海洋环境中的厌氧氨氧化细菌与厌氧氨氧化作用

厌氧氨氧化是细菌在厌氧条件下将氨氮氧化成氮气的过程,主要应用在污水处理反应器中,最近几年发现在海洋环境中也广泛存在,并在海洋氮循环过程中发挥了重要作用,代表了海洋中一个巨大的氮汇,对碳循环和全球气候变化也有重要影响。梯烷膜脂结构独特,是厌氧氨氧化细菌的化学生物标志物,具有化学分类与古海洋学应用潜力。厌氧氨氧化作用及厌氧氨氧化细菌已成为海洋生物地球化学、微生物学、有机地球化学等研究领域的热点。     

厌氧氨氧化细菌物种组成对环境因子响应的研究进展

分析海洋水体及沉积物中厌氧氨氧化细菌所处的环境条件,比较海洋与淡水环境中其物种组成的差异,总结了其物种组成、丰度及厌氧氨氧化活性对环境因子的响应规律。认为厌氧氨氧化细菌物种组成受多种环境因子的影响。其中,盐度为影响其物种组成的最显著因素,如Candidatus Brocadia,Candidatus Kuenenia及Candidatus Scalindua分别适应淡水、寡盐和海水环境;同一地区,其物种组成在不同季节表现出明显差异,夏季的多样性高于冬季的;根据陆源厌氧氨氧化细菌多样性及海洋沉积物中浮霉菌门多样性随深度增加而减小的趋势,推断海洋沉积物中厌氧氨氧化细菌多样性与深度成反比。厌氧氨氧化细菌丰度、厌氧氨氧化活性与温度、NO-2及NO-3浓度之间成正相关;丰度与季节的关系,在不同地区的研究结果存在差异。未来厌氧氨氧化细菌研究工作重点应在其对氧浓度的耐受范围、海洋中厌氧氨氧化细菌多样性随深度的变化规律、导致其物种组成、丰度及厌氧氨氧化活性季节性差异的具体环境因子。     

用于海洋沉积物中厌氧氨氧化细菌分子生态学研究的引物比较

为比较厌氧氨氧化细菌16SrRNA基因的2对特异性引物(Amx368F/Amx820R、Brod541F/Amx820R)在其分子生态学研究方面的优点与不足,本研究采用高通量测序和qPCR技术对长江口及其邻近海域沉积物中厌氧氨氧化细菌细菌的群落组成、多样性和丰度进行比较分析,结果表明,引物Amx368F/Amx820R特异性较高,能够较为完整地反映沉积物中厌氧氨氧化细菌的多样性信息,且其覆盖的厌氧氨氧化细菌丰度与功能基因hzo丰度正相关(P<0.01);而引物Brod541F/Amx820R特异性较低,覆盖部分厌氧氨氧化细菌及其他多个门的细菌,其量化的厌氧氨氧化细菌与功能基因hzo丰度无明显相关关系(P>0.05)。因此,引物Amx368F/Amx820R能够更加真实地反映厌氧氨氧化细菌在海洋沉积物中的群落特征,在其分子生态学研究中更具优势。     

珠江口沉积柱中厌氧氨氧化活动的梯烷脂记录

为了反演珠江口过去环境的厌氧氨氧化活动状况,本文分析了来自珠江口3个站位的柱状沉积物中梯烷脂的含量,在所有的样品中均检测到了梯烷脂,表明珠江口地区存在厌氧氨氧化活动。P03沉积柱的梯烷脂总含量为0.16~4.13μg/g,L1为0.15~1.3μg/g,C2T为0.07~2.19μg/g。梯烷脂的垂直分布特征表明3个沉积柱中厌氧氨氧化细菌的来源可能不同,P03沉积柱中梯烷脂可能主要来源于沉积物中的厌氧氨氧化细菌,而L1梯烷脂可能来自上层水体和沉积物,C2T的厌氧氨氧化活动可能主要发生在底层缺氧水体中。将梯烷脂与总有机碳(TOC)作相关性分析,结果显示在TOC含量高的柱状样中,两者相关性好,TOC校准后的梯烷脂与原梯烷脂的分布趋势的一致性,表明梯烷脂含量能够指示厌氧氨氧化活动。对珠江口过去的厌氧氨氧化活动的重建表明P03站位沉积物中厌氧氨氧化活动自1970—1990增强,L1站位的厌氧氨氧化活动近十年来亦逐年增强,C2T站位上覆水体的厌氧氨氧化活动则呈现近几年快速增强的趋势。虽然本研究仅初步探讨了近些年来珠江口厌氧氨氧化活动的变化规律,但为进一步研究珠江口氮循环过程提供了良好的基础。     

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.     

