Abstract

Acid mine drainage (AMD) is a wastewater caused by mining activities and sulfide minerals oxidation. It occurs generally as a result of the relation of water, oxygen and microorganisms. In this study, the pH and the temperature of samples were determined around 2 and 20 °C, respectively. The chemical analysis was made by - ICP-MS. Sulfur concentration was found in higher amounts to be 21,892 ppm, 6155 ppm, 7893 ppm, and 31,305 ppm and the amounts of iron were also high with concentrations ranging from 14,318 ppm, 2181 ppm, 2715 ppm, and 13,801 ppm in samples including K2, K3, K4, and K5, respectively. Prokaryotic community fingerprints were obtained by DGGE and 16S rRNA gene clone library. High-throughput sequencing of 16S rRNA gene fragments and qPCR were performed by environmental DNAs. FISH was carried out by interacting probes. The extremely acidic four mine ponds contained high concentrations of sulfate and iron. Archaeal genera Acidiplasma sp. and Ferroplasma sp. with high abundance were determined in sulfate-rich sampling area according to metagenomic approach. Acidithiobacillus, Leptospirillum and Acidiphilium were often found in acid mine ponds. Otherwise, Sulfobacillus sp., Ferrithrix sp., Ferrimicrobium sp., Acidimicrobium sp., and Aciditerrimonas sp. were also obtained at the end of analyses. As a consequence, 16S clone library, metagenomic approach, qPCR and DGGE analysis were combined collaboratively, complementarily and comparatively for acidophilic prokaryotic diversity.

摘要

酸性矿山排水 (AMD) 是由采矿活动和硫化矿物氧化引起的废水。它通常是由于水、氧和微生物的关系而发生的。在本研究中，样品的 pH 值和温度分别确定在 2 和 20 °C 左右。通过-ICP-MS进行化学分析。发现硫浓度较高，分别为 21,892 ppm、6155 ppm、7893 ppm 和 31,305 ppm，铁含量也很高，样品中的浓度范围为 14,318 ppm、2181 ppm、2715 ppm 和 13,801 ppm，包括 K分别为 K3、K4 和 K5。通过DGGE和16S rRNA基因克隆文库获得原核生物群落指纹图谱。环境 DNA 对 16S rRNA 基因片段和 qPCR 进行高通量测序。FISH 是通过相互作用的探针进行的。极度酸性的四个矿池含有高浓度的硫酸盐和铁。古菌属酸浆菌属 和铁质体。根据宏基因组学方法在富含硫酸盐的采样区确定了高丰度。酸性矿池中经常发现酸性硫杆菌、钩端螺旋体和嗜酸菌。否则，硫化杆菌属，Ferrithrix属，Ferrimicrobium属，Acidimicrobium属，Aciditerrimonas SP。也在分析结束时获得。因此，将 16S 克隆文库、宏基因组方法、qPCR 和 DGGE 分析协同、互补和比较结合起来，以实现嗜酸性原核生物多样性。