应用Illumina MiSeq高通量测序技术分析活性银离子处理饮用水微生物多样性

Microbiota Diversity Analysis of Potable Water Treated with Silver Ion Using Illumina Miseq Sequencing Technology

为观察作为载人航天消毒剂的银离子消毒水所呈现的微生物多样性,采用Illumina MISeq高通量测序技术,对添加不同浓度的活性银离子的市售桶装水和对照水样进行了分析,分组为CK(对照)、L(0.1 mg/L)、M(0.3 mg/L)和H(0.5 mg/L)组,每隔30 d定期取样,同时以GB 4789-2010进行了菌落总数和大肠菌群数测定和16S r DNA V3-V4区高通量测序,以及生物信息学分析,包括群落结构、多样性、主成分及物种丰度分析。结果表明:CK组30 d菌落计数即达557.3±58.2,活性银离子有较为显著的抑菌效果(L组60 d后1.7±0.6、H组90 d后1.7±0.6);高通量测序数据显示,其群落结构比例最大的是变形菌门,其次是厚壁菌门,然后是放线菌门;beta多样性和主成分分析结果显示鞘氨醇单胞菌属和乳球菌属丰度较高;活性银离子对饮用水有显著的抑菌作用,且可减少细菌群落。

Silver ion is commonly used in water purification in human space flight missions.To ana-lyze and compare the microbiota diversity in potable water treated or untreated with silver ion,bar-relled water were treated with different concentrations of silver ion and analyzed by Illumina MISeq sequencing.The experiments were divided into the CK(Con),L(0.1mg/L),M(0.3mg/L)and H(0.5mg/L)groups and sampling was made once every30days.Genomic DNA of the samples was extracted and the16S rDNA V3-V4region was amplified by PCR and identified by Illumina.The bioinformatic analysis included the community composition,the beta diversity,the principal compo-nents,and species abundance analyses.The bacteria concentrations were lower in water treated with silver ion than that of the untreated.Protecobacteria was the predominant phylums.The two second most commonly identi?ed genera were Firmicutes and Actinobacteria.Beta diversity and principal components analysis showed that lower variations in community composition were presented in water treated with silver ion analyzed,while Sphingomonas and Lactococcus might be the top two species according to the abundance analysis.The data showed that silver ions could effectively suppress the reproduction and simplify the community composition of bacteria.