好氧和缺氧条件下游离亚硝酸对氨氧化菌和亚硝酸盐氧化菌的选择性抑制

【背景】稳定短程硝化是实现城市污水厌氧氨氧化技术的瓶颈,目前国内外关于游离亚硝酸(Free nitrous acid,FNA)对硝化菌活性的影响大多是在曝气条件下进行研究,鲜有关于缺氧条件下FNA对硝化菌活性影响的报道。【目的】探究好氧和缺氧下FNA对氨氧化菌(Ammonia oxidizing bacteria,AOB)和亚硝酸盐氧化菌(Nitrite oxidizing bacteria,NOB:Nitrospira和Nitrobacter)活性的抑制影响。【方法】采用序批式反应器(Sequencing batch reactor,SBR),基于混合液悬浮固体浓度(Mixed liquid suspended solids,MLSS)为8 300 mg/L的全程硝化污泥条件,通过批次试验分别考察好氧和缺氧下FNA(初始浓度为1.16 mg/L)处理48 h后,AOB和NOB活性的变化情况。【结果】好氧FNA处理活性污泥48 h后,FNA浓度维持在1.16-1.17 mg/L,游离氨(Free ammonia,FA)浓度小于0.017 mg/L,AOB、Nitrospira、Nitrobacter丰度均未发生明显变化;过曝气至99 h时,与空白组相比,比氨氮氧化速率(r~+_(NH4-N))、比亚硝酸盐氮氧化速率(r_(NO2-N))均出现小幅下降,分别由3.5、4.828 mg N/(g VSS·h)降至3.3、4.668 mg N/(g VSS·h),且亚硝酸盐氮累积率(Nitrite accumulation rate,NAR)始终低于33.2%。缺氧FNA处理活性污泥48 h后,FNA浓度维持在0.64-1.16 mg/L,FA浓度低于0.039 mg/L,AOB丰度变化较小,而Nitrospira、Nitrobacter丰度均明显下降,分别由3.002 9×10~9、4.245×10~8 copies/g VSS降至1.666 5×10~8、5.163 8×10~7 copies/g VSS;过曝气至99 h时,与空白组相比,r~+_(NH4-N)值下降幅度较小,而r_(NO2-N)值明显降低,由4.828 mg N/(g VSS·h)降至0.007 mg N/(g VSS·h),且在过曝气0-292 h内,NAR均大于94%。【结论】好氧FNA处理活性污泥48 h后对AOB和NOB无明显抑制作用,但缺氧FNA处理活性污泥48 h后对AOB具有轻微抑制作用,而对NOB具有强烈的抑制作用,可以实现稳定的短程硝化。     

Effect of nitrite concentration and pH on most probable number enumerations of non-growing Nitrobacter spp.

Abstract Enumerations of nitrite-oxidizing bacteria in soil samples by a Most Probable Number technique, often showed relatively high cell numbers at a low nitrite concentration compared with the numbers of ammonium-oxidizing bacteria. It was hypothesized that the high numbers enumerated at low nitrite concentration would represent non-growing or organotrophically growing cells of nitrite-oxidizing species. In this paper, the sensitivity of non-growing Nitrobacter species to high nitrite concentrations as well as to low pH was examined. Different Nitrobacter species were pre-cultured at 0.5 mM nitrite. Non-growing cells differing in age were enumerated at different nitrite concentrations and pH values. The incubation period lasted for 5 months at 20°C. However, during the incubation periods of the older non-growing cells, it appeared that a period of 5 months might have been too short for reaching constant numbers. Early stationary cells of all species that were studied appeared not to be affected by high nitrite concentrations or low pH. Eight- and 18-month-old non-growing cells of Nitrobacter hamburgensis were also insensitive to 5 mM nitrite. The numbers of 8- and 18-month-old resting cells of N. vulgaris were only repressed by a combination of 5 mM nitrite and a low pH. Eight-month-old non-growing cells of N. winogradskyi were sensitive to 5 mM irrespective of pH, but 18-month-old cells only to 5 mM nitrate at low pH. The numbers of 8- and 18-month-old resting cells of N. winogradskyi serotype agilis were repressed by low pH rather than high nitrite concentration. Hence, it was concluded that the large differences in numbers of nitrite-oxidizing bacteria obtained with low and high nitrite concentrations in the incubation medium, was not likely to be due to the presence of non-growing Nitrobacter species in soil samples, but rather to the existence of organotrophically growing Nitrobacter cells.     

