好氧反硝化细菌碳氮代谢特点及途径的研究进展

好氧反硝化作用的发现打破了反硝化只能在严格厌氧条件下进行的传统认知,为生物脱氮提供了一条新的途径,已成为近些年的研究热点。碳源可为好氧反硝化过程提供能量和电子供体,其代谢难易程度直接影响着好氧反硝化细菌的脱氮效率,因此有必要明确碳源在好氧反硝化脱氮过程中的代谢机理。基于此,本文阐述了好氧反硝化细菌的种类及其对硝态氮与亚硝态氮的代谢途径;系统分析了不同好氧反硝化细菌对碳氮源代谢的差异与代谢机理;综合分析了碳代谢差异对好氧反硝化脱氮过程的影响,并对未来的研究方向进行了展望,旨在深入理解好氧反硝化细菌同时去除碳氮的机理,为提高废水生物脱氮除碳效率提供理论依据。     

好氧反硝化细菌的筛选及反硝化特性研究

目的：筛选好氧反硝化细菌,减少水体的亚硝态氮污染。方法：通过BTB平板初筛及反硝化培养基复筛得到目的菌,并探讨了不同溶解氧浓度、自然水体环境对该菌株反硝化作用的影响。结果：分离到1株高效的好氧反硝化细菌A1,该菌的反硝化作用主要发生在菌体的对数生长期,在溶解氧浓度为5mg/L时,对亚硝酸盐氮的降解率达到99％,在自然水体环境中当碳氮摩尔比为10：1时,对亚硝酸盐氮降解率达99％。结论：筛选到一株高效好氧反硝化细菌A1,将其应用于治理养殖水体的亚硝态氮污染有广阔的前景。     

一株脱氮细菌鉴定及其效果研究

针对传统人工湿地治理污染河湖水工艺中存在的脱氮效率低、环境安全性差等问题，从污水处理厂生化池污泥中分离纯化获得菌株TY（CGMCC1.18865，该菌种已于2020年在中国普通微生物菌种保藏中心入库），通过配水试验，验证菌株脱氮特性，并进一步以火山岩为填料，采用湿地模拟系统装置对比附配TY菌的湿地系统与传统湿地系统对污染水体的脱氮效果的差异；通过16S rRNA基因序列分析，鉴定获得的菌株TY，同时对其表现出的反硝化作用进行分析研究。结果表明：分离获得的TY菌株为假单胞菌属（Pseudomonas sp.），配水试验中对氨氮和总氮的去除率分别为84.2%和93.6%。通过对模拟湿地系统出水氨氮、总氮和化学需氧量（COD）监测，发现30 d平均去除率分别可提升至94.3%、88.0%和82.3%，较传统湿地分别提高27.9%、28.8%和7.3%。研究表明，TY菌脱氮过程中有少量的硝态氮和亚硝态氮的积累，可降低对环境的不利影响、有利于脱氮反应连续进行，从而提高人工湿地脱氮效率，具有进一步探究的前景。     

新型异养硝化-好氧反硝化菌Paracoccus sp. QD-19的分离及脱氮特性研究

从沈阳市南部污水处理厂活性污泥中分离获得同时具备异养硝化和好氧反硝化能力的新型菌株,研究其脱氮特性,为改善污水厂的脱氮处理工艺奠定基础。对菌株进行形态学观察和16S rRNA基因鉴定;分别以NH₄Cl、NaNO₂、KNO₃为唯一氮源探究菌株的脱氮能力;以碳源、C/N比、pH值、温度、转速、接种量(V∶V)等因素对菌株脱氮效果的影响进行研究。获得一株新型异养硝化-好氧反硝化菌株,经16S rRNA基因序列比对为副球菌属(Paracoccus),命名为Paracoccus sp. QD-19。菌株对初始氨氮浓度在300 mg/L以下的低浓度氨氮去除率能够达到100%,去除速率为8.707 mg/(L·h)且在脱氮过程中几乎没有亚硝态氮和硝态氮的积累。以亚硝态氮和硝态氮作为唯一氮源时,对此两种氮源的去除率36 h内均能达到99%,去除速率分别为4.944和5.666mg/(L·h)。确定了去除氨氮的最佳脱氮条件：琥珀酸钠为碳源,C/N比为10,pH值为7,接种量(V:V)为1%,温度为30℃,转速为140 r/min。菌株Pa...     

