Effect of free ammonia and free nitrous acid concentration on the anabolic and catabolic processes of an enriched Nitrosomonas culture

The effects of free ammonia (FA; NH(3)) and free nitrous acid (FNA; HNO(2)) concentrations on the metabolisms of an enriched ammonia oxidizing bacteria (AOB) culture were investigated using a method allowing the decoupling of growth and energy generation processes. A lab-scale sequencing batch reactor (SBR) was operated for the enrichment of an AOB culture. Fluorescent in-situ hybridization (FISH) analysis showed that 82% of the bacterial population in the SBR bound to the NEU probe specifically designed for Nitrosomonas europaea. Batch tests were carried out to measure the oxygen and ammonium consumption rates by the culture at various FA and FNA levels, in the presence or absence of inorganic carbon (CO(2), HCO(3) (-), and CO(3) (2-)). It was revealed that FA of up to 16.0 mgNH(3)-N . L(-1), which was the highest concentration used in this study, did not have any inhibitory effect on either the catabolic or anabolic processes of the Nitrosomonas culture. In contrast, FNA inhibited both the growth and energy production capabilities of the Nitrosomonas culture. The inhibition on growth initiated at approximately 0.10 mgHNO(2)-N . L(-1), and the data suggested that the biosynthesis was completely stopped at an FNA concentration of 0.40 mgHNO(2)-N . L(-1). The inhibition on energy generation initiated at a slightly lower level but the Nitrosomonas culture was still oxidizing ammonia at half of the maximum rate at an FNA concentration of 0.50-0.63 mgHNO(2)-N . L(-1). The affinity constant of the Nitrosomonas culture with respect to ammonia was determined to be 0.36 mgNH(3)-N . L(-1), independent of the presence or absence of inorganic carbon.     

Ammonia-oxidizing bacteria dominates over ammonia-oxidizing archaea in a saline nitrification reactor under low DO and high nitrogen loading

A continuous nitrification reactor treating saline wastewater was operated for almost 1 year under low dissolved oxygen (DO) levels (0.15-0.5 mg/L) and high nitrogen loadings (0.26-0.52 kg-N/(m(3) day)) in four phases. The diversity and abundance of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) were analyzed by cloning, terminal restriction fragment length polymorphism (T-RFLP) and quantitative polymerase chain reaction (qPCR). The results showed that there were only one dominant AOA species and one dominant AOB species in the reactor in all of the four experimental phases. The amoA gene of the dominant AOA only had a similarity of 89.3% with the cultured AOA species Nitrosopumilus maritimus SCM1. All of the AOB species detected in the reactor belong to Nitrosomonas genus and it was found that the AOB populations changed with the ammonium loadings and DO levels. The abundance of AOB in the reactor was ～40 times larger than that of AOA, and the ratio of AOB to AOA increased significantly up to ～2,000 to ～4,000 with the increase of ammonium loading, indicating that AOB are much more competitive than AOA in high ammonium environments and probably AOA play a less important role than AOB in the nitrification reactors.     

Effect of oxic and anoxic conditions on nitrous oxide emissions from nitrification and denitrification processes

A lab-scale sequencing batch reactor fed with real municipal wastewater was used to study nitrous oxide (N(2)O) emissions from simulated wastewater treatment processes. The experiments were performed under four different controlled conditions as follows: (1) fully aerobic, (2) anoxic-aerobic with high dissolved oxygen (DO) concentration, (3) anoxic-aerobic with low DO concentration, and 4) intermittent aeration. The results indicated that N(2)O production can occur from both incomplete nitrification and incomplete denitrification. N(2)O production from denitrification was observed in both aerobic and anoxic phases. However, N(2)O production from aerobic conditions occurred only when both low DO concentrations and high nitrite concentration existed simultaneously. The magnitude of N(2) O produced via anoxic denitrification was lower than via oxic denitrification and required the presence of nitrite. Changes in DO, ammonium, and nitrite concentrations influenced the magnitude of N(2)O production through denitrification. The results also suggested that N(2)O can be produced from incomplete denitrification and then released to the atmosphere during aeration phase due to air stripping. Therefore, biological nitrogen removal systems should be optimized to promote complete nitrification and denitrification to minimize N(2)O emissions.     

