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    Chromate resistance mechanisms in Leucobacter chromiiresistens
    Chromate is one of the major anthropogenic contaminants on Earth. Leucobacter chromiiresistens is a highly chromate-resistant strain, tolerating chromate concentrations in LB medium of up to 400 mM. In response to chromate stress, L. chromiiresistens forms biofilms, which are held together via extracellular DNA. Inhibition of biofilm formation leads to drastically decreased chromate tolerance. Moreover, chromate is reduced intracellularly to the less-toxic Cr(III). The oxidation status and localization of chromium in cell aggregates were analyzed by energy-dispersive X-ray spectroscopy coupled to scanning transmission electron microscopy and X-ray absorption spectroscopy measurements. Most of the heavy metal is localized as Cr(III) at the cytoplasmic membrane. As a new cellular response to chromate stress, we observed an increased production of the carotenoid lutein. Carotenoid production could increase membrane stability and reduce the concentration of reactive oxygen species. Bioinformatic analysis of the L. chromiiresistens genome revealed several gene clusters that could enable heavy-metal resistance. The extreme chromate tolerance and the unique set of resistance factors suggest the use of L. chromiiresistens as a new model organism to study microbial chromate resistance.
    Publicationtype: Journal Article
    Scopus© Citations 30  45
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    Arsenic-rich acid mine water with extreme arsenic concentration: Mineralogy, geochemistry, microbiology, and environmental implications
    (American Chemical Society (ACS), 2014-11-14) ; ; ; ;
    Extremely arsenic-rich acid mine waters have developed by weathering of native arsenic in a sulfide-poor environment on the 10th level of the Svornost mine in Jáchymov (Czech Republic). Arsenic rapidly oxidizes to arsenolite (As2O3), and there are droplets of liquid on the arsenolite crust with high As concentration (80,000-130,000 mg·L-1), pH close to 0, and density of 1.65 g·cm-1. According to the X-ray absorption spectroscopy on the frozen droplets, most of the arsenic is As(III) and iron is fully oxidized to Fe(III). The EXAFS spectra on the As K edge can be interpreted in terms of arsenic polymerization in the aqueous solution. The secondary mineral that precipitates in the droplets is kaatialaite [Fe3+(H2AsO4)3·5H2O]. Other unusual minerals associated with the arsenic lens are běhounekite [U4+(SO4)2·4H2O], štěpite [U4+(AsO3OH)2·4H2O], vysokýite [U4+[AsO2(OH)2]4·4H2O], and an unnamed phase (H3O)+2(UO2)2(AsO4)2·nH2O. The extremely low cell densities and low microbial biomass have led to insufficient amounts of DNA for downstream polymerase chain reaction amplification and clone library construction. We were able to isolate microorganisms on oligotrophic media with pH ∼ 1.5 supplemented with up to 30 mM As(III). These microorganisms were adapted to highly oligotrophic conditions which disabled long-term culturing under laboratory conditions. The extreme conditions make this environment unfavorable for intensive microbial colonization, but our first results show that certain microorganisms can adapt even to these harsh conditions.
    Publicationtype: Journal Article
    Scopus© Citations 52  10
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    Oxygen-dependent niche formation of a pyrite-dependent acidophilic consortium built by archaea and bacteria
    (Nature Publishing Group, 2013-04-25) ; ; ; ;
    Biofilms can provide a number of different ecological niches for microorganisms. Here, a multispecies biofilm was studied in which pyrite-oxidizing microbes are the primary producers. Its stability allowed not only detailed fluorescence in situ hybridization (FISH)-based characterization of the microbial population in different areas of the biofilm but also to integrate these results with oxygen and pH microsensor measurements conducted before. The O 2 concentration declined rapidly from the outside to the inside of the biofilm. Hence, part of the population lives under microoxic or anoxic conditions. Leptospirillum ferrooxidans strains dominate the microbial population but are only located in the oxic periphery of the snottite structure. Interestingly, archaea were identified only in the anoxic parts of the biofilm. The archaeal community consists mainly of so far uncultured Thermoplasmatales as well as novel ARMAN (Archaeal Richmond Mine Acidophilic Nanoorganism) species. Inductively coupled plasma analysis and X-ray absorption near edge structure spectra provide further insight in the biofilm characteristics but revealed no other major factors than oxygen affecting the distribution of bacteria and archaea. In addition to catalyzed reporter deposition FISH and oxygen microsensor measurements, microautoradiographic FISH was used to identify areas in which active CO 2 fixation takes place. Leptospirilla as well as acidithiobacilli were identified as primary producers. Fixation of gaseous CO 2 seems to proceed only in the outer rim of the snottite. Archaea inhabiting the snottite core do not seem to contribute to the primary production. This work gives insight in the ecological niches of acidophilic microorganisms and their role in a consortium. The data provided the basis for the enrichment of uncultured archaea.
    Publicationtype: Journal Article
    Scopus© Citations 51  14
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    A mineralogical, geochemical, and microbiogical assessment of the antimony- and arsenic-rich neutral mine drainage tailings near Pezinok, Slovakia
    The mineralogical composition of mining wastes deposited in voluminous tailing impoundments around the world is the key factor that controls retention and release of pollutants. Here we report a detailed mineralogical, geochemical, and microbiological investigation of two tailing impoundments near the town of Pezinok, Slovakia. The primary objective of this study was the mineralogy that formed in the impoundment after the deposition of the tailings (so-called tertiary minerals). Tertiary minerals include oxyhydroxides of Fe, Sb, As, Ca and are present as grains and as rims on primary ore minerals. X-ray microdiffraction data show that the iron oxyhydroxides with abundant As are X-ray amorphous. The limiting (lowest) Fe/As (wt/wt%) ratio in this material is 1.5; beyond this ratio, the hydrous ferric oxide does not retain arsenic. The grains with less As and little to moderate amounts of Sb are goethite; the grains where Sb dominates over Fe are poorly crystalline tripuhyite (FeSbO4). Even the most heavily contaminated samples (up to 29 wt% As2O5) are populated with diverse communities of microorganisms including typical arsenic-resistant heterotrophic species as well as iron reducers and sulfur oxidizers. Several recovered clones cluster within phylogenetic groups that are solely based on environmental sequences and do not contain a single cultivated species, thus calling for more work on such extreme environments.
    Publicationtype: Journal Article
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    Matrix composition and community structure analysis of a novel bacterial pyrite leaching community
    Here we describe a novel bacterial community that is embedded in a matrix of carbohydrates and bio/geochemical products of pyrite (FeS2) oxidation. This community grows in stalactite-like structures - snottites - on the ceiling of an abandoned pyrite mine at pH values of 2.2-2.6. The aqueous phase in the matrix contains 200 mM of sulfate and total iron concentrations of 60 mM. Micro-X-ray diffraction analysis showed that jarosite [(K,Na,H 3O)Fe3(SO4)2(OH)6] is the major mineral embedded in the snottites. X-ray absorption near-edge structure experiments revealed three different sulfur species. The major signal can be ascribed to sulfate, and the other two features may correspond to thiols and sulfoxides. Arabinose was detected as the major sugar component in the extracellular polymeric substance. Via restriction fragment length polymorphism analysis, a community was found that mainly consists of iron oxidizing Leptospirillum and Ferrovum species but also of bacteria that could be involved in dissimilatory sulfate and dissimilatory iron reduction. Each snottite can be regarded as a complex, self-contained consortium of bacterial species fuelled by the decomposition of pyrite.
    Publicationtype: Journal Article
    Scopus© Citations 42  14