤氮素循环的认识。     

Removal of nitrogen by heterotrophic nitrification–aerobic denitrification of a novel halotolerant bacterium Pseudomonas mendocina TJPU04

Excess inorganic nitrogen in water poses a severe threat to enviroment. Removal of inorganic nitrogen by heterotrophic nitrifying–aerobic denitrifying microorganism is supposed to be a promising and applicable technology only if the removal rate can be maintained sufficiently high in real wastewater under various conditions, such as high concentration of salt and wide range of different nitrogen concentrations. Here, a new heterotrophic nitrifying–aerobic denitrifying bacterium was isolated and named as Pseudomonas mendocina TJPU04, which removes NH₄⁺-N, NO₃⁻-N and NO₂⁻-N with average rate of 4.69, 5.60, 4.99 mg/L/h, respectively. It also maintains high nitrogen removal efficiency over a wide range of nitrogen concentrations. When concentration of NH₄⁺-N, NO₃⁻-N and NO₂⁻-N was up to 150, 150 and 50 mg/L, 98%, 93%, and 100% removal efficiency could be obtained, respectively, after 30-h incubation under sterile condition. When it was applied under non-sterile condition, the ammonia removal efficiency was slightly lower than that under sterile condition. However, the nitrate and nitrite removal efficiencies under non-sterile condition were significantly higher than those under sterile condition. Strain TJPU04 also showed efficient nitrogen removal performance in the presence of high concentration of salt and nitrogen. In addition, the removal efficiencies of NH₄⁺-N, NO₃⁻-N and TN in real wastewater were 91%, 52%, and 75%, respectively. These results suggest that strain TJPU04 is a promising candidate for efficient removal of inorganic nitrogen in wastewater treatment.

     

Effects of slow-release urea fertilizers on urease activity,microbial biomass,and nematode communities in an aquic brown soil

A field experiment was carried out at the Shenyang Experimental Station of Ecology (CAS) in order to study the effects of slow-release urea fertilizers high polymer-coated urea (SRU1), SRU1 mixed with dicyandiamide DCD (SRU2), and SRU1 mixed with calcium carbide CaC₂ (SRU3) on urease activity, microbial biomass C and N, and nematode communities in an aquic brown soil during the maize growth period. The results demonstrated that the application of slow-release urea fertilizers inhibits soil urease activity and increases the soil NH₄ ⁺-N content. Soil available N increment could promote its immobilization by microorganisms. Determination of soil microbial biomass N indicated that a combined application of coated urea and nitrification inhibitors increased the soil active N pool. The population of predators/omnivores indicated that treatment with SRU2 could provide enough soil NH₄ ⁺-N to promote maize growth and increased the food resource for the soil fauna compared with the other treatments.     

Screening and characterization of an aerobic nitrifying-denitrifying bacterium from activated sludge

Taking advantage of the good biocompatibility and high efficiency of nitrogen removal with microbes, nitrifying and denitrifying bacteria, are becoming increasingly more widely used for wastewater treatment and prevention of eutrophication. In this research, an aerobic nitrifying-denitrifying bacterium was successfully screened from activated sludge and identified as Pseudomonas sp. (CCTCC No M2010209) by the 16S rDNA sequence. The activity verification confirmed its nitrifying-denitrifying capability of removing ammonium, nitrate and nitrite nitrogen. The types of carbon sources and carbon-nitrogen ratio greatly influenced the removal efficiency of NH₄ ⁺-N and NO₃ ⁻-N. When the initial concentrations of NH₄ ⁺-N and NO₃ ⁻-N in synthetic wastewater were less than 70 and 50 mg/L, the nitrogen removal rates reached 94 and 90% in 9 h, respectively. Preliminary comparisons of nitrogen removal capacity between this isolate and other commercial preparations in the treatment of synthetic wastewater revealed its promising potential to be used in the actual wastewater treatment.     

