黄河三角洲湿地硝化作用强度及影响因素研究

采用现场调查和实验室培养的方法,研究了黄河三角洲滨海湿地不同环境氨氧化细菌分布、硝化强度及影响因素,结果表明:5个不同生态环境中氨氧化细菌数量和硝化强度差别较大。氨氮浓度、盐度、温度、pH对硝化强度均有明显影响。硝化强度随着氨氮浓度的增加而增大;低盐度条件下,含盐量的变化对土壤的硝化强度影响不明显,高盐度条件下,硝化作用明显受到抑制;硝化作用最适宜的温度是28℃左右,低温限制硝化作用的进行;在偏碱性的环境中硝化强度较强,较低的pH严重抑制硝化作用。     

渤海和南黄海沉积物中氨氧化微生物对硝化潜势的相对贡献

硝化作用是海洋氮循环的核心过程。作为硝化过程关键步骤的氨氧化过程的主要参与者,氨氧化古菌和氨氧化细菌对硝化作用的相对贡献是海洋氮循环关注的热点问题之一。本文选取渤海和南黄海20个站位的表层沉积物,通过微宇宙培养实验研究了沉积物中氨氧化古菌和氨氧化细菌对硝化潜势的相对贡献。结果表明,渤海和南黄海海域表层沉积物中潜在硝化速率(以氮计,下同)为0.004 6～0.283 1μmol/(g·d),其中氨氧化古菌潜在硝化速率为0.004 3～0.274 3μmol/(g·d),氨氧化细菌潜在硝化速率为0.000 4～0.056 0μmol/(g·d)。氨氧化古菌是硝化潜势的主要贡献者,在渤海海域的贡献率为59.79%～97.95%,在南黄海海域的贡献率为18.47%～94.26%。渤海海域潜在硝化速率显著高于南黄海海域。此外,本研究海域中盐度是影响潜在硝化速率的关键环境因子,对渤海海域的分析则表明越高的NO₃^-浓度可能指示着越高的硝化潜势。在河口及近海沉积物中,氨氧化古菌在硝化过程中起着更加重要的作用;河口和近岸沉积物硝化潜势总体高于远海。本研究为进一...     

过硫酸氢钾复合物对对虾养殖底泥硝化作用、氨氧化微生物丰度和群落结构的影响

采用在模拟池塘中投放过硫酸氢钾复合物（KMPS）进行对比实验的方法,探究KMPS对养殖底质硝化作用的影响。通过对氨氮和亚硝态氮含量的检测,探究对不同时期氮素转化的影响,低频率高剂量投放组中的氨氮和亚硝态氮含量显著降低,而高频率低剂量组中氨氮和亚硝态氮的含量显著上升。高频率低剂量KMPS的投放使氨氧化古菌（AOA）和氨氧化细菌（AOB）丰度降低,而低频率高剂量KMPS的投放不会造成丰度降低,而且还表现出部分时期AOB丰度的上升。进一步对群落结构进行分析发现KMPS的投放使AOA群落中Nitrosopumilu属相对丰度降低,Nitrososphaera属的相对丰度上升,这种相对丰度的变化与KMPS的投放方式无关;但AOB群落受到KMPS投放方式的影响,低频率高剂量的KMPS投放下AOB群落优势属相对丰度显著提高。以上结果均说明低频率高剂量KMPS的投放起到了促进底质硝化作用的效果。同时,可为KMPS用于对虾养殖池塘底质改良开辟一个新的途径。     

