源自生物絮团产絮凝剂的异养硝化-好氧反硝化菌xt1的鉴定及其脱氮特性

为了高效进行水体脱氮,本实验从形成于凡纳滨对虾(Litopenaeus vannamei)养殖水体的生物絮团中分离到一株具产絮能力的脱氮菌xt1,经16S r RNA基因测序与生理生化分析确定菌株xt1为短小芽孢杆菌(Bacillus pumilus)。在此基础上,本文研究了该菌的脱氮特性。结果表明:菌株xt1最佳碳源为葡萄糖,以其为底物对氨氮、硝态氮去除率分别达95.56%和57.40%。以蔗糖为碳源亦具较高脱氮率,对氨氮、硝态氮去除率分别达69.95%和49.50%;该菌能利用有机氮加速生长,添加0.25%、0.5%、1%和2%的蛋白胨能促进OD600,分别达到0.925、1.034、1.103和1.314,均高于未加蛋白胨下的生长,且氨氮去除率均超过90%,硝态氮去除率均超过88%;该菌能适应20—200mg/L无机氮浓度;该菌能以NH4+-N、NO2–-N或NO3–-N为唯一氮源进行异养硝化-好氧反硝化,反应84h去除率分别达到94.16%、47.60%和91.17%。其中,该菌的硝化形式是将氨氮转化为气态氮脱除,其硝态氮反硝化形式是先将硝态氮转化为亚硝氮,再以气态氮脱除。在进行异养硝化-好氧反硝化同时,菌株xt1体现絮凝特性,絮凝率最高分别达到82.28%、73.15%和75.60%;此外,添加该菌于养殖水体中能加速生物絮团形成,同时提高脱氮率。各项结果表明,菌株xt1可作为水产养殖水体脱氮的备选菌株。     

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.     

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.     

一株卓贝尔氏菌F13-1好氧反硝化特性及其反硝化基因的初步研究

为获得高效好氧反硝化细菌,并研究这些细菌的脱氮特性以及好氧反硝化机理,采集了近海表层沉积物进行富集培养,分离获得一株具有高效好氧反硝化能力的菌株F13-1.结合生理生化特征及16S rRNA基因序列分析,初步鉴定菌株F13-1为卓贝尔氏菌(Zobellella sp.).在以葡萄糖为碳源,C/N比值为15,盐度为30,摇床转速160 r/min,p H值为7,28℃的最优脱氮条件下菌株F13-1脱氮效果较好,分别能将100 mg/dm3硝酸盐和亚硝酸盐在24和16 h内彻底脱除,脱除速率分别为:5.87 mg/(dm3·h)和5.97 mg/(dm3·h).并且菌株F13-1对高浓度亚硝酸盐具有较好的耐受性,能在高浓度亚硝酸盐存在条件下正常生长及脱氮,156 h内能将400 mg/dm3亚硝酸盐脱除完全.基因组分析表明,该菌株具有完整的反硝化基因簇,包括nap、nir、nor和nos等共17个基因.研究结果表明该菌株具有高效好氧反硝化特性,在养殖废水处理中具有较好的应用潜力.     

The effects of semi-lunar spring and neap tidal change on nitrification, denitrification and N2O vertical distribution in the intertidal sediments of the Yangtze estuary, China

To explore the influences of semi-lunar spring and neap tidal changes on nitrogen cycling in intertidal sediments, a comparative study among waterlogged, desiccated and reflooded systems was carried out in August 2005 and February 2006 by analyzing nitrification, denitrification and N₂O depth profiles in the intertidal flats of the Yangtze estuary. Laboratory experiments showed that alternating emersion and inundation resulted in the significant changes in nitrification and denitrification rates in the intertidal sediment systems. Due to the desiccation-related effects, lowest nitrification and denitrification rates were observed in the desiccated sediment cores. Highest nitrification and denitrification rates were however detected in the waterlogged and reflooded systems, respectively. It is hypothesized that the highest nitrification rates in the waterlogged sediments were mainly attributed to higher nitrifier numbers and NH₄⁺ being more available, whereas the availability of NO₃⁻ might dominate denitrification in the reflooded sediments. In addition, the highest N₂O concentrations were detected in the reflooded sediment cores, and the lowest found in the dried sediment cores. It was also shown that N₂O in the intertidal sediments was mainly from nitrification under the desiccated condition. In contrast, N₂O in the intertidal sediments was produced mainly via denitrification under the waterlogged and reflooded conditions. It is therefore concluded that the semi-lunar tidal cycle has a significant influence on nitrification, denitrification and N₂O production in the intertidal sediment systems.     