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。     

深海沉积物高效脱氮菌筛选分离及其脱氮特性研究

【背景】深海海域具有高压、低温、无光等环境条件,蕴含着丰富而独特的微生物资源。【目的】从深海沉积物中定向分离、筛选脱氮效率高的好氧脱氮菌株资源,并揭示其脱氮特性,为开发水体脱氮微生物技术提供物质基础。【方法】以东太平洋、南大西洋、西南印度洋共10个站位的深海沉积物为研究材料,在28°C下使用无机氮源连续进行两轮富集培养,然后定性筛选可以脱除氨氮、亚硝态氮和硝态氮的菌株,并通过形态学和16S rRNA基因序列分析进行初步分类鉴定;对优选得到的功能菌株,分别采用以氨氮、亚硝态氮、硝态氮为唯一氮源的培养基定量研究其生长和脱氮性能。【结果】从10份大洋深海沉积物样品中共分离得到49株好氧反硝化菌,其中3株在有氧条件下反硝化效率较高,分别命名为Pseudomonassp.G111、Pseudomonassp.G112和Dietziamaris W023a,其中菌株G111和G112与模式菌株博岑假单胞菌Pseudomonas bauzanensis BZ93T的16S rRNA基因序列相似度为99.2%,菌株W023a与模式菌株海洋迪茨氏菌DietziamarisATCC35013T的16SrRNA基因序列相似度为99.9%。菌株G111、G112和W023a培养48h后,对氨氮的脱除率分别为98.0%、85.2%和97.6%;对亚硝态氮的脱除率分别为71.9%、67.5%和34.7%;对硝态氮的脱除率分别为66.0%、52.6%和56.3%。菌株G111、G112和W023a均为异养硝化-好氧反硝化菌,可通过好氧反硝化作用将亚硝态氮和硝态氮还原为含氮气体,也可通过异养硝化-好氧反硝化作用将氨氮转化为含氮气体。【结论】从深海沉积物中分离筛选得到3株高效好氧反硝化菌,所获得的菌株在水体净化、污水处理、生态系统修复等领域具有应用潜力。     

脱氮功能菌去除市政废水中氮素的研究

为了探讨实验室筛选获得的氨氧化细菌CM-NRO14和反硝化细菌CM-NRD3联合去除市政废水中氮素的应用价值,采用了两级A/O工艺进行菌株去除废水中氮素的小试实验,最后将菌株用于废水脱氮工程中。结果表明,脱氮功能菌实现了短程硝化-反硝化,氨氮去除率在98%以上,总氮去除率在75%以上, COD (化学需氧量)去除率大于90%,出水各项指标均低于城镇污水处理厂污染物排放一级(A)标准。脱氮功能菌在去除市政废水中氮素方面有很高的应用价值,可用于城镇污水处理厂提标改造等。     

异养硝化微球菌的分离及性能评价

利用亚硝化细菌分离培养基从湿地浅层底泥中分离到一株具有硝化反硝化能力的异养硝化细菌,经过常规的分析和部分长度的16S rDNA序列分析表明,分离所得HB-1菌株与多株Micrococcus sp.16S rDNA具有99%的相似性,初步鉴定为Micrococcus sp.。分别采用乙酸钠和氯化铵作为碳源和氮源进行硝化特性研究,结果表明,8d的好氧培养,铵态氮去除率为49.2%,并且具有产生亚硝态氮的硝化性能。以硝酸钠为氮源进行好氧培养,总氮没有去除效果;而6d的缺氧静置培养,总氮的去除率达90.7%,表现明显的反硝化作用。     

以亚硝氮为唯一氮源生长的海洋紫色硫细菌去除无机三态氮

【目的】揭示以亚硝氮为唯一氮源生长的海洋紫色硫细菌去除水体中无机三态氮的特征和规律。【方法】在光照厌氧环境下,以乙酸盐为唯一有机物,在分别以氨氮、亚硝态氮、硝态氮为唯一氮源和三氮共存的模拟水体中,采用Nessler’s试剂分光光度法、N-(1-萘基)-乙二胺分光光度法和紫外分光光度法分别测定水体中氨氮、亚硝态氮和硝态氮的含量,比浊法测定菌体生物量。【结果】随着时间的延长,海洋紫色硫细菌Marichromatium gracile YL28分别在氨氮、亚硝态氮和硝态氮为唯一氮源的水体中对三氮的去除量增加,生物量增大,水体pH升高,并逐渐趋于平衡;YL28对氨氮的最大去除量和最大耐受浓度分别为9.64 mmol/L和36.64 mmol/L,当氨氮浓度低于3.21 mmol/L时,去除率可达97.61%以上;与氨氮相比,以亚硝态氮和硝态氮为唯一氮源,菌体的生长速率、生物量和水体最终pH较低,但对亚硝态氮和硝态氮的去除速率和去除量仍然很高,当亚硝态氮和硝态氮浓度分别达13.50 mmol/L和22.90 mmol/L时,YL28仍能够完全去除。在三氮共存的水体中,YL28也能良好的去除无机三态氮,对亚硝态氮和硝态氮去除能力更强。【结论】在模拟水体中,海洋紫色硫细菌YL28能够分别以氨氮、亚硝态氮和硝态氮为唯一氮源生长,具有良好的耐受和去除无机三态氮的能力,尤其对亚硝态氮具有良好的去除能力。本研究为进一步开发高效脱氮,尤其是去除亚硝态氮的不产氧光合细菌水质调节剂奠定了基础,也为微生物制剂的合理应用提供参考。     