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。     

生物标志物梯烷脂指示胶州湾厌氧氨氧化活动

厌氧氨氧化是重要的氮脱除途径,对海洋中的氮循环,尤其是人类活动影响下海湾的氮循环起重要作用。梯烷脂作为厌氧氨氧化的生物标志物,能指示不同生态系统中的厌氧氨氧化活动,但其在海湾环境中的来源以及对人类活动影响的响应尚未明确。本文对人类活动影响下的典型海湾——山东半岛的胶州湾海域进行研究,通过分析胶州湾表层沉积物中梯烷脂及其他有机质指标,来研究海湾中有机质及厌氧氨氧化菌的来源及梯烷脂对环境因子的响应。研究显示厌氧氨氧化活动在胶州湾海域普遍存在。对总有机质参数(TOC,δ~(13)C和TOC/TN)与有机质来源贡献指标(TMBR和BIT)的分析显示胶州湾海域表层沉积物有机质以海源为主(BIT指示为83%、TMBR指示为54%),但各有机质来源指标与海湾中梯烷脂含量无明显相关。对梯烷脂总量及不同种类梯烷脂占比的分析显示,梯烷脂主要来自夏季水体中的颗粒物,且在梯烷脂总量较高的区域,梯烷脂可能主要受河流输入及污水排放的影响。主成分分析显示海湾沉积物中梯烷脂与营养盐及溶解氧含量受相同因子所调控,表明梯烷脂可以在胶州湾指示人类活动对海水环境质量的影响。这也为沉积物中梯烷脂在海湾环境,乃至其他边缘海环境中对人类活动的指示作用提供了新思路。     

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.     

九龙江口肥水-缢蛏养殖塘厌氧氨氧化微生物活性研究

缢蛏串联养殖经由饵料藻类养殖塘(俗称肥水塘)投放适量含氮化合物以提高生产效率,了解此过程中的沉积物氮转化对调控和管理河口养殖塘氮的收支具有重要意义。以福建九龙江口肥水-缢蛏养殖塘为研究对象,运用同位素示踪技术和高通量测序技术探究缢蛏塘和肥水塘沉积物厌氧氨氧化速率及微生物群落结构,并分析其环境调控机制。结果表明,肥水塘沉积物厌氧氨氧化速率为(1.76±0.11) nmol/(g·h),相对贡献为9.16%±0.40%,显著高于缢蛏塘(P<0.05)。缢蛏塘和肥水塘厌氧氨氧化微生物主要由Candidatus Scalindua、Ca.Kuenenia、Ca.Brocadia和Ca.Anammoximicrobium属组成。其中,Ca.Kuenenia属为养殖塘的优势菌属。沉积物厌氧氨氧化微生物多样性和丰富度呈显著的空间分布特征,肥水塘中生物多样性更高(P<0.05)。厌氧氨氧化微生物活性和群落结构差异主要受养殖土地利用方式、溶解氧(DO)、NH₄⁺和NO₃^-含量等环境因子的调控。该研究在一定...     

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...     

Anammox bacteria and the anaerobic oxidation of ammonium in the oxygen minimum zone off northern Chile

Anammox is the anaerobic oxidation of ammonium by nitrite or nitrate to yield N₂. This process, along with conventional denitrification, contributes to nitrogen loss in oxygen-deficient systems. Anammox is performed by a special group of bacteria belonging to the Planctomycetes phylum. However, information about the distribution, activity, and controlling factors of these anammox bacteria is still limited. Herein, we examine the phylogenetic diversity, vertical distribution, and activity of anammox bacteria in the coastal upwelling region and oxygen minimum zone off northern Chile. The phylogeny of anammox bacteria was studied using primers designed to specifically target 16S rRNA genes from Planctomycetes in samples taken during a cruise in 2004. Anammox bacteria-like sequences affiliated with Candidatus “Scalindua spp.” dominated the 16S rRNA gene clone library. However, 62% of the sequences subgrouped separately within this cluster and together with a single sequence retrieved from the suboxic zone of the freshwater Lake Tanganyika. The vertical distribution and activity of anammox bacteria were explored through CARD-FISH (fluorescence in situ hybridization with catalyzed reporter deposition) and ¹⁵N labeling incubations, respectively, at two different open-ocean stations during a second cruise in 2005. Anammox bacterial CARD-FISH counts (up to 3000 cells ml⁻¹) and activity (up to 5.75 nmol N₂ L⁻¹ d⁻¹) were only detected at the station subjected directly to the upwelling influence. Anammox cell abundance and activity were highest at 50 m depth, which is the upper part of the OMZ. In this layer, a high abundance of cyanobacteria and a marked nitrogen deficit were also observed. Thus, our results show the presence of a new subcluster within the marine anammox phylogeny and indicate high vertical variability in the abundance and activity of anammox bacteria that could be related to an intensification of carbon and nitrogen cycling in the upper part of the OMZ.     