不同施肥方式对土壤氨挥发和氧化亚氮排放的影响

采用密闭室间歇通气法和静态箱法对不同施肥方式(撒施后翻耕、条施后覆土、撒施后灌水)下的土壤氨挥发和氧化亚氮排放进行了研究.结果表明:不同施肥方式显著影响了土壤中的氨挥发和氧化亚氮排放.撒施后灌水处理明显促进了氨挥发,其最大氨挥发速率明显高于其它处理,氨挥发累计达2.465 kg N·hm-2.不同施肥方式下氧化亚氮排放通量存在显著差异(P《0.05),且峰值出现时间也不同.施肥后第2天,撒施后灌水处理达到峰值,为193.66 μg·m-2·h-1,而条施后覆土处理在施肥后第5天才出现峰值,为51.13 μg·m-2·h-1,且其排放峰值在3种施肥方式中最低.撒施后灌水处理的氧化亚氮累积净排放量达121.55 g N·hm-2,显著大于撒施后翻耕和条施后覆土处理.撒施后翻耕和条施后覆土处理能有效抑制氨挥发和氧化亚氮排放损失,是较为合理的施肥方式.     

Fluxes of nitrous oxide from boreal peatlands as affected by peatland type,water table level and nitrification capacity

Peat soils with high nitrogen content are potential sources of nitrous oxide (N₂O). Fluxes of nitrous oxide were measuredin situ on nine virgin and ten drained peatlands of different hydrology and nutrient status. Numbers of nitrifying bacteria were estimated in different layers of the peat profiles with a most-probable-number technique. Nitrification potentials were determined in soil slurries of pH 4 and 6 from the profiles of six peat soils. Many virgin peatlands showed low N₂O uptake. Lowering of the water table generally increased the average fluxes of N₂O from the soils, although more in minerotrophic (nutrient rich) than in ombrotrophic (nutrient poor) sites. Ammonium oxidizing bacteria were found on only two sites but nitrite oxidizers were detected in almost all peat profiles. More nitrite oxidizers were found in drained than in virgin peat profiles. Nitrification was enhanced after lowering of the water table in minerotrophic peat but not in ombrotrophic peat. The N₂O fluxes correlated positively with the numbers of nitrite oxidizers, nitrification potential, N, P and Ca content and pH of the soil and negatively with the level of water table (expressed as negative values) and K content of the soil.     

Multi‐species nitrifying biofilm model (MSNBM) including free ammonia and free nitrous acid inhibition and oxygen limitation

A multi‐species nitrifying biofilm model (MSNBM) is developed to describe nitrite accumulation by simultaneous free ammonia (FA) and free nitrous acid (FNA) inhibition, direct pH inhibition, and oxygen limitation in a biofilm. The MSNBM addresses the spatial gradient of pH with biofilm depth and how it induces changes of FA and FNA speciation and inhibition. Simulations using the MSNBM in a completely mixed biofilm reactor show that influent total ammonia nitrogen (TAN) concentration, bulk dissolved oxygen (DO) concentration, and buffer concentration exert significant control on the suppression of nitrite‐oxidizing bacteria (NOB) and shortcut biological nitrogen removal (SBNR), but the pH in the bulk liquid has a weaker influence. Ammonium oxidation increases the nitrite concentration and decreases the pH, which together can increase FNA inhibition of NOB in the biofilm. Thus, a low buffer concentration can accentuate SBNR. DO and influent TAN concentrations are efficient means to enhance DO limitation, which affects NOB more than ammonia‐oxidizing bacteria (AOB) inside the biofilm. With high influent TAN concentration, FA inhibition is dominant at an early phase, but finally DO limitation becomes more important as TAN degradation and biofilm growth proceed. MSNBM results indicate that oxygen depletion and FNA inhibition throughout the biofilm continuously suppress the growth of NOB, which helps achieve SBNR with a lower TAN concentration than in systems without concentration gradients. Biotechnol. Bioeng. 2010;105: 1115–1130. © 2009 Wiley Periodicals, Inc.     

