Analysis of the Bacterial Community of Metal Scrap Using an Enrichment Culture Approach

Hironaga Akita, Y. Shinto, Z. Kimura
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Abstract

Microbiologically influenced corrosion (MIC) of metal alloys is promoted by biofilms formed on metal surfaces. In the marine environment, MIC causes serious metal infrastructure problems, which lead to significant economic losses. In this study, we used an enrichment culture approach to examine the bacterial community that grows on metal surface at levels below the detection limit as a preliminary study for developing guidelines to prevent biofilm formation. An enrichment culture approach was employed to analyze the bacterial community on metal surface without biofilms and corrosion. Genomic DNA was extracted from culture sample after incubation in the enrichment culture with a metal piece, and then the V3–V4 variable regions of the bacterial 16S rRNA gene were amplified using the extracted genomic DNA as the template. Subsequently, using a next-generation sequencing approach, the amplified V3–V4 regions were sequenced, and the bacterial community was analyzed using the QIIME 2 microbiome bioinformatics platform. Using this enrichment culture approach, more than 80 bacterial genera were detected with Sphingomonas bacteria exhibiting the highest relative abundance (44%). These results demonstrated that this method could be useful for bacterial community analysis for bacteria below detection limits, and will serve as a basis for the development of the guidelines.
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用富集培养法分析金属废料细菌群落
金属合金的微生物影响腐蚀(MIC)是由金属表面形成的生物膜促进的。在海洋环境中,MIC会造成严重的金属基础设施问题,造成重大的经济损失。在这项研究中,我们使用富集培养方法来检查在低于检测限的水平下生长在金属表面的细菌群落,作为制定防止生物膜形成指南的初步研究。采用富集培养法对金属表面无生物膜和腐蚀的细菌群落进行了分析。用金属片富集培养后,从培养样品中提取基因组DNA,以提取的基因组DNA为模板扩增细菌16S rRNA基因的V3-V4可变区。随后,采用新一代测序方法,对扩增的V3-V4区进行测序,并使用QIIME 2微生物组生物信息学平台分析细菌群落。利用这种富集培养方法,检测到80多个细菌属,其中鞘氨单胞菌的相对丰度最高(44%)。这些结果表明,该方法可用于检测限以下细菌的细菌群落分析,并将为制定指南提供基础。
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