辽河口芦苇湿地氨氧化菌群落结构与土壤盐分的关系

采用最大或然数(MPN)法和变性梯度凝胶电泳技术(DGGE)对辽河口芦苇湿地不同盐分土壤中氨氧化细菌(Ammonia-oxidizing Bacteria,AOB)的数量分布和群落变化进行了解析,旨在探究辽河口芦苇湿地土壤中AOB群落结构与土壤盐分的关系,为河口湿地的生物修复提供理论依据和技术支撑。结果表明:辽河口芦苇湿地土壤中盐分含量越高,AOB数量越少,表现为土壤盐分含量为10g/kg的S10组AOB数量大于土壤盐分含量为15g/kg的S15组AOB数量,大于土壤盐分含量为20g/kgS20;其中S10组AOB数量多于对照组,S15组AOB数量与对照组相当,S20组AOB数量小于对照组,即土壤盐分较低时,盐分的升高能促进AOB的增长,当土壤盐分超过一定范围时,盐分的增加则抑制AOB数量的增加;不同盐度胁迫下AOB菌群群落结差异很大,而聚类结果没有表现出很大的差异性;盐度对AOB的影响体现在低、中高、高盐度土壤中氨氧化菌的种类和群落结构显著不同;不同时期低盐度土壤中的AOB群落结构和优势种群均发生了变化;中高盐度土壤中的AOB群落结构相似,不同时期优势菌比例不同;高盐度土壤中AOB数量明显减小,种群多样性明显降低。     

过硫酸氢钾复合物对养殖底泥硝化作用、氨氧化微生物丰度和群落结构的影响

采用在模拟池塘中投放KMPS进行对比实验的方法,探究KMPS对养殖底质硝化作用的影响。通过对氨氮和亚硝态氮含量的检测,探究对不同时期氮素转化的影响,低频率高剂量投放组中的氨氮和亚硝态氮含量显著降低,而高频率低剂量组中氨氮和亚硝态氮的含量显著上升。高频率低剂量KMPS的投放使AOA和AOB丰度降低,而低频率高剂量KMPS的投放不会造成丰度降低,而且还表现出部分时期AOB丰度的上升。进一步对群落结构进行分析发现KMPS的投放使AOA群落中Nitrosopumilu属相对丰度降低,Nitrososphaera属的相对丰度上升,这种相对丰度的变化与KMPS的投放方式无关;但AOB群落受到KMPS投放方式的影响,低频率高剂量的KMPS投放下AOB群落优势属相对丰度显著提高。以上结果均说明低频率高剂量KMPS的投放起到了促进底质硝化作用的效果。同时,可为KMPS用于对虾养殖池塘底质改良开辟一个新的途径。     

海水反硝化和厌氧氨氧化速率同步测定的15N示踪法及其应用

反硝化与厌氧氨氧化作用是海洋结合态氮迁出的主要途径,15N示踪法是定量反硝化和厌氧氨氧化作用的新方法,它具有灵敏度高、可同步测量反硝化与厌氧氨氧化速率的优点.本研究开展了一系列方法学实验,以确定高纯氦气驱赶水样所含气体的时间、超声赶气时间、同位素比值质谱仪测量N2含量的精度、N2含量及其同位素组成测定的工作曲线以及示踪剂的最佳添加量等,从而确立起15N示踪同步测定海水反硝化和厌氧氨氧化速率的方法.所建立方法测量N2含量的相对标准偏差小于8%,可满足海洋反硝化与厌氧氨氧化速率测定的需要.应用该方法实测了厦门筼筜湖富氧与缺氧水体的反硝化与厌氧氨氧化速率,结果表明,缺氧底层水的反硝化速率[4.10～5.47μmol/（dm3·d）]比富氧表层水[0.43～0.46μmol/（dm3·d）]明显来得高,但缺氧底层水的厌氧氨氧化速率[0.00～0.12μmol/（dm3·d）]则比富氧表层水的速率[0.52～0.67μmol/（dm3·d）]低得多.     

