Community dynamics of ammonia oxidizing bacteria in a full-scale wastewater treatment system with nitrification stability

To determine whether the functional stability of nitrification was correlated to a stable community structure of ammonia oxidizing bacteria (AOB) in a full-scale wastewater treatment plant, the AOB community dynamics in a wastewater treatment system was monitored over one year. The community dynamics were investigated using specific PCR followed by terminal restriction fragment length polymorphism (T-RFLP) analysis of the amoA gene. The T-RFLP results indicated that during the period of nitrification stability, the AOB community structure in the full-scale wastewater treatment system was relatively stable, and the average change rate every 15 d of the system was 6.6%±5.8%. The phylogenetic analysis of the cloned amoA gene showed clearly that the dominant AOB in the system was Nitrosomonas spp. The results of this study indicated that throughout the study period, the AOB community structure was relatively stable in the full-scale wastewater treatment system with functional stability of nitrification.     

Differences in distributions,assembly mechanisms,and putative interactions of AOB and NOB at a large spatial scale

● Nitrifiers in WWTP were investigated at large spatial scale. ● AOB populations varied greatly but NOB populations were similar among cities. ● Drift dominated both AOB and NOB assembling processes. ● DO did not show a significant effect on NOB. ● NOB tended to cooperate with AOB and non-nitrifying microorganisms. Ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) play crucial roles in removing nitrogen from sewage in wastewater treatment plants (WWTPs) to protect water resources. However, the differences in ecological properties and putative interactions of AOB and NOB in WWTPs at a large spatial scale remain unclear. Hence, 132 activated sludge (AS) samples collected from 11 cities across China were studied by utilizing 16S rRNA gene sequencing technology. Results indicated that Nitrosomonas and Nitrosospira accounted for similar ratios of the AOB community and might play nearly equal roles in ammonia oxidation in AS. However, Nitrospira greatly outnumbered other NOB genera, with proportions varying from 94.7% to 99.9% of the NOB community in all WWTPs. Similar compositions and, hence, a low distance–decay turnover rate of NOB (0.035) across China were observed. This scenario might have partly resulted from the high proportions of homogenizing dispersal (~13%). Additionally, drift presented dominant roles in AOB and NOB assembling mechanisms (85.2% and 81.6% for AOB and NOB, respectively). The partial Mantel test illustrated that sludge retention time and temperature were the primary environmental factors affecting AOB and NOB communities. Network results showed that NOB played a leading role in maintaining module structures and node connections in AS. Moreover, most links between NOB and other microorganisms were positive, indicating that NOB were involved in complex symbioses with bacteria in AS.     

Characterization of CANON reactor performance and microbial community shifts with elevated COD/N ratios under a continuous aeration mode

COD/N at low ratios (0–0.82) improved N removals of CANON. CANON performance decreased after COD/N up to 0.82. The relative abundance of AOB decreased continuously with increasing COD/N. AOB outcompeted at a high COD load led to CANON failure. The relative abundance of AnAOB decreased and increased with increasing COD/N. The effects of increasing COD/N on nitrogen removal performance and microbial structure were investigated in a SBR adopting a completely autotrophic nitrogen removal over nitrite process with a continuous aeration mode (DO at approximately 0.15–0.2 mg/L). As the COD/N increased from 0.1 to≤0.59, the nitrogen removal efficiency (NRE) increased from 88.7% to 95.5%; while at COD/N ratios of 0.59–0.82, the NRE remained at 90.7%–95.5%. As the COD/N increased from 0.82 to 1.07, the NRE decreased continuously until reaching 60.1%. Nitrosomonas sp. (AOB) and Candidatus Jettenia (anammox bacteria) were the main functional genera in the SBR. As the COD/N increased from 0.10 to 1.07, the relative abundance of Nitrosomonas decreased from 13.4% to 2.0%, while that of Candidatus Jettenia decreased from 35% to 9.9% with COD/N<0.82 then increased to 45.4% at a COD/N of 1.07. Aerobic heterotrophic bacteria outcompeted AOB at high COD loadings (650 mg/L) because of oxygen competition, which ultimately led to deteriorated nitrogen removal performance.     