Estimation of nitrous oxide, nitric oxide and ammonia emissions from croplands in East, Southeast and South Asia

Agricultural activities have greatly altered the global nitrogen (N) cycle and produced nitrogenous gases of environmental significance. More than half of all chemical N fertilizer produced globally is used in crop production in East, Southeast and South Asia, where rice is central to nutrition. Emissions of nitrous oxide (N₂O), nitric oxide (NO) and ammonia (NH₃) from croplands in this region were estimated by considering background emission and emissions resulting from N added to croplands, including chemical N, animal manure, biologically fixed N and N in crop residues returned to fields. Background emission fluxes of N₂O and NO from croplands were estimated to be 1.22 and 0.57 kg N ha^?1 yr^?1, respectively. Separate fertilizer‐induced emission factors were estimated for upland fields and rice fields. Total N₂O emission from croplands in the study region was estimated to be 1.19 Tg N yr^?1, with 43% contributed by background emissions. The average fertilizer‐induced N₂O emission, however, accounts for only 0.93% of the applied N, which is less than the default IPCC value of 1.25%, because of the low emission factor from paddy fields. Total NO emission was 591 Gg N yr^?1 in the study region, with 40% from background emissions. The average fertilizer‐induced NO emission factor was 0.48%. Total NH₃ emission was estimated to be 11.8 Tg N yr^?1. The use of urea and ammonium bicarbonate and the cultivation of rice led to a high average NH₃ loss rate from chemical N fertilizer in the study region. Emissions were displayed at a 0.5° × 0.5° resolution with the use of a global landuse database.     

Mechanisms and kinetics of nitric and nitrous oxide production during nitrification in agricultural soil

Laboratory experiments were conducted with three California agricultural soils to examine substrate and process controls over temporal variability of NO and N₂O production during nitrification, and to quantify the kinetics of HNO₂‐mediated chemical reactions. Gross NO production rates were highly correlated (r² = 0.93–0.97) with calculated concentrations of HNO₂, which were shown to originate from autotrophic microbial oxidation of NH₄ ⁺ to NO₂ ^? Production of NO was not correlated with NH₄ ⁺ or NO₃^–, or with the overall nitrification rate. Distinct periods of high NO₂^– accumulation occurred below critical pH values in each soil, apparently due to inhibition of microbial NO₂^– oxidation. Data suggest that even during periods of relatively low NO₂^– accumulation and rapid overall nitrification, HNO₂‐mediated reactions may have been the primary source of NO. Rate coefficients (k_PNO) relating NO production to HNO₂ concentrations were determined for sterile (λ‐irradiated) soils, and were similar to k_PNO values in 2 of 3 nonsterile soils undergoing nitrification. Production of N₂O was correlated with HNO₂ (r² = 0.88–0.99) in sterile soils, and with NO₂^– and NO₃^– (R² = 0.72–0.91) in nonsterile soils. Experiments using ¹⁵N confirmed that dissimilatory NO₃^– reduction contributed to N₂O production even under primarily aerobic conditions. Sterile kPNO and kPN2O values were correlated (r² = 0.90 and 0.82) with soil organic matter content. Overall, the results demonstrate that both steps of the nitrification sequence, together with abiotic reactions involving NO₂^–/HNO₂ need to be considered in developing improved models of NO and N₂O emissions from soils.     

