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Microbial Community Composition in Acid Mine Drainage Lake of Xiang This article was downloaded by: [Zhejiang University] On: 11 April 2013, At: 17:31 Publisher: Taylor & Francis Informa Ltd Registered in England and Wales Registered Number: 1072954 Registered office: Mortimer House, 37-41 Mortimer Street, London W1T 3JH, UK Geomicrobiology Journal Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/ugmb20 Microbial Community Composition in Acid Mine Drainage Lake of Xiang Mountain Sulfide Mine in Anhui Province, China Chunbo Hao a b , Lina Zhang a b , Lihua Wang a b , Siyuan Li a b & Hailiang Dong a c a Geomicrobiology Laboratory, State Key Laboratory of Geobiology and Environmental Geology, China University of Geosciences, Beijing, China b School of Water Resources and Environment, China University of Geosciences, Beijing, China c Department of Geology and Environmental Earth Science, Miami University, Oxford, Ohio, USA Accepted author version posted online: 18 Apr 2012.Version of record first published: 14 Sep 2012. To cite this article: Chunbo Hao , Lina Zhang , Lihua Wang , Siyuan Li & Hailiang Dong (2012): Microbial Community Composition in Acid Mine Drainage Lake of Xiang Mountain Sulfide Mine in Anhui Province, China, Geomicrobiology Journal, 29:10, 886-895 To link to this article: http://dx.doi.org/10.1080/01490451.2011.635762 PLEASE SCROLL DOWN FOR ARTICLE Full terms and conditions of use: http://www.tandfonline.com/page/terms-and-conditions This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. The publisher does not give any warranty express or implied or make any representation that the contents will be complete or accurate or up to date. The accuracy of any instructions, formulae, and drug doses should be independently verified with primary sources. The publisher shall not be liable for any loss, actions, claims, proceedings, demand, or costs or damages whatsoever or howsoever caused arising directly or indirectly in connection with or arising out of the use of this material. Geomicrobiology Journal, 29:886–895, 2012 Copyright © Taylor & Francis Group, LLC ISSN: 0149-0451 print / 1521-0529 online DOI: 10.1080/01490451.2011.635762 Microbial Community Composition in Acid Mine Drainage Lake of Xiang Mountain Sulfide Mine in Anhui Province, China Chunbo Hao,1, 2 Lina Zhang,1, 2 Lihua Wang,1, 2 Siyuan Li,1, 2 and Hailiang Dong1, 3 1Geomicrobiology Laboratory, State Key Laboratory of Geobiology and Environmental Geology, China University of Geosciences, Beijing, China 2School of Water Resources and Environment, China University of Geosciences, Beijing, China 3Department of Geology and Environmental Earth Science, Miami University, Oxford, Ohio, USA INTRODUCTION Acid mine drainage (AMD) lake of Xiang Mountain in An- Metalliferous mining activity has produced large quantities hui Province, China, was characterized by acidic waters (pH of sulfidic mineral wastes (such as pyrite or arsenopyrite) all around 2.8) containing high concentrations of soluble metals and over the world (Rowe et al. 2007). These mine wastes are con- sulfate. To investigate the function and dynamics of this ex- treme ecosystem, four water samples were collected from the tinuing sources of environmental contamination, mostly in the lake in the fall of 2010. The acidophilic community structure form of acid mine drainage (AMD), which is usually character- was analyzed by molecular approaches, and bacterial and ar- ized by low pH (below 3.0). Such waste waters generally contain chaeal clone libraries of 16S rRNA genes were constructed. In high levels of iron, toxic metals such as aluminum, manganese, contrast to dominance of chemolithotrophic acidophiles in typi- lead, cadmium, and zinc, as well as metalloids, of which ar- cal AMD environments, autotrophic iron/sulfur-oxidizing bacte- ria were detected in only one sample with low abundance. Un- senic is generally of the greatest concern (Bruneel et al. 2011). expectedly, the Cyanobacteria group was the predominant in all AMD is generated when waste sulfide ores come into contact four samples (54.9%−77%). Chemoheterotrophs Acidiphilium and with oxygenated water. The process can be greatly enhanced by Acidisphaera were also abundant. These two heterotrophic groups microbial activity because many acidophilic microorganisms contain bacteriochlorophyll that can perform photosynthesis, an can accelerate dissolution of sulfide minerals through oxidation advantage to grow and survive in such oligotrophic acidic envi- ronments. Only two clone sequences related to Legionella (2.8% of of ferrous iron and/or reduced forms of sulfur (Johnson and the total clones) were recovered from one sample in sharp contrast