模拟人工湿地中植物多样性配置对硝态氮去除的影响

为检验植物多样性对人工湿地脱氮功能的影响,在模拟人工湿地试验系统中设置了植物单种和混种处理并定期供给氮形态仅为硝态氮的模拟污水。结果表明:混种系统的出水硝态氮浓度显著低于单种(P<0.05);混种与单种系统在基质氮含量和植物氮积累量上无统计差异;质量平衡分析表明混种促进系统反硝化强度;菩提子单种系统中的硝态氮移除能力显著高于香蒲、芦苇和菖蒲单种系统,后3种硝态氮移除能力则无显著差异。本研究可为人工湿地选择高效物种、多样性配置以提高氮去除率提供依据。     

Distinctive microbial ecology and biokinetics of autotrophic ammonia and nitrite oxidation in a partial nitrification bioreactor

Biological nitrogen removal (BNR) based on partial nitrification and denitrification via nitrite is a cost-effective alternate to conventional nitrification and denitrification (via nitrate). The goal of this study was to investigate the microbial ecology, biokinetics, and stability of partial nitrification. Stable long-term partial nitrification resulting in 82.1 +/- 17.2% ammonia oxidation, primarily to nitrite (77.3 +/- 19.5% of the ammonia oxidized) was achieved in a lab-scale bioreactor by operation at a pH, dissolved oxygen and solids retention time of 7.5 +/- 0.1, 1.54 +/- 0.87 mg O(2)/L, and 3.0 days, respectively. Bioreactor ammonia oxidizing bacteria (AOB) and nitrite oxidizing bacteria (NOB) populations were most closely related to Nitrosomonas europaea and Nitrobacter spp., respectively. The AOB population fraction varied in the range 61 +/- 45% and was much higher than the NOB fraction, 0.71 +/- 1.1%. Using direct measures of bacterial concentrations in conjunction with independent activity measures and mass balances, the maximum specific growth rate (micro(max)), specific decay (b) and observed biomass yield coefficients (Y(obs)) for AOB were 1.08 +/- 1.03 day(-1), 0.32 +/- 0.34 day(-1), and 0.15 +/- 0.06 mg biomass COD/mg N oxidized, respectively. Corresponding micro(max), b, and Y(obs) values for NOB were 2.6 +/- 2.05 day(-1), 1.7 +/- 1.9 day(-1), and 0.04 +/- 0.02 mg biomass COD/mg N oxidized, respectively. The results of this study demonstrate that the highly selective partial nitrification operating conditions enriched for a narrow diversity of rapidly growing AOB and NOB populations unlike conventional BNR reactors, which host a broader diversity of nitrifying bacteria. Further, direct measures of microbial abundance enabled not only elucidation of mixed community microbial ecology but also estimation of key engineering parameters describing bioreactor systems supporting these communities.     

Effects of vegetation,limestone and aeration on nitritation,anammox and denitrification in wetland treatment systems

Integration of partial nitrification (nitritation) and anaerobic ammonium oxidation (anammox) in constructed wetlands creates a sustainable design for nitrogen removal. Three wetland treatment systems were operated with synthetic wastewater (60 mg NH₃–N L^?1) in a batch mode of fill – 1-week reaction – drain. Each treatment system had a surface flow wetland (unplanted, planted, and planted plus aerated, respectively) with a rooting substrate of sandy loam and limestone pellets, followed by an unplanted subsurface flow wetland. Meanwhile, three surface flow wetlands with a substrate of sandy loam and pavestone were operated in parallel to the former surface flow wetlands. Influent and effluent were monitored weekly for five cycles. Aeration reduced nitrogen removal due to hindered nitrate reduction. Vegetation maintained pH near neutral and moderate dissolved oxygen, significantly improved ammonia removal by anammox, and had higher TN removal due to coexistence of anammox and denitrification in anaerobic biofilm layers. Nitrite production was at a peak at the residence time of 4–5 d. Relative to pavestone, limestone increased the nitrite mass production peak by 97%. The subsurface flow wetlands removed nitrogen via nitritation and anammox, having an anammox activity of up to 2.4 g N m^?3 d^?1 over a startup operation of two months.     

