Acid Mine Drainage Lakes Research Articles

Metagenomic insight into the acidophilic functional communities driving elemental geochemical cycles in an acid mine drainage lake

Acidophiles play a key role in the generation, evolution and attenuation of acid mine drainage (AMD), which is characterized by strong acidity (pH<3.5) and high metal concentrations. In this study, the seasonal changes of acidophilic communities and their roles in elemental cycling in an AMD lake (pH∼3.0) in China were analyzed through metagenomics. The results showed eukaryotic algae thrived in the lake, and Coccomyxa was dominant in January (38.1%) and May (33.9%), while Chlorella in July (9.5%). The extensive growth of Chlamydomonas in December (22.7%) resulted in an ultrahigh chlorophyll a concentration (587 μg/L), providing abundant organic carbon for the ecosystem. In addition, the iron-oxidizing and nitrogen-fixing bacterium Ferrovum contributed to carbon fixation. Ammonia oxidation likely occurred in the acidic lake, as was revealed by archaea Ca. Nitrosotalea. To gain a competitive advantage in the nutrient-poor environment, some acidophiles exhibited facultative characteristics, e.g. the most abundant bacterium Acidiphilium utilized both organic and inorganic carbon, and obtained energy from organic matter, inorganic sulfur, and sunlight simultaneously. It was suggested that sunlight, rather than chemical energy of reduced iron-sulfur was the major driver of elemental cycling in the AMD lake. The results are beneficial to the development of bioremediation strategies for AMD.

New Insights into Microbial Interactions with Dissolved Organic Matter in Acid Mine Drainage with the Integration of Microbial Community and Chemical Composition Analysis

Acid mine drainage (AMD) environments are one of the most extreme ecosystems on earth, and their biogeochemical cycling has been widely studied. However, the carbon cycling underlying microbial interactions with dissolved organic matter (DOM) is poorly understood in these environments. Here, we evaluated the relationships between microbial populations and DOM pools in an AMD lake by combining high-throughput microbial and chemical compositional analyses. The results demonstrated that microalgae were the predominant primary producers in the system and significantly contributed to the total DOM concentration in the water column, whereas DOM chemodiversity was correlated with bacterial community diversity. Network analyses further revealed that DOM associated with microalgae had low diversity but high relative abundance, suggesting microalgal selective organic matter contribution to the DOM pool. Conversely, diverse DOM molecules with generally low abundance were associated with heterotrophic acidophiles, including members of Alphaproteobacteria, Actinobacteria, Firmicutes, and Acidobacteria. Moreover, microalgae and bacteria exhibited predominantly positive co-occurrences with potential metabolic interactions via DOM exchange. Taken together, this study provides new insights into the microbe–DOM interactions in extreme AMD ecosystems and has significant implications for understanding the carbon cycle and microbial metabolism in extreme environments.

Vertical environmental gradient drives prokaryotic microbial community assembly and species coexistence in a stratified acid mine drainage lake

Acid mine drainage (AMD) lakes are typical hydrologic features caused by open pit mining and represent extreme ecosystems and environmental challenges. Little is known about microbial distribution and community assembly in AMD lakes, especially in deep layers. Here, we investigated prokaryotic microbial diversity and community assembly along a depth profile in a stratified AMD lake using 16S rRNA gene sequencing combined with multivariate ecological and statistical methods. The water column in the AMD lake exhibited tight geochemical gradients, with more acidic surface water. Coupled with vertical hydrochemical variations, prokaryotic microbial community structure changed significantly, and was accompanied by increased diversity with depth. In the surface water, heterogeneous selection was the most important assembly process, whereas stochastic processes gained importance with depth. Meanwhile, microbial co-occurrences, especially positive interactions, were more frequent in the stressful surface water with reduced network modularity and keystone taxa. The pH was identified as the key driver of microbial diversity and community assembly along the vertical profile based on random forest analysis. Taken together, environmental effects dominated by acid stress drove the community assembly and species coexistence that underpinned the spatial scaling patterns of AMD microbiota in the lake. These findings demonstrate the distinct heterogeneity of local prokaryotic microbial community in AMD lake, and provide new insights into the mechanism to maintain microbial diversity in extreme acidic environments.

