Characterization of microbial communities from coastal waters using microarrays

Molecular methods, including DNA probes, were used to identify and enumerate pathogenic Vibrio species in the Chesapeake Bay; our data indicated that Vibrio vulnificus exhibits seasonal fluctuations in number. Our work included a characterization of total microbial communities from the Bay; development of microarrays that identify and quantify the diversity of those communities; and observation of temporal changes in those communities. To identify members of the microbial community, we amplified the 16S rDNA gene from community DNA isolated from a biofilm sample collected from the Chesapeake Bay in February, 2000. The resultant 75 sequences were 95% or more similar to 7 species including two recently described Shewanella species, baltica and frigidimarina, that have not been previously isolated from the Chesapeake. When the genera of bacteria from biofilm after culturing are compared to those detected by subcloning amplified 16S fragments from community DNA, the cultured sample exhibited a strong bias. In oysters collected in February, the most common bacteria were previously unknown. Based on our 16S findings, we are developing microarrays to detect these and other microbial species in these estuarine communities. The microarrays will detect each species using four distinct loci, with the multiple loci serving as an internal control. The accuracy of the microarray will be measured using sentinel species such as Aeromonas species, Escherichia coli, and Vibrio vulnificus. Using microarrays, it should be possible to determine the annual fluctuations of bacterial species (culturable and non-culturable, pathogenic and non-pathogenic). The data may be applied to understanding patterns of environmental change; assessing the health of the Bay; and evaluating the risk of human illness associated with exposure to and ingestion of water and shellfish.     

Molecular behavior and interactions with microbes during anaerobic degradation of bio-derived DOM in waste leachate

It is the key to control bio-derived dissolved organic matters (DOM) in order to reduce the effluent concentration of wastewater treatment, especially for waste leachate with high organic contaminants. In the present study, the anaerobic degradation of aerobically stabilized DOM was investigated with DOM substrate isolated through electrodialysis. The degradation of bio-derived DOM was confirmed by reduction of 15% of total organic carbon in 100 days. We characterized the molecular behavior of bio-derived DOM by coupling molecular and biological information analysis. Venn based Sankey diagram of mass features showed the transformation of bio-derived DOM mass features. Occurrence frequency analysis divided mass features into six categories so as to distinguish the fates of intermediate metabolites and persistent compounds. Reactivity continuum model and machine learning technologies realized the semi-quantitative determination on the kinetics of DOM mass features in the form of pseudo-first order, and confirmed the reduction of inert mass features. Furthermore, network analysis statistically establish relationship between DOM mass features and microbes to identify the active microbes that are able to utilize bio-derived DOM. This work confirmed the biological technology is still effective in controlling recalcitrant bio-derived DOM during wastewater treatment.     

含纤维素废水生物水解的应用研究

提取植物或动物纤维所产生的高浓有机废水，含短小纤维、木质素、纤维素、半纤维素、蜡质、果胶及其分解产物(糖、脂肪酸、醇等)，较难生物处理，属含纤维素废水。试验研究和生产应用表明，生物水解对含纤维素废水中高分子有机物，可予分解并提高度水可生化性：在常温、PH约9．5时，水解池污泥浓度达6．5～16．0g/L，容积负荷达2．9kgCOD／m^3．d，COD去除率为24—36％，VFA提高3．9—5．1倍，BOD／COD提高12～16％。分析认为：中温、酸性，纤维素、木素含量低时，水解效果应更好。     

Characterisation of Organic Matter and Carbon Cycling in Rehabilitated Lignite-rich Mine Soils

Open-cast lignite mining in the Lusatian mining district resulted in rehabilitated mine soils containing up to four organic matter types: (1) recent plant litter, (2) lignite deposited by mining activity, (3) carbonaceous ash particles deposited during amelioration of the lignite-containing parent substrate and (4) airborne carbonaceous particles deposited during contamination. The influence of lignite-derived carbon types on the organic matter development and their role in the soil carbon cycle was unknown. This paper presents the findings obtained during a six year project concerning the impact of lignite on soil organic matter composition and the biogeochemical functioning of the ecosystem. The organic matter development after rehabilitation was followed in a chronosequence of rehabilitated mine soils afforested in 1966, 1981 and 1987. A differentiation of the organic matter types and an evaluation of their role within the ecosystem was achieved by the use of ¹⁴C activity measurements, ¹³C CPMAS NMR spectroscopy and wet chemical analysis of plant litter compounds. The results showed that the amount and degree of decomposition of the recent organic matter derived from plant material of the 30 year old mine soil was similar to natural uncontaminated forest soil which suggests complete rehabilitation of the ecosystem. The decomposition and humification processes were not influenced by the presence of lignite. On the other hand it was shown that lignite, which was thought to be recalcitrant because of its chemical structure, was part of the carbon cycle in these soils. This demonstrates the need to elucidate further the stabilisation mechanisms of organic matter in soils.     