生产性短程硝化-厌氧氨氧化装置处理制药废水的启动性能

为拓展新型生物脱氮技术的应用领域,研究了生产性短程硝化-厌氧氨氧化装置处理制药废水的启动性能。制药废水氨氮浓度为(430.40±55.43)mg/L时,氨氮去除率达(81.75±9.10)%,实现了短程硝化-厌氧氨氧化工艺对制药废水的生物脱氮。制药废水短程硝化系统的启动时间约为74 d,亚硝氮积累率达(52.11±9.13)%,证明了结合模拟废水和实际废水的"两步法"模式对短程硝化系统启动的适用性。制药废水厌氧氨氧化系统的启动时间约为145 d,最大容积氮去除速率达6.35 kg N/(m3·d),容积效能为传统硝化-反硝化工艺的数十倍,证明了结合菌种自繁和菌种流加的模式对厌氧氨氧化系统启动的适用性。     

Influence of tree species and ground vegetation on nitrification in an acid forest soil

Soil N transformations were studied at Ironhill, near Liphook, UK as part of a forest fumigation experiment. Nitrification potential was measured in a humoferric podzol soil, of pH 3 (in 0.01 M CaCl₂). An initial experiment into nitrogen mineralisation potential indicated that nitrification was linked strongly to the species of coniferous tree growing in the soil. Transfer of soil solution between soils had no influence on mineralisation potential and allelopathic effects of the trees were not demonstrated. The initial finding was attributed subsequently to the type of ground vegetation and its management. Attempts to reproduce soil conditions, which promoted nitrification, were partially successful.Soil, from the Ironhill site, was incubated with various nitrogenous substrates and other nutrients and sources of carbon to test whether heterotrophs were responsible for nitrification. Organic N (which was ammonified) promoted nitrification, but the addition of ammonium was inhibitory unless supplied with a readily available carbon source such as acetate. Nitrification potential was unaffected when soils were incubated with an inhibitor of autotrophic nitrification. The results of these experiments supported strongly the hypothesis that heterotrophic organisms were responsible for nitrification in this soil.     

Activity and composition of ammonia oxidizing bacterial communities and emission dynamics of NH3 and N2O in a compost reactor treating organic household waste

Aims:  To monitor emissions of NH₃ and N₂O during composting and link these to ammonia oxidation rates and the community structure of ammonia oxidizing bacteria (AOB).
Methods and Results:  A laboratory-scale compost reactor treating organic household waste was run for 2 months. NH₃ emissions peaked when pH started to increase. Small amounts of N₂O and CH₄ were also produced. In total, 16% and less than 1% of the initial N was lost as NH₃-N and N₂O-N respectively. The potential ammonia oxidation rate, determined by a chlorate inhibition assay, increased fourfold during the first 9 days and then remained high. Initially, both Nitrosospira and Nitrosomonas populations were detected using DGGE analysis of AOB specific 16S rRNA fragments. Only Nitrosomonas europaea was detected under thermophilic conditions, but Nitrosospira populations re-established during the cooling phase.
Conclusions:  Thermophilic conditions favoured high potential ammonia oxidation rates, suggesting that ammonia oxidation contributed to reduced NH₃ emissions. Small but significant amounts of N₂O were emitted during the thermophilic phase. The significance of different AOBs detected in the compost for ammonia oxidation is not clear.
Significance and Impact of Study:  This study shows that ammonia oxidation occurs at high temperature composting and therefore most likely reduces NH₃ emissions.     

污水短程硝化体系氨氧化菌与亚硝酸盐氧化菌的生态学研究进展

短程硝化(partial nitrification, PN)是一种绿色低碳的生物脱氮创新技术,伴随厌氧氨氧化(anaerobic ammonia oxidation, Anammox)污水脱氮技术的进一步推广,短程硝化作为提供其电子受体的重要环节,已成为了污水脱氮领域的研究热点。氨氧化菌(ammonia-oxidizing bacteria,AOB)和亚硝酸盐氧化菌(nitrite-oxidizing bacteria, NOB)是该技术的核心竞争微生物,掌握这两类微生物的生态学特征,借助生态学理论和手段调控AOB淘汰NOB,提高种群的可预测性,对于实现稳定高效的短程硝化具有重要意义。本文基于生态学角度介绍了AOB和NOB基础分类、生理性能及生态位分离,重点综述了短程硝化系统中AOB和NOB的生长动力学、群落构建、环境因素和相互作用,最后对这两类微生物的未来研究重点和研究方法进行了展望,为短程硝化工艺的快速启动和稳定运行提供理论指导。     

