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利用颗粒状活性炭和蒙脱石黏土设计和验证被动式环境 DNA 采样器。

Design and Validation of Passive Environmental DNA Samplers Using Granular Activated Carbon and Montmorillonite Clay.

机构信息

Department of Civil, Structural, and Environmental Engineering, The State University of New York at Buffalo, Buffalo, New York 14228, United States.

Department of Engineering, University of San Francisco, San Francisco, California 94117, United States.

出版信息

Environ Sci Technol. 2020 Oct 6;54(19):11961-11970. doi: 10.1021/acs.est.0c01863. Epub 2020 Sep 9.

Abstract

Environmental DNA (eDNA) analysis is gaining prominence as a tool for species and biodiversity monitoring in aquatic environments. eDNA shed by organisms is captured in grab samples, concentrated by filtration, extracted, and analyzed using molecular methods. Conventional capture and filtration methods are limited because (1) filtration does not capture all extracellular DNA, (2) eDNA can degrade during sample transport and storage, (3) filters often clog in turbid waters, reducing the eDNA captured, and (4) grab samples are time sensitive due to pulse eDNA inputs. To address these limitations, this work designs and validates Passive Environmental DNA Samplers (PEDS). PEDS consist of an adsorbent-filled sachet that is suspended in water to collect eDNA over time. Both extracellular and cellular DNA are captured, and the extracellular DNA is protected from degradation. The eDNA captured over time may be more representative than a grab sample. Two adsorbents, Montmorillonite Clay (MC) and Granular Activated Carbon (GAC), are tested. In laboratory experiments, MC-PEDS adsorbed five times more extracellular DNA and desorbed up to four times more than GAC-PEDS (despite high levels of eDNA loss during desorption). In microcosm and field experiments, GAC-PEDS captured over an order of magnitude more eDNA than MC-PEDS. Field results further validated PEDS as an effective eDNA capture method compared to conventional methods.

摘要

环境 DNA(eDNA)分析作为一种在水生环境中监测物种和生物多样性的工具,正变得越来越重要。生物体释放的 eDNA 在抓取样本中被捕获,通过过滤浓缩,然后使用分子方法进行提取和分析。传统的捕获和过滤方法存在局限性,原因如下:(1) 过滤无法捕获所有细胞外 DNA;(2) eDNA 在样本运输和储存过程中会降解;(3) 过滤器在浑浊的水中经常堵塞,减少了捕获的 eDNA;(4) 由于脉冲 eDNA 的输入,抓取样本具有时间敏感性。为了解决这些限制,本工作设计并验证了被动环境 DNA 采样器(PEDS)。PEDS 由一个装有吸附剂的小袋组成,该小袋被悬挂在水中,以随时间收集 eDNA。细胞外和细胞内 DNA 都被捕获,并且细胞外 DNA 受到保护,不会降解。随时间捕获的 eDNA 可能比抓取样本更具代表性。两种吸附剂,蒙脱石粘土(MC)和颗粒状活性炭(GAC),进行了测试。在实验室实验中,MC-PEDS 吸附的细胞外 DNA 是 GAC-PEDS 的五倍,尽管在解吸过程中 eDNA 大量损失,但解吸量是 GAC-PEDS 的四倍。在微宇宙和现场实验中,GAC-PEDS 捕获的 eDNA 比 MC-PEDS 多一个数量级。现场结果进一步验证了 PEDS 作为一种有效的 eDNA 捕获方法,与传统方法相比具有优越性。

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