Uncovering Viable Microbiome in Anaerobic Sludge Digesters by Propidium Monoazide (PMA)-PCR
Uncovering Viable Microbiome in Anaerobic Sludge Digesters by Propidium Monoazide (PMA)-PCR
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DOI：
10.1007/s00248-019-01449-w
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发表时间：
2019-11
影响因子：
3.6
通讯作者：
Jialing Ni;Shingo Hatori;Yin Wang;Yuyou Li;K. Kubota
中科院分区：
文献类型：
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作者：
Jialing Ni;Shingo Hatori;Yin Wang;Yuyou Li;K. Kubota
使用厌氧污泥消化器是世界各地利用城市污水污泥进行固体消化和产生甲烷的常见做法。了解微生物群落结构对于更好地了解厌氧消化过程和获得更好的过程控制至关重要。然而，对活微生物的选择性分析受到基于DNA的分析的限制。本研究采用单叠氮丙酸丙二钠(PMA)-聚合酶链式反应(PMA)-16S rRNA基因测序技术，以细胞膜完整性为基础，对活菌和死菌进行了鉴定。比较了PMA处理前后厌氧消化池污泥的微生物群落结构。定量聚合酶链式反应显示，5-30%的rRNA基因来自厌氧污泥消化器中的灭活或死亡细胞。与未经PMA处理的污泥相比，这导致可操作分类单元的数量以及Chao1和Shannon指数显著降低。微生物群落分析表明，大多数活的微生物群由Eurya chaeota，Bacteroidtes，DeltaProteacteria，Chloroflei，Firmicents，WWE1，螺旋藻，协生菌和Caldiserica组成。另一方面，经PMA治疗后，数字ofAlphaproteobacteriaandBetaproteobacteriadeclined.这些被认为是残留的微生物成员。网络分析还揭示了属于WWE1的OTUS与拟杆菌科之间的关系。基于PMA-PCR的16S rRNA基因测序分析是发现复杂环境样品中活性微生物群的有效工具。
Use of anaerobic sludge digester is a common practice around the world for solids digestion and methane generation from municipal sewage sludge. Understanding microbial community structure is vital to get better insight into the anaerobic digestion process and to gain better process control. However, selective analysis of viable microorganisms is limited by DNA-based assays. In this study, propidium monoazide (PMA)-PCR with 16S rRNA gene sequencing analysis was used to distinguish live and dead microorganisms based on cell membrane integrity. Microbial community structures of PMA-treated and PMA-untreated anaerobic digester sludge samples were compared. Quantitative PCR revealed that 5–30% of the rRNA genes were derived from inactive or dead cells in anaerobic sludge digesters. This caused a significant decrease in the numbers of operational taxonomic units and Chao1 and Shannon indices compared with that of the PMA-untreated sludge. Microbial community analysis showed that majority of the viable microbiome consisted ofEuryarchaeota,Bacteroidetes,Deltaproteobacteria,Chloroflexi,Firmicutes, WWE1,Spirochaetes,Synergistetes, andCaldiserica. On the other hand, after the PMA treatment, numbers ofAlphaproteobacteriaandBetaproteobacteriadeclined. These were considered residual microbial members. The network analysis also revealed a relationship among the OTUs belonging to WWE1 andBacteroidales. PMA-PCR-based 16S rRNA gene sequencing analysis is an effective tool for uncovering viable microbiome in complex environmental samples.