一株海洋聚磷菌YSR-3的分离与鉴定

采用平板分离法,从黄海海域分离到一株聚磷菌,编号为YSR-3,对其进行了部分生理生化特性以及分子特性的研究。结果表明,YSR-3菌体呈杆状,大小为3·5μm×1μm,革兰氏染色阴性,好氧生长;显微镜下观察能运动。最适培养条件为:氯化钠浓度3%,生长温度为24℃,起始pH值为6·5。该菌能在细胞内累积多磷酸盐形成异染粒。16SrDNA鉴定结果表明,YSR-3属于γ-变形菌纲、盐单胞菌属(Halomonas)。与盐单胞菌属中的其他已知种比较,YSR-3具有其特性:对氯霉素和卡那霉素敏感、淀粉水解呈阳性、反硝化和几丁质降解呈阴性、能将葡萄糖作为唯一碳源和能源。     

深海来源芳烃类降解菌筛选与盐单胞菌芘降解能力测定

海洋来源的菌株在高盐污水类修复处理中有重要的应用前景,而烃类物质是污水中的重要组成部分,其中含有多个苯环的芳香烃族化合物,对人类和环境具有较大的毒性,而目前关于多环芳烃(PAHs)厌氧代谢的相关报道相对较少。本研究以PAHs(萘、菲和芘等)为唯一碳源和能源,通过厌氧富集对太平洋深海沉积物中的PAHs降解菌展开研究。富集物细菌菌群分析结果表明,盐单胞菌属(Halomonas)、海旋菌属(Thalassospira)、海杆菌属(Marinobacter)、海洋杆菌属(Oceanobacter)和食烷菌属(Alcanivorax)等是主要的功能类群;其中盐单胞菌属是最主要的功能类群。通过筛选分离获得一高效烃降解盐单胞菌株,鉴定并命名为泰坦尼克盐单胞菌(Halomonas titanicae) PA16-9,该菌与模式菌株Halomonas titanicae BH1^T的16S rRNA相似性为99.52%。基于16S rRNA基因序列比对搜寻NCBI数据库,发现Halomonas titanicae广泛存在于废水、活性污泥、油田、湿地等有机质复杂区域,生态位分布极广,可...     

新型生物脱氮技术及其相关酶系的研究进展

水中氮素浓度过高会导致水体富营养化, 而新型生物脱氮技术有多种耦合手段, 可以针对不同的氨氮浓度进行使用, 因此在水体氮素污染处理领域具有广阔的应用前景。本文系统介绍了新型生物脱氮技术的反应通路、研究进展及优缺点; 并对不同反应路径中生物脱氮酶的作用机制、酶活测定和研究现状等进行了系统的阐述, 最后对新型生物脱氮通路上的生物脱氮酶进行总结归纳, 对生物脱氮酶的未来提出展望。本综述旨在为提高新型生物脱氮技术在水体氮素处理中的效率及相关生物脱氮酶的研究提供理论依据。     