海水反硝化和厌氧氨氧化速率同步测定的~(15)N示踪法及其应用

反硝化与厌氧氨氧化作用是海洋结合态氮迁出的主要途径,15N示踪法是定量反硝化和厌氧氨氧化作用的新方法,它具有灵敏度高、可同步测量反硝化与厌氧氨氧化速率的优点.本研究开展了一系列方法学实验,以确定高纯氦气驱赶水样所含气体的时间、超声赶气时间、同位素比值质谱仪测量N2含量的精度、N2含量及其同位素组成测定的工作曲线以及示踪剂的最佳添加量等,从而确立起15N示踪同步测定海水反硝化和厌氧氨氧化速率的方法.所建立方法测量N2含量的相对标准偏差小于8%,可满足海洋反硝化与厌氧氨氧化速率测定的需要.应用该方法实测了厦门筼筜湖富氧与缺氧水体的反硝化与厌氧氨氧化速率,结果表明,缺氧底层水的反硝化速率[4.10~5.47μmol/(dm3·d)]比富氧表层水[0.43~0.46μmol/(dm3·d)]明显来得高,但缺氧底层水的厌氧氨氧化速率[0.00~0.12μmol/(dm3·d)]则比富氧表层水的速率[0.52~0.67μmol/(dm3·d)]低得多.     

竹环填料生物滤器在两种海水鱼养殖废水处理中的运行效果及微生物群落分析

以点带石斑鱼(Epinephelus malabaricas)、半滑舌鳎(Cynoglossus semilaevis)养殖排放水为处理对象,在生产现场研究了竹环填料生物滤器处理高盐度、低氨氮负荷海水养殖废水的运行效果以及挂膜阶段和稳定运行阶段微生物群落变化规律。结果表明,点带石斑鱼养殖废水进水氨氮质量浓度为0.93～1.33 mg/L,氨氮去除率达到27%～39%,挂膜时间需45 d;半滑舌鳎养殖废水进水氨氮质量浓度为0.38～0.52 mg/L,氨氮去除率达到20%～30%,挂膜时间需65 d。另外,对生物滤器挂膜阶段和稳定运行阶段的氨氧化细菌数量和亚硝酸氧化细菌数量进行了统计,氨氧化细菌的数量在点带石斑鱼和半滑舌鳎养殖排放水处理系统中分别达到104～105 CFU/mL和103 CFU/mL,亚硝酸氧化细菌数量则分别达到104 CFU/mL和103 CFU/mL,并分析了温度、进水氨氮负荷、反硝化作用对半滑舌鳎养殖废水生物滤器亚硝酸盐积累现象的影响。     

湛江湾沉积物中反硝化和厌氧氨氧化细菌的丰度、多样性及分布特征

采用实时荧光定量PCR、高通量测序等方法对湛江湾沉积物中四个月份的反硝化细菌与厌氧氨氧化细菌的多样性和丰度进行了分析。结果表明:湛江湾沉积物中反硝化细菌和厌氧氨氧化细菌丰度在四个月份的变化和空间分布趋势为:nirS型反硝化细菌在二月份最高,四月份最低,且其平均丰度有从湛江湾湾内向湾口附近呈现先升高再降低的趋势;nirK型反硝化细菌丰度在九月份最高,十一月份最低;nosZ型反硝化细菌在四月份最高,其余月份变化不大;厌氧氨氧化细菌丰度在九月份最高,二月份最低。通过相关性分析结果表明,亚硝酸盐、铵盐等共同调控着湛江湾沉积物中反硝化和厌氧氨氧化细菌丰度变化。系统发育分析表明:湛江湾中存在着一些广泛分布的反硝化细菌,但也生活着一些新奇的nirK型和nosZ型反硝化细菌。对于厌氧氨氧化细菌而言,其主要属于浮霉菌门及Candidatus Scalindua属,具有较高的耐盐性,另外湛江湾海区的厌氧氨氧化细菌也生活着一类在其他地方没有的新分支。典范对应分析分析结果表明:硝酸盐显著影响湛江湾反硝化细菌和厌氧氨氧化细菌的群落结构。湛江湾沉积物中反硝化细菌和厌氧氨氧化细菌存在特殊的竞争与共存的关系,且由亚硝酸盐、硝酸盐、pH等多种环境因子共同驱动。     