自组装膜进样质谱系统及其在砂质沉积物异化硝酸盐还原研究中的应用

沉积物中的异化硝酸盐还原过程对于海洋氮循环起着至关重要的作用。基于15N标记的培养技术是目前测定沉积物异化硝酸盐还原的主要手段。准确快速测定15N标记的产物(29N2、 30N2)是量化异化硝酸盐还原各个过程速率的关键。本研究自行组装膜进样质谱系统用于29N2和30N2的测定,对其测量条件进行了优化。结果表明,进样蠕动泵进样流速0.80 mL/min,进样时间3~3.5 min,恒温槽温度20~25℃,同时铜还原炉温度在300~600℃的条件下,^29N2/^28N2和^30N2/^28N2的测试精密度分别可以控制在0.1%和1%以内,比较适合29N2和30N2的测定。利用自组装的膜进样质谱系统结合15N标记的培养技术研究了青岛石老人沙滩沉积物中的异化硝酸盐还原过程。石老人沙滩沉积物不存在将硝酸盐完全还原为氮气好氧的反硝化。厌氧铵氧化、厌氧反硝化和异化硝酸盐还原为铵(Dissimilatory Nitrate Reduction to Ammonium,DNRA)的潜在速率(以湿沉积物N计)分别为(0.05±0.01) nmol/(cm^3·h),(2.32±0.21) nmol/(cm^3·h)和(1.02±0.15) nmol/(cm^3·h)。厌氧反硝化是硝酸盐异化还原主要的贡献者,其比例接近70%,其次是DNRA,比例可达30%,而厌氧铵氧化的贡献最低,仅为1%。在N2产生过程中,主要贡献者是反硝化,厌氧铵氧化的贡献仅为2%。     

Determination of phospholipid fatty acid structures and stable carbon isotope compositions of deep-sea sediments of the Northwest Pacific, ODP site 1179

Sediment samples ranging from 0.05 to 278 m below sea floor (mbsf) at a Northwest Pacific deep-water (5564 mbsl) site (ODP Leg 191, Site 1179) were analyzed for phospholipid fatty acids (PLFAs). Total PLFA concentrations decreased by a factor of three over the first meter of sediment and then decreased at a slower rate to approximately 30 mbsf. The sharp decrease over the first meter corresponds to the depth of nitrate and Mn(IV) reduction as indicated by pore water chemistry. PLFA-based cell numbers at site 1179 had a similar depth profile as that for Acridine orange direct cell counts previously made on ODP site 1149 sediments which have a similar water depth and lithology. The mole percentage of straight chain saturated PLFAs increases with depth, with a large shift between the 0.95 and 3.95 mbsf samples. PLFA stable carbon isotope ratios were determined for sediments from 0.05 to 4.53 mbsf and showed a general trend toward more depleted δ¹³C values with depth. Both of these observations may indicate a shift in the bacterial community with depth across the different redox zones inferred from pore water chemistry data. The PLFA 10me16:0, which has been attributed to the bacterial genera Desulfobacter in many marine sediments, showed the greatest isotopic depletion, decreasing from − 20 to − 35‰ over the first meter of sediment. Pore water chemistry suggested that sulfate reduction was absent or minimal over this same sediment interval. However, 10me16:0 has been shown to be produced by recently discovered anaerobic ammonium oxidizing (anammox) bacteria which are known chemoautotrophs. The increasing depletion in δ¹³C of 10me16:0 with the unusually lower concentration of ammonium and linear decrease of nitrate concentration is consistent with a scenario of anammox bacteria mediating the oxidation of ammonium via nitrite, an intermediate of nitrate reduction.     

Methane-derived carbonates and chemosymbiotic communities of Piedmont (Miocene,northern Italy): An update

Stable carbon isotope values of authigenic carbonate rocks in the Miocene terrigenous sediments of Piedmont indicate a methane-related origin. Some of these methane-derived carbonates (Lucina limestone) are characterized by the presence of abundant lucinid remains. Carbonate dissolution/precipitation and development of lucinid communities were related to bacterial methane oxidation, both aerobic and anaerobic. Anaerobic oxidation led to carbonate precipitation and production of sulfide, which sustained lucinid communities through chemosynthetic symbiotic bacteria. Aerobic oxidation of methane likely resulted in dissolution of carbonate skeletal grains. Several phases of carbonate precipitation, characterized by slightly different isotopic compositions, are recognizable in the limestones.     