Analysis of factors affecting the performance of partial nitrification in a sequencing batch reactor

The objective of this study was to analyze the factors affecting the performance of partial nitrification in a sequencing batch reactor. During a 140-day long-term operation, influent pH value, dissolved oxygen (DO), and chemical oxygen demand/nitrogen (COD/N) ratio were selected as operating factors to evaluate the maintenance and recovery of nitrite accumulation. Results showed that high DO concentration (2–4 mg/L) could damage nitrite accumulation immediately. However, nitrite accumulation ratio (NAR) could be increased from 1.68?±?1.51 to 35.46?±?7.86 % when increasing the pH values from 7.5 to 8.3 due to the increased free ammonia concentration. Afterwards, stable partial nitrification and high NAR could be recovered when the reactor operated under low DO concentration (0.5–1.0 mg/L). However, it required a long time to recover the partial nitrification of the reactor when the influent COD/N ratios were altered. Fluorescence in situ hybridization analysis implied that ammonium oxidizing bacteria were completely recovered to the dominant nitrifying bacteria in the system. Meanwhile, sludge volumetric index of the reactor gradually decreased from 115.6 to 56.6 mL/g, while the mean diameter of sludge improved from74.57 to 428.8 μm by using the strategy of reducing settling time. The obtained results could provide useful information between the operational conditions and the performance of partial nitrification when treating nitrogen-rich industrial wastewater.     

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。     

Changes in Diversity and Functional Gene Abundances of Microbial Communities Involved in Nitrogen Fixation, Nitrification, and Denitrification in a Tidal Wetland versus Paddy Soils Cultivated for Different Time Periods

In many areas of China, tidal wetlands have been converted into agricultural land for rice cultivation. However, the consequences of land use changes for soil microbial communities are poorly understood. Therefore, we investigated bacterial and archaeal communities involved in inorganic nitrogen turnover (nitrogen fixation, nitrification, and denitrification) based on abundances and relative species richness of the corresponding functional genes along a soil chronosequence ranging between 50 and 2,000 years of paddy soil management compared to findings for a tidal wetland. Changes in abundance and diversity of the functional groups could be observed, reflecting the different chemical and physical properties of the soils, which changed in terms of soil development. The tidal wetland was characterized by a low microbial biomass and relatively high abundances of ammonia-oxidizing microbes. Conversion of the tidal wetlands into paddy soils was followed by a significant increase in microbial biomass. Fifty years of paddy management resulted in a higher abundance of nitrogen-fixing microbes than was found in the tidal wetland, whereas dominant genes of nitrification and denitrification in the paddy soils showed no differences. With ongoing rice cultivation, copy numbers of archaeal ammonia oxidizers did not change, while that of their bacterial counterparts declined. The nirK gene, coding for nitrite reductase, increased with rice cultivation time and dominated its functionally redundant counterpart, nirS, at all sites under investigation. Relative species richness showed significant differences between all soils with the exception of the archaeal ammonia oxidizers in the paddy soils cultivated for 100 and 300 years. In general, changes in diversity patterns were more pronounced than those in functional gene abundances.     

人工湿地的氮去除机理

湖泊等水环境的富营养化给人类带来诸多损害,如环境、生态和经济等方面的损害。富营养化的原因和控制途径引起了包括中国在内的很多国家的关注。我国针对水环境的富营养化问题开展了大量的工作。氮是引发水环境富营养化的主要营养物之一。外源氮负荷（分点源和非点源两部分）是水环境污染负荷的重要组成部分。传统污水处理技术应用于收集系统欠缺的非点源污染的治理时成本过高。人工湿地是有效削减水环境中外源氮负荷的重要技术手段,在处理非点源污染源带来的氮负荷时更是如此。人工湿地具有氮去除效果好、耐冲击负荷能力强、投资低和生态环境友好等优点。因此人工湿地非常适合于水环境富营养化的防治。阐明人工湿地中氮的去除机理对水环境的富营养化等具有重要的意义。防渗人工湿地的氮去除机理主要包括挥发、氨化、硝化／反硝化、植物摄取和基质吸附。未防渗的人工湿地中,周围水体与人工湿地的氮交换影响着人工湿地中氮的去除。一般情况下,人工湿地中硝化／反硝化是最主要的氮去除机理。pH值小于7．5时,氨挥发可忽略。pH值在9．3以上时,氨挥发很显著。处理生活污水的人工湿地中氮的去除主要是依靠微生物的硝化／反硝化作用。在进水负荷低、气候适宜、植物物种适宜和收割频率与时机适宜的条件下,植物收割可能成为主要的去氮途径。人工合理导向的湿地的氮去除效果通常优于天然湿地。合理的设计（填料的搭配、植物物种的配置以及布水和集水的优化）对人工湿地系统中氮去除的改善有重要影响。合理的运行,如有效的水位控制,正确的植物培育、合理的植物收割等,能有效地改善湿地中的氮去除。     