超高盐度对耐盐活性污泥脱氮、微生物活性和群落结构的影响

采用能耐受3%盐度的活性污泥处理高盐度废水,探究了盐度升至4%～7%对污染物去除的影响,考察了微生物活性和群落结构随盐度升高的变化。结果表明,盐度提升到4%、5%和6%对COD、NH~+_4-N和总无机氮(TIN)去除几乎没有影响,而7%盐度时三者的去除率均明显下降;盐度提高到4%对污泥的氨氧化活性和亚硝酸盐氧化活性有刺激作用,使两者提高,而盐度提高到5%、6%和7%时氨氧化活性受到明显抑制,亚硝酸盐氧化活性在6%和7%盐度条件下明显降低;盐度提升使硝酸盐和亚硝酸盐还原活性均受到明显抑制;耗氧速率测试结果表明,盐度提升对自养硝化菌的负面影响较异养好氧菌更大。微生物群落结构随盐度升高发生了明显变化,微生物群落丰富度和多样性均在6%盐度时最大,3%盐度时Roseovarius为优势菌属,而盐度提高至4%～7%时Azoarcus成为优势菌属。氨氧化菌(AOB)只在3%、4%和6%盐度下被检出,亚硝酸盐氧化菌(NOB)在所有盐度下均未检出,短程硝化反硝化(PND)为主要脱氮途径;自养反硝化菌、好氧反硝化菌和厌氧氨氧化菌的存在说明脱氮途径不局限于传统的自养硝化-异养反硝化。盐度驯化提高了活性污泥的抗盐能力,使生物法处理超高盐废水成为可能。     

小清河口春季亚硝酸盐偏高问题探讨

2008年春季对莱州湾小清河口的走航调查发现,调查区域大部分水体的DOC,CODMn,NH4+-N,PO43--P高于4类海水的限定值,处于严重污染状态。水体CHla的浓度范围在1.02~37.92μg/L之间,但DO饱和度却在23%~96%之间,盐度小于15水体的DO饱和度低于50%,水体缺氧严重。河口存在NO2--N含量偏高现象。由河口向外,NO2--N浓度逐渐增加,至盐度24.4的水体达到0.27 mg/L的高值,NO2--N增加主要是NH4+-N的硝化反应产生的。NH4+-N生成NO2--N主要受DO和NH4+-N比例与浓度的影响,DO饱和度在30%~70%,NH4+-N大于3.33 mg/L的水体中NO2--N生成速率最快。     

长江口海域悬浮颗粒有机物的稳定氮同位素季节分布与关键生物地球化学过程

对2010年2、5、8、11月份长江口海域水体中的悬浮颗粒物进行稳定氮同位素分析。根据不同季节、不同区域内悬浮颗粒有机物的稳定氮同位素组成(δ15 Np)值的变化研究水体中氮的迁移、转化等生物地球化学过程,揭示其环境行为,从而对该海域的氮循环机制进行探索。研究发现:长江口海域悬浮颗粒物的稳定氮同位素组成具有较宽的分布范围,δ15 Np值分布范围为-1.1‰~8.6‰,具有明显的时空分布特点,反映了不同程度的陆源输入和氮的生物地球化学过程的影响。其中,2月份生物反应较弱,δ15 Np分布体现了陆源和海源的混合特征;5月和11月份上层水体δ15 Np随叶绿素a升高而降低,指示了生物的同化吸收作用;8月和11月δ15 Np和总溶解无机氮呈现极显著正相关关系,说明该海域发生了氮的矿化再生。     

Nitrogen isotopic discrimination by water column nitrification in a shallow coastal environment

Temporal changes in nitrogen isotopic composition (δ¹⁵N) of the NO₃ ⁻ pool in the water column below the pycnocline in Ise Bay, Japan were investigated to evaluate the effect of nitrification on the change in the δ¹⁵N in the water column. The δ¹⁵N of NO₃ ⁻ in the lower layers varied from −8.5‰ in May to +8.4‰ in July in response to the development of seasonal hypoxia and conversion from NH₄ ⁺ to NO₃ ⁻. The significantly ¹⁵N-depleted NO₃ ⁻ in May most likely arose from nitrification in the water column. The calculated apparent isotopic discrimination for water column nitrification (ɛ_nit = δ¹⁵N_substrate − δ¹⁵N_product) was 24.5‰, which lies within the range of previous laboratory-based estimates. Though prominent deficits of NO₃ ⁻ from hypoxic bottom waters due to denitrification were revealed in July, the isotopic discrimination of denitrification in the sediments was low (ɛ_denit = ∼1‰). δ¹⁵N_NO3 in the hypoxic lower layer mainly reflects the isotopic effect of water column nitrification, given that water column nitrification is not directly linked with sedimentary denitrification and the effect of sedimentary denitrification on the change in δ¹⁵N_NO3 is relatively small.     

