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墨西哥东南部腐蚀油管道生物膜相关细菌群落的特征分析

Characterization of bacterial community associated to biofilms of corroded oil pipelines from the southeast of Mexico.

作者信息

Neria-González Isabel, Wang En Tao, Ramírez Florina, Romero Juan M, Hernández-Rodríguez César

机构信息

Departamento de Microbiología, Escuela Nacional de Ciencias Biológicas-IPN, Mexico DF 11340, Mexico.

出版信息

Anaerobe. 2006 Jun;12(3):122-33. doi: 10.1016/j.anaerobe.2006.02.001. Epub 2006 May 2.

Abstract

Microbial communities associated to biofilms promote corrosion of oil pipelines. The community structure of bacteria in the biofilm formed in oil pipelines is the basic knowledge to understand the complexity and mechanisms of metal corrosion. To assess bacterial diversity, biofilm samples were obtained from X52 steel coupons corroded after 40 days of exposure to normal operation and flow conditions. The biofilm samples were directly used to extract metagenomic DNA, which was used as template to amplify 16S ribosomal gene by PCR. The PCR products of 16S ribosomal gene were also employed as template for sulfate-reducing bacteria (SRB) specific nested-PCR and both PCR products were utilized for the construction of gene libraries. The V3 region of the 16S rRNA gene was also amplified to analyse the bacterial diversity by analysis of denaturing gradient gel electrophoresis (DGGE). Ribosomal library and DGGE profiles exhibited limited bacterial diversity, basically including Citrobacter spp., Enterobacter spp. and Halanaerobium spp. while Desulfovibrio alaskensis and a novel clade within the genus Desulfonatronovibrio were detected from the nested PCR library. The biofilm samples were also taken for the isolation of SRB. Desulfovibrio alaskensis and Desulfovibrio capillatus, as well as some strains related to Citrobacter were isolated. SRB consists in a very small proportion of the community and Desulfovibrio spp. were the relatively abundant groups among the SRB. This is the first study directly exploring bacterial diversity in corrosive biofilms associated to steel pipelines subjected to normal operation conditions.

摘要

与生物膜相关的微生物群落会促进输油管道的腐蚀。输油管道中形成的生物膜内细菌的群落结构是了解金属腐蚀的复杂性和机制的基础知识。为了评估细菌多样性,在正常运行和流动条件下暴露40天后,从X52钢试片上获取生物膜样本。生物膜样本直接用于提取宏基因组DNA,该DNA用作模板通过PCR扩增16S核糖体基因。16S核糖体基因的PCR产物也用作硫酸盐还原菌(SRB)特异性巢式PCR的模板,两种PCR产物都用于构建基因文库。16S rRNA基因的V3区域也通过变性梯度凝胶电泳(DGGE)分析进行扩增,以分析细菌多样性。核糖体文库和DGGE图谱显示细菌多样性有限,主要包括柠檬酸杆菌属、肠杆菌属和嗜盐厌氧菌属,而从巢式PCR文库中检测到阿拉斯加脱硫弧菌和脱硫嗜盐弧菌属内的一个新分支。还采集生物膜样本用于分离SRB。分离出了阿拉斯加脱硫弧菌和毛细管脱硫弧菌,以及一些与柠檬酸杆菌相关的菌株。SRB在群落中所占比例非常小,脱硫弧菌属是SRB中相对丰富的群体。这是第一项直接探索与正常运行条件下的钢质管道相关的腐蚀性生物膜中细菌多样性的研究。

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