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High Throughput Analysis of Integron Gene Cassettes in Wastewater Environments

机译:废水环境中整合子基因盒的高通量分析

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摘要

Integrons are extensively targeted as a proxy for anthropogenic impact in the environment. We developed a novel high-throughput amplicon sequencing pipeline that enables characterization of thousands of integron gene cassette-associated reads, and. applied, it toacquire-a comprehensive overview of gene,. cassette composition in effluents from wastewater treatment facilities across Europe. Between 38 100 and 172995 reads per- sample were generated and functionally characterized by screening against nr, SEED, ARDB and/?-lactamase databases. Over 75% of the reads were characterized as hypothetical, but thousands were associated with toxin-antitoxin systems, DNA repair, cell membrane function, detoxification and aminoglyco- side and/Mactam resistance. Among the reads characterized as fi- lactamases, the carbapenemase bla_(OXA) was dominant in most of the effluents, except for Cyprus and Israel where bla_(GES) was also abundant. Quantitative PCR assessment of bla_(OXA) and bla_(GES) genes in the European effluents revealed similar trends to those displayed in the integron amplicon sequencing pipeline described above, corroborating the robustness of this method and suggesting that these integron-associated genes may be excellent targets for source tracking of effluents in downstream environments. Further application of the above analyses revealed several order-of-magnitude reductions in effluent-associated β- lactamase genes in effluent-saturated soils, suggesting marginal persistence in the soil microbiome.
机译:整数被广泛用作人类对环境影响的替代指标。我们开发了一种新颖的高通量扩增子测序管线,可表征成千上万个与整合子基因盒相关的读数,并且。应用后,它获得了基因的全面概述。整个欧洲废水处理设施的废水中的卡式盒组成。通过对nr,SEED,ARDB和β-内酰胺酶数据库进行筛选,每个样品产生了38 100至172995个读数,并在功能上进行了表征。超过75%的读数被认为是假设的,但成千上万的与毒素-抗毒素系统,DNA修复,细胞膜功能,排毒以及氨基糖苷和/或Mactam耐药有关。在以半乳糖苷酶为特征的读物中,碳青霉烯酶bla_(OXA)在大多数废水中占主导地位,除了塞浦路斯和以色列,bla_(GES)含量也很高。欧洲废水中bla_(OXA)和bla_(GES)基因的定量PCR评估显示了与上述整合子扩增子测序管线中显示的趋势相似的趋势,从而证实了该方法的稳健性,并表明这些与整合子相关的基因可能是出色的下游环境中废水来源追踪的目标。上述分析的进一步应用揭示了污水饱和土壤中与污水相关的β-内酰胺酶基因的数量级降低,表明土壤微生物组中存在一定程度的持久性。

著录项

  • 来源
    《Environmental Science & Technology》 |2016年第21期|11825-11836|共12页
  • 作者单位

    The Institute of Soil, Water and Environmental Sciences, The Volcani Center, Agricultural Research Organization, Bet-Dagan, Israel ,The Department of Soil and Water Sciences, The Robert H. Smith Faculty of Agriculture, Food and Environment, The Hebrew University of Jerusalem, Rehovot, Israel;

    The Department of Soil and Water Sciences, The Robert H. Smith Faculty of Agriculture, Food and Environment, The Hebrew University of Jerusalem, Rehovot, Israel;

    DNA Services Facility, Research Resources Center, University of Illinois at Chicago, Chicago, Illinois 60612, United States;

    Escola Superior de Biotecnologia, Universidade Catolica Portuguesa, Lisboa, Portugal;

    Faculty of Environmental Sciences, Technische Universitaet Dresden, Dresden, Germany;

    Faculty of Environmental Sciences, Technische Universitaet Dresden, Dresden, Germany;

    CNRS, Laboratoire de Chimie Physique et Microbiologie pour l'Environnement (LCPME), UMR 7564, Institut Jean Barriol, 15 Avenue du Charmois, 54500 Vandoeuvre-les-Nancy, France ,Universite de Lorraine, LCPME, UMR 7564, 15 Avenue du Charmois, 54500 Vandoeuvre-les-Nancy, France;

    Institute for Water Quality, Resources and Waste Managment, Technische Universitaet Wien, Wien, Austria;

    Karlsruhe Institute of Technology, Eggenstein-Leopoldshafen, Germany;

    Department of Civil and Environmental Engineering and Nireas, International Water Research Center, University of Cyprus, P.O. Box 20537, 1678 Nicosia, Cyprus;

    Department of Civil Engineering, University of Salerno, Salerno, Italy;

    Norwegian Institute for Water Research, Oslo, Norway;

    School of Science and Technology, Middlesex University, London, U.K.;

    The Department of Plant Pathology and Microbiology, The Robert H. Smith Faculty of Agriculture, Food and Environment, The Hebrew University of Jerusalem, Rehovot, Israel;

    The Institute of Soil, Water and Environmental Sciences, The Volcani Center, Agricultural Research Organization, Bet-Dagan, Israel;

  • 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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