印度洋深海热液区一株盐单胞菌Halomonas sp.YD-7的分离鉴定及研究

从印度洋深海热液区沉积物中筛选到一株耐盐菌,菌株形态呈杆状,长1.5～2.0 μm,宽0.5～0.7 μm,属革兰氏阴性菌,生长适温为4～55 ℃,最适为35 ℃;pH测试范围为4.0～10.5,最适为7.0;测试NaCl浓度为0～5.13 mol/L,最适为0.86 mol/L,该菌属于兼性厌氧菌.经16S rDNA鉴定该菌属于γ-变形菌纲(γ-Proteobacteria)盐单胞菌属(Halomonas),命名为Halomonas sp. YD-7.该菌与菌株Halomonas sp. ANT9086在进化位置上最为接近,同源性达98.1%,有可能是新种.对该菌株的生理生化特性进行了研究,发现该菌具有很强的适应能力,对温度、盐度和氧气的适应范围广.该菌株还具有很强的耐受和去除Mn2+的能力及一定的耐受和去除Cr6+的能力.表明该菌在污水处理与生物修复方面可能具有重要的应用价值.     

微型异养鞭毛虫摄食选择性的研究进展

微型异养鞭毛虫(HNF)是海洋微微型浮游生物的重要摄食者,通过摄食作用对后者的种类(或类群)组成、粒径分布、数量结构和营养价值等属性具有重要的影响,而这与HNF的摄食选择性有直接关系。对HNF摄食选择性的研究有助于深入了解HNF在海洋微食物环乃至整个海洋生态系统中的作用。就国际上已开展的HNF摄食选择性相关研究进行了回顾,分析和总结了影响HNF选择性摄食的关键因素,如食物大小、游动性、营养价值及食物细胞表面的生化结构特征等,并重点介绍了HNF摄食选择性形成的主要机制以及HNF的选择性摄食在调节海洋微微型浮游生物群落结构中的作用。     

Nitrous oxide concentration and nitrification and denitrification in Zhujiang River Estuary, China

1 Introduction Estuaries have been subject to intense anthro- pogenic influences over recent decades by inputs of ni- trogen as a consequence of fertilizer usage and sewage/waste input (Howarth et al., 1996). Thus, the capacity of an estuary to buffer or alleviate additional nutrients is of importance in modifying the load of ni- trogen in the marine ecosystem. The role of microor- ganisms in regulating fluxes of nitrogen from land to sea through estuaries has received increased attention (Ni…     

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。     

A seasonal comparison of prokaryote numbers,biomass and heterotrophic productivity in waters of the KwaZulu-Natal Bight,South Africa

The KwaZulu-Natal Bight is a shallow indentation of the eastern seaboard of South Africa, characterised by a narrow (45 km wide) extension of the continental shelf, with a shelf break at about 100 m. It has a complex hydrography: the waters of the bight are derived from the fast-flowing, southward-trending Agulhas Current, which is fed mostly by the tropical and subtropical surface waters of the South-West Indian Ocean subgyre, which are generally oligotrophic in nature, notably depleted in reduced nitrogen and phosphate except at river mouths and during periodic upwelling of deeper nutrient-rich water. Despite this, the bight is believed to be relatively productive, and it is suggested that efficient nutrient recycling by prokaryotes may sustain primary productivity efficiently, even in the absence of new nutrient inputs. Here we have measured bacterial numbers, biomass and heterotrophic productivity during summer and winter in conjunction with phytoplankton standing stock and factors that influence it. Bacterial distribution closely matched phytoplankton distribution in surface waters, and was highest close to the coast. Bacterial standing stocks were similar to those of oligotrophic systems elsewhere (0.5–5.0 × 10⁵ cells ml^–1; 1 × 10^–8 to 1.25 × 10^–7 g C ml^–1) and increased in association with the development of phytoplankton blooms offshore and with inputs of allochthonous material by rivers at the coast. Heterotrophic productivity in summer was lowest in the far south and north of the bight (0.5 × 10^–10 g C ml^–1 h^–1) but higher close to the shore, over shallow banks, and in association with increased phytoplankton abundance over the midshelf (1.0–3.5 × 10^–9 g C ml^–1 h^–1). There were marked seasonal differences with lower bacterial standing stocks (5 × 10⁴ to 2 × 10⁵ cells ml^–1; 4–5 × 10^–9 to 1–2 × 10^–8 g C ml^–1) and very low bacterial productivity (4 × 10^–11 to 1 × 10^–10 g C ml^–1 h^–1) in winter, probably resulting from lowered rates of primary productivity and dissolved organic matter release as well as reduced riverine allochthonous inputs during the winter drought.     