Response of soil protozoa to acid mine drainage in a contaminated terrace

Acid mine drainage (AMD) system represents one of the most unfavorable habitats for microorganisms due to its low pH and high concentrations of metals. Compared to bacteria and fungi, our understanding regarding the response of soil protozoa to such extremely acidic environments remains limited. This study characterized the structures of protozoan communities inhabiting a terrace heavily contaminated by AMD. The sharp environmental gradient of this terrace was generated by annual flooding from an AMD lake located below, which provided a natural setting to unravel the environment-protozoa interactions. Previously unrecognized protozoa, such as Apicomplexa and Euglenozoa, dominated the extremely acidic soils, rather than the commonly recognized members (e.g., Ciliophora and Cercozoa). pH was the most important factor regulating the abundance of protozoan taxa. Metagenomic analysis of protozoan metabolic potential showed that many functional genes encoding for the alleviation of acid stress and various metabolic pathways were enriched, which may facilitate the survival and adaptation of protozoa to acidic environments. In addition, numerous co-occurrences between protozoa and bacterial or fungal taxa were observed, suggesting shared environmental preferences or potential bio-interactions among them. Future studies are required to confirm the ecological roles of these previously unrecognized protozoa as being important soil microorganisms.

Responses of Acidophilic Communities in Different Acid Mine Drainages to Environmental Conditions in Nanshan Mine, Anhui Province, China

Acidophiles play a vital catalytic role in the formation of acid mine drainage (AMD). In this study, acidophilic communities in a stream, two lakes and five ponds in Nanshan Mine, Anhui Province, China were comparatively investigated. Among the three types of AMDs, the stream which originated from a rock wall had the strongest acidity (pH 2.30–2.42), while the ponds (pH 2.35–2.60) formed in mine dump and the two large-scale AMD lakes (pH 2.66–3.05) were less acidic. The chemolithotrophic iron-oxidizer Leptospirillum dominated at the stream starting point with the lowest pH and highest Fe content. However, with streamflow, pH increased and Fe concentration decreased dramatically, and the dominance of Leptospirillum was replaced by the less acid-resistant iron-oxidizer Ferrovum. In contrast to the stream, prokaryotic communities in the ponds were mostly dominated by chemoheterotrophic Acidiphilium because of the abundant organic matter from surrounding mine waste. Both chemolithotrophic Ferrovum and chemoheterotrophic Acidiphilium were abundant in the AMD lakes, which may form a symbiotic relationship around the carbon cycle. Statistical analyses indicated that pH was the predominant environmental factor influencing the prokaryotic communities. As for the eukaryotic communities, the microeukaryotes in the stream were mostly uncultured groups. Both photoautotrophic algae Chrysonebula and chemoheterotrophic fungi were abundant in the ponds, while the two AMD lakes were only dominated by photoautotrophic algae Coccomyxa and Ochromonas.

The Stratified Distribution of Dissolved Organic Matter in an AMD Lake Revealed by Multi-sample Evaluation Procedure.

As a typical extreme environment, acid mine drainage (AMD) has been extensively studied for its biogeochemical cycle, but little is known about the quality of dissolved organic matter (DOM) in AMD. In this study, DOM molecules in an AMD lake were detected with Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS), and the change of DOM molecules in the stratified water column was analyzed with a multi-sample evaluation procedure. The results demonstrate that DOM quality is highly stratified and can be linked with severe biogeochemical gradients. In the surface layer, DOM molecules can be distinguished by low quantities and intensities, as well as potential photodegradation products. Oxygen-poor and oxygen-rich molecules alternately dominate the chemocline, which can be explained by the redox-dependent adsorption/desorption of DOM on metastable secondary minerals. A rich and abundant DOM pool with a high proportion of heteroatoms exists at the bottom which can be significantly influenced by material exchange with sediments. These findings emphasize the active role of DOM in extreme AMD environments and expand the understanding of the carbon cycle in the hydrosphere.