从典型硝化细菌到全程氨氧化微生物：发现及研究进展

生物硝化过程在全球氮循环中起关键性作用,被认为由氨氮氧化成亚硝酸盐和亚硝酸盐氧化成硝酸盐两个步骤组成,分别由氨氧化微生物(Ammonia oxidizing microorganisms,AOM)和硝化细菌(Nitrite oxidizing bacteria,NOB)催化完成。AOM包括氨氧化细菌(Ammonia oxidizing bacteria,AOB)和氨氧化古菌(Ammonia oxidizing archaea,AOA),AOB与AOA分布广泛,两者的相对丰度和氨氮浓度密切相关。2015年底,3个硝化螺菌属(Nitrospira)谱系Ⅱ的NOB被证实含有AOM的特征功能酶,包括氨单加氧酶(AMO)和羟胺脱氢酶(HAO),并证明NOB同时具有氨氧化和亚硝酸盐氧化的能力,命名为全程氨氧化微生物(Complete ammonia oxidizer,Comammox)。根据AMO的α亚基基因amoA的相似性将Comammox分为两大分支clade A和clade B。它们广泛分布于自然环境和人工系统,包括土壤(稻田、森林)、淡水(湿地、河流、湖泊沉积物、蓄水层)、污水处理厂和自来水厂等。本文综述了Comammox的发现及其最新的研究进展,并展望了Comammox作为氮循环关键功能菌群的研究方向和应用前景。     

Effective and robust partial nitrification to nitrite by real-time aeration duration control in an SBR treating domestic wastewater

Achieving sustainable partial nitrification to nitrite has been proven difficult in treating low strength nitrogenous wastewater. Real-time aeration duration control was used to achieve efficient partial nitrification to nitrite in a sequencing batch reactor (SBR) to treat low strength domestic wastewater. Above 90% nitrite accumulation ratio was maintained for long-term operation at normal condition, or even lower water temperature in winter. Partial nitrification established by controlling aeration duration showed good performance and robustness even though encountering long-term extended aeration and starvation period. Process control enhanced the successful accumulation of ammonia oxidizing bacteria (AOB) and washout of nitrite oxidizing bacteria (NOB). Scanning electron microscope observations indicated that the microbial morphology showed a shift towards small rod-shaped clusters. Fluorescence in situ hybridization (FISH) results demonstrated AOB were the dominant nitrifying bacteria, up to 8.3 ± 1.1% of the total bacteria; on the contrary, the density of NOB decreased to be negligible after 135 days operation since adopting process control.     

pH对5种臂尾轮虫后代存活率和混交雌体率及种群内禀增长率的影响

水生生态系统中,pH是重要的非生物因子之一.pH可以通过影响水生生物的存活和繁殖,进而影响其在水体中的分布和多度,最终决定水生群落的物种组成和丰富度[1].     

Optimization of free ammonia concentration for nitrite accumulation in shortcut biological nitrogen removal process

A shortcut biological nitrogen removal (SBNR) utilizes the concept of a direct conversion of ammonium to nitrite and then to nitrogen gas. A successful SBNR requires accumulation of nitrite in the system and inhibition of the activity of nitrite oxidizers. A high concentration of free ammonia (FA) inhibits nitrite oxidizers, but unfortunately decreases the ammonium removal rate as well. Therefore, the optimal range of FA concentration is necessary not only to stabilize nitrite accumulation but also to achieve maximum ammonium removal. In order to derive such optimal FA concentrations, the specific substrate utilization rates of ammonium and nitrite oxidizers were measured. The optimal FA concentration range appeared to be 5–10 mg/L for the adapted sludge. The simulated results from the modified inhibition model expressed by FA and ammonium/nitrite concentrations were shown very similar to the experimental results.     