温度、盐度和体长对西藏拟溞（Daphniopsis tibetana Sars）排氨率的影响

在实验室内研究了温度(T=3、8、14、20 ℃和22 ℃)、盐度(S=5、10、15、20 和 25)和体长(0.80、1.23、1.51、1.84 mm和2 38 mm)对西藏拟溞排氨率(NR)的影响.结果表明,在试验温度范围内,西藏拟溞幼溞排氨率随温度(T)的升高而升高,其回归方程为 NR=0.0359Ln(T)-0.0185 (n=5, r=0.9881, P<0.05),数值范围为 0.022~0.092 mg/(g·h).在盐度(S)试验中,西藏拟溞幼溞排氨率有同样的趋势,回归方程为NR = 0.0535e0.0213S(n=5, r=0.9884),其数值范围为0.061~0.092mg/(g·h).而体长试验中,西藏拟溞排氨率随体长(L)的增加而降低,回归方程为NR=0.0568L-1.4154(n=5, r=0.9938, P<0.05),数值范围为0.017~0.083mg/(g·h).     

Salicylic acid,hydrogen peroxide and calcium-induced saline tolerance associated with endogenous hydrogen peroxide homeostasis in naked oat seedlings

This study investigates the role of salicylic acid (SA), hydrogen peroxide (H₂O₂) and calcium chloride (CaCl₂) singly or in combination, in inducing naked oat plant tolerance to sodium chloride (NaCl). Two-week-old naked oat plants were pretreated with both single and double of 0.5 mM SA, 0.5 mM H₂O₂ and 5 mM CaCl₂ by adding them to the culture solution for 24 h. At the end of the pretreatment, the plants were subjected to 200 mM NaCl exposure for 7 days. Data were collected on plant biomass, H₂O₂ level, antioxidant enzyme activity, non-enzymatic antioxidant content and malondialdehyde (MDA) content. Results showed that exposure to salt significantly inhibited plant growth, and the shoot and root dry weights were reduced 47.5% and 63.4%, and the H₂O₂ levels elevated 5.8 and 2.4 times in comparison with those in the control, respectively. Under the saline stress, the activities of superoxide dismutase (SOD) and catalase (CAT) were induced, but the contents of ascorbic acid (AA) and glutathione (GSH) decreased, and MDA largely accumulated. The various pretreatments efficiently counteracted the salt-caused growth inhibition, especially with H₂O₂ + CaCl₂ the shoot and root dry weights reduced only 9.4% and 24.4% of the non salt-stressed plants. The determination of endogenous H₂O₂ level demonstrated that the pretreatments induced H₂O₂ accumulation, with H₂O₂ + CaCl₂ being most efficient, but the effect was transient. After 7 days of saline stress, the H₂O₂ contents in the pretreated shoots and roots accounted for 23.7–41.8% and 31.7–57.3% of the non-pretreated plants, varying according to the different pretreatments. Under saline stress, SOD and CAT further increased, AA and GSH maintained higher levels and MDA decreased in the pretreated plants compared to the untreated plants. With application of diphenylene iodonium (DPI) during the pretreatment, which inhibited the accumulation of H₂O₂, the ameliorative effect of the pretreatment on salt-caused plant growth inhibition was reduced. However, applied DPI at the immediate end of the pretreatment did not alter its favorable role, indicating a H₂O₂ peak formed at the early time of saline stress might play an important role in regulating plant tolerance to saline stress.     