微藻异养培养技术的研究进展

微藻作为水产动物的活铒料,在水产养殖中一直有着广泛的应用.很多微藻营养价值高,且含有丰富的生物活性物质,既是海洋药物的潜在来源, 也是人类优质的保健食品(Vincent,1969,Schwarz et al,1995).     

海洋聚磷菌中核苷二磷酸激酶基因的克隆及序列分析

以海洋聚磷菌Halomonas YSR-3的总DNA为模板,用PCR法扩增核苷二磷酸激酶基因,将扩增片段克隆到pGM．T载体,转化Escherichia coli TOP10菌株,经蓝白斑筛选、菌落PCR得到阳性克隆,测序后对序列进行Blast比对分析。得到的基因序列长度为420bp,翻译后的序列与Loktanella vestfoldensis SKA53,Jannaschia sp．CCS1,Roseobacter sp．CCS2的核苷二磷酸激酶蛋白序列相似性分别为89％,86％,85％。     

南海北部大鹏湾中DOP的分布、滞留时间、自养生产和异养降解

The importance of dissolved organic phosphorus(DOP) as a potential nutrient source for primary producers in marine systems has been recognized for up to eight decades, but currently, the understanding of the biogeochemistry of DOP is in its infancy. In the present study, monthly data between 2000 and 2014 were used to analyze the temporal and spatial distributions of DOP in the Mir Bay, the northern South China Sea. The DOP residence time(T_(DOP)) was also investigated using a simple regression analysis in combination with chlorophyll a(Chl a) measurements while excess DOP(ΔDOP), produced by the biogeochemical processes of autotrophic production and heterotrophic removal, was determined using a two-component mixing mass-balance model in combination with salinity measurements. The results showed that the DOP concentration was(0.017±0.010) mg/L higher in the surface-water compared with the bottom-water and higher in the inner Tolo Harbour and waters adjacent to Shatoujiao compared with the main zone of the bay. Although seasonal changes and annual variability in the DOP were small, the surface DOP concentration was higher in the wet season(April–September)than in the dry season(October–March) due to the impacts of seaward discharges and atmospheric deposition into the bay. Measurement and regression results showed that the DOP release rate from phytoplankton production was about 1.83(gP)/(gChl a) and the T_(DOP) was about 7 d, which implied that the DOP cycle in the bay was rapid. The ΔDOP was calculated from the model to be about 0.000 mg/L in the main zone of the bay and about 0.002 mg/L in the inner Tolo Harbour and waters adjacent to Shaotoujiao, suggesting that the autotrophic production of DOP was almost balanced by the heterotrophic removal in the main zone of the bay and dominated in the inner Tolo Harbour and waters adjacent to Shaotoujiao. In conclusion, the Mirs Bay is very productive and fairly heterotrophic.     

龙门口水库滨岸带硝化作用研究

以龙门口水库滨岸带湿地为研究对象,以农业径流中氮元素为主要的农业面源污染物,2012年7~9月通过设置植被类型和氮浓度不同的3个采样区进行对比试验,对滨岸带土壤中氮含量、硝化细菌数量及其硝化速率进行了研究。结果表明,表层土壤硝化速率和硝化细菌数量水平分布特征为有农田径流且有水草区有农田径流且无水草区无农田径流且无水草区,且均随着土壤深度的增加明显降低。从月份变化来看氨氧化细菌(Ammonia-Oxidizing Bacteria,AOB)的数量分布表现为7月8月9月,与氨氮含量变化趋势基本一致,硝化速率也呈现出夏季大于秋季的特点。对滨岸带土壤硝化作用有显著影响的因子为AOB的数量、氨氮、总氮、总磷含量、含水率和温度,从而进一步证实了氨氧化是硝化作用的限制性步骤。