Identification of novel denitrifying bacteria Stenotrophomonas sp. ZZ15 and Oceanimonas sp. YC13 and application for removal of nitrate from industrial wastewater

Two novel denitrifying bacteria were successfully isolated from industrial wastewater and soil samples. Using morphological, biochemical/biophysical and 16S rRNA gene analyses, these two bacteria were identified as Stenotrophomonas sp. ZZ15 and Oceanimonas sp. YC13, respectively. Both of these two bacteria showed efficient NO₃ ⁻-N removing abilities under a semi-anaerobic condition without obvious accumulation of NO₂ ⁻-N, N₂O-N and NH₄ ⁺-N. NO₃ ⁻-N removal from paper mill wastewater was also successful by treatments with either a denitrifier or an immobilization method. Therefore, this study provides valuable denitrifying bacteria in biotreatment of industrial wastewater and other environmental pollution caused by NO₃ ⁻/NO₂ ⁻.     

晋西北不同年限小叶锦鸡儿灌丛土壤氮矿化和硝化作用

利用PVC管顶盖埋管法研究了晋西北黄土高原区小叶锦鸡儿人工灌丛不同定植年限(5,10,20,30,40a)土壤氮矿化与硝化速率的动态和净矿化与硝化总量。结果表明,⑴小叶锦鸡儿灌丛土壤无机氮主要以NO_-~3-N形式存在,不同生长年限相同月份的土壤硝态氮(NO-3-N)含量分别是铵态氮(NH+4-N)含量的1.5—15.4倍;⑵土壤氮素硝化速率和矿化速率随生长年限延长而加快,30年生时达到高峰,数值达40.2,44.1 mg m~(-2)d~(-1)。从季节性变化看,7—8月份是硝化速率和矿化速率快速增长期,30年生小叶锦鸡儿灌丛土壤硝化速率和矿化速率分别达到86.9,93.1 mg m~(-2)d~(-1),显著高于其它生长年限(P0.05);(3)土壤氮素硝化与矿化总量同样随小叶锦鸡儿生长年限延长而增加,30年生时达到最高,与5年生相比,分别增加了3.7和3.1倍。(4)5—10月份小叶锦鸡儿生长期内,各年限土壤全氮量的2.3%被矿化成无机氮,其中87%最终被转化成NO-3-N形式存在于土体中。     

Effects of slow-release urea fertilizers on urease activity,microbial biomass,and nematode communities in an aquic brown soil

A field experiment was carried out at the Shenyang Experimental Station of Ecology (CAS) in order to study the effects of slow-release urea fertilizers high polymer-coated urea (SRU1), SRU1 mixed with dicyandiamide DCD (SRU2), and SRU1 mixed with calcium carbide CaC₂ (SRU3) on urease activity, microbial biomass C and N, and nematode communities in an aquic brown soil during the maize growth period. The results demonstrated that the application of slow-release urea fertilizers inhibits soil urease activity and increases the soil NH₄ ⁺-N content. Soil available N increment could promote its immobilization by microorganisms. Determination of soil microbial biomass N indicated that a combined application of coated urea and nitrification inhibitors increased the soil active N pool. The population of predators/omnivores indicated that treatment with SRU2 could provide enough soil NH₄ ⁺-N to promote maize growth and increased the food resource for the soil fauna compared with the other treatments.

     

Integrated soil and plant phosphorus management for crop and environment in China. A review

In semi-arid grassland ecosystems, soil biogeochemical processes are controlled by seasonal and inter-annual rainfall variation and temperature, which may override the long-term impact of grazers on N availability and N dynamics. In a three-year (2004?C2006) case study of an Inner Mongolian grassland, we analysed time-integrated (ion-exchange resins) and instantaneous (soil mineral N extractions) inorganic N availability at three sites of varying grazing intensities and combined these data with information on soil water content (SWC), aboveground net primary productivity (ANPP) and plant N uptake. Additionally, the effects of rainfall and grazing on N-form availability (NO ₃ ^? -N, NH ₄ ⁺ -N) were considered. Grazing had less impact on N availability compared to seasonal and annual rainfall distribution. One of the three study years (2004) showed a grazing effect with higher resin-N availability at the ungrazed site compared to the heavily grazed site. Inorganic N availability was low in the driest year (2005) and highest in a year of average rainfall amount and favourable distribution (2004). In general, we found a positive relationship between inorganic N availability and both plant productivity and plant N uptake. Rainfall also controlled the plant available NO ₃ ^? -N and NH ₄ ⁺ -N pools; NH ₄ ⁺ -N dominated the available inorganic N-form in times of low SWC, while the available NO ₃ ^? -N increased with SWC. We observed N availability and plant productivity in a temporal synchronized pattern. Increased rainfall variability and land-use practices affecting SWC will likely alter N availability dynamics (and the relation of N-forms) and, therefore, important processes of semi-arid natural grassland carbon and N cycling.     