不同盐度对闽江河口沉积物硝化作用的影响

选择闽江河口鳝鱼滩湿地与道庆洲湿地为对象,采集湿地沉积物在室内进行沉积物硝化培养,分析不同盐度水平对湿地沉积物硝化作用的影响.结果表明:闽江河口湿地沉积物硝化速率普遍较低,鳝鱼滩湿地沉积物最高硝化速率只有0.193 mg·kg^-1·d^-1,而道庆洲湿地沉积物硝化速率最高不超过0.050 mg·kg^-1·d^-1.盐度的升高会抑制闽江河口湿地沉积物的硝化作用.低盐度时(5),硝化速率下降的主要原因是硝化细菌活性受到抑制;随着盐度的升高(10),硝化速率略有上升但仍低于初始值,这是由于随着盐度的升高,盐度对好氧氨化细菌活性的抑制程度有所加强,导致系统产生NH_4^+-N的速率下降,从而造成好氧氨化细菌对表观硝化速率下降的贡献减少.沉积物硝化活性对于盐度的响应存在地域差异.咸水湿地(鳝鱼滩湿地)沉积物中的微生物对盐度的变化适应性较强,从而使该湿地沉积物硝化活性在高盐度条件下仍然较高.淡水湿地(道庆洲湿地)沉积物对盐度变化的适应性较弱,导致其沉积物硝化活性在高盐度条件下低于中等盐度.闽江河口鳝鱼滩与道庆洲的短叶茳芏湿地沉积物硝化作用较低的主要原因是沉积物呈酸性及淹水状态下的缺氧条件弱化了沉积物的硝化作用.两处湿地的硝化速率与硝化活性随时间的变化趋势为先升高后下降,这是由初始NH_4^+-N浓度、氧气含量和反硝化共同作用造成的.     

Effects of urine on soil microbial biomass,methanogenesis, nitrification and denitrification in grassland soils

Urine was added under controlled conditions to intact turfs taken from long-term permanent pasture on clay loam and sandy loam soils in South West England. Methane exchanges were small (<+/−0.03 μg CH₄ m^-2 min^-1) and overall absorption equalled or exceeded emission in both soils. On the clay loam, wetting with water or urine increased soil microbial biomass C and N contents by about 20% but there was no specific effect of urine. Urine, however, caused an increase in soil respiration of >50% and the average increase was greater for cow's urine (30.8 mg CO₂ m^-2 min^-1) than for an artificial urine (20.1 mg CO₂ m^-2 min^-1). Emissions of nitric and nitrous oxides following urine application were substantial (on average 0.36 μg NO-N and 29 μg N₂O-N m^-2 min^-1) but short lived (<40 days). The high levels of ammonium found in the urine treated soils (>200 mg NH₄ ⁺-N kg^-1) were nitrified to nitrate over a period of 42 days. Qualitative changes in the soil microbial biomass were evidently not related to biomass size. Relationships between trace gas emissions and soil processes are discussed. ei]Section editor: R Merckx     

盐分对生物脱氮工艺中硝化反应的影响与机理

高盐废水来源广泛,在利用生物脱氮法处理高盐含氮废水时,盐分会对生物脱氮产生抑制作用.硝化反应是生物脱氮工艺中的关键过程,研究盐分对硝化反应的影响机理具有重要意义.本文概述了盐分对废水生物脱氮过程中硝化反应影响的研究进展,总结了盐胁迫对好氧氨氧化过程、亚硝酸盐氧化过程中硝化效率和反应特性的影响规律,并分析了盐分对硝化微生物细胞形态、生物絮体结构和胞外聚合物特性变化以及菌群结构的影响,系统阐述了盐胁迫下的硝化反应机理,为高盐分高铵氮废水生物脱氮工艺设计提供理论指导.
     

Design of 16S rRNA-targeted oligonucleotide probes and microbial community analysis in the denitrification process of a saline industrial wastewater treatment system

Three 16S rRNA-targeted oligonucleotide probes, namely, PSMg437 targeting several members of the genus Pseudomonas, Hlm474 targeting several members of the genus Halomonas, and Clw844 targeting several members of the genus Colwellia, were designed. The microbial community structure and nitrogen removal ability of nitrate-containing saline wastewater treatment systems with anaerobic packed bed and fluidized bed were monitored. Direct cell counting using fluorescence in situ hybridization (FISH) images revealed that various phylogenetic groups were evenly distributed in the anaerobic packed bed whereas members of the genus Halomonas were dominant particularly in the anaerobic fluidized bed. These results suggest that the microbial communities produced by different flow conditions correlated with denitrification ability in saline industrial wastewater treatment systems.