沣河水系脱氮微生物群落结构研究

河流水体氮素的超负荷不仅破坏了水体生态环境,也严重威胁着人类的生存和发展.水体中有机氮、无机氮（氨氮、亚硝氮、硝氮）和分子氮之间的转化（氮循环）有赖于水体中大量的氮循环微生物（固氮细菌、硝化细菌和反硝化细菌）,然而这些氮循环微生物的生长繁殖也受到包括氮素的形态和浓度在内的多种环境因子的影响,这些因素也通过影响氮循环微生物的生长繁殖进而使得水体中氮素的转化速率发生变化,对水体氮污染的防治有不可忽视的作用.本研究通过在沣河设置不同的研究断面,采集水体样品,进行水质分析,并通过现代分子生物学技术（PCR-DGGE）方法对研究断面水体中氮循环微生物（固氮细菌、硝化细菌和反硝化细菌）的群落结构进行分析.再通过统计学软件对所得分子生物学信息与水质环境因子的相关性进行统计学分析,发现沣河水体中氮循环微生物群落结构受到多种环境因子共同影响,且在枯水期和丰水期表现出不同的特征.在丰水期沣河水体中,硝化细菌群落在中游表现出较高的多样性和丰富性,这与沣河中上游农业COD（化学需氧量）、BOD（生化需氧量）氨氮及有机氮污染物排放量较大,沣河水体DO（溶解氧）高有关.水体中的氨氮、亚硝氮、温度的增加是促进水体中硝化细菌的均匀性和丰富度的增高的主要因子,而pH 值的升高,使得水体中硝化细菌的均匀性和丰富度降低.反硝化微生物在中游和下游的多样性和丰富度较高,与有机物及硝酸盐含量相关.水体中的BOD、COD、TP（总磷）、硝氮的增加是促进水体中反硝化细菌的均匀性和丰富度的增高主要相关因子,而DO 的增多则会对部分反硝化细菌产生不利影响,使得水体中反硝化细菌的均匀性和丰富度降低.本研究结果为沣河以及其他河流的污染控制以及基于微生物的生态修复提供了科?     

Overlooked nitrogen-cycling microorganisms in biological wastewater treatment

• AOA and comammox bacteria can be more abundant and active than AOB/NOB at WWTPs. • Coupled DNRA/anammox and NO_x-DAMO/anammox/comammox processes are demonstrated. • Substrate level, SRT and stressors determine the niches of overlooked microbes. • Applications of overlooked microbes in enhancing nitrogen removal are promising. Nitrogen-cycling microorganisms play key roles at the intersection of microbiology and wastewater engineering. In addition to the well-studied ammonia oxidizing bacteria, nitrite oxidizing bacteria, heterotrophic denitrifiers, and anammox bacteria, there are some other N-cycling microorganisms that are less abundant but functionally important in wastewater nitrogen removal. These microbes include, but not limited to ammonia oxidizing archaea (AOA), complete ammonia oxidation (comammox) bacteria, dissimilatory nitrate reduction to ammonia (DNRA) bacteria, and nitrate/nitrite-dependent anaerobic methane oxidizing (NO_x-DAMO) microorganisms. In the past decade, the development of high-throughput molecular technologies has enabled the detection, quantification, and characterization of these minor populations. The aim of this review is therefore to synthesize the current knowledge on the distribution, ecological niche, and kinetic properties of these “overlooked” N-cycling microbes at wastewater treatment plants. Their potential applications in novel wastewater nitrogen removal processes are also discussed. A comprehensive understanding of these overlooked N-cycling microbes from microbiology, ecology, and engineering perspectives will facilitate the design and operation of more efficient and sustainable biological nitrogen removal processes.     

饮用原水中微量NH_3-N的生物膜脱除

针对NH3-N浓度为1mg.l-1的饮用原水,分别以硅锰砂石和聚胺脂海绵作为硝化生物膜载体,考察了自制固定床生物膜反应器的脱氨效果.30d的连续运行结果表明,砂石载体系统达到95%—100%的NH3-N去除率,出水NH3-N浓度接近0,出水NO2--N浓度在0—0.02mg.l-1之间;而海绵载体系统虽然挂膜快,但运行阶段NH3-N去除率仅为10%—35%,出水NH3-N不能达标,且具有生物可降解性,反硝化过程优先导致NO2--N的积累.进一步考察了硅锰砂石生物膜系统稳定运行的最优参数,得出试验条件下,水力停留时间为6min,气水比为1:1,反冲洗周期为6d.硅锰砂石因其良好的孔隙率、粒径分布和密度等物理特性,以及化学稳定性和生物安全性,是含低浓度NH3-N原水生物膜净化的良好载体选择.     

