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[哥伦比亚里奥格兰德二号和拉费水库蓝藻检测的分子技术]

[Molecular techniques for cyanobacteria detection at Riogrande II and La Fe water reservoirs, Colombia].

作者信息

Hurtado-Alarcón Julio César, Polanía-Vorenberg Jaime

出版信息

Rev Biol Trop. 2014 Mar;62(1):403-19.

Abstract

In lentic water bodies as reservoirs occur eutrophication processes, originated mainly from human activities (i.e. agriculture, animal exploitation). This influx of nutrients in aquatic ecosystems could promote blooms of potentially toxic cyanobacteria. The purpose of this work is to detect the presence of cyanobacteria strains in water samples, using molecular techniques to help in preventive management of reservoirs dedicated to water purification. We used two molecular techniques to detect genes implied with the synthesis of hepatotoxic microcystins from potentially toxic cyanobacteria strains, and to evaluate the molecular diversity of cyanobacteria in water samples from two high-mountain reservoirs used for purification of drinking water for the metropolitan area of Medellin, Colombia. Between 2010-2011 collections of 12 water samples were taken and DNA extraction together with PCR and DGGE analyses where carried out. We amplified 22 sequences between 250-300bp of the genes mcyA and mcyE, and these sequences were related with several strains and cyanobacteria genera accessions from NCBI-GenBank databases. Moreover, sequence amplifications of the 16S small ribosomal RNA subunit - 16S rRNA- between 400-800bp were also performed in order to use them for the DGGE technique. The amplification products of DGGE were set in polyacrilamide gel with posterior denaturing electrophoresis, and the scanned images of the gel bands were analysed with the software GelCompar II. For Riogrande II and La Fe reservoirs we found 35 and 30 different DGGE bands, respectively, as a measurement of molecular diversity in these artificial ecosystems. Here, we demonstrated the utility of two molecular techniques for the detection of genes associated with toxicity and molecular diversity of cyanobacteria in reservoirs destined for drinking water in urban centers. We recommend strongly following with periodically molecular biology studies in these ecosystems combined with limnological and ecological data, as new tools for management of plants of water purification and for the prevention of potentially public health concerns.

摘要

在作为水库的静水水体中会发生富营养化过程,主要源于人类活动(即农业、畜牧业)。水生生态系统中这种营养物质的流入会促使具有潜在毒性的蓝藻大量繁殖。这项工作的目的是利用分子技术检测水样中蓝藻菌株的存在,以协助对用于水净化的水库进行预防性管理。我们使用两种分子技术来检测与潜在毒性蓝藻菌株合成肝毒素微囊藻毒素相关的基因,并评估来自哥伦比亚麦德林大都市区用于饮用水净化的两个高山水库水样中蓝藻的分子多样性。在2010 - 2011年期间采集了12份水样,并进行了DNA提取以及PCR和DGGE分析。我们扩增了基因mcyA和mcyE的22个250 - 300bp之间的序列,这些序列与NCBI - GenBank数据库中的多个菌株和蓝藻属登录号相关。此外,还进行了400 - 800bp之间的16S小核糖体RNA亚基 - 16S rRNA的序列扩增,以便用于DGGE技术。将DGGE的扩增产物置于聚丙烯酰胺凝胶中进行后续变性电泳,并使用GelCompar II软件分析凝胶条带的扫描图像。对于里奥格兰德二号水库和拉费水库,我们分别发现了35条和30条不同的DGGE条带,作为这些人工生态系统中分子多样性的一种衡量。在此,我们证明了两种分子技术在检测与城市中心用于饮用水的水库中蓝藻毒性和分子多样性相关基因方面的实用性。我们强烈建议结合湖泊学和生态学数据,定期对这些生态系统进行分子生物学研究,作为水净化厂管理和预防潜在公共卫生问题的新工具。

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