Prokaryotic Community Determination of Metal-Rich Acidic Environment by Comparative Methods

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 degrees 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.