近地层臭氧浓度升高对稻田土壤氨氧化与反硝化细菌活性的影响

利用中国唯一的稻麦轮作臭氧FACE（free-air O3 concentration enrichment,开放式空气臭氧浓度增高）试验平台,研究近地层臭氧浓度升高对稻田不同生育时期植株氮吸收量、土壤氮含量与脲酶活性、土壤氨氧化细菌与反硝化细菌数量以及成熟期土壤硝化与反硝化作用强度的影响。结果发现,臭氧浓度升高条件下,单株水稻（Oryza sativaL.）的氮吸收量趋于升高,土壤全氮含量趋于下降、脲酶活性趋于增强,在成熟期土壤全氮与对照相比平均下降9%,土壤脲酶活性与对照相比平均升高13%。在水稻整个生长季节,土壤氨氧化细菌和反硝化细菌的数量均呈现出先增多后减少的趋势,且均在开花期达到峰值,但臭氧熏蒸土壤与对照相比趋于升高;在水稻成熟期,土壤中单个氨氧化细菌和反硝化细菌的活性均趋于下降,土壤硝化强度和反硝化强度与对照相比也平均下降17%和24%。结果表明,近地层臭氧浓度升高条件下,稻田土壤氮素转化因为水稻氮素吸收增强而加快,土壤氨氧化细菌和反硝化细菌的数量均趋于增多,但它们的生理代谢活性均趋于下降。     

不同C/N值下亚硝酸盐氧化菌和异养菌混合体系的微生物多样性

采用PCR-RFLP技术研究了不同C/N比下亚硝酸盐氧化菌及异养菌混合体系的微牛物多样性,并探讨了微生物菌群结构与其功能(硝化件能)的关系.C/N=0时,混合体系主要由自养菌和寡营养菌(85.1%)组成,包括亚硝酸盐氧化菌(NOB)、拟杆菌门、α-变形菌纲、浮霉菌门和绿色非硫细菌中的一些菌株.C/N=0.44时,混合体系中的自养菌减少,异养菌(主要是γ-变形菌纲的成员)大量出现.C/N=8.82时,γ-变形菌纲的菌株尤其是反硝化菌Pseudomonas sp.占主导(93.8%),与此同时,随着C/N升高,该混合体系的硝化性能也由专一的亚硝酸盐氧化过程转变为同时硝化反硝化过程.微生物菌群结构的转变较好地解释了其硝化性能的改变.本研究揭示了微生物菌群结构与其功能的内在联系,同时表明PCR-RFLP技术与化学分析相结合是研究微生物菌群结构与功能的有力工具.图3表2参13     

低温下曝气生物滤池预处理污染河水的试验研究

采用两段曝气生物滤床串联工艺预处理入滇新运粮河河水,研究了其在冬天低温条件下对有机物和氨氮的去除效果,并考察了pH值的变化。结果表明,在进水流量为2.4 m3.d-1、水温为13-16℃、原水ρ（CODCr）为66.46-107.82 mg·L-1、ρ（氨氮）为22.15-30.68 mg·L-1的水质特征条件下,系统对CODCr和氨氮的去除率分别为36.08%-50.37%和76.98%-93.56%。其中,碳氧化段以去除有机物为主,硝化段以去除氨氮为主;系统中硝酸氮质量浓度明显升高,无亚硝氮的积累;装置对总氮的平均去除率为19.56%,可以认为总氮的去除是同步硝化反硝化的结果。系统中pH值有所变化但维持在7-8之间,其中碳氧化段pH值升高,硝化段pH值下降。系统对有机物和氨氮良好的去除效果为后续进一步的生物处理提供了条件。     