高效硝化与反硝化功能菌株的分离筛选及其性能研究

本研究利用选择培养基从鳗鲡精养殖池和循环水水处理系统中分离筛选出二株脱氮细菌,应用16S rRNA分子生物学鉴定,确定菌株的种属。菌株NB-1属于芽孢杆菌属（Bacillus）、菌株DB-1是恶臭假单胞菌（Pseudomonas putida）。对菌株硝化及反硝化作用性能的研究结果表明：菌株NB-1具有良好的亚硝化及硝化作用性能,在投菌量为0.025%时,氨氮降解率为69.5%,亚硝酸盐氮降解率为99.2%。菌株DB-1在厌氧条件下表现出良好的反硝化作用性能,最适投菌量为0.6%,最佳C/N为12,对硝酸盐氮的降解率为94.6%,对总氮的降解率为64.0%。因此本研究筛选出的二株脱氮细菌可以广泛应用于养殖水体水质调控,具有良好的市场应用前景。     

稳定同位素方法在海洋N_2O研究中应用展望

氧化亚氮(N2O)是大气中重要和微量的温室气体,且在平流层N2O形成NO自由基与O3发生反应破坏臭氧层.海洋是大气中N2O净源,但由于海洋中生物化学过程的复杂性,有关N2O形成机制至今是人们研究的重点和难点.应用稳定同位素分馏原理对海洋中N2O形成机制的研究,区分海洋中通过硝化和反硝化过程不同途径产生的N2O过程,为深入研究海洋中N2O循环、估算将来大气中N2O浓度变化提供有用工具.     

Nutrient regeneration at bottom after a massive spring bloom in a subarctic coastal environment,Funka Bay,Japan

Nutrient regeneration and oxygen consumption after a spring bloom in Funka Bay were studied on monthly survey cruises from February to November 1998 and from March to December 1999. A high concentration of ammonium (more than 4 μmol l⁻¹) was observed near the bottom (80–90 m) after April. Phosphate and silicate gradually accumulated and dissolved oxygen decreased in the same layer. Salinity near the bottom did not change until summer, leading to the presumption that the system in this layer is semi-closed, so regenerated nutrients were preserved until September. Nitrification due to the oxidation of ammonium to nitrate was observed after June. Nitrite, an intermediate product, was detected at 4–7 μmol L⁻¹ in June and July 1999. Assuming that decomposition is a first order reaction, the rate constant for decomposition of organic nitrogen was determined to be 0.014 and 0.008 d⁻¹ in 1998 and 1999, respectively. The ammonium oxidation rate increased rapidly when the ambient ammonium concentration exceeded 5 μmol L⁻¹. We also performed a budget calculation for the regeneration process. The total amount of N regenerated in the whole water column was 287.4 mmol N m⁻² in 4 months, which is equal to 22.8 gC m⁻², assuming the Redfield C to N ratio. This is 34% of the primary production during the spring bloom and is comparable to the export production of 25 gC m⁻² measured by a sediment trap at 60 m (Miyake et al., 1998).     