Contrasting seasonal variations of geochemistry and microbial community in two adjacent acid mine drainage lakes in Anhui Province, China.

Acid mine drainage (AMD) is generated by the bio-oxidation of sulfide minerals. To understand the AMD formation and evolution, it is necessary to determine the composition and variation of acidophilic community, and their role in AMD ecosystem. In this study, we compared seasonal variations of geochemistry and microbial composition of two adjacent AMD lakes with different formation histories in Anhui Province, China. Lake Paitu (PT) formed in 1970s near a mine dump and the pH was in the range of 3.01-3.16, with the lowest in spring and summer while the highest in winter. The main ions in PT were Al and SO42-, whereas Fe concentration was relatively low. The concentrations of these ions were the lowest in summer and the highest in winter. Lake Tafang (TF) formed in around 2013 in a pit was more acidic (pH 2.43-2.75), but the seasonal variation of pH was the same as PT. Compared with Lake PT, TF had higher Fe, lower Al and SO42- concentrations, and showed no significant seasonal changes. Despite salient seasonal variations of prokaryotic composition in Lake PT, Ferrovum was the major iron-oxidizing bacterium in most seasons. Furthermore, Lake PT was also rich in heterotrophic bacteria (48.6±15.9%). Both prokaryotic diversity and evenness of Lake TF were lower than PT, and chemolithotrophic iron-oxidizing bacteria (71.7±25.4%) were dominant in almost all samples. Besides Ferrovum, more acid tolerant iron-oxidizer Leptospirillum and Acidithiobacillus were also abundant in Lake TF. Chlamydomonas was the major eukaryote in Lake PT and it flourished repeatedly at the end of December, causing an extremely high chlorophyll a concentration (587μg/L) at one sampling site in 2016, which provided rich nutrients for heterotrophic bacteria. The main alga in Lake TF was Chrysonebula, but its concentration was low, apparently because of the strong acidity and dark red color of lake water.

Molecular Research of Acid-Generating Microbial Communities in Abandoned Ores in the Waste Dump of an Iron Mine in Anhui Province

The waste dump of an iron mine in Anhui Province has been abandoned for several decades. Pyrite in the exposed waste ores is oxidized by acidophiles and large amounts of metal ions and H2SO4 are released, resulting in the formation of an acid mine drainage (AMD) lake since 1970s. Besides the lake, there are also some small-scale AMD adjacent to the newly deposited waste ore. In order to study the acid generation potential of the waste ore and the related microbial communities, soil samples were taken from beside the AMD lake (1LL) and small-scale AMD (5J, 5Y, 6-1, 6-2, 6-3) and the physicochemical properties and microbial community of these samples were analyzed. The results reveal that all of samples were highly acidic and the pH of the 1LL sample was 2.77, while the other samples were even more acidic, at less than 2.6. The electrical conductivity (EC) (0.32 mS·cm-1) of the 1LL sample was obviously lower than the other samples (2.25-7.08 mS·cm-1), which indicates that the newly deposited waste ore contains higher ion concentrations. The Fe2+ concentration of the 1LL sample was only 0.80 mg·kg-1 but the other five samples were as high as 2.91-33.40 mg·kg-1. This suggests that most of the Fe2+ in the 1LL sample has been converted to Fe3+ after long-term oxidization. High-throughput sequencing results showed that most acidophiles in 1LL sample were Actinobacteria, Acidobacteria and Chloroflexi but the microbes in the remaining five samples were γ-Proteobacteria, Firmicutes and Nitrospira. The iron-sulfur oxidizing bacteria, such as Sulfobacillus, Leptospirillum, Acidithiobacillus, were scarce in the 1LL sample, while they highly abundant in the other five samples, which proves that the acid-generation process of the newly deposited waste ore is strong. However, the reduced iron and sulfur in the 1LL sample has nearly been depleted. Statistical analysis shows that the microbial composition of the 1LL sample is significantly different to that of the five newly deposited samples, illustrating that microbial community composition is remarkably influenced by physicochemical conditions.