Effects of hydraulic loading rate on pollutants removal by a deep subsurface wastewater infiltration system

The subsurface wastewater infiltration (SWI) system proved to be an effective and low-cost technique for decentralized sewage treatment in areas without adequate domestic treatment facilities. Field-scale experiments were conducted through a deep SWI system, with effective depth of 1.5 m, under hydraulic loading rates of 0.040, 0.065, 0.081 and 0.10 m³/m² d. Taking the hydraulic and treatment efficiencies into consideration, the hydraulic loading rate of 0.081 m³/m² d was recommended. Under this condition, NH₃-N, TN, and COD removal efficiencies were 86.2 ± 3.0, 80.7 ± 1.9 and 84.8 ± 2.1%, respectively. In the effluent, NH₃-N concentration declined to 2.3-4.4 mg/L, accounting for 63.2-65.6% of TN. NO₃-N concentration increased from 0.2 to 0.3 mg/L in the influent to 2.0-2.5 mg/L in the effluent. The nitrifying bacteria number declined with increased depth, while the amount of denitrifying bacteria increased. The analysis of results about the nitrifying and denitrifying bacteria distribution indicated that the most effective ranges for nitrification and denitrification process were 0.3-0.7 m and 0.7-1.5 m, respectively.     

Immobilization of nitrifying bacteria in porous pellets of urethane gel for removal of ammonium nitrogen from waste-water

Summary The effects of immobilizing materials on the activity of nitrifying bacteria and removal of ammonium nitrogen (NH₄-N) from waste-water by immobilized nitrifying bacteria were investigated using six urethane prepolymers. With a urethane prepolymer containing 2.27% free isocyanate, a high activity yield of nitrifying bacteria was obtained. There was a drastic improvement over the conventional method of immobilization by acrylamide in the activity yield. Inorganic synthetic waste-water was treated at a high loading rate of 0.24 kg N·m^–3·day^–1. The NH₄-N concentration of the effluent could be reduced to 2 mg·1^–1 or less and the removal was 90%. The life of the pellets in terms of activity was at least 120 days. Offprint requests to: T. Sumino     

Effect of influent substrate ratio on anammox granular sludge: performance and kinetics

Effect of influent substrate ratio on anammox process was studied in sequencing batch reactor. Operating temperature was fixed at 35 ± 1 °C. Influent pH and hydraulic retention time were 7.5 and 6 h, respectively. When influent NO₂ ^?-N/NH₄ ⁺-N was no more than 2.0, total nitrogen removal rate (TNRR) increased whereas NH₄ ⁺-N removal rate stabilized at 0.32 kg/(m³ d). ΔNO₂ ^?-N/ΔNH₄ ⁺-N increased with enhancing NO₂ ^?-N/NH₄ ⁺-N. When NO₂ ^?-N/NH₄ ⁺-N was 4.5, ΔNO₂ ^?-N/ΔNH₄ ⁺-N was 1.98, which was much higher than theoretical value (1.32). The IC₅₀ of NO₂ ^?-N was 289 mg/L and anammox activity was inhibited at high NO₂ ^?-N/NH₄ ⁺-N ratio. With regard to influent NH₄ ⁺-N/NO₂ ^?-N, the maximum NH₄ ⁺-N removal rate was 0.36 kg/(m³ d), which occurred at the ratio of 4.0. Anammox activity was inhibited when influent NH₄ ⁺-N/NO₂ ^?-N was higher than 5.0. With influent NO₃ ^?-N/NH₄ ⁺-N of 2.5–6.5, NH₄ ⁺-N removal rate and NRR were stabilized at 0.33 and 0.40 kg/(m³ d), respectively. When the ratio was higher than 6.5, nitrogen removal would be worsened. The inhibitory threshold concentration of NO₂ ^?-N was lower than NH₄ ⁺-N and NO₃ ^?-N. Anammox bacteria were more sensitive to NO₂ ^?-N than NH₄ ⁺-N and NO₃ ^?-N. TNRR would be enhanced with increasing nitrogen loading rate, but sludge floatation occurred at high nitrogen loading shock. The Han-Levenspiel could be applied to simulate nitrogen removal resulting from NO₂ ^?-N inhibition.