海洋沉积物中硝化和反硝化过程研究进展

硝化和反硝化过程是氮循环的中心环节。水生态环境中氮的转化受诸多因素的制约,了解这些控制要素的作用机理,对预防和消除水体富营养化或缓解初级生产的氮限制有重要意义。综述了硝化和反硝化的研究现状,着重介绍了硝化-反硝化影响因素及测量方法的研究进展,并预测了进一步研究重点。     

The nitrogen cycle of an East Coast, U.K. saltmarsh: II. Nitrogen fixation, nitrification, denitrification, tidal exchange

Measurements of nitrogen fixation (acetylene reduction) showed greatest rates in the saltmarsh pans with a benthic layer of cyanobacteria present. The smallest amount of nitrogen fixation occurred on the marsh surface where a Puccinellia maritima/Halimione portulacoides plant association shaded the underlying sediment. Phototrophic nitrogen fixation was always greater than dark, chemotrophic, bacterial fixation.Only a small proportion of the total amount of ammonium, which was formed during detrital breakdown, was nitrified to nitrate. Although there is a high capacity for bacterial nitrate reduction in these sediments, the process is limited by low nitrate availability and most nitrate upon reduction is converted to ammonium rather than being denitrified to gaseous products. Denitrification does not, therefore, result in any great loss of nitrogen from the saltmarsh.There was little net import or export of nitrogen on an annual basis, although nitrate and organic-N in small particulate material was removed from tidal water by the marsh, and there was net annual export of ammonium, dissolved organic-N and organic-N in large particulate material. Losses of nitrogen by the small net tidal export and by denitrification were approximately balanced by nitrogen fixation. It was concluded that the nitrogen cycle of the Colne Point saltmarsh was balanced on an annual basis, with most nitrogen being recycled within the marsh. The saltmarsh did not apparently act as a net source of nitrogen for the adjacent estuary, although it may act as an important processor of nitrogen, removing some forms of nitrogen such as nitrate from tidal water while exporting other forms of nitrogen such as dissolved organic-N.     

Eutrophication and the rate of net nitrification in a coastal marine ecosystem

Rates of net nitrification were calculated for four large (13 m³) estuarine-based microcosms that had been subjected to inorganic nutrient enrichment. Calculated rates were based on two years of weekly nitrate and nitrite measurements and ranged from a maximum of 0·55 μmol NO₂₊₃⁻ produced l⁻¹ day⁻¹ in the control tank (no enrichment) to over 13 μmol NO₂₊₃⁻ produced l⁻¹ day⁻¹ in the most enriched tank (receiving 18·6 μmol NH₄ l⁻¹ day⁻¹). Almost all NO₂₊₃⁻ production was pelagic, little was benthic. Net NO₃⁻ production or net NO₂⁻ production dominated the net nitrification rates during different seasons. Good correlations were found between various oxidation rates and substrate concentrations. The calculated net nitrite production rates were 10 to 1000 times higher than previously reported rates for open ocean systems, demonstrating the potential importance of nitrification to estuarine systems.     

Simultaneous nitrification and denitrification conducted by Halomonas venusta MA-ZP17-13 under low temperature conditions

Nitrification is a key step in the global nitrogen cycle.Compared with autotrophic nitrification,heterotrophic nitrification remains poorly understood.In this study,Halomonas venusta MA-ZP17-13,isolated from seawater in shrimp aquaculture (Penaeus vannamei),could simultaneously undertake nitrification and denitrification.With the initial ammonium concentration at 100 mg/L,the maximum ammonium-nitrogen removal rate reached98.7%under the optimal conditions including C/N concentration ratio at 5.95,p H at 8.93,and Na Cl at 2.33%.The corresponding average removal rate was 1.37 mg/(L·h)(according to nitrogen) in 3 d at 11.2°C.By whole genome sequencing and analysis,nitrification-and denitrification-related genes were identified,including ammonia monooxygenase,nitrate reductase,nitrite reductase,nitric oxide dioxygenase and nitric oxide synthase;while no gene encoding hydroxylamine oxidase was identified,it implied the existence of a novel nitrification pathway from hydroxylamine to nitrate.These results indicate heterotrophic bacterium H.venusta MA-ZP17-13 can undertake simultaneous nitrification and denitrification at low temperature and has potential forNH_4~+-N/NH3-N removal in marine aquaculture systems.