再生水补给型景观水体微生物健康风险评价——以圆明园为例

再生水作为景观水体的非常规补给水源被全球各个国家广泛使用,而其内病原微生物的存在可能会通过不同传输途径对人体健康产生一定的威胁.因此,本文以粪大肠菌群(Fecal Coliform,FC)、大肠杆菌(Escherichia coli,EC)、肠球菌(Enterococcus,ENT)为研究对象,系统调查了圆明园景观水体...     

利用果胶培养基分离番茄根际细菌

采用平板稀释和PCR-DGGE相结合的方法,比较了4种培养基(果胶富营养培养基PM,只含果胶一种营养的培养基PA,添加果胶的寡营养培养基YPP以及与YPP营养成分一致但不添加果胶的寡营养培养基YPG)分离番茄根际细菌的能力.结果显示:PA培养基能够分离到42种形态的细菌;YPP培养基可以分离到最高的细菌菌落数,分离获得的种类比PA少;YPP培养基分离获得细菌的种类和菌落数量比YPG培养基要多.聚类分析也显示只有果胶一种营养的PA培养基能分离到最多种类,分离获得的菌群与自然环境中微生物群落最相似.研究结果表明添加番茄根际主要分泌物——果胶到培养基中可以提高培养基分离细菌的能力.     

退化草原碱蓬土壤微生物生物量的季节动态模型

研究了东北退化草原土壤微生物生物量的季节变化,生态因子的季节变化以及生态因子对土壤微生物生物量的综合作用规律,微生物生物量W和呼吸速率W1的季节动态呈单峰上凸式曲线变化,8月份的有明显的高峰值出现,其值为：w/mg g^-1=4.9,w1/mg g^-1d^-1=28.1,同时利用IRM模型研究了生态因子对土壤微生物生物量变化的综合作用规律;Mt △Mto[1 （0.21rn-w1)△t],生态因子对土壤微生物生长的综合作用指数rn在整个生长季节的变化范围为：0.0273-0.7264,其中7-8月份rn较大,对微生物生长的限制作用较小,微生物生长较快,5月、6月、9月和10月rn较小,对微生物生长的限制作用较大,微生物生长较慢。     

厌氧条件下微生物对芳香族化合物分解的研究进展

本文总结了国内外芳香族化合物厌氧微生物分解的代谢类型和途径,指出厌氧条件下微生物对芳香族化合物的分解,包括光和代谢作用、硝酸盐还原作用、发酵作用、硫酸盐还原作用和产甲烷作用五种类型.厌氧条件下芳环的生物裂解包括还原性和非还原性两种类型,讨论了参与芳环裂解过程的一些酶类.     

环境微生物群落分析的T-RFLP技术及其优化措施

末端限制性酶切片段长度多态性分析(terminal restriction fragment length polymorphism, T-RFLP)是近年来发展起来的、不依赖于培养的微生物群落分析方法之一.由于其在微生物群落结构分析方面的特点,包括分辨率高、易于实现自动化及互联网海量数据共享等优势,自1997年最先被报道以来得到了广泛的应用,成为环境微生物群落分析的最强有力的工具之一.本文详细介绍了T-RFLP技术的原理,并从环境样品群落DNA的提取、引物设计和PCR扩增、限制性酶切、电泳分离检测和T-RFLP图谱解析等5个方面讨论了用该技术解析环境微生物群落的方法和技巧,简述了近8 a来国外T-RFLP技术在群落分析中的研究进展.类似于其他的分子微生物生态学技术, T-RFLP也有自身的缺陷,因此重点分析了该技术的局限性及相应的解决办法.图2表1参62