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Molecular analysis of microbial community in a groundwater sample polluted by landfill leachate and seawater*

机译:垃圾渗滤液和海水污染的地下水样品中微生物群落的分子分析*

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摘要

Seashore landfill aquifers are environments of special physicochemical conditions (high organic load and high salinity), and microbes in leachate-polluted aquifers play a significant role for intrinsic bioremediation. In order to characterize microbial diversity and look for clues on the relationship between microbial community structure and hydrochemistry, a culture-independent examination of a typical groundwater sample obtained from a seashore landfill was conducted by sequence analysis of 16S rDNA clone library. Two sets of universal 16S rDNA primers were used to amplify DNA extracted from the groundwater so that problems arising from primer efficiency and specificity could be reduced. Of 74 clones randomly selected from the libraries, 30 contained unique sequences whose analysis showed that the majority of them belonged to bacteria (95.9%), with Proteobacteria (63.5%) being the dominant division. One archaeal sequence and one eukaryotic sequence were found as well. Bacterial sequences belonging to the following phylogenic groups were identified: Bacteroidetes (20.3%), β, γ, δ and ε-subdivisions of Proteobacteria (47.3%, 9.5%, 5.4% and 1.3%, respectively), Firmicutes (1.4%), Actinobacteria (2.7%), Cyanobacteria (2.7%). The percentages of Proteobacteria and Bacteroides in seawater were greater than those in the groundwater from a non-seashore landfill, indicating a possible influence of seawater. Quite a few sequences had close relatives in marine or hypersaline environments. Many sequences showed affiliations with microbes involved in anaerobic fermentation. The remarkable abundance of sequences related to (per)chlorate-reducing bacteria (ClRB) in the groundwater was significant and worthy of further study.
机译:沿海垃圾掩埋含水层是具有特殊物理化学条件(高有机负荷和高盐度)的环境,渗滤液污染的含水层中的微生物在固有的生物修复中起着重要作用。为了表征微生物多样性并寻找微生物群落结构与水化学之间关系的线索,通过对16S rDNA克隆文库的序列分析,对从海岸垃圾填埋场获得的典型地下水样品进行了与培养无关的检查。使用两组通用的16S rDNA引物扩增从地下水中提取的DNA,从而可以减少引物效率和特异性引起的问题。从文库中随机选择的74个克隆中,有30个包含独特的序列,其分析表明它们中的大多数属于细菌(95.9%),而变形杆菌(63.5%)是显性片段。还发现了一个古细菌序列和一个真核序列。鉴定了属于以下系统发生组的细菌序列:细菌杆菌(20.3%),变形杆菌的β,γ,δ和ε细分(分别为47.3%,9.5%,5.4%和1.3%),硬毛菌(1.4%),放线菌(2.7%),蓝细菌(2.7%)。海水中Proteobacteria和Bacteroides的百分比高于非沿海垃圾填埋场的地下水中的百分比,表明可能存在海水影响。在海洋或高盐环境中,有不少序列具有近亲。许多序列显示与厌氧发酵有关的微生物的隶属关系。地下水中与(减少)氯酸盐还原菌(ClRB)有关的序列非常丰富,具有重要意义,值得进一步研究。

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