无机氮对土壤甲烷氧化作用的影响

无机氮输入(施氮肥和大气N沉降)对土壤CH4氧化作用的影响取决于甲烷氧化菌类型、输N种类和量以及土壤状况．这种作用既有抑制作用,又有刺激作用,但文献报道的抑制作用多于刺激作用,NH4^ 对CH4氧化的抑制作用多于NO3^-．随着全球N输入的增加,应在广泛的土壤类型和气候带观测和评价无机氯对土壤CH4氧化作用的影响．无机氮对土壤CH4氧化的抑制作用表现为立即或直接抑制、延迟抑制以及缺乏抑制等多种模式．尽管目前—些学者用酶基质竞争、增高的阈值、盐作用和离子交换、N转化率和N浓度等来解释抑制现象,但抑制机理依旧不完全清楚．因此,抑制机理是本领域未来研究的主要目标之一．     

Effect of nitrate and nitrite on sulfide production by two thermophilic,sulfate-reducing enrichments from an oil field in the North Sea

Thermophilic sulfate-reducing bacteria (tSRB) can be major contributors to the production of H₂S (souring) in oil reservoirs. Two tSRB enrichments from a North Sea oil field, NS-tSRB1 and NS-tSRB2, were obtained at 58°C with acetate-propionate-butyrate and with lactate as the electron donor, respectively. Analysis by rDNA sequencing indicated the presence of Thermodesulforhabdus norvegicus in NS-tSRB1 and of Archaeoglobus fulgidus in NS-tSRB2. Nitrate (10 mM) had no effect on H₂S production by mid-log phase cultures of NS-tSRB1 and NS-tSRB2, whereas nitrite (0.25 mM or higher) inhibited sulfate reduction. NS-tSRB1 did not recover from inhibition, whereas sulfate reduction activity of NS-tSRB2 recovered after 500 h. Nitrite was also effective in souring inhibition and H₂S removal in upflow bioreactors, whereas nitrate was similarly ineffective. Hence, nitrite may be preferable for souring prevention in some high-temperature oil fields because it reacts directly with sulfide and provides long-lasting inhibition of sulfate reduction.     

Effect of salicylic acid on nitrite reductase and glutamate dehydrogenase activities in maize roots

The effects of 0.01 to 5 m M salicyclic acid on the increase in nitrite reductase or glutamate dehydrogenase activities in maize roots by nitrate or ammonium respectively, were examined. Nitrite reductase activity was inhibited by the highest concentration of the acid. The activity of NADH-glutamate dehydrogenase was stimulated slightly (but consistently) by the lowest concentration and was inhibited by higher concentrations. Total protein content was also inhibited at high concentrations. When the crude enzyme extract was stored at 25°C in light, the glutamate dehydrogenase activity in the control decreased after 4 h of incubation. Low concentrations of the acid had no effect on this decrease but higher concentration accelerated the process. The divalent cations Caz²⁺, Mn²⁺, Mg²⁺ and Zn²⁺ protected against loss of enzyme activity during storage, both in the absence and presence of the acid. The inhibitory effect of 5 m M salicylic acid on glutamate dehydrogenase activity is apparent due to interference with the activity of the enzyme rather than with its synthesis.     

Effect of varying salinity, temperature, ammonia and nitrous acid concentrations on nitrification of saline wastewater in fixed-bed reactors

Nitrification under changing salinities (0-9%), temperatures (6-50 °C), ammonia (0-5 g N L⁻¹) and nitrite concentrations (0-0.4 g N L⁻¹) was investigated in fixed-bed reactors. For all conditions ammonia oxidation rates (AOR) were lower than nitrite oxidation rates (NOR). AORs and NORs increased from 12.5 to 40 °C and were very low at 6 °C and almost zero at 50 °C. No recovery of nitrification was obtained after incubation at 50 °C, whereas nitrification was restorable after incubation at 6 °C. Ammonia concentrations of 5 g N L⁻¹ or nitrite concentrations up to 0.125 g N L⁻¹ decreased AOR to almost zero. AORs and NORs recovered if ammonia or nitrite was removed. At concentrations of 1 and 5 g N L⁻¹ ammonia AOR and NOR were inhibited by 50%, whereas 27 mg N/L nitrite inhibited AOR by 50%.     