Seasonal variations in abundance of nitrifying bacteria in fish pond ecosystem

Seasonal changes in abundance of nitrifiers (ammonia-oxidizing and nitrite-oxidizing bacteria) in surface and bottom water of freshwater ponds were examined with respect to temperature, DO, pH as well as concentration of ammonia and nitrite. The most probable number (MPN) of ammonia-oxidizers in different ponds varied from 1297 +/- 3.6 to 1673.23 +/- 0.36 ml(-1) in bottom and 720.5 +/- 8.1 to 955.3 +/- 10.8 ml(-1) in surface water during the rainy season while the MPN ranged from 1074 +/- 1.07 to 1372.17 +/- 4.6 ml(-1) in bottom and 515 +/- 10.1 to 678 +/- 11.8 ml(-1) in surface water in winter. However, the MPN were greatly reduced in summer and ranged from 435.05 +/- 15.7 to 547.54 +/- 2.12 ml(-1) in bottom and 218.7 +/- 7.3 to 368.4 +/- 9.32 ml(-1) in surface water. Similar seasonal trends were also observed in MPN of nitrite-oxidizers. Among all the physico-chemical parameters, abundance of nitrifiers was more positively correlated with ammonia and nitrite concentration in all the seasons. The abundance of nitrifiers in surface and bottom water was highest in rainy season followed by winter and modest in summer. The potential nitrification activities and oxidation rates were shown to be linear and activity of ammonia-oxidizing and nitrite-oxidizing bacteria was highest during rainy season.     

Nitrification model with an inhibitory effect of sea water

A dynamic model of nitrification including bacterial growth equations was developed for the understanding of nitrification in estuaries. Sensitivity analysis based on the model revealed that not only maximum specific growth rates of nitrifiers but also yield constants and initial bacterial concentrations have great effect on the nitrification process.Batch experiments were carried out in order to investigate nitrification rates and an inhibitory effect on nitrification by sea water. The maximum specific growth rate of ammonia oxidizing bacteria in fresh water was estimated as 0.04 (h^?1) using a nonlinear regression method. The sea water content of 10%, 25%, 50% and 100% reduced the maximum specific growth rate of ammonia oxidizing bacteria to 0.027, 0.016, 0.0085 and 0.0028 (h^?1) respectively. The effect of sea water on nitrification was interpreted as noncompetitive inhibition. The computed results of inhibition of nitrification based on the noncompetitive inhibition model are presented.     

硝化作用微生物的分子生物学研究进展

回顾了硝化作用微生物的分子生物学研究进展 ,主要介绍了硝化作用微生物的种类 ,包括氨氧化菌、亚硝酸氧化菌、异养氨氧化菌和厌氧氨氧化菌 .这些微生物的系统发育分析表明亚硝化菌的系统发育相对简单 ,而硝化细菌则要复杂许多 .还介绍了在硝化作用微生物生态学研究中应用的分子生物学技术 ,如PCR、变性梯度胶电泳 (DGGE)、原位荧光杂交 (FISH)等 ,以及不同类群细菌中与硝化作用相关的酶类及其基因 .文章的最后还提出了一些研究展望 .图 3参 35     

三种载体上生物膜硝化作用动力学初步研究

通过测定水中COD、：NH4^ —N、NO2^-—N及NO3^-—N的浓度变化，研究了淹没式废水处理装置中沸石、活性炭和沙粒3种载体上硝化作用生物膜的动力学过程和反硝化作用。结果表明，3种载体上生物膜降解有机物(以COD表示)的过程可用一级动力学方程描述，反应速率常数分别为：沙粒0．0848h^-1、活性炭0．1187h^-1、沸石0．1334h^-1。3种载体上生物膜去除氨态氮的过程则可用零级动力学方程描述，反应速率常数分别为：沙粒-0．7743h^-1、活性炭-0．9886h^-1、沸石-1．0714h^-1．附着于沙上的生物膜去除亚硝酸盐氮的过程也可用零级动力学方程描述，反应速率常数为-0．6057h^-1，水中硝酸盐氮浓度较高时，载体沸石和活性炭上可能附着生长反硝化菌。图5表2参15。     