Significant seasonal variations of microbial community in an acid mine drainage lake in Anhui Province, China

Acid mine drainage (AMD),characterized by strong acidity and high metal concentrations, generates from the oxidative dissolution of metal sulfides, and acidophiles can accelerate the process significantly. Despite extensive research in microbial diversity and community composition, little is known about seasonal variations of microbial community structure (especially micro eukaryotes) in response to environmental conditions in AMD ecosystem. To this end, AMD samples were collected from Nanshan AMD lake, Anhui Province, China, over a full seasonal cycle from 2013 to 2014, and water chemistry and microbial composition were studied. pH of lake water was stable (∼3.0) across the sampling period, while the concentrations of ions varied dramatically. The highest metal concentrations in the lake were found for Mg and Al, not commonly found Fe. Unexpectedly, ultrahigh concentration of chlorophyll a was measured in the extremely acidic lake, reaching 226.43–280.95 μg/L in winter, even higher than those in most eutrophic freshwater lakes. Both prokaryotic and eukaryotic communities showed a strong seasonal variation. Among the prokaryotes, “Ferrovum”, a chemolithotrophic iron-oxidizing bacterium was predominant in most sampling seasons, although it was a minor member prior to September, 2012. Fe2+ was the initial geochemical factor that drove the variation of the prokaryotic community. The eukaryotic community was simple but varied more drastically than the prokaryotic community. Photoautotrophic algae (primary producers) formed a food web with protozoa or flagellate (top consumers) across all four seasons, and temperature appeared to be responsible for the observed seasonal variation. Ochromonas and Chlamydomonas (responsible for high algal bloom in winter) occurred in autumn/summer and winter/spring seasons, respectively, because of their distinct growth temperatures. The closest phylogenetic relationship between Chlamydomonas species in the lake and those in Arctic and Alpine suggested that the native Chlamydomonas species may have been both acidophilic and psychrophilic after a long acclimation time in this extreme environment.

Microbial Community Composition in Acid Mine Drainage Lake of Xiang Mountain Sulfide Mine in Anhui Province, China

Acid mine drainage (AMD) lake of Xiang Mountain in Anhui Province, China, was characterized by acidic waters (pH around 2.8) containing high concentrations of soluble metals and sulfate. To investigate the function and dynamics of this extreme ecosystem, four water samples were collected from the lake in the fall of 2010. The acidophilic community structure was analyzed by molecular approaches, and bacterial and archaeal clone libraries of 16S rRNA genes were constructed. In contrast to dominance of chemolithotrophic acidophiles in typical AMD environments, autotrophic iron/sulfur-oxidizing bacteria were detected in only one sample with low abundance. Unexpectedly, the Cyanobacteria group was the predominant in all four samples (54.9%−77%). Chemoheterotrophs Acidiphilium and Acidisphaera were also abundant. These two heterotrophic groups contain bacteriochlorophyll that can perform photosynthesis, an advantage to grow and survive in such oligotrophic acidic environments. Only two clone sequences related to Legionella (2.8% of the total clones) were recovered from one sample in sharp contrast to its higher abundance (12.7%) in the summer of 2009. All archaeal sequences were affiliated to the phylum Crenarchaeota. The results of statistical analysis suggested that the water chemistry of the AMD lake controlled microbial composition of the AMD ecosystem.