Importance of the operating pH in maintaining the stability of anoxic ammonium oxidation (anammox) activity in moving bed biofilm reactors

Two bench-scale parallel moving bed biofilm reactors (MBBR) were operated to assess pH-associated anammox activity changes during long term treatment of anaerobically digested sludge centrate pre-treated in a suspended growth partial nitrification reactor. The pH was maintained at 6.5 in reactor R1, while it was allowed to vary naturally between 7.5 and 8.1 in reactor R2. At high nitrogen loads reactor R2 had a 61% lower volumetric specific nitrogen removal rate than reactor R1. The low pH and the associated low free ammonia (FA) concentrations were found to be critical to stable anammox activity in the MBBR. Nitrite enhanced the nitrogen removal rate in the conditions of low pH, all the way up to the investigated level of 50 mg NO₂-N/L. At low FA levels nitrite concentrations up to 250 mg NO₂-N/L did not cause inactivation of anammox consortia over a 2-days exposure time.     

Modelling of inhibition of nitrite oxidation in biological nitritation processes by free ammonia

To understand nitrite build-up in the nitritation process, the inhibition kinetics of free ammonia (FA) on nitrite oxidation were investigated. FA inhibition on nitrite oxidation was the key factor of nitrite build-up and its type was mixed inhibition. Therefore, the mixed inhibition model should be adapted to a Michaelis–Menten equation to set up a process rate equation for nitrite oxidation in the nitritation process.     

Effect of pH on domoic acid toxicity in mice

Domoic acid is a shellfish toxin which produces gastrointestinal distress, followed by neurological symptoms such as headache, confusion, disorientation and severe deficits in short-term memory. Domoic acid is an amino acid which contains three carboxylic groups, and one imino group, and its solubility, rate of absorption, and elimination would vary depending on the protonation of these groups at different pH's. We propose that domoic acid toxicity varies with pH of administered domoic acid solution. Domoic acid toxicity was measured in mice as the onset times for scratching behaviour, seizure activity, and death, after the intraperitoneal administration of domoic acid at different pH's. Results of the present study show that the scratching behaviour, seizure activity, and death, occurred at 12, 40, and 55 min, after intraperitoneal administration of domoic acid at pH 3.7. Apparently, the onset times for three types of behaviours were relatively long, and well separated from each ot her. Domoic acid toxicity was lowest at pH 3.7, and highest at pH 7.4, with intermediate toxicity at other pH's. The onset time of scratching behaviour was not influenced by pH of domoic acid solution at three different doses. In contrast, the onset times for seizure activity, and death were significantly affected by pH of domoic acid, toxicity being higher at pH 7.4 than at pH 3.7. The pH effect on domoic acid toxicity diminished as the dose of domoic acid was increased. In fact, at 14.5 mg/kg domoic acid toxicity was similar at both pH's of 3.7 and 7.4. It is concluded that in vivo toxicity of domoic acid varies depending on pH of the administered solution. The differential toxicity of domoic acid at different pH may be related to its solubility, rate of absorption, and elimination, depending on the degree of protonation of domoic acid molecule. Domoic acid toxicity would also vary depending on the age of animal, receptor sensitivity and density in different regions of brain. (Mol Ce ll Biochem 167: 179-185, 1997)     

温度、pH对圆背角无齿蚌滤水率的影响

圆背角无齿蚌（Anodonta woodianapa cifica）是我国重要的经济贝类,广泛分布于各大水系;营底栖生活,通过滤水作用摄食水中的浮游生物和有机碎屑;其肉多壳薄,是优良的动物饵料;在水体生态系统的物质循环和能量流动及水质净化中起着十分重要的作用。     

Effects of medium glucose concentration and pH on docosahexaenoic acid content of heterotrophic Crypthecodinium cohnii

The effects of medium glucose concentration and pH on growth and docosahexaenoic acid (DHA, C22:6 ω-3) content of Crypthecodinium cohnii were investigated. Over a range of glucose concentrations (5–40 g l⁻¹) investigated, the highest specific growth rate (0.12 h⁻¹), highest cell dry weight concentration (3.13 g l⁻¹) and highest growth yield on glucose (0.6 g g⁻¹) were obtained at 20 g l⁻¹ glucose. However, the highest degree of fatty acid unsaturation (3.2) and highest DHA proportion (53.4% of total fatty acids) were achieved at 5 g l⁻¹ glucose. Low glucose concentrations enhanced the degree of fatty acid unsaturation and DHA formation. Medium pH also affected cell growth, fatty acid unsaturation and DHA proportion. When medium pH was 7.2, the highest specific growth rate (0.089 h⁻¹), highest cell dry weight concentration (2.73 g l⁻¹), highest growth yield on glucose (0.564 g g⁻¹), highest degree of fatty acid unsaturation (3.4) and highest DHA proportion (56.8% of total fatty acids) were obtained. Results suggest that glucose concentration and pH value could be effectively manipulated to achieve maximum DHA production by C. cohnii.     