The impact of ultrasonic treatment on activity of ammonia-oxidizing bacteria and nitrite-oxidizing bacteria in activated sludge

Conditions for ultrasonic treatment to achieve partial nitritation are optimized. Ultrasound reduces metabolic activity and releases intracellular metabolites. Mechanical shearing is essential to inhibit nitrite oxidation. The ultrasonic treatment of sludge has been considered as an effective method to facilitate the partial nitritation of municipal sewage. This study aims to reveal the effects of ultrasound on ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB). The impact factors including ultrasonic irradiation time and intensity, sludge concentration, thermal effect and released free radicals were studied. The maximized difference between the changes in AOB and NOB activities were obtained with 10 g mixed liquor suspended solids (MLSS)/L, using 0.9 kJ/mL ultrasonic energy density and 12 h interval time. The increased ultrasonic intensity destroyed the floc structure of activated sludge, increased the microbial death, and decreased the cellular ATP level. Further, the mechanism exploration indicated that the mechanical shearing could be a critical factor in achieving the nitritation with inhibitory effect on nitrite oxidation.     

养殖水体复合功能菌的分离及其性能

针对养殖水体中因氨态氮、硫化氢和小分子有机酸富营养化引起的污染问题,分离筛选出硝化细菌、反硝化细菌、光合细菌、硫化细菌和生物絮凝菌等具有不同生理功能的污染物治理菌株,经优化配伍制备出性能优良的复合功能菌,结果表明:硝化细菌对氨态氮的去除率达97.8%,亚硝态氮的去除率达95.7%,反硝化细菌对硝态氮的去除率为96.4%,光合细菌和硫化细菌对硫化氢的去除率为55%,微生物絮凝菌的絮凝效率为83%;复合功能菌对CODCr、NH4+-N,总氮、硫化物的去除率分别可达94.3%,89.6%,88.7%和71.3%。     

亚硝酸盐急性胁迫对中华绒螯蟹幼体相关免疫指标和应激蛋白(HSP70)表达的影响

为探讨亚硝酸盐急性胁迫对中华绒螯蟹机体免疫力的影响,以及机体在胁迫过程中的应对策略,分别测定了幼蟹(平均体重3.25 g)在10、20、30、40 mg L-1亚硝酸盐暴露组和对照组中血细胞总数(Total hemocyte counts,THC)、超氧化物歧化酶(Superoxide dismutase,SOD)活力、丙二醛(Malodiadehyde,MDA)含量以及热休克蛋白70(Heatstress protein 70,HSP70)表达量在受胁迫2 d内的动态变化.结果表明:1)低浓度亚硝酸盐(10 mg L-1)胁迫组THC在胁迫3 h时显著高于对照组,肝胰腺中SOD活性在胁迫3 h和6 h时亦显著提高,表现出一定的"毒物兴奋效应".2)但随着暴露时间的延长和亚硝酸盐浓度的升高,幼蟹THC减少,肝胰腺SOD活性下降,而MDA含量升高.3)对照组中有少量的HSP70表达.幼蟹经10、20 mg L-1亚硝酸盐处理后,鳃组织中HSP70表达量在胁迫6 h后达到最大,而经30、40 mg L-1亚硝酸盐处理后,HSP70表达量在胁迫3 h后达到最大.各浓度组所诱导的HSP70最大表达量与水中亚硝酸盐的浓度呈正相关.以上结果说明亚硝酸盐胁迫可通过加剧组织中脂质过氧化的方式导致机体免疫力下降,肝胰腺中MDA的含量变化可作为亚硝酸盐胁迫的有效评价指标之一,而HSP70表达量的变化可作为早期预警指标应用于亚硝酸盐胁迫评价中.图2表1参36     

Autotrophic denitrification using hydrogen oxidizing bacteria in continuous flow biofilm reactor

Autotrophic denitrification was investigated in five bench‐scale upflow attached growth reactors using hydrogen oxidizing bacteria under anoxic conditions. The performance of sand, granular activated carbon (GAC), crushed pumice, crushed volcanic rock, and plastic media were evaluated as the support material. The reactors were inoculated with acclimated cultures obtained from domestic sewage treatment plant. A synthetic solution containing nitrate was used as the influent. The reactor performance was evaluated by measuring influent and effluent nitrate concentration. The design parameters demonstrated that the effectiveness of autotrophic denitrification is comparable to that of the heterotrophic process and may be utilized economically for drinking water treatment either as the main process or as a supplemental process for ion exchange regenerant treatment.     