Analysis of the characteristics of short-cut nitrifying granular sludge and pollutant removal processes in a sequencing batch reactor

Aerobic granular sludge is a new type of microbe auto-immobilization technology; in this paper, short-cut nitrification and denitrification were effectively combined with the granular sludge technology. Simultaneous nitrification and denitrification granules were developed in a sequencing batch reactor (SBR) using synthetic wastewater with a high concentration of ammonia nitrogen at 25 °C with a dissolved oxygen concentration above 2.0 mg/L and a 15 days sludge retention time. The characteristics of the sludge and the removal efficiency were studied, and the removal mechanisms of the pollutants and the process of short-cut nitrification were analyzed. The average granule diameter of the granular sludge was 704.0 μm. The removal rates of pollutants and the accumulation rate of nitrite in the SBR were studied. During treatment of wastewater with a high concentration of ammonia nitrogen, simultaneous nitrification, and denitrification and the stripping process could contribute to the removal of total nitrogen. The high pH value, the high concentration of free ammonia, and the delamination of granular sludge were the main factors contributing to the short-cut nitrification property of granular sludge in the reaction process.     

几种湿地植物净化生活污水COD、总氮效果比较

以无植被、基质为河砂的潜流型人工湿地为对照,研究了石菖蒲、灯心草和蝴蝶花3种类型植被、基质均为河砂的潜流型人工湿地净化生活污水COD、总氮的效果．结果表明,在污水COD浓度小于200mg·L^-1、总氮浓度小于30mg·L^-1的低浓度范围里,无植被的人工湿地和有植被的人工湿地对污水中COD、总氮均有很好的去除效果,两者差异不大,其COD去除率均达90％以上,总氮的去除率达80％以上．随着污水中COD和总氮浓度的增加,无植被人工湿地和有植被人工湿地去除COD和总氮的效果均有不同程度下降,两者差异明显,有植被的人工湿地能维持较高的COD、总氮的去除效果,无植被的人工湿地COD和总氮去除效果下降很快,植被在人工湿地系统去除污水COD和总氮过程中起着重要的作用．在整个试验阶段,石菖蒲植被人工湿地COD和总氮平均净化效率分别为80．46％和77．77％、灯心草人工湿地分别为75．53％和71．17％、蝴蝶花人工湿地分别为70．50％和66．38％,无植被人工湿地分别为61．39％和55．81％．同无植被人工湿地COD和总氮净化效果相比,石菖蒲植被人工湿地净化效果最好;其次为灯心草植被人工湿地,再次为蝴蝶花植被人工湿地．不同类型植被的人工湿地净化污水中COD和总氮的效果与其生物量关系密切,这与植被系统吸收同化有机物质和总氮数量、根际微生物分解有机物质和硝化-反硝化作用有关。     

Soil amendments promote denitrification in restored wetlands

Wetlands perform important ecosystem functions, including improvement of water quality through the process of denitrification. To offset the negative environmental impact of replacing wetlands with agriculture and development, the United States has a policy requiring that losses in wetland area are compensated for through wetland restoration elsewhere. However, these restored wetlands may require decades to achieve functional equivalency to natural wetlands. We evaluated the efficacy of using carbon amendments during restoration to promote denitrification potential in four restored wetlands in central New York State, United States. The amendments were straw, topsoil, and biochar, chosen to range along a gradient of carbon lability. Soil samples collected 6 years after restoration were analyzed for denitrification potential and associated soil properties, including soil carbon and nitrogen, pH, microbial biomass carbon and nitrogen, carbon lability, and potential net nitrogen mineralization and nitrification. Compared to unamended control plots, denitrification potential was approximately 3 times higher in straw‐amended plots, 8 times higher in topsoil‐amended plots, and 11 times higher in biochar‐amended plots. Denitrification potential positively correlated with both soil organic carbon and microbial biomass nitrogen, suggesting that the use of soil amendments in restorations can help stimulate the development of denitrification potential by facilitating the suite of carbon and nitrogen cycling processes that underlie this function. However, denitrification potential in a nearby natural reference wetland was at least 50 times higher than it was in the restored wetland plots, highlighting the limitations of using wetland restoration to compensate for the loss of natural wetlands.