Hallberg 2003). to its higher abundance (12.7%) in the summer of 2009. All ar- The culture-independent molecular techniques for studying chaeal sequences were affiliated to the phylum Crenarchaeota.The microbial ecology has been well established and applied world- results of statistical analysis suggested that the water chemistry wide in the last decade. Many researchers have employed molec- of the AMD lake controlled microbial composition of the AMD Downloaded by [Zhejiang University] at 17:31 11 April 2013 ecosystem. ular techniques to analyze acidophile populations and have described microbial community structures in various acidic en- Keywords acid mine drainage, acidophile, 16S rRNA, clone library, vironments (Demergasso et al. 2005; Gonzalez-Toril et al. 2003; Cyanobacteria Hao et al. 2010a; Rzhepishevska et al. 2005). The power of these techniques lies in their ability to identify and quantify microorganisms without cultivation, which may lead to biased conclusions about the structure and dynamics of microbial com- Received 19 July 2011; accepted 21 September 2011. munities. By the application of these cultivation-independent This research was supported by grants from the National Sci- molecular techniques, our understanding of the biodiversity of ence Foundation of China (40802059, 41030211), the Specialized Re- search Funds for the Doctoral Program of Higher Education of China acidophiles has been rapidly expanding, which has given more (200804911008), and the Fundamental Research Funds for the Central comprehensive insights into the role of acidophilic microorgan- Universities (2010ZD03, 2011YXL035). The authors are grateful to isms in mediating elemental cycling in acidic ecosystems (Bond two anonymous reviewers whose comments improved the quality of et al. 2000; Bruneel et al. 2006; Hallberg et al. 2006; He et al. the manuscript. 2007). Address correspondence to Hailiang Dong, Geomicrobiology Lab- oratory, State Key Laboratory of Geobiology and Environmental Geol- In a previous study, we characterized the planktonic aci- ogy, China University of Geosciences, Beijing 100083, China. E-mail: dophilic community in a water sample collected from the AMD [email protected] lake, Anhui Province, China, in the summer of 2009 using 886 MICROBIAL COMMUNITY IN ACID MINE DRAINAGE LAKE 887 the molecular approaches, and found that heterotrophic aci- The geochemical samples were kept at 4◦C and shipped to dophiles were dominant (>50% of total bacterial population) the Geomicrobiology Laboratory of China University of Geo- while autotrophic organisms were scarce. Paucity of ferrous sciences in Beijing on the same day. The following analyses iron and turbid water may have accounted for low abundance were conducted within 2 days of collection. For sulfate and of chemoautotrophic and photosynthetic organisms. Abundant metal ions, the water samples were filtered through 0.45 µm heterotrophic acidophiles may be due to input of allochthonous filter prior to analyses. Sulfate concentration was determined dissolved organic carbon from soils and grass community in by ion chromatography; dissolved metals by inductively cou- the bank of the AMD lake (Hao et al. 2010b). Moreover, it pled plasma-atomic emission spectroscopy (ICP-AES, JYl- was unexpected that many sequences of Legionella, which can ULTIMA, France); dissolved ferrous iron titrated with ceric cause Legionnaires’ disease, were also detected in the AMD lake sulfate. Total organic carbon (TOC) was measured by an or- (Hao et al. 2010b). The high abundance of Legionella-related ganic carbon analyzer (Shimadzu, Japan). Total nitrogen (TN), + sequences was ascribed to the presence of a large number of total phosphorous (TP), and NH4 -N were determined in ac- protozoa in eukaryotic population. However, it was unclear if cordance with standard methods (Ministry of Environmental this community structure was transient in nature or stable over Protection 2002). long periods of time. The microbial samples (2.5 liter in volume in sterile plastic The objective of this study was therefore to examine if the bottles) were kept on regular ice for one day before they were community structure observed in the summer of 2009 remained filtered through a Millex-GS Millipore filter (pore size, 0.22 µm; stable over time. If the microbial structure was different, what diameter, 50 mm) in the lab. The filters were used immediately were the environmental conditions that were responsible for the for extraction of microbial genomic DNA. change? Similar molecular methods as those used in our earlier study (Hao et al. 2010b) were employed in this study. Our results DNA Extraction and Purification showed that the
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