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Live-dead discrimination analysis, qPCR assessment for opportunistic pathogens, and population analysis at ozone wastewater treatment plants

机译：活死判别分析，机会病原体的qPCR评估以及臭氧废水处理厂的种群分析

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In respect to direct and indirect water reuse, the microbiological quality of treated wastewater is highly important. Conventional wastewater treatment plants are normally not equipped with advanced technologies for the elimination of bacteria. Molecular biology analyses were combined with live-dead discrimination analysis of wastewater population using Propidium monoazide (PMA) to study population shifts during ozonation (1 g ozone/g DOC) at a municipal wastewater treatment plant. Escherichia coli, enterococci, and Pseudomonas aeruginosa were quantified by polymerase chain reaction (qPCR) and the whole wastewater population was analyzed by metagenomic sequencing. The PMA-qPCR experiments showed that the abundances of P. aeruginosa didn't change by ozone treatment, whereas a reduction was observed for E. coli and enterococci. Results comparing conventional cultivation experiments with PMA-qPCR underlined the presence of viable but not culturable cells (VBNC) and their regrowth potential after ozone treatment. alumina HiSeq sequencing results with and without PMA treatment demonstrated high population similarities in water samples originating from ozone inflow sampling sides. Upon using PMA treatment after ozonation, population shifts became visible and also underlined the importance of PMA treatment for the evaluation of elimination and selection processes during ozonation at WWTPs. Amongst a number of 14 most abundant genera identified in the inflow samples, 9 genera were found to be reduced, whereas 4 genera increased in relative abundance and 1 genus almost remained constant. The strongest increase in relative abundance after ozonation was detected for Oscillatoria spp., Microcoleus spp. and Nitrospira spp. Beside this, a continuous release of Pseudomonas spp. (including P. aeruginosa) to the downstream receiving body was confirmed. Regrowth experiments demonstrated a high prevalence of P. aeruginosa as part of the surviving bacterial population. Summing up, molecular biology analyses in combination with live-dead discrimination are comprehensive methods to evaluate the elimination processes targeting specific species and/or whole microbial populations. (C) 2017 Elsevier Ltd. All rights reserved.
机译：对于直接和间接的水回用，处理后的废水的微生物质量非常重要。传统的废水处理厂通常不配备用于消除细菌的先进技术。使用单叠氮化丙锭（PMA）将分子生物学分析与活泼的废水人口判别分析相结合，以研究市政废水处理厂在臭氧化过程中的人口迁移（1 g臭氧/ g DOC）。通过聚合酶链反应（qPCR）对大肠杆菌，肠球菌和铜绿假单胞菌进行定量，并通过宏基因组测序对整个废水种群进行分析。 PMA-qPCR实验表明，通过臭氧处理，铜绿假单胞菌的丰度没有变化，而大肠杆菌和肠球菌的丰度却降低了。将常规培养实验与PMA-qPCR进行比较的结果强调了存在但不能培养的细胞（VBNC）的存在及其在臭氧处理后的再生潜力。使用和不使用PMA处理的氧化铝HiSeq测序结果表明，源自臭氧流入采样侧的水样品具有很高的群体相似性。在臭氧化后使用PMA处理后，种群转移变得明显，这也突出了PMA处理对于污水处理厂臭氧化过程中消除和选择过程的评估的重要性。在流入样品中鉴定的14个最丰富属中，有9个属减少了，而4个属的相对丰度增加了，而1个属几乎保持不变。臭氧处理后，Oscillatoria spp。，Microcoleus spp的相对丰度增加最强。和硝化螺旋菌属。除此之外，Pseudomonas spp的持续释放。 （包括铜绿假单胞菌）到达下游接收体。再生实验表明，铜绿假单胞菌作为幸存细菌种群的一部分很高。总而言之，分子生物学分析与活死人识别相结合是评估针对特定物种和/或整个微生物种群的消除过程的综合方法。 （C）2017 Elsevier Ltd.保留所有权利。

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  • 来源
    《Environmental Pollution》 |2018年第1期|571-579|共9页
  • 作者

    Jaeger Thomas; Alexander Johannes; Kirchen Silke; Doetsch Andreas; Wieland Arne; Hiller Christian; Schwartz Thomas;

  • 作者单位

    KIT, IFG, Dept Bioengn & Biosyst, Campus North,POB 3640, D-76021 Karlsruhe, Germany;

    KIT, IFG, Dept Bioengn & Biosyst, Campus North,POB 3640, D-76021 Karlsruhe, Germany;

    KIT, IFG, Dept Bioengn & Biosyst, Campus North,POB 3640, D-76021 Karlsruhe, Germany;

    KIT, IFG, Dept Bioengn & Biosyst, Campus North,POB 3640, D-76021 Karlsruhe, Germany;

    Xylem Serv GmbH, Boschstr 4-14, D-32051 Herford, Germany;

    Zweckverband Klarwerk Steinhaule, Reinzstr 1, D-89233 Neu Ulm, Germany;

    KIT, IFG, Dept Bioengn & Biosyst, Campus North,POB 3640, D-76021 Karlsruhe, Germany;

  • 收录信息 美国《科学引文索引》(SCI);美国《工程索引》(EI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
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