Effect of ATP concentration and pH on rigor tension development and dissociation of rigor complex in glycerinated rabbit psoas muscle fiber

Isometric rigor tension development of glycerinated rabbit psoas muscle fibers in a medium, due to the formation of rigor complexes, was estimated at varying ATP concentrations from 0 to 2.5 mM and pH values from 6.75 to 8.20. The dissociation of rigor complexes was also estimated under the same conditions. When muscle fibers developed rigor tension from the relaxed and rigor states, the magnitude of rigor tension increased with increasing concentration of ATP. Transition between rigor and relaxation in single fibers occurred discontinuously at constant levels (critical levels) of ATP which were determined by pH. The critical concentrations of ATP necessary for inducing the transitions between rigor and relaxed states were also increased exponentially with increased pH. Incomplete repetition of tension development by the same fiber was also observed. This incomplete reversibility was divided into two types: one which showed a decay in rigor tension and another which showed no decay. The reason for the incomplete reversibility was discussed.     

Influence of pH and micellar systems on the sensitized photo‐oxidation of bovine serum albumin

Photosensitized oxidation of bovine serum albumin (BSA), by using perinaphtenone as a sensitizer, has been studied at pH 7.4 and 11. The selected sensitizer does not present ground‐state complexation with BSA and ensures that the mechanism is mediated by O₂(¹△_g). Strong dependence between BSA–O₂(¹△_g) photo‐oxidation and the pH of the medium has been found. The relative oxygen uptake rate (v_? △ O2 ) and the total quenching rate constant (k_t) values are higher at pH 11 than pH 7.4. The enhancement in the alkaline condition is due to conformational changes in the protein and the reactivity of tyrosinate anion with O₂(¹△_g). Even when the tendency with the pH in the presence of sodium dodecyl sulfate (SDS) micelles is similar to that observed in homogeneous media, an increment on the k_t value is detected. This effect may be attributable to the strong interaction of BSA–SDS, which leads to the protein unfolding and could leave more exposed photo‐oxidizable amino acids. A protective effect against the O₂(¹△_g)‐mediated photo‐oxidation was observed in reverse micelles (RMs) of sodium bis(2‐ethylhexyl)sulfosuccinate (AOT) by comparing the k_t values obtained at W = 10 with respect to the one obtain in homogeneous media. The latter could be mainly explained by the modification in the solvent polarity. Also, another important observation was found, the internal pH inside RMs of AOT sensed through tyrosine absorption was independent of the one used for the formation of the water pool. Hence, the k_t values observed at both pH, are quite similar.     

一株嗜盐嗜碱硫氧化菌的筛选、鉴定及硫氧化特性

【背景】沼气和天然气等清洁能源中往往会含有一定量的硫化氢,硫化氢的存在不仅污染环境,而且对人类危害很大。【目的】以硫代硫酸钠为唯一硫源从巴丹吉林沙漠盐碱湖岸边沉积物中分离筛选得到一株硫氧化菌BDL05,并研究其硫氧化特性。【方法】通过形态观察、生理生化特征及16S rRNA基因序列分析对硫氧化菌BDL05进行鉴定。【结果】菌株BDL05为革兰氏阴性菌,弧状,其16S rRNA基因序列与Thiomicrospira microaerophila ASL 8-2的相似性达99.8%,将其命名为Thiomicrospira microaerophila BDL05。该菌氧化硫代硫酸盐的最适pH为9.3,最适总钠盐浓度为0.8mol/L,在以硫化钠为硫源的气升式反应器中单质硫的生成率为94.7%,生成速率为3.0 mmol/(L·h)。【结论】菌株Thiomicrospira microaerophila BDL05为嗜盐嗜碱硫氧化菌,其耐盐耐碱性较强,比生长速率快,硫化钠氧化能力较强,是一株在气体生物脱硫方面具有应用价值的菌株。