大生活用海水的厌氧生物处理过程研究

文章研究了大生活用海水厌氧生物处理过程中海水盐度、水温、有机负荷、水力停留时间等因素对污染物去除率的影响。结果表明,厌氧塘对大生活用海水中COD,TKN,NH3-N,TP的净化效果分别可以达到50％、30．4％、24．9％和22．5％。温度对厌氧塘的去除率有较大的影响,随着温度的上升,厌氧塘对污染物的去除率提高。厌氧塘能够经受一定盐度变化的冲击和有机负荷的冲击,其水力停留时间以10d为宜。在污水中海水比例为36％时,经过培养驯化,厌氧塘中的优势耐盐细菌以假单胞菌属、索氏甲烷杆菌属、硝酸盐还原菌、硫酸盐还原菌为主。     

Review of inorganic nitrogen transformations and effect of global climate change on inorganic nitrogen cycling in ocean ecosystems

Inorganic N transformations (nitrification, anaerobic ammonium oxidation, denitrification, and dissimilatory nitrate reduction to ammonium) are regulated by various biogeochemical factors linked either by the supply of electron acceptors and donors or by competition for electron acceptors. This review considers both the microbial community related to each process and the technical methods used to measure each process rate. With this background knowledge, this article summarizes how global climate change through increased pCO2, ocean acidification, deoxygenation and anthropogenic N deposition will alter oceanic N cycling, and finally emphasizes the need for comprehensive research on inorganic N transformation in marine ecosystems.     

Analysis of cultivable aerobic bacterial community composition and screening for facultative sulfate-reducing bacteria in marine corrosive steel

Anaerobic, aerobic, and facultative bacteria are all present in corrosive environments. However, as previous studies to address corrosion in the marine environment have largely focused on anaerobic bacteria, limited attention has been paid to the composition and function of aerobic and facultative bacteria in this process. For analysis in this study, ten samples were collected from rust layers on steel plates that had been immersed in seawater for diff erent periods (i.e., six months and eight years) at Sanya and Xiamen, China. The cultivable aerobic bacterial community structure as well as the number of sulfate-reducing bacteria (SRB) were analyzed in both cases, while the proportion of facultative SRB among the isolated aerobic bacteria in each sample was also evaluated using a novel approach. Bacterial abundance results show that the proportions are related to sea location and immersion time;abundances of culturable aerobic bacteria (CAB) and SRB from Sanya were greater in most corrosion samples than those from Xiamen, and abundances of both bacterial groups were greater in samples immersed for six months than for eight years. A total of 213 isolates were obtained from all samples in terms of CAB community composition, and a phylogenetic analysis revealed that the taxa comprised four phyla and 31 genera. Bacterial species composition is related to marine location;the results show that Firmicutes and Proteobacteria were the dominant phyla, accounting for 98.13% of the total, while Bacillus and Vibrio were the dominant genera, accounting for 53.06% of the total. An additional sixfacultative SRB strains were also screened from the isolates obtained and were found to encompass the genus Vibrio (four strains), Staphylococcus (one strain), and Photobacterium (one strain). It is noteworthy that mentions of Photobacterium species have so far been absent from the literature, both in terms of its membership of the SRB group and its relationship to corrosion.     

东南太平洋热液区硫化物样品富集培养物中Fe3＋还原细菌的多样性分析

采集了东南太平洋热液区DYl15-20V一$35一TVGl7站位的沉积物硫化物样品,其含有76．717％（m／m）强还原性的磁性良好的黄铁矿．以乳酸钠为电子供体,不溶性的氢氧化铁为电子受体,对该样品进行Fe¨还原细菌的定向富集培养．通过构建该茵群的16SrDNA基因文库,随机挑选82个阳性克隆子进行群落结构多样性分析．研究结果表明：82个克隆代表的6种基因型分别属于4个主要类群,其中ε一变形杆菌占有绝对优势,达到55．2％．此外其还包括厚壁菌门、β一变形杆菌和仅．变形杆菌,克隆子比例依次为27．6％、10．3％、6．9％．在富集产物中还原氢氧化铁的功能菌群主要是具有硫还原作用的ε一变形菌中的硫还原菌．