Carbon isotope fractionation in phospholipid fatty acid biomarkers of bacteria and fungi native to an acid mine drainage lake

This study identifies isotope signatures associated with autotrophic and heterotrophic microbial communities that may provide a means to determine carbon cycling relationships in situ for acid mine drainage (AMD) sites. Stable carbon isotope ratios (δ13C) of carbon sources, bulk cells, and membrane phospholipids (PLFA) were measured for autotrophic and heterotrophic microbial enrichment cultures from a mine tailings impoundment in northern Ontario, Canada, and for pure strains of the sulfur oxidizing bacteria Acidithiobacillus ferrooxidans and Acidithiobacillus thiooxidans. The autotrophic enrichments had indistinguishable PLFA distributions from the pure cultures, and the PLFA cyc-C19:0 was determined to be a unique biomarker in this system for these sulfur oxidizing bacteria. The PLFA distributions produced by the heterotrophic enrichments were distinct from the autotrophic distributions and the C18:2 PLFA was identified as a biomarker for these heterotrophic enrichments. Genetic analysis (16S, 18S rRNA) of the heterotrophic cultures indicated that these communities were primarily composed of Acremonium fungi.Stable carbon isotope analysis revealed that bulk cellular material in all autotrophic cultures was depleted in δ13C by 5.6–10.9‰ relative to their atmospheric CO2 derived carbon source, suggesting that inorganic carbon fixation in these cultures is carbon limited. Individual PLFA from these autotrophs were further depleted by 8.2–14.6‰ compared to the bulk cell δ13C, which are among the largest biosynthetic isotope fractionation factors between bulk cell and PLFA reported in the literature. In contrast, the heterotrophic bulk cells were not significantly fractionated in δ13C relative to their carbon source and heterotrophic PLFA ranged from 3‰ enriched to 4‰ depleted relative to the isotopic composition of their total biomass. These distinct PLFA biomarkers and isotopic fractionations associated with autotrophic and heterotrophic activity in this laboratory study provide potential biomarkers for delineating autotrophic and heterotrophic carbon cycling in AMD environments.

Response of phytoplankton to ecological engineering remediation of a Canadian Shield Lake affected by acid mine drainage

Multivariate analyses were used to relate changes in water chemistry with those in the composition of the phytoplankton in two adjacent northern Ontario shield Lakes in Canada. One of the lakes is affected by both acid mine drainage (AMD) and a succession of remediation measures. The other, much larger lake is pristine and was monitored for both water quality and phytoplankton. The AMD lake, already contaminated at the start of the 17-year project, experienced further increases in Zn concentrations from around 10 mg L −1 to 35 mg L −1 and a pH decrease from 5 to 3 during the project. Over the same period, phytoplankton diversity declined from 52 taxa in 59 samples to 42 taxa in 74 samples. Only Ochromonas spp. and Chlamydomonas spp., apparently tolerant of a wide range of physical and chemical conditions, were consistently present in both lakes. Other taxa ( Peridinium, Pinnularia and Euglena) were frequent in the AMD lake in the early stages of the project but then declined. Asterionella and Botrycoccus (although common in the pristine lake), were rarely observed in the AMD lake. Lepocinclis proliferated as metal concentrations increased and pH declined in the AMD lake but was rarely found in the clean lake. Such gradual adaptation or changes in phytoplankton populations have not been reported previously for such extreme chemical conditions. The multivariate statistical analysis (PCA, CCA, DCA) revealed that the highest number of taxa were found to be controlled by pH and Ni in the years 1986–1992. One-third of the taxa in that period were more closely associated with pH alone. In the late stages of the study (1999–2003) the predominant controlling factors appeared to be metals and conductivity. Most of the taxa followed the time/pH/metal axis, suggesting that occurrence changed with slowly changing water conditions. Those taxa which fell outside the time/pH/Zn/Cu/Al axis included mostly Chrysophytes, Cryptophytes, and Chlorophytes, although there were some Euglenophyceae and diatoms in this group. By the end of the study in 2003, the combined effect of the restoration measures brought about the development of an extensive periphytic underwater meadow, along with a slight improvement in water quality and an increase in the number of phytoplankton taxa.