余氯对再生水中铜绿微囊藻(Microcystis aeruginosa)的生长抑制作用

以北京市污水处理厂二级出水为对象,考察了余氯对再生水中铜绿微囊藻(Microcystis aeruginosa)生长的影响.实验表明,0.2 mg·l~(-1)以上的余氯对铜绿微囊藻的生长有明显抑制效果,抑制率在培养第2天达60%-80%,第8天达99%,抑制效果持续60d.再生水中的氨氮浓度越高,余氯对铜绿微囊藻的抑制效果越差.氯的不同形态对铜绿微囊藻的生长抑制效果不同,氯对铜绿微囊藻的抑制效果优于氯胺.     

Applicability of the Arrhenius model for ammonia oxidizing bacteria subjected to temperature time gradients

The aim of this work is to identify the range of applicability of Arrhenius type temperature dependence for Ammonia Oxidizing Bacteria (AOB) subjected to temperature time gradients through continuous titrimetric tests. An innovative online differential titrimetric technique was used to continuously monitor the maximum biologic ammonia oxidation rate of the biomass selected in a pilot scale membrane bioreactor, as a function of temperature time gradients. The monitoring technique is based on the measurement of alkalinity and hydrogen peroxide consumption rates in two parallel reactors operated in non-limiting substrate conditions for AOB; both reactors were continuously fed with mixed liquor and in one of them AOB were inhibited with allylthiourea. The effects of temperature decrease rates in the range 1 to 4°C·h⁻¹ were evaluated by controlling the titrimetric reactor in the temperature range 10°C–20°C. The dependence of growth kinetics on temperature time gradients and the range of applicability of Arrhenius model for temperature dependency of AOB growth kinetics were assessed. The Arrhenius model was found to be accurate only with temperature gradients lower than 2°C·h⁻¹. The estimated Arrhenius coefficients (θ) were shown to increase from 1.07 to 1.6 when the temperature decrease rate reached 4°C·h⁻¹.     

Biological production of nitrite in seawater

The relative importance of 3 different sources for biological production of nitrite in seawater was studied. Decomposition of fecal pellets of the copepod Calanus helgolandicus (at a concentration of approximately 12 g-at N/l), in seawater medium, released small amounts of ammonia over a 6 week period. It nitrifying bacteria were added to the fecal pellets nitrite was barely detectable over the same period. Decomposition of phytoplankton (present at a concentration of about 8 g-at particulate plant N/l) with added heterotrophic bacteria, released moderate amounts of ammonia over a 12 week period. If the ammonia-oxidizing bacterium Nitrosocystis oceanus was added to the decomposing algae, nitrite was produced at a rate of 0.2 g-at N/l/week. Heterotrophic nitrification was not observed when 7 open-ocean bacteria were tested for their ability to oxidize ammonia. The diatom Skeletonema costatum, either non-starved or starved of nitrogen, produced nitrite when growing with 150 or 50 g-at NO ₂ ^- -N/l at a light intensity of about 0.01 ly/min. When nitrate in the medium was exhausted, S. costatum assimilated nitrite. If starved of vitamin B₁₂, both non-N-starved and N-starved cells of S. costatum produced nitrite in the medium with 150 g-at NO ₃ ^- -N/l. Nitrate was not exhausted and cell densities reached 2x10⁵/ml due to vitamin B₁₂ deficiency. If light intensity was reduced to 0.003 ly/min under otherwise similar conditions, cells did not grow due to insufficient light, and nitrite was not produced. In the sea, it appears that, in certain micro-environments, decomposition of particulate matter releases ammonia with its subsequent oxidation to nitrite. The amounts of these nutrients and the rate at which they are produced are dependent upon the nature of the materials undergoing decomposition and the associated bacteria. In certain other areas of the sea, where phytoplankton standing stock is high and nitrate is non-limiting, excretion by these organisms is a major source of nitrite.